GREEN. Silylates and Surface Modified Siloxysilicates

Transcription

1 GREEN Silylates and Surface Modified Siloxysilicates CIR EXPERT PANEL MEETING MARCH 34, 2011

2 February 3, 2011 MEMORANDUM To: From: Subject: CIR Expert Panel and Liaisons Lillian C. Becker, M.S. Scientific Analyst and Writer Draft Report for Silylates The Cosmetic Ingredient Review (CIR) announced the Scientific Literature Review (SLR) for silylates in December, Comments from industry have been addressed and unpublished data have been incorporated. A few of the larger unpublished data documents that were submitted to the EPA had more detail than necessary (e.g., raw data, informed consent forms). These extra pages have been removed in the print version of the Panel book. If these data are necessary for evaluation, you may examine the removed pages in the online version. The Panel should review the Draft Report and decide whether any additional data are needed in order to reach a safety conclusion. If so, an insufficient data announcement should be issued. If no additional data are required, then the Panel should develop a conclusion and a rationale/discussion then issue a Tentative Report.

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4 Report History for Silylates December, 2010 SLR issued for public comment for silica silylate, silica dimethyl silylate, trimethylsiloxysilicate, and trifluoropropyldimethyl/trimethylsiloxysilicate. Unpublished data and technical comments were received from industry. March, 2011 Panel reviews Draft Report. Panel Book Page 2

5 Search Strategy for Silylates Lillian Becker PUBMED (July, 2010): Trimethylsiloxysilicate OR OR OR Silicic acid, trimethylsilyl ester OR Hexamethyldisiloxane tetraethyl orthosilicate copolymer OR Trimethylsilyl silicate OR OR OR OR OR OR OR OR OR OR OR Siloxane Polyalkyleneoxide Copolymer The following terms were not found in PubMed: , , Hexamethyldisiloxanetetraethyl, , , , , , , , , , , , , , , , , Silicic acid, trimethylsilyl ester 9 hits; none useful. Trimethylsilyl silicate 12 hits; none useful. Trifluoropropyldimethyl/trimethylsiloxysilicate 1 hit Silica Dimethyl Silylate OR OR Silica, [(dimethylsilyl)oxy] modified OR OR dichlorodimethylsilane treated fumed silica OR OR hydrophobic dichlorodimethylsilane treated fumed silica OR OR OR silica dimethyl silylate The following terms were not found in PubMed: , , dichlorodimethylsilanetreated, , , , , , "dichlorodimethylsilane"[substance Name] OR "dichlorodimethylsilane"[all Fields] 40 hits; none useful hydrophobic dichlorodimethylsilane treated fumed silica No hits. silica dimethyl silylate no hits. Silica Silylate OR OR dichlorodimethylsilanetreated fumed silica OR OR OR OR OR OR OR The following terms were not found in PubMed: , , dichlorodimethylsilanetreated, , , , , , , , , , , dichlorodimethylsilanetreated fumed silica No hits. Studies discovered at (July, 2010): HPV Chemical Database Studies discovered at EPA (July, 2010): 16 studies Panel Book Page 3

6 ADME Silylates Data Profile for March, Writer Lillian Becker Repeated dose Acute toxicity Irritation Sensitization toxicity Phototoxicity Carcinogenicity Genotoxicity Repro/Devel toxicity Sensitization Human Sensitization Animal Dermal Irr Human Dermal Irr. Animal Ocular Irritation Inhale Dermal Oral Inhale Dermal Oral Use Log Kow Dermal Penetration Chem/Phys Prop silica silylate silica dimethyl silylate Trimethylsiloxysi licate trifluoropropyldi methyl/trimethyls iloxysilicate X x X X X X x x X X x x X X X X X X X x x X Panel Book Page 4

7 Draft Report Silylates and Surface Modified Siloxysilicates March 3, 2011 The 2011 Cosmetic Ingredient Review Expert Panel members are: Chairman, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; Ronald A Hill, Ph.D. James G. Marks, Jr., M.D.; Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is F. Alan Andersen, Ph.D. This report was prepared by Lillian C. Becker, Scientific Analyst/Writer. Cosmetic Ingredient Review th Street, NW, Suite 412 " Washington, DC " ph " fax " cirinfo@cirsafety.org Panel Book Page 5

8 TABLE OF CONTENTS TABLE OF CONTENTS... i INTRODUCTION... 1 Chemistry... 1 Definition and Structure... 1 Physical and Chemical Properties... 2 UV Absorption... 2 Method of Manufacture... 2 USE... 2 Cosmetic... 2 NonCosmetic Use... 3 TOXICOKINETICS... 3 Absorption, Distribution, Metabolism, and Excretion... 3 Oral... 3 Inhalation... 3 TOXICOLOGICAL STUDIES... 3 Acute Toxicity... 4 Dermal NonHuman... 4 Oral NonHuman... 4 Intraperitoneal NonHuman... 4 Repeated Dose Toxicity... 4 Oral NonHuman... 4 Inhalation NonHuman... 5 REPRODUCTIVE AND DEVELOPMENTAL TOXICITY... 5 Genotoxicity... 6 In Vitro... 6 In Vivo... 7 CARCINOGENICITY... 7 Irritation and sensitization... 7 Irritation... 7 Dermal NonHuman... 7 Ocular NonHuman... 7 Irritation Human... 8 Sensitization... 8 NonHuman... 8 Human... 8 SUMMARY... 8 TABLES REFERENCES Panel Book Page 6

9 INTRODUCTION This is a safety assessment for silica silylate, silica dimethyl silylate, trimethylsiloxysilicate, and trifluoropropyldimethyl/trimethylsiloxysilicate. The functions of these ingredients in cosmetics include: antifoaming agents, anticaking agents, bulking agents, binders, skinconditioning agentsemollient, skinconditioning agentsocclusive, slip modifiers, suspension agentsnonsurfactant, and viscosity increasing agentsnonaqueous. Silica, which is the core of silica silylate and silica dimethyl, has been reviewed by the Cosmetic Ingredient Review Expert Panel and was found to be safe as cosmetic ingredients in the practices of use and concentrations as described in this safety assessment. 1 The silica materials in that assessment and in this one are amorphous (synthetic amorphous silica and silicates), not crystalline silica. The ingredients in this safety assessment are organosilane hybrid materials, modified to have desired properties for their use in cosmetics. Data on silane, dichlorodimethyl, reaction products with silica (CAS No ) and treated fumed silica dust (as CABOSIL TS720) were also included in this literature review, since these chemicals are the same as silica dimethyl silylate. Data from 2 mixtures: siloxanes and silicones, dime (dimethyl silicones and siloxanes) 93% (CAS No ) with dimethyl silicones and siloxanes, reaction product with silica 7% (as Antifoam A; CAS No ) and siloxanes and silicones, dime, hydroxylterminated (as Antifoam M) were also included because the data may also be relevant to the individual ingredients. CHEMISTRY Definition and Structure The cosmetic ingredient definitions, functions in cosmetics, and structures of silica silylate, silica dimethyl silylate, trimethylsiloxysilicate, and trifluoropropyldimethyl/trimethylsiloxysilicate are provided in Table 1. These four hybrid silica materials can be divided into two distinct types, grafted and cocondensed. Grafted silica materials These materials consist of silica particles that are surface modified by organosilanes. For example, dichlorodimethylsilane can be used to produce dimethyl silyl groups on the surface of a particle of fumed silica. 2 Silica silylate and silica dimethyl silylate are grafted silica materials. Silica silylate consists of fumed silica, surface modified with trimethylsilyl groups (Figure 1). Figure 1. Cocondensed silica materials In contrast to grafted materials, cocondensed silica materials are not surface modified silica particles. Instead, cocondensed materials are prepared by the simultaneous reaction of condensable inorganic silica and silylated organic compounds. 2 This process is similar to random copolymer synthesis, but is nonlinear. Trimethylsiloxysilicate and trifluoropropyldimethyl/trimethylsiloxysilicate are cocondensed silica materials. Trimethylsiloxy silicate is the cohydrolysis product of a tetraalkoxysilane and a trimethylalkoxysilane. 3 The chemical structure of trimethylsiloxy silicate can be visualized as a threedimensional network of polysilicic acid units (resultant from the tetraalkoxysilane), which are endblocked with trimethylsilyl groups (Figure 2). Panel Book Page 7

10 Figure 2. Trifluoropropyldimethyl/trimethylsiloxysilicate differs from trimethylsiloxy silicate only by the replacement of some of the methyl groups with trifluoropropyl groups. Physical and Chemical Properties These ingredients are amorphous solids, with virtually no water solubility. The water solubility of grafted silica materials is below 10e 6 g/l. Chemical and physical properties for silica dimethyl silylate are provided in Table 2. One manufacturer reported that less than 0.83% of silica dimethyl silylate had a particle size of < 125 µm and none were < 90 µm. 4 Physical and chemical properties were not discovered for silica silylate, trimethylsiloxysilicate, and trifluoropropyldimethyl/trimethylsiloxysilicate. UV Absorption While no data were available, the ingredients included in this review would not be expected to have any significant ultraviolet (UV) absorption because these materials do not contain any of the functional groups commonly associated with UV absorption. Method of Manufacture Grafted material such as silica silylate, and silica dimethyl silylate can be manufactured via reaction of a fumed silica particle with one of an alkoxysilane (e.g. (CH 3 O) 3 SiCH 3 ), a halosilane (e.g., ClSi(CH 3 ) 3 ) or an alkylsilazane (e.g., NH[Si(CH 3 ) 3 ] 2 ). 2 For example, amorphous silica can be modified by reaction with hexamethyldisilazane (HMDS), in hexanes at 275 o C and 30 atm, to manufacture silica silylate. 5 The degree of surface modification can be adjusted by varying the concentration of the silylating agent (e.g., increasing the amount of HMDS). Cocondensed silica materials can be manufactured via the cohydrolysis of a tetraalkoxysilane (e.g., tetraethoxysilane; which result in the inorganic silane groups in the reaction product) and a trialkylalkoxysilane (e.g., trimethylethoxysilane; which will result in the organosilane groups in the reaction product). 2 Some residual alkoxyl (i.e. leaving groups that did not leave; SiOR) and hydroxyl (SiOH) functional groups are likely to be present. 3 The average molecular weight can be adjusted by varying the ratio of the silanes. USE Cosmetic According to the Voluntary Cosmetic Registration Program (VCRP) administered by the Food and Drug Administration (FDA), the total number of uses of silica dimethyl silylate was 734 (593 leaveon and 141 rinseoff products). 6 A survey conducted by the Personal Care Products Council (Council) found that silica dimethyl silylate was used at % 10% in leaveon products (highest concentration in lipsticks) and % 4% in rinseoff products (highest concentration in personal cleanliness products; Table 3). 7 There were 633 uses reported of trimethylsiloxysilicate at % 30% in leaveon products (highest in lipsticks) and 0.002% 5% in rinseoff products (highest in hair straighteners and eye lotions). There were no uses reported of trifluoropropyldimethyl/trimetylsiloxysilicate by FDA but the Council reported use at 2% 20% in leaveon products (highest in eyeliner). No uses were reported for silica silylate; concentration of use information may be forthcoming. Panel Book Page 8

11 Silica dimethyl silylate is used in perfumes and may be aerosolized. Most of the particles are > 125 µm and none were < 90 µm. 4 Therefore, effects on the lungs that may be induced by aerosolized products containing these ingredients are of concern. The aerosol properties that determine deposition in the respiratory system are particle size and density. The parameter most closely associated with deposition is the aerodynamic diameter, da, defined as the diameter of a sphere of unit density possessing the same terminal settling velocity as the particle in question. In humans, particles with an aerodynamic diameter of 10µm are respirable. Particles with a d a from µm settle in the upper respiratory tract and particles with a d a < 0.1 µm settle in the lower respiratory tract. 8,9 Particle diameters of 6080 µm and 80 µm have been reported for anhydrous hair sprays and pump hairsprays, respectively. 10 In practice, aerosols should have at least 99% of their particle diameters in the µm range and the mean particle diameter in a typical aerosol spray has been reported as ~38 µm. 11 Therefore, most aerosol particles are deposited in the nasopharyngeal region and are not respirable. NonCosmetic Use FDA has approved silicon dioxide (a compound similar to silica dimethyl silylate) to be used as a direct food additive as an anticaking agent up to 2% and in the manufacture of materials that come in direct contact with food in various production, manufacturing, packaging, preparing, transporting, and holding operations. 12 Silica dimethyl silylate is generally recognized as safe (GRAS) in dietary supplements. 13 TOXICOKINETICS Absorption, Distribution, Metabolism, and Excretion Orally administered silica dimethyl silylate was eliminated from the body primarily in the feces. Inhaled silica dimethyl silylate collected in the lungs and lymph nodes of rats. Oral SILICA DIMETHYL SILYLATE Antifoam A (siloxanes and silicones, dime [dimethyl silicones and siloxanes] 93% with dimethyl silicones and siloxanes, reaction product with silica 7%; 0.5 ml/kg), Antifoam M (siloxanes and silicones, dime, hydroxylterminated; 0.5 mg/kg), or silica (6 mg/kg) were orally administered in sesame oil to male Buckberg mice (n = 12) after fasting. 14 Controls were administered sesame oil (0.5 ml). There was an increase in urinary and biliary silicon in the Antifoam A group but not the others. The authors suggested that the source was organosoluble silicon rather than inorganic silica from the absorption of silica. Antifoam A (1.15, 13.7, or 18.0 mg/kg) and Antifoam M (21.8 or 41.8 mg/kg) were randomly radiolabeled and orally administered to rhesus monkeys (n = 5) that were then observed for 7 days. For Antifoam A, there was 0.5% 0.9% of the dose expired in the breath and 2.0% 2.5% in the urine with a half life of 24 h. There was 0.1% to 0.9% in the bile in the first 24 h after dosing. Over 92 h, 80% to 90% of the dose was recovered in the feces. For Antifoam M, there was 0.1% 0.2% expired in the breath and 0.22% in the urine with a half life of 24 h. There was 0.1% to 0.9% in the bile in the first 24 h after dosing. Over 92 h, 93% to 97% of the dose was recovered in the feces. There was < 0.01% detected in ~40 tissues examined in the 1 monkey necropsied after 7 days. A range of 93% to 98% was recovered in the feces. Human subjects were orally administered Antifoam M (100 mg/kg; n = 6) after 5 days of a consistent diet (that continued through the rest of the experiment) on day 6 and data collected through day Antifoam A (100 mg/kg; n = 5) was administered on day 8 and data collected through day 9. Increased silicon levels were not found in urine, feces, or expelled air after oral administration of Antifoam M. Silicon levels in urine, feces and expelled air were increased after oral Antifoam A. The authors suggest that it is probable that the increased silicon levels represent organosoluble silicon rather than inorganic silicon (silica). The average percent of dose found in the urine over six days was 1.8% for Antifoam A with a halflife of 24 hours. At least 0.5% of the dose was found in expired air with a halflife of 8 hours, and has been identified as low molecular weight cyclosiloxanes (octamethylcyclotetrasiloxane and decamethylcyclotetrasiloxane). The organosoluble silicon in urine was not identified. Inhalation SILICA DIMETHYL SILYLATE Rats (n = 40) were exposed to aerosolized silica dimethyl silylate (200 mg/m 3 ; particle size not provided) for 5 h/d for 3 days. 15 At 24 h there was 0.91 mg test substance in the lung and none at 1 month post exposure. There was 0.383, 0.239, and mg in the mediastinal lymph nodes at 1, 2, and 3 months, respectively. At 3 months, 81% of the test substance had been eliminated. Female SpragueDawley rats (n = 50) were exposed to aerosolized silica dimethyl silylate (50 mg/m 3 ; <7 µm) for 5 h. 3 At necropsy, silica deposited in lungs (0.156, 0.034, mg) and mediastinal lymph nodes (0, 0.003, 0.004) at 20 h, 1 and 3 months, respectively. Test substance eliminated at 78% and 85% at 1 and 3 months, respectively. Female SpragueDawley rats (n = 30) were exposed to aerosolized silica dimethyl silylate (50 mg/m 3 ; <7 µm) for 5 h for 3 days. 3 At necropsy, silica deposited in lungs (0.34, 0.085, 0.30 mg) and mediastinal lymph nodes (0.34, 0.085, 0.30 mg) at 20 h, 1 and 3 months, respectively. Test substance eliminated at 75% and 92% at 1 and 3 months, respectively. TOXICOLOGICAL STUDIES 14 Panel Book Page 9

12 Acute Toxicity Dermally administered silica dimethyl silylate up to 2000 mg/kg and trimethylsiloxysilicate up to 2 g/kg was not toxic to rats. The oral LD 50 of silica dimethyl silylate was >7900 mg/kg for rats; trimethylsiloxysilicate had no effects at 5 g/kg. There was no mortalities from the inhalation of silica dimethyl silylate up to 520 mg/m 3. Intraperitoneally administered silica dimethyl silylate caused thickening of the liver and spleen capsules. The test substance was in observed in the abdominal cavity. Dermal NonHuman SILICA DIMETHYL SILYLATE Silica dimethyl silylate (2000 mg/kg in propylene glycol) applied in a single dose to the skin of Wistar rats (n = 5/sex) for 24 h caused no mortality. 16 No clinical signs were observed. Necropsies were unremarkable. TRIMETHYLSILOXYSILICATE Trimethylsiloxysilicate (100%; 0.5 g) was administered to the intact skin of New Zealand White rabbits (n = 6) under occlusion for 4 h. 17 All rabbits survived and gained weight during the study. There were no signs of toxicity. Trimethylsiloxysilicate (2 g/kg) was administered to the shaved skin of New Zealand White rabbits (n = 10) under occlusion for 24 h. 17 The patch was then removed and the skin rinsed in corn oil. The rabbits were observed for 14 days. All rabbits gained weight. There were no signs of toxicity. There were no abnormalities observed at necropsy. Oral NonHuman SILICA DIMETHYL SILYLATE The oral LD 50 of silica dimethyl silylate was > 5000 mg/kg for SpragueDawley rats. 18,19 LD 50 to be > 7900 mg/kg. 20 TRIMETHYLSILOXYSILICATE Trimethylsiloxysilicate (5 g/kg in corn oil) was orally administered to SpragueDawley rats (n = 5/sex). 17 clinical signs. All rats gained weight. There were no lesions at necropsy. The observation time was not provided. Another study in rats found the oral There were no Inhalation NonHuman SILICA DIMETHYL SILYLATE Inhalation studies using rats were conducted. The results are presented in Table 4. There was no mortality up to 520 mg/m 3. Intraperitoneal NonHuman SILICA DIMETHYL SILYLATE Silica dimethyl silylate (up to 30 mg in water with Tween 80) was administered intraperitoneally (i.p.) to mice (n = 120; strain and sex not provided). 21 All mice survived treatment. The observation period was not provided, but the report stated that at necropsy fibrosis was not observed, although thickening of the liver and spleen capsules were observed. Histopathology showed that the test substance was found in the abdominal cavity in a tight network of reticulin and collagen. Slight phagocyte accumulations and necrosis were observed. Histopathology of the liver showed some evidence of the test substance there, also in a tight network of reticulin and collagen. Silica dimethyl silylate (up to 200 mg in water with 0.5% Tween) was administered i.p. to female rats (n = 100; strain not provided) as described above. 21 All rats survived treatment. At necropsy, there was no fibrosis observed. Histopathology showed that the test substance was found in the abdominal cavity in a tight network of reticulin and collagen. Slight phagocyte accumulations and necrosis were observed. Histopathology of the liver showed some evidence of the test substance there, also in a tight network of reticulin and collagen. Other NonHuman Silica dimethyl silylate ( g; 0.1 ml) was applied to one eye of New Zealand white rabbits (n = 3/sex). 22 Three of the treated eyes were not rinsed and 3 rinsed with saline after sec. Two females had reduced feed consumption and of these had soft stool, anogenital staining, and reduced fecal volume. Repeated Dose Toxicity Antifoam A (dimethyl silicones and siloxanes, reaction product with silica) in feed did not cause any adverse effect in rabbits up to 1% and rats up to 1000 mg/kg. The oral NOAEL for silica dimethyl silylate was 500 mg/kg for 6 months and 100 mg/kg for 24 months. Rats that inhaled treated fumed silica dust for up to 4 weeks were observed to have crusty eyes, muzzle, and nose; closed eyes; irregular breathing; irritable disposition; lacrimation and salivation; scabs; and red and yellow/brown stained fur. The inhalation NOAEL was up to 500 mg/m 3 and the LOAEL up to 1000 mg/m 3. Oral NonHuman SILICA DIMETHYL SILYLATE Adult albino rabbits (n = 3/sex) were fed a powdered basic diet plus 1% of Antifoam A; basic diet plus 1% of Antifoam A and 0.8% cholesterol; 0.8% cholesterol; or just the basic diet (n = 6/sex) for 8 months. 23 One rabbit in the antifoam A group died, Panel Book Page 10

13 possibly due to pulmonary irritation and congestion. Two rabbits died in the antifoam A/cholesterol group, both having extensive hepatic injuries. One rabbit died in the cholesterol only group with similar hepatic findings. There were no deaths in the control group. There were no behavioral changes in any group. There were no weight changes or variation in leukocytes among groups. The test substance had no effect on the anemia and elevated cholesterol in both groups administered cholesterol. Erythrocytes were in normal ranges. Liver and biliary functions and urine findings were normal in all groups. Examination of the organs in animals administered the test substance were unremarkable. Necropsy and histopathological examinations found no effects by the test substance. The authors concluded that there were no adverse effects from the ingestion of Antifoam A at 1%. The above feeding study was repeated using weanling FDRL rats (n = 5/sex) and extended to 52 weeks. 24 Controls (n = 10/sex) were fed the base diet. At the end of the treatment period, the rats were killed and necropsied. There were 1 or 2 deaths/group in the final 10 weeks of the study that were not attributed to the test substance. There were no clinical signs during the study. Weight gains were greater than controls by the males in the antifoam group. Feed intake was similar in all groups. Blood chemistry and urine analysis were similar in all groups. Necropsies were unremarkable. The authors concluded that this combination of chemicals in the diet for a year did not have any adverse effects on rats. Silica dimethyl silylate (500 or 1000 mg/kg) was orally administered to Wistar rats (n = 40/sex) by gavage every other day 25 for 19 or 39 days. Rats were killed and necropsied at the end of the treatment period or after 4 weeks recovery. There were no clinical signs or treatment effects observed. The NOAEL was 1000 mg/kg. Silica dimethyl silylate (0, 500, 1000, 2000 mg/kg; the high dose groups was gradually increased to 4000, 8000, and mg/kg) was orally administered to Wistar rats (n = 40/sex) in feed daily for 5 or 8 weeks for the high dose. Rats were killed and necropsied at the end of the treatment period. Two males and 2 females in the high dose group died after 9 and 13 days of exposure to mg/kg. Clinical signs in the high dose group after increasing the dose to mg/kg were apathy and decreased grooming activity. Cachexia and hemorrhagic mucosa of the nose and eyes were observed prior to death. There was severe body weight decrease in males and females following 1 week exposure to 8000 mg/kg and exposure to mg/kg. Food consumption severely decreased following exposure to mg/kg in males and females. There was hemorrhage in the mucous membranes of the eyes and nose in animals exposed to mg/kg. In 2 females of the mid dose group and 8 animals of the high dose group, atrophic hepatocytes with decreased appearance [sic] and decreased glycogen contents of the cytoplasm were observed. The NOAEL was 500 mg/kg and the LOAEL was 1000 mg/kg. Silica dimethyl silylate (0, 500 mg/kg) was orally administered to Wistar rats (n = 40/sex) in feed daily for 6 months. 27 Rats were killed and necropsied at the end of the treatment period or after 3 weeks recovery. There were no clinical signs or treatment effects observed. The NOAEL was 500 mg/kg. Silica dimethyl silylate (100 mg/kg) was orally administered to Wistar rats (n = 20/sex) in feed daily for 24 months. 28 Rats were killed and necropsied at the end of the treatment period or after 3 weeks recovery. There were no clinical signs or treatment effects observed. The NOAEL was 100 mg/kg. Inhalation NonHuman SILICA DIMETHYL SILYLATE Repeated dose inhalation studies from 1 week to 1 year are presented in Table 4. In rats, silica dimethyl silylate was lethal at 0.06 mg/l. Clinical signs included crusty eyes, muzzle, and nose; crust around ear tags; closed eyes; irregular breathing; irritable disposition; lacrimation and salivation; scabs; and red and yellow/brown stained fur. An increase in lymphocytes and neutrophils, as well as reduced body weights was observed. Silica was deposited in the lungs and lymph nodes, but the deposits cleared over time. A LOAEL of 31 mg/m3 was concluded in one study. REPRODUCTIVE AND DEVELOPMENTAL TOXICITY Silica dimethyl silylate did not cause any reproductive or developmental toxicity to rats up to 3.8 g/kg/d, rabbits up to 1600 mg/kg, hamsters up to 1600 mg/kg, or mice up to 1340 mg/kg. SILICA DIMETHYL SILYLATE Pregnant Wistar SPF MW3rats (n = 17 19) were orally administered Antifoam A (0.3, 1.20, 3.8 g/kg/d) on days 6 to 15 of gestation. 29 Controls were administered water. The high dose is comparable to a therapeutic dose used in humans as an antacid. On day 20 of gestation, the dams were anesthetized and the fetuses examined. There were no differences in viability of fetuses, resorptions, gross external anomalies, soft tissue anomalies, skeletal anomalies, or fetal weight compared to controls at any dose level. Pregnant New Zealand white rabbits (n = 3) were administered Antifoam A (0, 0.5%, 1.0%, and 2.5%; 500, 1000, and fold higher than normal human consumption) in their feed days 6 19 of gestation. Cesarean sections and necropsies were performed at day 29 of gestation. No clinical signs were observed during treatment. Two dams died due to unrelated causes. There were no differences in body weights and feed consumption among groups. There were no signs of toxicity to the dams. No were no differences in incidences of external versceral or skeletal abnormalities to the fetuses. The authors concluded that Antifoam A is not embryotoxic or teratogenic in rabbits at dietary levels up to 2.5%. Silica dimethyl silylate (0, 13.5, 62.7, 292, 1350 mg/kg/d) was administered to pregnant Wistar rats (n = not provided) by gavage on days 6 15 of gestation. 31 There were no adverse effects on nidation or on maternal or fetal survival. There were no Panel Book Page 11

14 increases in skeletal or soft tissue abnormalities in the offspring. The maternal and fetal NOAEL was 1350 mg/kg/d. Silica dimethyl silylate (0, 16.0, 74.3, 345., 1600 mg/kg/d) was administered to pregnant Wistar rats (n = not provided) by gavage on days 6 15 of gestation. 31 There were no adverse effects on nidation or on maternal or fetal survival. There were no increases in skeletal or soft tissue abnormalities in the offspring. The maternal and fetal NOAEL was 1600 mg/kg/d. Silica dimethyl silylate (0, 13.4, 62.3, 289, 1340 mg/kg/d) was administered to pregnant CD1 mice (n = not provided) by 31 gavage on days 6 15 of gestation. There were no adverse effects on nidation or on maternal or fetal survival. There were no increases in skeletal or soft tissue abnormalities in the offspring. The maternal and fetal NOAEL was 1340 mg/kg/d. Silica dimethyl silylate (0, 16.0, 74.3, 345., 1600 mg/kg/d) was administered to pregnant Dutch rabbits (n = not provided) by 31 gavage on days 6 18 of gestation. There were no adverse effects on nidation or on maternal or fetal survival. There were no increases in skeletal or soft tissue abnormalities in the offspring. The maternal and fetal NOAEL was 1600 mg/kg/d. Silica dimethyl silylate (0, 16.0, 74.3, 345., 1600 mg/kg/d) was administered to pregnant Syrian hamsters (n = not provided) by gavage on days 6 10 of gestation. 31 There were no adverse effects on nidation or on maternal or fetal survival. There were no increases in skeletal or soft tissue abnormalities in the offspring. The maternal and fetal NOAEL was 1600 mg/kg/d. Silica dimethyl silylate (0, 500 mg/kg/d) was administered in feed to male (n = 2) and female (n = 10) Wistar rats for 6 months, during which the rats were mated twice, followed by a 3 week recovery period. 32 The offspring were observed through the 4 week lactation period then killed and necropsied. There were no mortalities attributable to treatment. There were no effects observed during treatment or at necropsy in the adults or the offspring. NOAEL was 500 mg/kg. Silica dimethyl silylate (0, 497, 509 mg/kg/d) was administered to Wistar rats (n = 40/sex) in feed for 6months, after which 33 the rats were mated (1 male to 5 females). The adult rats were killed and necropsied and the offspring were observed for external appearance and development. No abnormalities were observed in either generation. The NOAEL was 497 mg/kg/d for parental generation. Silica dimethyl silylate (0, 100 mg/kg/d) was administered to Wistar rats (n = 20/sex) in feed for 24 months, after which the rats were mated (1 male to 5 females). 28 The offspring were adjusted to 5/sex in each litter and allowed to mature. After 7 months, they were mated and their litters were also adjusted to 5/sex. Both sets of offspring were killed and necropsied. There were no reproductive toxicity effects observed. GENOTOXICITY Silica dimethyl silylate was not genotoxic in several Ames test and a mammalian chromosome aberration test. A product containing siloxanes and silicones, dime (dimethyl silicones and siloxanes (18%) and dimethyl silicones and siloxane, reaction products with silica (2%) was not genotoxic in an Ames test. Trimethylsiloxysilicate was not genotoxic in an Ames test. There were no in vivo genotoxicity studies discovered. In Vitro SILICA DIMETHYL SILYLATE In an Ames test of a toluene extract of silica dimethyl silylate ( µg/plate) using Salmonella. typhimurium (TA98, TA100, TA1537) and Escherichia coli (WP2urvA) with and without metabolic activation, no mutagenicity was observed. 34 Controls had the expected results. In an Ames test of a product made up of Antifoam A using S. typhimurium (TA98, TA100, TA1535, TA1537, TA1538), 35 there was no evidence of genetic activity with or without metabolic activation. Controls had the expected results. An Ames test of a product ( µg/plate) containing silica dimethyl silylate (27%) was conducted using S. typhilmurium (TAI535, TA1537, TA98, TA100) and E. coli (WP2 trp, WP2 trp uvra) with and without metabolic activation. 36 The concentration was calculated to be 1250 µg/plate dimethyl silicones and siloxanes and 100 µg/plate dimethyl silicones and siloxanes reaction products with silica. The test substance was not mutagenic. An Ames test of silica dimethyl silylate ( µg/plate) was conducted using S. typhilmurium (TA98, 100, TA1535, TA1538) with and without metabolic activation. 37 There was no evidence of mutagenicity. The controls had the expected results. An Ames test of silica dimethyl silylate ( µg/plate) was conducted using S. typhilmurium (TA98, 100, TA1537, TA1538) and E. coli (WP2 trp uvra) with and without metabolic activation. 38 There was no evidence of mutagenicity. The controls had the expected results. An in vitro mammalian chromosome aberration test of silica dimethyl silylate (63, 125, 250, 500 µg/ml) using Chinese hamster ovary (CHO) cells with and without metabolic activation was conducted. The frequency of effects without S9 were 0, 1%, 0, 0 at 63, 125, 250 and 500 µg/ml, respectively, and 3, 1, 1, 3 % with S9, respectively. 39 The author concluded that there was no evidence of genotoxicity. The controls had the expected results. TRIMETHYLSILOXYSILICATE An Ames test (0, 30, 80, 250, 700, 2000, 5000 µg/plate) using S. typhimurium (TA98, TA100, TA1535, TA1537) and E. coli (WP3uvrA) with and without metabolic activation was conducted. 17 There were no toxic effects and the revertant frequencies were similar to controls. Positive controls had the expected results. The authors concluded that trimethylsiloxysilicate was nonmutagentic in this assay. An Ames test of a mixture of trimethlsiloxysilicate (60%) and isododecane (40%; µg/plate) using S. typhimurium 40 ( TA98, TA100) was negative. Panel Book Page 12

15 In Vivo There were no in vivo genotoxicity studies discovered for any of the ingredients in this safety assessment. CARCINOGENICITY Silica dimethyl silylate (100 mg/kg) was orally administered to Wistar rats (n = 20/sex) in feed daily for 24 months. 28 Rats were killed and necropsied at the end of the treatment period or after 3 weeks recovery. There were no carcinogenic effects obsevered. The nature and incidence of tumors were comparable with the historical control data. IRRITATION AND SENSITIZATION Irritation Silica dimethyl silyate and Trimethylsiloxysilicate were not demally irritating to rabbits up to 100%. In a human patch test, a mixture containing trimethylsiloxysilicate at 60% resulted in irritation in 2 out of 19 subjects. Silica dimethyl silylate was slightly or not irritating to the rabbit eye. Trimethylsioloxysilicate was practically or nonirritating to the rabbit eye. Dermal NonHuman SILICA DIMETHYL SILYLATE Silica dimethyl silylate (0.5 g moistened with tap water) was applied to the shaved skin of New Zealand white rabbits (n = 3; 1 male, 2 female) under occlusion for 4 h. 41 There was mild erythema in one rabbit at 1 h after removal. The irritation score was 0.2. Silica dimethyl silylate (0.5 g; 100%) was applied to the intact skin of New Zealand white rabbits (n = 3/sex) for 4 h under semiocclusion. 42 After removal, the skin was scored at 1, 24, 48, and 72 h. There were no signs of irritation at any observation period. Silica dimethyl silylate (6% in aqueous methylhydroxethylcelluslosegel) was applied to the intact and abraded skin of New Zealand white rabbits (n = 3/sex) for 24 h under occlusion. 43 The skin was scored at removal, 48 h, and daily for 14 days. There were no signs of irritation at any observation period. Silica dimethyl silylate (50% in olive oil) was applied to the intact and abraded skin of New Zealand white rabbits (n = 3/sex) for 24 h under occlusion. 43 After removal, the skin was scored at removal, 48 h, and daily for 14 days. There were no signs of irritation at any observation period. A product (0.5 ml) containing Silica dimethyl silyate (27 wt.%) was tested for dermal irritation using male New Zealand white rabbits. 44 The test substance was administered to the clipped skin under semiocclusion for 4 h then washed. There was no dermal irritation observed at 1, 24, 48, and 72 h. TRIMETHYLSILOXYSILICATE Trimethylsiloxysilicate (100%; 0.5 g) was administered to the intact skin of New Zealand White rabbits (n = 6) under occlusion for 4 h. 17 There were no signs of erythema or edema observed. The authors rated trimethylsiloxysilicate a dermal nonirritant. Trimethylsiloxysilicate (2 g/kg) was administered to the shaved skin of New Zealand White rabbits (n = 10) under occlusion for 24 h. 17 The patch was then removed and the skin rinsed in corn oil. The rabbits were observed for 14 days. There were slight signs of grade 1 irritation in 4 rabbits which were resolved by day 2. Ocular NonHuman SILICA DIMETHYL SILYLATE Silica dimethyl silylate administered into the eyes of New Zealand white rabbits (n = 3) caused only slight conjunctivae redness at 1 h after instillation. 45 In an ocular irritation test using New Zealand white rabbits (n = 6), Silica dimethyl silyate (27 wt.%) was administered to the eye and examined at 1, 24, 48, and 72 h. 46 There was a diffuse crimson coloration of the conjunctivae and slight swelling of the eyelids observed in 1 rabbit. Slight redness of the conjunctivae alone was seen in the remaining 5 animals. The mean conjunctival redness was 0.6. Ocular reactions had resolved completely in all animals by 1, 3, or 7 days after instillation. All corneal and iridial scores for all animals at all observation times were zero. The authors concluded that the test substance was a nonirritant. Silica dimethyl silylate ( g; 0.1 ml) was applied to one eye of New Zealand white rabbits (n = 3/sex). 22 Three of the treated eyes were not rinsed and 3 rinsed with saline after sec. The eyes were scored at 1, 24, 48, and 72 h. There were no signs of irritation at any observation period. Silica dimethyl silylate ( g) was applied to one eye of New Zealand white rabbits (n = 8; sex not provided). 47 Five of the rabbits eyes were not rinsed and 3 rinsed with saline after sec. The eyes were scored at 1, 24, 48, and 72 h and 7 days. There were no signs of irritation at any observation period. Silica dimethyl silylate (50%; 0.1 ml in olive oil) was applied to one eye of New Zealand white rabbits (n = 8; sex not provided). 47 Five of the rabbits eyes were rinsed with saline after 5 min and 3 rinsed after 24 h. The eyes were scored at 1, 24, 48, and 72 h. Conjutiva redness was scored at 1.0 at the first 3 observations and was resolved at 72 h. There were no other signs of irritation at any observation period. Silica dimethyl silylate (0.1 ml; g undiluted) was applied to one eye of New Zealand white rabbits (n = 5 males, 4 females). There was mild conjunctivae redness at 1 and 24 h which was resolved at 48 h. Panel Book Page 13

16 An EpiOcular Human Cell Construct assay was conducted of a product containing silica dimethyl silylate (2%). 48 There was no sensitization predicted. 49 Female subjects (n = 31) used an eyeliner containing silica dimethyl silylate (2%) for 13 or 14 consecutive days. There were no adverse reactions. TRIMETHYLSILOXYSILICATE Trimethylsiloxysilicate (100%; 0.1 ml) was administered to the right eye of New Zealand White rabbits (n not provided). 17 The eyes were examined at 0, 1, 24, 48, and 72 h. There were no clinical signs or signs of irritation at any observation period. A mixture of trimethylsiloxysilicate (60%) and isododecane (50% in olive oil) was reported to be practically nonirritating in 40 rabbits (n = 3). Irritation Human In a human patch test (n = 19), a mixture of trimethylsiloxysilicate (60%) and isododecane (40%) in petrolatum resulted in irritation in 2 subjects at 40% in petrolatum. 40 Sensitization The results were negative in a LLNA of trimethylsiloxysilicate up to 60%. HRIPTs of silica dimethyl silylate up to 100% were negative. NonHuman TRIMETHYLSILOXYSILICATE In as local lymph node assay (LLNA), trimethylsiloxysilicate (15%, 30%, 60% in acetone/olive oil) was dermally administered to the entire dorsal surface of each ear of mice (strain and n not provided) for 3 consecutive days. 17 The stimulation indexes were 1.0, 1.1, and 0.8 at 15%, 30%, and 60%, respectively. The authors concluded that trimethylsiloxysilicate had no reaction that was identified as sensitization. Human SILICA DIMETHYL SILYLATE A human repeated insult patch test (HRIPT) of 2 products containing silica dimethyl silylate (1.4%; 0.2 g) was conducted. 50 The products were applied neat to the back or the upper arm of subjects (n = 99) under occlusion for 24 h on alternate days for 9 applications. After a 10 15day rest, a new patch was applied to a naïve site for 24 h and observed at 24 and 48 h after removal. The authors reported that there was no evidence of sensitization by either of these products. 50 A HRIPT of a product containing silica dimethyl silylate (1.4%; 0.2 g) was conducted. The product was applied neat to the back of subjects (n = 102) under occlusion for 24 h on alternate days for 9 applications. After a 10 15day rest, a new patch was applied to a naïve site for 24 h and observed at 24 and 48 h after removal. The authors reported that there was no evidence of sensitization by this product. 51 A HRIPT of a product containing silica dimethyl silylate (2%; 0.2 g) was conducted. The product was applied neat to (n = 107) under occlusion; details of induction not provided. After a 10 15day rest, a new patch was applied to a naïve site for 24 h. The authors reported that there was no evidence of sensitization by this product. A HRIPT of a product containing silica dimethyl silylate (7%; 0.2 g) was conducted. 52 The product was applied neat to the back of subjects (n = 100) under occlusion for 24 h 3 days/week for 9 applications. After a 2week rest, a new patch was applied to a naïve site for 24 h and observed at 24 and 48 h after removal. The authors reported that there was no evidence of sensitization by this product. SUMMARY This is safety assessment of silica silylate, silica dimethyl silylate, trimethylsiloxysilicate, and trifluoropropyldimethyl/trimethylsiloxysilicate. The functions of these ingredients include: antifoaming agents, anticaking agents, bulking agents, binders, skinconditioning agentsemollient, skinconditioning agentsocclusive, slip modifiers, suspension agentsnonsurfactant, and viscosity increasing agentsnonaqueous. These grafted and cocondensed hybrid materials are amorphous and practically insoluble in most common solvents, much like unmodified silica. Silica dimethyl silylate was reported to be used in 734 cosmetic products (593 leaveon and 141 rinseoff products) at % 10%; up to 10% in leaveon products; and up to 4% in rinseoff products. There were 633 uses reported of trimethylsiloxysilicate at % 30% ; up to 30% in leaveon products; and up to 5% in rinseoff products. There were no uses reported of trifluoropropyldimethyl/trimetylsiloxysilicate but concentration of use was reported to be 2% 20% in leaveon products. No uses or concentrations of use were reported for silica silylate. Orally administered silica dimethyl silylate was eliminated from the body primarily in the feces. Inhaled silica dimethyl silylate collected in the lungs and lymph nodes of rats. In acute studies, dermally administered silica dimethyl silylate up to 2000 mg/kg and trimethylsiloxysilicate up to 2 g/kg was not toxic to rats. The oral LD 50 of silica dimethyl silylate was >7900 mg/kg for rats; trimethylsiloxysilicate had no effects at 5 g/kg. There was no mortalities from the inhalation of silica dimethyl silylate up to 520 mg/m 3. Intraperitoneally administered silica Panel Book Page 14

17 dimethyl silylate caused thickening of the liver and spleen capsules. The test substance was in observed in the abdominal cavity. Antifoam A (dimethyl silicones and siloxanes, reaction product with silica) in feed did not cause any adverse effect in rabbits up to 1% and rats up to 1000 mg/kg. Antifoam A (dimethyl silicones and siloxanes, reaction product with silica) in feed did not cause any adverse effect in rabbits up to 1% and rats up to 1000 mg/kg. The oral NOAEL for silica dimethyl silylate was 500 mg/kg for 6 months and 100 mg/kg for 24 months. Rats that inhaled treated fumed silica dust for up to 4 weeks were observed to have crusty eyes, muzzle, and nose; closed eyes; irregular breathing; irritable disposition; lacrimation and salivation; scabs; and red and yellow/brown stained fur. The inhalation NOAEL was up to 500 mg/m 3 and the LOAEL up to 1000 mg/m 3. Aerosolized silica dimethyl silylate caused mortality at 209 mg/m and respiratory distress at lower doses. There was a dosedependent reduction in weight gain, reduction in feed consumption, increase in lung weight, decrease in relative liver weights, and decrease in absolute kidney weights. There was an increase in red blood cell counts, packed cell volume and hemoglobin in the middose males and highdose males and females. Silica dimethyl silylate did not cause any reproductive or developmental toxicity to rats up to 3.8 g/kg/d, rabbits up to 1600 mg/kg, hamsters up to 1600 mg/kg, or mice up to 1340 mg/kg. Silica dimethyl silylate was not genotoxic in several Ames test and a mammalian chromosome aberration test. A product containing siloxanes and silicones, dime (dimethyl silicones and siloxanes) (18%) and dimethyl silicones and siloxane, reaction products with silica (2%) was not genotoxic in an Ames test. Trimethylsiloxysilicate was not genotoxic in an Ames test. There were no in vivo genotoxicity studies discovered. Silica dimethyl silylate at 100 mg/kg was not carcinogenic to rats. Silica dimethyl silyate and Trimethylsiloxysilicate were not demally irritating to rabbits up to 100%. In a human patch test, a mixture containing trimethylsiloxysilicate at 60% resulted in irritation in 2 out of 19 subjects. Silica dimethyl silylate was slightly or not irritating to the rabbit eye. Trimethylsioloxysilicate was practically or nonirritating to the rabbit eye. The results were negative in a LLNA of trimethylsiloxysilicate up to 60%. HRIPTs of silica dimethyl silylate up to 100% were negative. 3 Panel Book Page 15

18 Ingredient CAS No. Silica Silylate From trade name ingredients: Silica Dimethyl Silylate From trade name ingredients: Trimethylsiloxysilicate From trade name ingredients: Trifluoropropyldi methyl/ trimethylsiloxysil icate TABLES Table 1. Definitions, functions, and structures of siloxysilicates and silylates assessment. Definition Function(s) Other Names Formula/structure Silica Silylate is a hydrophobic silica derivative where some of the hydroxyl groups on the surface of the fumed silica have been replaced by trimethylsiloxyl groups. Silica Dimethyl Silylate is a silica derivative in which the surface of the fumed silica has been modified by the addition of dimethyl silyl groups. Trimethylsiloxysilicate is a variable network of polysilicic acid units, which are endblocked with trimethylsilyl groups. Trifluoropropyldimethyl / trimethylsiloxysilicate is a variable network of polysilicic acid units, which are endblocked with a mixture of trimethylsilyl groups and trifluoropropyldimethylsilyl groups. Antifoaming Agent; Bulking Agent; Skin Conditioning Agent Emollient; Suspending Agent Nonsurfactant Anticaking Agent; Bulking Agent; Slip Modifier; Suspending Agent Nonsurfactant; Viscosity Increasing Agent Nonaqueous Antifoaming Agent; Skin Conditioning Agent Occlusive Binder; Skin Conditioning Agent Emollient Hydrophobic silica; a silylated silica Silica, [(dimethylsilyl)oxy] modified; dichlorodimethylsilane treated fumed silica; hydrophobic dichlorodimethylsilane treated fumed; dichlorodimethylsilanetreated silica; silane, dichlorodimethyl, reaction products with silica; treated fumed silica dust Silicic acid, trimethylsilyl ester; hexamethyldisiloxanetetraethyl orthosilicate copolymer; poly(trimethylsiloxysilicate); tetraethoxysilanehexamethyldisiloxane copolymer; trimethylsilyl silicate See Figure 1 See Figure 1 Panel Book Page 16

19 Table 2. Physical and chemical properties of silica dimethyl silylate. Property Value Reference Silica dimethyl silylate Physical Form Fluffy powder Color White Density/Specific Gravity g/cm 20 o C 2.2 Melting Point o C >520 Water Solubility o C & ph No log K ow Does not dissolve in either solvent Table 3. Current Frequency and Concentration of Use According to Duration and Type of Exposure Provided in ,7 Silica dimethyl silylate # of Uses Concentration (%) Trimethylsiloxysilicate # of Uses Concentration (%) Trifluoropropyldimethyl/ trimethylsiloxysilicate # of Concentration Uses (%) Exposure Type Eye NR 20 Possible ingestion NR NR Inhalation NR NR NR Dermal contact NR 220 Deodorant (underarm) NR NR NR NR Hair non coloring 2 NR 13 5 NR NR Hair coloring 130 NR NR NR Nail NR NR Mucous Membrane NR NR Bath products NR NR NR NR NR NR Baby products 1 NR NR NR NR NR Duration of Use Leaveon NR 220 Rinseoff NR NR Totals***/Conc. range NR 220 NR = Not Reported; Totals = Rinseoff + Leaveon Product Uses. Note: Because each ingredient may be used in cosmetics with multiple exposure types, the sum of all exposure type uses may not equal the sum total uses. Panel Book Page 17

20 Species (n) Dose(s); duration; particle size CRL: Cd (SD)BR rats (5/sex) 0, 2280 mg/m 3 / 1 h; 0.15 µm Cpb;WU Wistar rat 0, 477 mg/m 3 / 4 h; 2.9 (5/sex) µm Wistar rats (5/sex) 210, 540, 2100 mg/m 3 ; 4 h; 0.81 µm Crl:[WI] WU BR rats (n = 5/sex; high dose group 7/sex) CD rats (n = 10/sex) Wistar rats (n = 40/sex) Wistar Cpb:WU rats (n = 10/sex) 520, 1120, 2790 mg/m 3; 4 h; 0.81 µm As CABOSIL TS720; 0, mg/l; 5 d/wk, to 1, 2, and 4 weeks; particle size not provided 0, 31, 87, 209 mg/m 3 ; for 6 h/d, 5 d/wk, 2 weeks (high dose started at 420 mg/m 3 and reduced over 4 d); particle size not provided 0, 35 mg/m 3 (calculated mg/m 3 ); 6 h/d, 5 d/wk, 13 weeks with 13, 26, 39 and 52 weeks recovery; particle size not provided Table 4. Inhalation studies of silica dimethyl silylate. Results Acute/single dose studies Clinical signs during and after treatment: irregular breathing. After treatment: poor coat quality and alopecia in females. No mortalities. Clinical signs: restless, half closed eyes. Body weights reduced first days, weight increased afterwards. No mortalities at the low dose group, 7 in the mid dose group, all the rats died in the high dose group. During exposure, clinical signs: closed eyes, labored breathing, respiratory distress, and hunched posture. Low dose group had few feces; mid dose group had lethargy, dyspnea, ptosis, piloerection, and few feces. High dose group had crusting/lacramating eyes, opaque eyes, redstained nose/mouth area, wet anogenital area, and an unkempt appearance. Necropsy: lungs were discolored at all concentrations and the mid and high dose groups had opaque eyes and white material in the nasal turbinates. None died in the low dose group; body weight gain was normal in the low dose group. All of the rats in the mid and high dose groups died. During exposure, the rats exhibited decreased, irregular breathing at all doses. After exposure, the low dose group exhibited increased breathing rates, labored breathing, and blepharospasm, all of which resolved in 4 days. At necropsy, the lungs in the low dose group were filled with foam. The mid dose group had hemorrhage and reduced elasticity in the lungs, soiled fur, and white powder in the nasal cavity. The high dose group had petechia on the lungs, blocking lumps of white particles and slime in the nose and and hemorrhage in the nasopharynx. Histopathology revealed erythrocytes and edema in alveoli, epithelial lining interrupted or flattened, and scarce goblets cells. The lumina of the nasopharynx, larynx, and brochi/bronchioli contained large quantities of paleeosinophilic material mixed with nucleated cells and erythrocytes. The material filled the entire lumen in the smaller bronchioli. Multiple dose Studies The original dose was 0.06 mg/l air, but was reduced after 9 male rats died on the first day of treatment. All treated animals were observed to have crusty eyes, muzzle, and nose; crust around ear tags; closed eyes; irregular breathing; irritable disposition; lacrimation and salivation; scabs; and red and yellow/brown stained fur. There were no differences in body weights among groups and the recovery animals gained weight at the same rate as the controls. There was an increase in lymphocytes and neutrophils after 2 and 4 weeks and an increase in leukocytes at 4 weeks. There were no differences observed in serum chemistry. Histological examination results were consistent with chronicactive pulmonary inflammatory process. 4 males and 2 females died within 2 days in highdose group. Rats in this group had severe respiratory distress and apathy. After reduction to 209 mg/m 3, clinical signs: slight to moderate respiratory distress and poor general health. Mid dose group: Dyspnea. Dosedependent reduction in weight gain, reduction in feed consumption, increase in lung weight, decrease in relative liver weights, and decrease in absolute kidney weights. Increase in red blood cell counts, packed cell volume and hemoglobin in the middose males and highdose males and females. Lungs had focal bronchiolar mucus proliferation, intralunimal mucus deposition, granulomata, focal increased septal celluarlaity, and accumulation of alveolar macrophages. In rats that died, perivascular edema, alveolar edema, and hemorrhages along with slight bronchiolar necrosis. The LOAEL was 31 mg/m 3. An additional group of 50 rats were treated and allowed post recovery times of 12, 26, 39, and 52 weeks. 3 rats died for unrelated reasons during treatment. Decreased body weights in males weeks 6 9. At 13 weeks, males had increased red blood cells, hemoglobin content, packed cell volumes and prothrombine time. At the end of treatment and 13 weeks recovery, females had increased neutrophils and decreased lymphocytes. Females decreased urinary volume with associated increased density. Increased lung weights (absolute and relative) in males and females until week 39 postexposure and at week 13 of exposure. Increased absolute thymus weights in males. At week 13, males and females had lesions of the lungs (including spotted, irregular or gray surface and spongy tissue) and enlarged mediastinal lymph nodes. Reference Panel Book Page 18

21 Female Sprague Dawley rats (80) Rats (n = 340) Female rats (n = 235) 50 mg/m 3 ; 5 h/d, twice/wk, 8 or 12 months with 0 5 months recovery; <7 µm 100 mg/m 3 ; 5 h/d, 5 d/wk for 1 yr, 3 or 6 months recovery; particle size not provided 0, 80 mg/m 3 ; 4 h/d 1 yr; particle size not provided At week 13 of exposure, lungs had granulomalike lesions, accumulations of alveolar macrophages, alveolar spaces filled with granular material, debris and polymorphonuclear leucocytes, increased septal cellularity, alveolar bronchiolization and interstitial fibrosis. Mediastinal lymph nodes characterised by accumulation of macrophages. Slight necrosis or atrophy of the olfactory in the nose. Changes in the lungs and mediastinal lymph nodes decreased in incidence and severity at 13 weeks post exposure or had completely disappeared at 52 weeks post exposure. Nose [sic] was recovered at week 13 post exposure. No effects on male and female gonads. Collagen content of the lungs increased in males and females at week 13 of exposure and, at 13 and 39 weeks post exposure. Silicon levels in the lungs as well as in the mediastinal lymph nodes increased in males and females at week and 12 Month post exposure: Interstitial white dust deposits and slightly enlarged lymph nodes. Lung had many dust cells in alveoli, locally perivascular and peribronchiolar dust cell deposits with slight to moderate formation of fibrous tissue. Lymph nodes increased in number of granular phagocytes and local fibrosis. Post recovery period: Interstitial greywhite dust deposits, more at 5 months; moderately enlarged greyblack lymph nodes after 1 month, smaller after 3 and 5 months. Lungs had slight epithelial desquamation up to 1 month; locally perivascular and peribronchiolar dust cell deposits with slight to moderate formation of fibrous tissue; thinkening of part of the alveolar wall. Lymph nodes had increased number of granular phagocytes and local fibrosis. Signs of recovery from 1 5 months. All treated rats had small greywhite dust foci under the lung surface, particularly in the upper lung lobes. Mediastinal lymph nodes were slightly to moderately enlarged after a period of 3 months exposure. After 9 months of exposure these lymph nodes also had a greyblack appearance. In rats found dead, adhesion of the pleura, inflamatory cell infiltrations and lung abscesses. After 3 or 6 month post exposure period a timedependent reduction of the number of greywhite dustfoci was observed in the lungs; mediastinal lymph nodes were reduced in size (compared to during the exposure period) and had a grey/black and soft appearance. At 3, 6 and 12 months of exposure increasing incidences of desquamous alveolar cells (with and without dust content), foci of dust cells (in bronchioli, peribrochiolar and perivascular) with increasing (in time) number of dust granulas and cell detritus in the alveolar space (12 months) in the lungs. Mediastinal lymph nodes had increasing number of dust cells containing higher numbers of dust granulas (312 months exposure). A reticulin network developed with increased exposure times in lungs. No signs of proliferation, fibrosis, or necrosis in the lungs or mediastinal lymph nodes. At 3 or 6 month post exposure period, lungs had groups of alveoli containing accumulations of dust cells, but no desquamous alveolar cells. Peribronchiolar and perivascular small nodules were noted, without proliferation, necrosis or fibrosis. Number and size of the dust cell foci were reduced. Mediastinal lymph nodes contained large amounts of dust cells after 3 and 6 months. Fine reticulin network was visible with no connective tissue. 60 rats in the treatment group died during treatment due to bronchiopneumonia, bronchoectasia, and abscess in the lungs but were not considered substancespecific. At 3 months, the rats had dust cell granulomata in the lungs and alveolar spaces filled with dust cells and desquamous alveolar cells. Mediastinal lymph nodes were enlarged and filled with dust cells. In the control group, 41 of 120 rats died spontaneously. At 1 year, small greywhite foci under the pleura and moderately enlarged mediastinal lymph nodes were observed. Dust accumulation did not cause fibrotic responses during the 3, 5, and 8 month recovery period and the dust amount decreased over time. There were no indications of silicosis Panel Book Page 19

22 REFERENCES 1. Becker LC, Bergfeld WF, Belsito DV, Hill RA, Klaassen CD, Liebler DC, Marks Jr.JG, Shank RC, Slaga TJ, and Snyder PW. Safety assessment of silica and related cosmetic ingredients. Washington, DC, USA, Cosmetic Ingredient Review pp Unpublished report by the Cosmetic Ingredient Review. 2. Hoffmann F, Cornelius M, Morell J, and Fröba M. SilicaBased Mesoporous Organic Inorganic Hybrid Materials. Angew.Chem.Int.Ed. 2010; WackerChemie GmbH. Gewerbehygienisches Gutachten über die ochdisperse "reaction products of dichlorodimethyl silane with silica" (inhalation study rat) , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 4. WackerChemie GmbH. Particle analysis of pyrogenic (fumed) silicas at technical concentrations and under technical handling conditions Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Fuji M, Iwata H, Takei T, Watanabe T, and Chikazawa M. The change in the water vapor affinity of fine silica particles loaded with trimethylsilyl groups. Advanced Powder Technol. 1997;8(4): Food and Drug Administration (FDA). Frequency of use of cosmetic ingredients. FDA Database Washington, DC: FDA.Data obtained by FOIA request from FDA. 7. Personal Care Products Council Concentration of use by FDA product category silica demethyl silylate, trimethylsiloxysilicate and trifluoropropyldimethyl/trimethylsiloxysilicate. Unpublished data submitted by the Personal Care Products Council. 8. James, A. C., Stahlhofen, W, Rudolf, G, Kobrich, R, Briant, J. K., Egan, M. J., Nixon, W, and Birchall, A. Annexe D. Deposition of inhaled particles. Annals of the ICRP. 1994;24(13): Oberdorster, G, Oberdorster, E, and Oberdorster, J. Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles. Environmental Health Perspectives. 2005;113(7): Bower, D Information on hair spray particle sizes provided at the September 9, 1999 CIR Expert Panel meeting. Unpublished data from CIR Expert Panel Meeting. 11. Johnson, M. A. The Influence of Particle Size. Spray Technology and Marketing. 2004;November Food and Drug Administration (FDA). Anticaking agents: Silicon dioxide U.S. Code of Federal Regulations:21 CFR : Food and Drug Administration (FDA). Substances Generally Recognized as Safe: Silica aerogel CFR : Dow Corning Corp. Pharmacokinetic and metabolic studies on Dow Corning Antifoams A and M in mice, monkeys and humans with cover letter dated 04/20/ Report No. 1974I pp Unpublished data submitted to the Environmental Protection Agency. 15. Degussa AG. Gewerbehygienischtoxicologische Untersuchung der Wesselinger hydrophoben "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 16. Research & Consulting Co, AG. Acute dermal toxicity (LD50) in rats with cover letter dated Report No pp Unpublished data submitted to the Environmental Protection Agency. 17. Momentive. Toxicology summary for SR1000 (trimethylsiloxysilicate) pp. 14. Unpublished data submitted by the Personal Care Products Council. 18. Cabot Corporation. Reaction products of dichlorodimethyl silane with silica, Lot # 6C264 Acute toxicity limit test , , , , , Report No. Study 1. Data submitted to High Product Volume Information System of the US EPA. 19. Degussa AG. Prüfung der akuten Toxizität von "reaction products of dichlorodimethyl silane with silica" an SpragueDawley Ratten bei peroraler Verabreichung Panel Book Page 20

23 2, , , , , Report No. Study 2. Study submitted to the High Production Volume Information System of the US EPA. 20. Degussa AG. Prüfung der akuten Toxizität von "reaction products of dichlorodimethyl silane with silica" bei peroraler Verabreichung an SpragueDawley Ratten , , , , , Report No. Study 3. Study submitted to the High Production Volume Information System of the US EPA. 21. Degussa AG. Gewerbehygienischexperimentelle Untersuchungen mit "reaction products of dichlorodimethyl silane with silica" , , , , , Study submitted to the High Production Volume Information System of the US EPA. 22. Cabot Corporation. Primary Eye Irritation Unpublished data submitted to the High Production Volume Information System of the US EPA. 23. Dow Corning Corp. Chronic (8month) feeding studies with Antifoam A in rabbtis with cover letter dated Report No. 1965I pp Unpublished data submitted to the Environmental Protection Agency. 24. Food & Drug Research Labs, Inc. Chronic (1year) feeding studies with Antifoam A in rats with cover letter dated Report No. 1966I pp Unpublished data submitted to the Environmental Protection Agency. 25. Degussa AG. Gewerbehygienischexperimentelle Untersuchungen mit "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 26. Degussa AG. Uber die subakute Toxizitat von "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 27. Degussa AG. Uber die chronische Vertraglichkeit von "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 28. Degussa AG. Betrift der Ergebnisse der langfristigen oralen Verabreichung von Kieselsaure (reaction products of dichlorodimethyl silane with silica) der Firma DEGUSSA, entsprechend dem product "R." unseres Institutes , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 29. Atlas Chemical Indus, Inc. Dow Corning Antifoam A (medical grade): A teratogenic potential study in rats with cover letter dated 04/20/ Report No. 1970I pp Unpublished data submitted to the Environmental Protection Agency. 30. Dow Corning Corp. Teratogenic potential of Dow Corning Antifoam A compound, food grade in rabbtis, with cover letter dated 4/20/ Report No. 1984I pp Unpublished data submitted to the Environmental Protection Agency. 31. Food & Drug Research Labs, Inc. Teratologic Evaluation of FDA 7148 (Syloid; silica aerogel). Maspeth NY, Report No. Reports perpared under DHEW Contract No. FDA pp Study submitted to the High Production Volume Information System of the US EPA. 32. Lewinson J. Characterization and toxicological behavior of synthetic amorphous hydrophobic silica. Regulatory Toxicology and Pharmacology. 1994; Degussa AG. Uber die chronische Toxizität von AEROSIL Report No. USIT No DKT. Study submitted to the High Production Volume Information System of the US EPA. 34. Degussa Corp. Bacterial mutagenicity test on a toluene extract from Aerosil R 202 with cover letter dated 08/30/ Report No. SP 598. pp Unpublished data submitted to the Environmental Protection Agency. 35. Dow Corning Corp. Genetic evaluation of Dow Corning 1520 Silicone antifoam inthe Ames bacterial assay, with cover letter dated 4/20/ Report No. 1983I pp Unpublished data submitted to the Environmental Protection Agency. 36. Huntingdon Life Sciences LTD. Bacterial reverse mutation assay of Dow Corning AF Emulsion food grade, with cover letter dated 10/30/ Report No. 1998I pp Unpublished data submitted to the Environmental Protection Agency. Panel Book Page 21

24 37. Cabot Corporation. Salmonella Plate Incorporation Mutagenicity Assay (Ames Test) reaction products of dichlorodimethyl silane with silica Study submitted to the High Production Volume Information System of the US EPA. 38. Degussa AG. Bacterial Mutagenicity Test on a Toulene Extract from reaction products of dichloromethyl silane with silica Study submitted to the High Production Volume Information System of the US EPA. 39. Cabot Corporation. Chromosome Aberrations in Chinese Hamster Ovary (CHO) Cells reaction products of dichlorodimethyl silane with silica Study submitted to the High Production Volume Information System of the US EPA. 40. Personal Care Products Council. Summary of information on a mixture containing trimethylsiloxysilcate pp. 11. Unpublished data submitted by the Personal Care Products Council. 41. Research & Consulting Co, AG. Primary skin irritation study with Aerosil R 202 in rabbits (4hr occlusive application) with ocver letter dated Report No pp Cabot Corporation. Primary skin irritation Unpublished data submitted to the High Production Volume Information System of the US EPA. 43. Degussa AG. Lokale Verträglichkeit von Testsubstanz an der Kaninchenhaut (Patchtest) Study submitted to the High Production Volume Information System of the US EPA. 44. Huntingdon Life Sciences LTD. Skin irritation study of Dow Corning AF emulsion food grade (contains 25 wt.% of dimethyl silicones and siloxanes) in the rabbit, with cover letter dated 08/04/ Report No. 1999I pp Unpublished data submitted to the Environmental Protection Agency. 45. Research & Consulting Co, AG. Primary irritation in rabbits with cover letter dated Report No pp Unpublished data submitted to the Environmental Protection Agency. 46. Huntingdon Life Sciences LTD. Eye irritation study of Dow Corning AF Emulsion food grade (contians 25wt.% of dimethyl silicones and siloxanes) in the rabbit, with cover letter dated 08/04/ Report No. 1998I pp Unpublished data submitted to the Environmental Protection Agency. 47. Degussa AG. Schleimhautverträglichkeit am Kaninchenauge von Testsubstanz bei einmaliger Applikation Study submitted to the High Production Volume Information System of the US EPA. 48. Institute for In Vitro Sciences, Inc. Topical application ocular irritation screening assay using the Epiocular hman cell construct (eyeliner containing 2% silica dimethyl silylate) Report No. 10AF14AF pp Unpublished data submitted by the Personal Care Products Council. 49. Consumer Product Testing Co. Ophthalmological inuse safety evaluation of an eyeliner containing 2% silica dimethyl silylate Report No. C pp Unpublished data submitted by the Personal Care Products Council. 50. TKL Research. Human repeated insult patch study on two antiperspirants containing 1.4% silica dimethyl silylate Report No. DS pp Unpublished data submitted by the Personal Care Products Council. 51. TKL Research. Repeated insult patch test of an eyeliner containing 2% silica dimethyl silylate Report No. DS pp Unpublished data submitted by the Personal Care Products Council. 52. Consumer Product Testing Co. Exclusive repeated insult patch test on a lipstick containing 7% silica dimethyl silylate Report No. C pp Unpublished data submitted by the Personal Care Products Council. 53. Cabot Corporation. Safety data sheet Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised WackerChemie GmbH. Determination of physiocochemical properties Wacker reaction products of dichlorodimethyl silane with silica Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Cabot Corporation. One hour acute dust inhalation toxicity study in rats of Degussa Surface treated Fumed Silica Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Degussa AG. Acute inhalation toxicity study of reaction products of dichlorodimethyl silane with silica in rats Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Panel Book Page 22

25 57. Cabot Corporation. Inhalation toxicity in rats, reaction products of dichlorodimethyl silane with silica Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Cabot Corporation. Acute (4hour) inhalation toxicity study with reaction products of dichlorodimethyl silane with silica in rats Submitted to International Uniform Chemical Information Database (IUCLID) Data Set for ID: ; revised Toxigenics, Inc. Initial submission: 28day dust inhalation toxicity study of CabOSil N70TS in albino rats with cover letter dated Report No pp Unpublished data submitted to the Environmental Protection Agency. 60. Degussa AG. A subacute (14day) inhalatin toxicity study of reaction products of dichlorodimethl silane with silica in rats , , , , , , Report No. Study 2. Study submitted to the High Production Volume Information System of the US EPA. 61. Degussa AG. Subchronic (13week) inhalation toxicity study of aerosols of "reaction products of dichlorodimethyl silane with silica" and Quartz in rats , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 62. Degussa AG. Gewerbehygienischtoxicologische Untersuchung der Wesselinger hydrophoben "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. 63. Degussa AG. Gewerbehygienischexperimentelle Untersuchungen mit "reaction products of dichlorodimethyl silane with silica" , , , , , , Study submitted to the High Production Volume Information System of the US EPA. Panel Book Page 23

26 Unpublished Data Unpublished data have been submitted by the Council and discovered on line. This information has been incorporated into the benzoate report. The following is a list of the types of data in the order that they are presented in this section. 1 Concentration of Use survey by PCPC 2 Ocular irritation, Ames assay, and human patch test submitted by PCPC 3 HRIPTS of an antiperspirant containing silica dimethyl silylate submitted by PCPC 4 Various tests of trimethylsiloxysilicate submitted by PCPC 5 Various tests of products containing silica dimethyl silylate submitted by PCPC 6 Various tests of silica dimethyl silylate submitted to the Office of pollution Prevention and Toxics of the EPA. These are abridged in the Panel books. Full versions containing raw data, consent forms, etc. are available in the on line version. Panel Book Page 24

27 Personal Care Memorandum Products Counci Committed to Safety, Quality & Innovation TO: FROM: F. Alan Andersen, Ph.D. Director COSMETIC 1]NTGREDIENT REVIEW (CW) John Bailey, Ph.D. Industry Liaison to the CIR Expert Panel DATE: August 11, 2010 SUBJECT: Updated Concentration of Use Silica Dimethyl Silyate and Trimethylsiloxysilicate th Street, N.W., Suite 300 Washington, D.C (fax) Panel Book Page 25

28 Concentration of Use by FDA Product Category Silica Dimethyl Silylate, Trimethylsiloxysilicate and TrifluoropropyldimethyllTrimethylsiloxysilicate Ingredient Product Category Concentration of Use Silica Dimethyl Silylate Eyeliner 12% Silica Dimethyl Silylate Eye shadow % Silica Dimethyl Silylate Eye lotion 0.03% Silica Dimethyl Silylate Mascara % Silica Dimethyl Silylate Colognes and toilet waters 0.5% Silica Dimethyl Silylate Perfumes 3% Silica Dimethyl Silylate Powders (dusting and talcum) 4% Silica Dimethyl Silylate Other fragrance preparations 3% Silica Dimethyl Silylate Blushers (all types) % Silica Dimethyl Silylate Face powders 0.022% Silica Dimethyl Silylate Foundations 0.36% Silica Dimethyl Silylate Lipstick 110% Silica Dimethyl Silylate Makeup bases 1% Silica Dimethyl Silylate Other makeup preparations 0.92% Silica Dimethyl Silylate Nail polish and enamel 2% Silica Dimethyl Silylate Other manicuring preparations 0.002% Silica Dimethyl Silylate Deodorants (underarm) % Silica Dimethyl Silylate Other personal cleanliness products % Silica Dimethyl Silylate Face and neck creams, lotions and 0.085% powders Silica Dimethyl Silylate Body and hand creams, lotions and 0.033% powders Silica Dimethyl Silylate Moisturizing creams, lotions and powders 12% Page 1 of 3 Panel Book Page 26

29 Silica Dimethyl Silylate Night creams, lotions and powders % Silica Dimethyl Silylate Skin fresheners 0.1% Silica Dimethyl Silylate Other skin care preparations % Silica Dimethyl Silylate Suntan gels, creams and liquids 0.5% Silica Dimethyl Silylate Indoor tanning preparations 0.9% Trifluoropropyldimethyl! Eyeliner 20% Trimethylsiloxysilicate Trifluoropropyldimethyl! Foundations 10% Trimethylsiloxysilicate Trifluoropropyldimethyl/ Suntan gels, creams and liquids 2% Trimethylsiloxysilicate Trimethylsiloxysilicate Eyebrow pencil 612% Trimethylsiloxysilicate Eyeliner 430% Trimethylsiloxysilicate Eye shadow 120% Trimethylsiloxysilicate Eye lotion 0.45% Trimethylsiloxysilicate Mascara 425% Trimethylsiloxysilicate Other eye makeup preparations 7% Trimethylsiloxysilicate Powders (dusting and talcum) % Trimethylsiloxysilicate Hair straighteners 5% Trimethylsiloxysilicate Hair dyes and colors (all types requiring 0.1% caution statement and patch test) Trimethylsiloxysilicate B lushers (all types) 0.77% Trimethylsiloxysilicate Face powders 319% Trimethylsiloxysilicate Foundations 0.513% Trimethylsiloxysilicate Leg and body paints 4% Trimethylsiloxysilicate Lipstick 230% Trimethylsiloxysilicate Makeup bases 2% Trimethylsiloxysilicate Other makeup preparations 320% Page 2 of 3 Panel Book Page 27

30 Trimethylsiloxysilicate B asecoats and undercoats (manicuring 0.02% preparations) Trimethylsiloxysilicate Nail polish and enamel % Trimethylsiloxysilicate Bath soaps and detergents % Trimethylsiloxysilicate Other shaving preparations 0.7% Trimethylsiloxysilicate Skin cleansing (cold creams, cleansing 0.5% lotions, liquids and pads) Trimethylsiloxysilicate Face and neck creams, lotions and 0.42% powders Trimethylsiloxysilicate Body and hand creams, lotions and 0.13% powders Trimethylsiloxysilicate Body and hand sprays 0.4% Trimethylsiloxysilicate Moisturizing creams, lotions and powders 0.6% Trimethylsiloxysilicate Night creams, lotions and powders 0.4% Tnmethylsiloxysilicate Other skin care preparations 0.213% Tnmethylsiloxysilicate Suntan gels, creams and liquids % Tnmethylsiloxysilicate Indoor tanning preparations % Information collected in 2010 Table prepared July 21, 2010 Updated August 11, 2010 Page 3 of 3 Panel Book Page 28

31 COSMETIC Personal Care Memorandum Products Council Committed to Safety, Quality & Innovation TO: FROM: DATE: SIJEJECT: F. Alan Andersen, Ph.D. Director RGRED1ENT REVIEW (CIR) John Bailey, Ph.DZZZ._O.. F ( J i Industry Liaison to the CW Expert Panel September 9, 2010 Summary of Information on a Mixture Containing Trimethylsiloxysilicate Trimethylsiloxysilicate 60% and Isododecane 40% (more than 99% 2,2,4,6,6 Pentamethyiheptane) Ocular irritation Practically non irritant: mixture tested at 50% in olive oil in 3 rabbits Ames assay Negative: tested in strains TA 100 and TA98 at doses of jig/plate Human patch Irritation observed in 2/19 in a 48 hour closed patch test of 40% in petrolatum th Street, N.W., Suite 300 Washington, D.C (fax) Panel Book Page 29

32 COSMETIC Personal Care Products Council Committed to Safety, Quality & Innovation Memorandum TO: FROM: F. Alan Andersen, Ph.D. Director 1NGREDIENT REViEW (CIR) John Bailey, Ph.D. Industry Liaison to the CIR Expert Panel DATE: September 24, 2010 SUBJECT: HRIPTs of Antiperspirants Containing Silica Dimethyl Silylate TKL Research Human repeated insult patch study on two antiperspirants containing 1.4% Silica Dimethyl Silylate. TKL Study No. DS TKL Research Human repeated insult patch study a solid antiperspirant containing 1.4% Silica Dimethyl Silylate. TKL Study No. DS th Street, N.W., Suite 3O0 Washington, D.C (fax) Panel Book Page 30

33 III RESEARCH HUMAN REPEATED INSULT PATCH STUDY TKL STUDY NO. DS )o cvu).p 1h OrFQpr $0 JL/Oj CONDUCTED FOR: DATE OF ISSUE: October 17, 2007 I TKL Clinical Trials Division TKL Clinical Site Division N TKL Recruitment Management Division Corporate Office: 365 W. Passaic Street, Rochelle Park, NJ Phone wwwtklresearch.com Panel Book Page 31

34 TKL Study No. DS TABLE OF CONTENTS SIGNATURES I STATEMENT OF QUALITY ASSURANCE I TITLE OF STUDY 2 SPONSOR 2 STUDY MATERIAL 2 DATE STUDY INITIATED 2 DATE STUDY COMPLETED 2 DATE OF ISSUE 2 INVESTIGATIVE PERSONNEL 2 CLINICAL SITE 2 SUMMARY OBJECTIVE RATIONALE STUDY DESIGN STUDY POPULATION Inclusion Criteria Exclusion Criteria Informed Consent DESCRIPTION OF STUDY Outline of Study Procedures Definitions Used for Grading Responses Evaluation of Responses NATURE OF STUDY MATERIAL STUDY MATERIAL SPECIFICATIONS STORAGE, HANDLING, AND DOCUMENTATION OF STUDY MATERIAL APPLICATION OF STUDY MATERIAL DESCRIPTION OF PATCH CONDITIONS INTERPRETATION DOCUMENTATION AND RETENTION OF DATA RESULTS & DISCUSSION CONCLUSION REFERENCES 9 APPENDICES I SUMMARY TABLES II DATA LISTINGS III INFORMED CONSENT DOCUMENT Panel Book Page 32

35 TKL Study No. DS SIGNATURES Jq4tan S. Dosik, MD Dermatologist Principal liwestigator Date Kathleen Georgeian, Clinical Research Coordlator and Director, Dermatologic Safety Testing Date STATEMENT OF QUALITY ASSURANCE This report has been reviewed by the TKL Research, Inc. (TKL) Corporate Quality Assurance Department and the report accurately reflects the raw data for this study. Clinical research studies are performed by TKL in accordance with all applicable federal regulations and proposed guidelines for Good Clinical Practices which include: 21 CFR Part 312, Investigational New Drug Application 21 CFR Part 50, Protection of Human Subjects 21 CFR Part 56, Institutional Review Boards Quality Assurance Date Panel Book Page 33

36 TKL Study No. DS TITLE OF STUDY Human Repeated Insult Patch Study SPONSOR STUDY MATERIAL Test Material B Description Antiperspirant (Ot F, 5 ): c C Antiperspirant (_, iici, r5 /l O/ ç1 lc i/4.? 9frky/ DATE STUDY INITIATED Si August 13, 2007 DATE STUDY COMPLETED September 21, 2007 DATE OF ISSUE October 17, 2007 INVESTIGATIVE PERSONNEL Jonathan S. Dosik, MD Dermatologist Principal Investigator Kathleen Georgeian Clinical Research Coordinator and Director, Dermatologic Safety Testing Michelle Medina Assistant Clinical Research Coordinator CLINICAL SITE TKL RESEARCH, INC. 169 Wythe Avenue Brooklyn, NY Panel Book Page 34

37 TKL Study No. DS SUMMARY Two test materials, B and C, were evaluated neat to determine their ability to sensitize the skin of normal volunteer subjects using an occlusive repeated insult patch study. Ninety nine (99) subjects completed the study. Under the conditions employed in this study, there was no evidence of sensitization to Test Materials B and C. Panel Book Page 35

38 TKL Study No. DS OBJECTIVE The objective of this study was to determine the ability of the study material to cause sensitization by repeated topical applications to the skin of humans under controlled patch study conditions. 2.0 RATIONALE Substances that come into contact with human skin need to be evaluated for their propensity to irritate and/or sensitize. Once an appropriate preclinical safety evaluation has been performed, a reproducible, standardized, quantitative patch evaluation procedure must be used to demonstrate that a particular material can be applied safely to human skin without significant risk of adverse reactions. The method herein employed is generally accepted for such a purpose. Repeated insult patch evaluation is a modified predictive patch study that can detect weak sensitizers that require multiple applications to induce a cellmediated (Type IV) immune response sufficient to cause an allergic reaction. Irritant reactions may also be detected using this evaluation method, although this is not the primary purpose of this procedure. Results are interpreted according to interpretive criteria based upon published works, as well as the clinical experience of TKL Research, Inc. These interpretive criteria are periodically reviewed and amended as new information becomes available. 3.0 STUDY DESIGN 3.1 STUDY POPULATION A sufficient number of subjects were to be enrolled to provide 100 completed subjects Inclusion Criteria Individuals eligible for inclusion in the study were those who: 1. were males or females, 18 years of age or older, in general good health; 2. were free of any systemic or dermatologic disorder which, in the opinion of the investigative personnel, would have interfered with the study results or increased the risk of adverse events; 3. were of any skin type or race providing the skin pigmentation would allow discernment of erythema; 4. had completed a medical screening procedure; and 5. had read, understood and signed an informed consent agreement. Panel Book Page 36

39 TKL Study No. DS Exclusion Criteria Individuals excluded from participation in the study were those who: 1. had any visible skin disease at the study site which, in the opinion of the investigative personnel, would have interfered with the evaluation; 2. were receiving systemic or topical drugs or medication which, in the opinion of the investigative personnel, would have interfered with the study results; 3. had psoriasis and/or active atopic dermatitis/eczema; 4. were females who were pregnant, planning to become pregnant during the study, or breastfeeding; and/or 5. had a known sensitivity to cosmetics, skin care products, or topical drugs as related to the material being evaluated Informed Consent A properly executed informed consent document in compliance with FDA regulations (21 CFR Part 50) was obtained from all subjects prior to entering the study. The signed informed consent document is maintained in the study file. In addition, the subjects were provided with a copy of the informed consent document (see Appendix III). 3.2 DESCRIPTION OF STUDY Outline of Study Procedures Subjects participated in the study over a 6week period involving 3 phases: (1) Induction, (2) Rest, and (3) Challenge. Prior to study entry, the subjects were screened to assure that they met the inclusion/exclusion criteria. Informed consent was obtained. Each subject was provided with a schedule of the study activities. All subjects were told to avoid wetting the patches and were asked not to engage in activities that caused excessive perspiration. They were instructed to notify the staff if they experienced any discomfort beyond mild itching or observed any adverse changes at the evaluation sites while on the study or within 2 weeks of completing the study. The Induction Phase consisted of 9 consecutive applications of the study material and subsequent evaluations of the application sites. Prior to application of the patches, the sites were outlined with a skin marker, eg, gentian violet. The subjects were required to remove the patches approximately 24 hours after application. They returned to the facility at 48hour intervals to have the sites evaluated and identical patches applied to the same sites. Patches applied on Friday were removed by subjects after 24 hours and sites were evaluated on the following Monday, ie, 72 hours after patch application. Following the ninth evaluation, the subjects were dismissed for a rest period of approximately 1015 days. * A Monday or Friday holiday could result in evaluation at 96 hours after patch application. Panel Book Page 37

40 TKL Study No. DS Subjects who were absent once during the 3week, 9patch induction phase received a makeup (MU) patch at the last induction visit. The MU applications were graded 48 hours later at the MU visit, or were recorded as N9G (no ninth grading). The Challenge Phase was initiated during the sixth week of the study. identical patches were applied to sites previously unexposed to the study material. The patches were removed by subjects after 24 hours and the sites graded after additional 24hour and 48hour periods (ie, 48 and 72 hours after application). Rechallenge was performed whenever there was evidence of possible sensitization. To be considered a completed case, a subject must have had 9 applications and no less than 8 subsequent readings during induction and a single application and 2 readings during challenge. Only completed cases were used to assess sensitization Definitions Used for Grading Responses The symbols found in the data listings accompanying this report were used to express the response observed at the time of examination: SYMBOL AND RESPONSE = No reaction? = Minimal or doubtful response, slightly different from surrounding normal skin + = Definite erythema, no edema ++ = Definite erythema, definite edema = Definite erythema, definite edema and vesiculation SPECIAL NOTATIONS F = Marked/severe erythema S = Spreading of reaction beyond patch site (ie, reaction where material did not contact skin) p = Papular response > 50% pv = Papulovesicular response> 50% D = Damage to epidermis: oozing, crusting and/or superficial erosions = Itching X = Subject absent PD = Patch dislodged NA = Not applied NP = Not patched (due to reaction achieved) N9G = No ninth grading Evaluation of Responses All responses were graded by a trained dermatologic evaluator meeting TKL s strict certification requirements to standardize the assignment of response grades. Panel Book Page 38

41 TKL Study No. DS NATURE OF STUDY MATERIAL 4.1 STUDY MATERIAL SPECIFICATIONS Identification : B antiperspirant Amount Applied : 0.2 g Special Instructions Patches used within 30 minutes of dosing. Identification : C antiperspirant Amount Applied : 0.2 g Special Instructions : Patches used within 30 minutes of dosing. 4.2 STORAGE, HANDLING, AND DOCUMENTATION OF STUDY MATERIAL Receipt of the material used in this study was documented in a general logbook, which serves as a permanent record of the receipt, storage, and disposition of all study material received by TKL. On the basis of information provided by the sponsor, the study material was considered reasonably safe for evaluation on human subjects. A sample of the study material was reserved and will be stored for a period of 6 months. All study material is kept in a locked product storage room accessible to clinical staff members only. At the conclusion of the clinical study, the remaining study material was discarded and the disposition documented in the logbook. 4.3 APPLICATION OF STUDY MATERIAL Study material was applied to patch as instructed and patch was applied to the infrascapular area of the back, either to the right or left of the midline, or to the upper arm. 4.4 DESCRIPTION OF PATCH CONDITIONS Material evaluated under occlusive patch conditions is applied to a 2 cm x 2 cm Webril pad attached to a nonporous, plastic film adhesive bandage (3M medical tape). The patch is secured with hypoallergenic tape (Micropore), as needed. Material evaluated under semiocclusive patch conditions is applied to a 2 cm x 2 cm Webril pad. pad is affixed to the skin with hypoallergenic tape (Micropore). The 5.0 INTERPRETATION Sensitization is characterized by an acute allergic contact dermatitis. Typical sensitization reactions begin with an immunologic response in the dermis resulting in erythema, edema formation, and secondary epidermal damage (vesiculation), sometimes extending beyond the patch site and often accompanied by itching. Sensitization reactions tend to be delayed. The reaction typically becomes evident between 24 and 48 hours, peaks at 4872 hours and subsequently subsides. The reaction is often greater at 72 hours than at 48 hours. The severity of the reaction is generally greater during the challenge phase of an RIPT than that seen during induction. Panel Book Page 39

42 TKL Study No. DS Irritant reactions are characterized as a nonimmunologic, localized, superficial, exudative, inflammatory response of the skin due to an externally applied material. The typical initial reaction does not develop much edema or vesiculation but results in scaling, drying, cracking, oozing, crusting, and erosions. The reaction is usually sharply delineated, not spreading beyond the patch site. Irritant reactions are typically evident by 24 hours and diminish over the next 4872 hours. Removal of the offending agent results in gradual improvement of the epidermal damage. The reaction seen at 72 hours is, therefore, less severe than that seen at 48 hours. Finally, the severity of the reaction experienced in the challenge phase is generally similar to that seen during induction. If the results of the study indicate the likelihood of sensitization, the recommended practice is to rechallenge the subjects who have demonstrated sensitizationlike reactions to confirm that these reactions are, indeed, associated with the product. Our preferred rechallenge procedure involves the application of the product to naïve sites, under both occlusive and semiocclusive patch conditions. Use of the semiocclusive patch condition helps to differentiate irritant and sensitization reactions. Generally speaking, if a product is a sensitizer it will produce a similar reaction under both occlusion and semiocclusion. Whereas, if the product has caused an irritant reaction, the reactions will be less pronounced under the semiocclusive condition. 6.0 DOCUMENTATION AND RETENTION OF DATA The case report forms (CRFs) were designed to identify each subject by subject number and initials and to record demographics, examination results, adverse events, and end of study status. Originals or copies of all case report forms, correspondence, study reports, and all source data will be kept on hardcopy file for a minimum of 5 years from completion of the study. Storage is maintained either at a TKL facility in a secured room accessible only to TKL employees, or at an offsite location which provides a secure environment with burglar/fire alarm systems, camera detection, and controlled temperature and humidity. Documentation will be available for the sponsor s review on the premises of TKL Research, Inc. 7.0 RESULTS & DISCUSSION One hundred six (106) subjects between the ages of 18 and 70 were enrolled and 99 subjects completed the study. (See Tables 1 and 2 in Appendix I and Data Listings 1 and 2 in Appendix II.) The following table summarizes subject enrollment and disposition. Number enrolled: 106 Number discontinued: 7 Lost to followup: 1 Voluntary withdrawal: 6 Number completed: 99 Source: Table 1, Appendix I There were no adverse events reported. Panel Book Page 40

43 TKL Study No. DS One protocol deviation occurred: 99 subjects completed the study, a deviation from the required 100 subjects. A summary of response data is provided in Table 3, Appendix I. grades are provided in Data Listing 3, Appendix II. Individual dermatological response 8.0 CONCLUSION Under the conditions employed in this study, there was no evidence of sensitization to Test Materials B and C. 9.0 REFERENCES Kligman AM. The identification of contact allergens by human assay II. J Invest Dermatol 1966; 47:369. A critique of standard methods. Kligman AM. The identification of contact allergens by human assay II. Factors influencing the induction and measurement of allergic contact dermatitis. J Invest Dermatol 1966; 47:375. Hardy J. Allergy hypersensitivity in cosmetics. J Soc Cosmet Chem 1973; 24:423. Marzulli FN, Maibach HI. Contact allergy: predictive testing in man. Contact Dermatitis 1976; 2:1. Marzulli FN, Maibach HI. Effects of vehicles and elicitation concentration in contact dermatitis testing I: experimental contact sensitization in humans. Contact Dermatitis 1976; 2:325. Marzulli FN, Maibach HI. Dermatotoxicology. 4th ed. New York:Hemisphere, Fisher AA. 3 ed. Contact Dermatitis. Philadelphia:Lea & Feiberger, Shelanski HA, Shelanski MV. A new technique of human patch tests. Proc Sci Sect Toilet Goods Assoc 1953; 204: Jordan WP, King SF. Related hypersensitivity in families. Contact Dermatitis 1977; 3:1926. Kligman AM, Epstein W. Updating the maximization test for identifying contact allergens. Contact Dermatitis 1975; 1: Stotts, J. Planning, conduct and interpretation of human predictive sensitization patch tests. In: Drill VA, Lazar P, eds. Current Concepts In Cutaneous Toxicity. New York:Academic Press, 1980:4153. Panel Book Page 41

44 APPENDIX I SUMMARY TABLES Panel Book Page 42

45 TKL STUDY NO. DS TABLE 1: SUMMARY OF SUBJECT ENROLLMENT AND DISPOSITION N (%) SUBJECTS ENROLLED 106 SUBJECTS COMPLETED INDUCTION PHASE 101 ( 95.3) SUBJECTS COMPLETED ALL PHASES 99 ( 93.4) TOTAL SUBJECTS DISCONTINUED 7 ( 6.6) LOST TO FOLLOWUP I ( 0.9) VOLUNTARY WITHDRAWAL 6 ( 5.7) NOTE: ALL PERCENTAGES ARE RELATIVE TO TOTAL SUBJECTS ENROLLED SEE DATA LISTING 1 FOR FURTHER DETAIL PROGRAM: DISPSMY.SAS/USES: FINAL/25SEPO7:12:26:31 Panel Book Page 43

46 TKL STUDY NO. DS TABLE 2: SUMMARY OF SUBJECT DEMOGRAPHICS ALL ENROLLED SUBJECTS AGE N (%) 18 TO ( 26.4) N () 45 TO ( 64.2) N (%) 65 AND UP 10 ( 9.4) MEAN (SD) 49.7 (13.2) MEDIAN 52.3 RANGE 18.2 TO 70.8 GENDER N () MALE 27 ( 25.5) N (%) FEMALE 79 ( 74.5) RACE N (%) BLACK 4 ( 3.8) N (%) CAUCASIAN 8 ( 7.5) N () HISPANIC 94 ( 88.7) SEE DATA LISTING 2 FOR FURTHER DETAIL PROGRAM: DEMOSMY.SAS/USES: DEMOGS/25SEP07:12:26:33 Panel Book Page 44

47 = TKL STUDY NO. DS TABLE 3: SUMMARY OF DERMATOLOGIC RESPONSE GRADES NUMBER OF SUBJECTS BY PRODUCT PRODUCT= B INDUCTION READING MAKE CHALLENGE PHASE RESPONSE UP 48HR 72HR 9GHR(*) TOTAL EVALUABLE NUMBER ABSENT NUMBER DISCONTINUED MAXIMUM ELICITED RESPONSE DURING INDUCTION ALL SUBJECTS COMPLETING INDUCTION (N=1O1) RESPONSE N(%) SUBJECTS (100.0%) (*) WHEN REQUIRED KEY TO SYMBOLS: NO REACTION? = MINIMAL OR DOUBTFUL RESPONSE, SLIGHTLY DIFFERENT FROM SURROUNDING NORMAL SKIN + = DEFINITE ERYTHEMA, NO EDEMA ++ = DEFINITE ERYTHEMA, DEFINITE EDEMA = DEFINITE ERYTHEMA, DEFINITE EDEMA AND VESICULATION D DAMAGE TO EPIDERMIS: OOZING, CRUSTING AND/OR SUPERFICIAL EROSIONS P = PAPULAR RESPONSE >50% PROGRAM: SUMMARY.SAS/USES: RESPONSE, PRODLIST, FINAL/25SEP07:12:26:46 Panel Book Page 45

48 = TKL STUDY NO. DS TABLE 3: SUMMARY OF DERMATOLOGIC RESPONSE GRADES NUMBER OF SUBJECTS BY PRODUCT PRODUCT= C INDUCTION READING MAKE CHALLENGE PHASE RESPONSE UP 48HR 72HR 96HR(*) TOTAL EVALUABLE NUMBER ABSENT NUMBER DISCONTINUED MAXIMUM ELICITED RESPONSE DURING INDUCTION ALL SUBJECTS COMPLETING INDUCTION (N=1O1) RESPONSE N(%) SUBJECTS (100.0%) (*) WHEN REQUIRED KEY TO SYMBOLS: NO REACTION? = MINIMAL OR DOUBTFUL RESPONSE, SLIGHTLY DIFFERENT FROM SURROUNDING NORMAL SKIN + = DEFINITE ERYTHEMA, NO EDEMA ++ = DEFINITE ERYTHEMA, DEFINITE EDEMA = DEFINITE ERYTHEMA, DEFINITE EDEMA AND VESICULATION D = DAMAGE TO EPIDERMIS: OOZING, CRUSTING AND/OR SUPERFICIAL EROSIONS P = PAPULAR RESPONSE >50% PROGRAM: SUMMARY.SAS/USES: RESPONSE, PRODLIST, FINAL/25SEP07:12:26:46 Panel Book Page 46

49 APPENDIX II DATA LISTINGS Panel Book Page 47

50 TKL STUDY NO. DS DATA LISTING 1: SUBJECT ENROLLMENT AND DISPOSITION PAGE 1 OF 3 SUBJECT STUDY DATES LAST COMPLETION DAYS ON NO. SCREENED 1ST APPLIC CHALL APPLIC ENDED READING # STATUS STUDY 1 08/13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 08/17/07 10 L /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 19 S /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 08/15/07 10 S /13/07 08/13/07 08/15/ KEY: LAST READING # (IINDUCTION PHASE, C=CHALLENGE PHASE) COMPLETION STATUS (C=COMPLETED, L=LOST TO FOLLOWUP, S=VOLUNTARY WITHDRAWAL V=PROTOCOL VIOLATION, AE=ADVERSE EVENT, O=OTHER) PROGRAM: DISPLIST.SAS/USES: DEMOGS, RESPONSE, FINAL/25SEPO7:12:26:10 Panel Book Page 48

51 TKL STUDY NO. DS DATA LISTING 1: SUBJECT ENROLLMENT AND DISPOSITION PAGE 2 OF 3 SUBJECT STUDY DATES LAST COMPLETION DAYS ON NO. SCREENED 1ST APPLIC CHALL APPLIC ENDED READING # STATUS STUDY 40 08/13/07 08/13/07 09/18/ /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/ /18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 08/15/07 10 S /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C 40 KEY: LAST READING # (I=INDUCTION PHASE, C=CHALLENGE PHASE) COMPLETION STATUS (CC0MPLETED, L=LOST TO FOLLOWUP, S=VOLUNTARY WITHDRAWAL VPROTOCOL VIOLATION, AE=ADVERSE EVENT, O=OTHER) PROGRAM: DISPLIST.SAS/USES: DEMOGS, RESPONSE, FINAL/25SEP07:12:26:10 Panel Book Page 49

52 TKL STUDY NO. DS DATA LISTING 1: SUBJECT ENROLLMENT AND DISPOSITION PAGE 3 OF 3 SUBJECT STUDY DATES LAST COMPLETION DAYS ON NO. SCREENED 1ST APPLIC CHALL APPLIC ENDED READING # STATUS STUDY 79 08/13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 08/31/07 17 S / /13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/l3/07 09/18/07 09/21/07 C2 C 40 los 08/13/07 08/13/07 09/18/07 09/21/07 C2 C /13/07 08/13/07 09/18/07 09/21/07 C2 C 40 KEY: LAST READING # (I=INDUCTION PHASE, CCHALLENGE PHASE) COMPLETION STATUS (C=COMPLETED, L=LOST TO FOLLOWUP, S=VOLUNTARY WITHDRAWAL V=PROTOCOL VIOLATION, AE=ADVERSE EVENT, O=OTHER) PROGRAM: DISPLIST.SAS/USES: DEMOGS, RESPONSE, FINAL/25SEP07:12:26:10 Panel Book Page 50

53 TKL STUDY NO. DS DATA LISTING 2: SUBJECT DEMOGRAPHICS PAGE 1 OF 3 SUBJECT NO. AGE GENDER RACE FEMALE HISPANIC MALE CAUCASIAN MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE CAUCASIAN MALE HISPANIC FEMALE CAUCASIAN FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC PROGRAM: DEMOLIST.SAS/USES: DEMOGS/25SEPO7: 12:26:11 Panel Book Page 51

54 TKL STUDY NO. DS1059O7 DATA LISTING 2: SUBJECT DEMOGRAPHICS PAGE 2 OF 3 SUBJECT NO. AGE GENDER RACE FEMALE HISPANIC FEMALE CAUCASIAN MALE BLACK FEMALE HISPANIC MALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE CAUCASIAN MALE CAUCASIAN FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE BLACK MALE BLACK FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE CAUCASIAN FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC PROGRAM: DEMOLIST.SAS/USES: DEMOGS/25SEPO7:12:26:11 Panel Book Page 52

55 TKL STUDY NO. DS DATA LISTING 2: SUBJECT DEMOGRAPHICS PAGE 3 OF 3 SUBJ ECT NO. AGE GENDER RACE FEMALE CAUCASIAN FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC MALE BLACK FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC MALE HISPANIC FEMALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC MALE HISPANIC MALE HISPANIC MALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC FEMALE HISPANIC PROGRAM: DEMOLIST SAS/USES: DEMOGS/25SEPO7: 12:26:11 Panel Book Page 53

56 2 N9G 17 TKL STUDY NO. DS DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= B PAGE 1 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) X X X X X X X X X X X X KEY TO SYMBOLS: = NO REACTION? = MINIMAL OR DOUBTFUL RESPONSE, SLIGHTLY DIFFERENT FROM SURROUNDING NORMAL SKIN + = DEFINITE ERYTHEMA, NO EDEMA ++ = DEFINITE ERYTHEMA, DEFINITE EDEMA +++ = DEFINITE ERYTHEMA, DEFINITE EDEMA AND VESICULATION N9G = NO NINTH GRADING NA=NOT APPLIED NP=NOT PATCHED DUE TO REACTION ACHIEVED X = READING NOT PERFORMED DUE TO MISSED VISIT OR SUBJECT DISCONTINUATION D = DAMAGE TO EPIDERMIS: OOZING, CRUSTING AND/OR SUPERFICIAL EROSIONS P = PAPULAR RESPONSE >5O6 NR=DATA NOT RECORDED MU = MAKEUP READING FOR MISSED INDUCTION VISIT (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEPO7:12:26:14 Panel Book Page 54

57 X X X X TKL STUDY NO. DS DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= B PAGE 2 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) X X X X X X X X X X X X X 39 X X X X X X X X X X X X N9G 49 X (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEP07:12:26:14 Panel Book Page 55

58 53 N9G N9G X TKL STUDY NO. DS DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= B PAGE 3 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) X X X X X X X X X X X X X X N9G X 84 (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEPO7:12:26:14 Panel Book Page 56

59 85 X N9G X X X N9G X x X X TKL STUDY NO. D51O59O7 DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= B PAGE 4 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO B 9 MU 48HR72HR96HR(*) 86 X B7 B8 89 X X X X X N9G (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEPO7:12:26:14 Panel Book Page 57

60 = 2 X TKL STUDY NO. DS DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= C PAGE 1 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) N9G X X X X X X X X X X X KEY TO SYMBOLS: NO REACTION = MINIMAL OR DOUBTFUL RESPONSE, SLIGHTLY DIFFERENT FROM SURROUNDING NORMAL SKIN + = DEFINITE ERYTHEMA, NO EDEMA ++ = DEFINITE ERYTHEMA, DEFINITE EDEMA +++ = DEFINITE ERYTHEMA, DEFINITE EDEMA AND VESICULATION N9G = NO NINTH GRADING NA=NOT APPLIED NP=NOT PATCHED DUE TO REACTION ACHIEVED X = READING NOT PERFORMED DUE TO MISSED VISIT OR SUBJECT DISCONTINUATION D = DAMAGE TO EPIDERMIS: OOZING, CRUSTING AND/OR SUPERFICIAL EROSIONS P = PAPULAR RESPONSE >50% NR=DATA NOT RECORDED MU = MAKEUP READING FOR MISSED INDUCTION VISIT (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEPO7:12:26:14 Panel Book Page 58

61 21 30 X X X TKL STUDY NO. DS1O59O7 DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= C PAGE 2 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) G X X X X X X X X X X X X 39 X X X X X X X X X X X X X X 49 X N9G (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/255EPO7:12:26:14 Panel Book Page 59

62 X X N9G 74 X TKL STUDY NO. DS1O59O7 DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= C PAGE 3 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) N9G X X X X X X X X X X X N9G 81 X X 84 (*) WHEN REQUIRED PROGRAM: DETAIL.SAS/USES: RESPONSE, PRODLIST/25SEP07:12:26:14 Panel Book Page 60

63 85 N9G X X X 97 N9G X X X TKL STUDY NO DATA LISTING 3: DERMATOLOGIC RESPONSE GRADES BY PRODUCT AND SUBJECT PRODUCT= C PAGE 4 OF 4 SUBJECT INDUCTION READING CHALLENGE PHASE NO MU 48HR 72HR 96HR(*) 8G X X 93 X N9G X X X X X (*) WHEN REQUIRED PROGRAM: DETAIL.SASIUSES: RESPONSE, PHODLIST/255EPO7:12:26:14 Panel Book Page 61

64 APPENDIX III INFORMED CONSENT DOCUMENT Panel Book Page 62

65 TKL RESEARCH, INC. 4 FOREST AVENUE PARAMUS, NJ STUDY NO.:DSI INFORMED CONSENT REPEATED INSULT PATCH STUDY PURPOSE You are invited to participate in this research study. The purpose of this research is to determine if the study materials can be applied to human skin without causing an allergic reaction. The study will involve a minimum of 100 participants. STUDY MATERIALS The study materials include or may be components of cosmetics, moisturizers, lipsticks, skin care products, shampoos, shower gel body wash, antiperspirants/deodorants, disinfectants, antibacterial, topical dngs, fragrances, soaps, sunscreens, fibers, adhesives, medications, insect repellents, antinjicrobial (an ingredient used as a preservative), and/or any other materials which are intended for and/or may come into contact with human skin. Included is SLS which is a soap solution used as a control for comparison. STUDY DURATION This study consists of 13 visits (14 visits, if required) over 6 weeks, most visits lasting approximately 10 minutes. You will receive a schedule of visit dates and instructions. PROCEDURE You will be required to have completed a medical screening procedure. If you agree to be in this study and meet all the study entrance requirements, a small amount of the study material will be put on your back with a patch (i.e., a snmll adhesive square with a cotton pad). The patches will remain on your back for 24 hours and/or 48 hours and, in some cases, patches applied on Friday will remain on your back for 72 hours. You will return to TKL Research, Inc., at specified times to have the patches removed and/or the materials reapplied. At each of these visits, a clinical evaluator will exarriine your back to see if you are reacting. If you have a strong reaction at the study site, the smdy material will not be applied to that site but may be applied to another site(s). If you are a female of childbearing potential (i.e., not surgically sterile or have not experienced menopause), you must agree to prevent pregnancy throughout this study by using an accepted form of birth control [e.g., oral/ injectable/transdermal contraceptive pill, IUD, condom/diaphragm with spermicide, abstinence (no sexual intercourse)). If you are breastfeeding a child, you will not be permitted to participate in this study. Pregnancy and breastfeeding are prohibited to prevent any unforeseen risk to an unborn child or breastfeeding child. POTENTIAL RISKS Some of the study materials maybe irritating under certain conditions but the degree of irritation is not expected to be greater than that described below. Individuals participating in this study may experience side effects such as redness, swelling, itching, cracking, peeling, or in rare cases, small blisters or sores. Reactions usually occur only where the study materials or patch materials (such as the patch tape adhesive) touch the skin. On rare occasions, the reactions may spread beyond the patch. A reaction may result in localized lightening or darkening of the skin, which may persist in an occasional individual. Reactions may be due to either skin irritation or allergy to either study materials or patch materials (e.g., patch tape adhesive). It may be necessary to do additional application (rechallenge) to determine if an allergic reaction has occurred. If you should prove to be allergic, you can expect to react to this material if you encounter it at a later date. Whenever possible, you will be informed as to the identity of the material in order that you may avoid contact with it in the future. For any significant reactions that may occur as a direct result of your participation in this study, appropriate and reasonable medical treatment will be provided by TKL Research, Inc. at no cost to you to resolve the immediate problem. Provision of such medical care is not an admission of legal liability or responsibility Panel Book Page 63

66 TKL RESEARCH, inc. 4 FOREST AVENUE PARAMUS, NJ STUDY NO.: DSI INFORMED CONSENT REPEATED INSULT PATCH STUDY for the condition being treated. If such reactions occur, TKL personnel should be contacted immediately at (201) day, night or weekends. Extended medical care will not be provided. POTENTIAL BENEFITS There is no personal benefit other than the satisfaction of participation in a clinical research study. SIGNIFICANT NEW FINDINGS You will be informed of any significant new findings that may affect your willingness to continue your participation. ALTERNATIVE TREATMENT Since this study is for research only, the only alternative is for you not to participate. WITHDRAWAL FROM STUDY Participation in the study is voluntary and you may refuse to participate or may withdraw at any time without penalty or loss of benefits, other than financial, to which you are otherwise entitled. Your participation may also be discontinued at any time without your consent by the study doctor, or the study sponsor(s) (the company(ies) that makes the product(s) being evaluated). If you fail to comply with study procedures, your participation may be terminated. COST Your participation in the study will not incur any cost to you. FINANCIAL INCENTIVE You will be paid a sum ofsl3o.00 upon completion of this study. If in the judgment of the investigating personnel, it is best to discontinue your participation in this study due to an adverse experience or severe reaction you will be paid in full for your participation. If you are dismissed for refusal to obey rules or follow instructions you will not be paid. CONFIDENTIALiTY AND AUTHORIZATION Reports prepared by TK.L Research will utilize statistical information only and at no time will your name be used. A new federal regulation called the Health Insurance Portability and Accountability Act (HIPAA), which went into effect on April 14, 2003, describes how your personal health information may be used, disclosed and made accessible to you. This privacy rule is designed to protect the confidentiality of your personal health information. The following information describes how the HIPAA rule applies to you and your rights. This study can be performed only by collecting and using your personal health information. Your study records will be kept as confidential as possible under local, state and federal laws. Personnel from the following organizations may examine your study records: the sponsor, personnel associated with this study, regulatory agencies, such as the Food and Drug Administration (FDA) or Environmental Protection Agency (EPA), and the Institutional Review Board (IRB), a committee that has reviewed this study to help ensure that your rights and welfare as a research participant are protected and that the study is carried out m an ethical manner. Because of the number of individuals who may see your records, absolute confidentiality cannot be guaranteed. Personal health information that may be used and disclosed includes that which is obtained to determine your eligibility to participate and that which is collected from the procedures that are carried out. It may identify you by name, address, telephone number, Social Security Number, study number, date of birth or other identifiers. Once the information is disclosed, it is possible that it may be redisclosed, at which time it is no longer protected by federal regulations, but may be by state laws. Panel Book Page 64

67 TKL RESEARCH, INC. 4 FOREST AVENUE PARAMUS, NJ STUDY NO.: DS INFORMED CONSENT REPEATED INSULT PATCH STUDY If tire final study data are prepared for publication and other reports, your identity will not be revealed. Under these federal privacy regulations, you have the right to see and copy any of the information gathered about you, until your study records are no longer kept by the study doctor. However, it may not be available until the study has been completed. There is no expiration date for this authorization. You may, by written notice to the study doctor, cancel your authorization to use or disclose your personal information at any time. If you withdraw your authorization, the information collected to that time may still be used to preserve the scientific integrity of the study. By signing this consent form, you authorize these uses and disclosures of your personal information. If you do not authorize these uses and disclosures, you will not be able to participate in the study. WHO TO CALL Additional information regarding this research is available either before or during the course of this study. If you have any questions or research related side effect or injury, you may contact Kathy Georgeian, Director, Derunatologic Safety Testing at (201) ext The emergency 24 hr. phone number is A signed copy of this consent form will be given to you. ** 1 have read and understand the information given in this consent form. I have had an opportunity to ask questions and my questions have been answered. I voluntarily consent to participate. By signing this form I have not given up any of my legal rights which I would otherwise have as a research subject. ENTRY NO. PRINT NAME SIGNATURE DATE SIGNATURE OF WITNESS DATE Panel Book Page 65

68 III RESEARCH HUMANREPEATED.INSULT PATCH TKL STUDY NO. DS So A + olfi 14 L/ 010 CONDUCTED FOR: 4Il) Icq / 5/y/ DATE OF ISSUE: July 18, 2008 TKL Clinical Trials Division TKL Clinical Site Division N TKL Recruitment Management Division TKL Research, Inc. 4 Forest Avenue, Paramus, NJ Phone: O0 www tkiresearch.c om Panel Book Page 66

69 STUDY TKL Study No. DS TABLE OF CONTENTS SIGNATURES I STATEMENT OF QUALITY CONTROL I TiTLE OF STUDY 2 SPONSOR.2 MATERIAL 2 DATE STUDY INITIATED 2 DATE STUDY COMPLETED 2 DATEOF issue 2 INVESTIGATIVE PERSONNEL 3 CLINICAL SITE 3 SUMMARY OBJECTIVE RATIONALE STUDY DESIGN STUDY POPULATION Inclusion Criteria Exclusion Criteria Informed Consent DESCRIPTION OF STUDY Outline of Study Procedures Definitions Used for Grading Responses Evaluation of Responses NATURE OF STUDY MATERIAL STUDY MATERIAL SPECIFICATIONS STORAGE, HANDLING, AND DOCUMENTATION OF STUDY MATERIAL APPLICATION OF STUDY MATERIAL DESCRIPTION OF PATCH CONDITIONS INTERPRETATION DOCUMENTATION AND RETENTION OF DATA RESULTS & DISCUSSION CONCLUSION REFERENCES 10 APPENDICES I SUMMARY TABLES II DATA LISTINGS III INFORMED CONSENT DOCUMENT Panel Book Page 67

70 ICR TKL Study No. DS SIGNATURES This study was conducted in compliance with the requirements of the protocol and TKL s Standard Operating Procedures, and in the spirit of GCP ICR Topic E6. The report accurately reflects the raw data for this study. Jonathk. Dosik, MD D e Dermatologist Principal Investigator Kathleen Georgeian, Cl4ical Research Coordinator and Director, Dermatol6ic Safety Testing Date STATEMENT OF QUALITY CONTROL The Quality Control Unit of the Dermatological Safety Department conducted a 100% review of all studyrelated documents. The protocol was reviewed prior to the start of the study, and the medical screening forms and informed consent documents were reviewed inprocess of the study. The regulatory binder and study data were reviewed poststudy to ensure accuracy. The study report was reviewed and accurately reflects the data for this study. ICH Topic 6 Note for guidance on Good Clinical Practices (CPMP!ICH/135/95) Harmonized Tripartite Guideline for Good Clinical Practices having reached Step 5 of the ICH Process at the ICH Steering Committee meeting on I May Panel Book Page 68

71 TKL Study No. DS TITLE OF STUDY Human Repeated Insult Patch Test SPONSOR STUDY MATERIAL Test Material Description Invisible Solid Antiperspirant DATE STUDY INITIATED May 12, 2008 DATE STUDY COMPLETED June 16, 2008 DATE OF ISSUE July 18, 2008 Panel Book Page 69

72 TKL Study No. DS INVESTIGATIVE PERSONNEL Jonathan S. Dosik, MD Dermatologist Principal hivestigator Kathleen Georgeian Clinical Research Coordinator and Director, Dermatologic Safety Testing Michelle Medina Assistant Clinical Research Coordinator CLINICAL SITE TKL RESEARCH, INC. I Palmer Terrace Carlstadt, NJ Panel Book Page 70

73 TKL Study No. DS SUMMARY One (1) test material, , was evaluated neat to determine its ability to sensitize the skin of volunteer subjects with normal skin using an occlusive repeated insult patch study. One hundred and two (102) subjects completed the study. Under the conditions employed in this study, there was no evidence of sensitization to Test Material Panel Book Page 71

74 TKL Study No. DS OBJECTIVE The objective of this study was to determine the ability of the study materials to cause sensitization by repeated topical applications to the skin of humans under controlled patch study conditions. 2.0 RATIONALE Substances that come into contact with human skin need to be evaluated for their propensity to irritate and/or sensitize. Once an appropriate preclinical safety evaluation has been performed, a reproducible, standardized, quantitative patch evaluation procedure must be used to demonstrate that a particular material can be applied safely to human skin without significant risk of adverse reactions. The method herein employed is generally accepted for such a purpose. Repeated insult patch evaluation is a modified predictive patch study that can detect weak sensitizers that require multiple applications to induce a cellmediated (Type IV) immune response sufficient to cause an allergic reaction. Irritant reactions may also be detected using this evaluation method, although this is not the primary purpose of this procedure. Results are interpreted according to interpretive criteria based upon published works, as well as the clinical experience of TKL Research, Inc. These interpretive criteria are periodically reviewed and amended as new information becomes available. 3.0 STUDY DESIGN 3.1 STUDY POPULATION A sufficient number of subjects were to be enrolled to provide 100 completed subjects. In the absence of any sensitization reactions in this sample size (100 evaluable subjects), a 95% upper confidence bound on the population rate of sensitization would be 3.5% Inclusion Criteria Individuals eligible for inclusion in the study were those who: 1. were males or females, 18 years of age or older, in general good health; 2. were free of any systemic or dermatologic disorder which, in the opinion of the investigative personnel, would have interfered with the study results or increased the risk of adverse events; 3. were of any skin type or race providing the skin pigmentation would allow discernment of erythema; 4. had completed a medical screening procedure; and 5. had read, understood, and signed an informed consent agreement. Panel Book Page 72

75 TKL Study No. DS1032O Exclusion Criteria Individuals excluded from participation in the study were those who: 1. had any visible skin disease at the study site which, in the opinion of the investigative personnel, would have interfered with the evaluation; 2. were receiving systemic or topical drugs or medication which, in the opinion of the investigative personnel, would have interfered with the study results; 3. had psoriasis and/or active atopic dermatitis/eczema; 4. were females who were pregnant, planning to become pregnant during the study, or breastfeeding; and/or 5. had a known sensitivity to cosmetics, skin care products, or topical drugs as related to the material being evaluated Informed Consent A properly executed informed consent document was obtained from all subjects prior to entering the study. The signed informed consent document is maintained in the study file. In addition, the subjects were provided with a copy of the informed consent document (see Appendix III). 3.2 DESCRIPTION OF STUDY Outline of Study Procedures Subjects participated in the study over a 6week period involving 3 phases: (1) Induction, (2) Rest, and (3) Challenge. Prior to study entry, the subjects were screened to assure that they met the inclusionlexclusion criteria. Informed consent was obtained. Each subject was provided with a schedule of the study activities. All subjects were told to avoid wetting the patches and were asked not to engage in activities that caused excessive perspiration. They were instructed to notifi the staff if they experienced any discomfort beyond mild itching or observed any adverse changes at the evaluation sites while on the study or within 2 weeks of completing the study. The Induction Phase consisted of 9 consecutive applications of the study material and subsequent evaluations of the application sites. Prior to application of the patches, the sites were outlined with a skin marker, eg, gentian violet. Patches were applied on Mondays, Wednesdays, and Fridays for 3 consecutive weeks and were removed by the subjects approximately 24 hours after application. Subjects returned to the facility at 48hour intervals to have the sites evaluated and identical patches applied to the same sites. Patches applied on Friday were removed by subjects after 24 hours and sites were evaluated on the following Monday, ie, 72 hours after patch application. Following the ninth evaluation, the subjects were dismissed for a rest period of approximately 1015 days. 2 2 A Monday or Friday holiday could result in evaluation at 96 hours after patch application. Panel Book Page 73

76 TKL Study No. DS Subjects who were absent once during the 3week, 9patch induction phase received a makeup (MU) patch at the last induction visit. The MU applications were graded 48 hours later at the MU visit, or were recorded as N9G (no ninth grading). Subjects who missed the ninth evaluation (N9G) but had 9 patch applications were considered to have completed the induction phase. The Challenge Phase was initiated during the sixth week of the study. Identical patches were applied to sites previously unexposed to the study material. The patches were removed by subjects after 24 hours and the sites graded after additional 24hour and 48hour periods (ie, 48 and 72 hours after application). Following a negative induction, a 48/72hour sequence of 1+,?/+, or +/+ resulted in an additional reading to be preformed at the 96hour interval. Rechallenge was performed whenever there was evidence of possible sensitization. To be considered a completed case, a subject must have had 9 applications of the study materials and no less than 8 subsequent readings during induction and a single application and 2 readings during challenge. Only completed cases were used to assess sensitization Definitions Used for Grading Responses The symbols found in the data listings accompanying this report were used to express the response observed at the time of examination: SYMBOL AND RESPONSE = No reaction = Minimal or doubtful response, slightly different from surrounding normal skin + = Definite erythema, no edema ++ = Definite erythema, definite edema = Definite erythema, definite edema and vesiculation SPECIAL NOTATIONS E = Marked/severe erythema S = Spreading of reaction beyond patch site (ie, reaction where material did not contact skin) p = Papular response > 50% pv = Papulovesicular response> 50% D = Damage to epidermis: oozing, crusting and/or superficial erosions I = Itching X = Subject absent PD = Patch dislodged NA = Not applied NP = Not patched (due to reaction achieved) N9G = No ninth grading Panel Book Page 74

77 I SL Study No. DS Evaluation of Responses All responses were graded by a trained dermatologic evaluator meeting TKL s strict certification requirements to standardize the assignment of response grades. 4.0 NATURE OF STUDY MATERIAL 4.1 STUDY MATERIAL SPECIFICATIONS Identification Invisible Solid Antiperspirant Amount Applied 0.2 g Special Instructions Patch used within 30 minutes of dosing. 4.2 STORAGE, HANDLING, AND DOCUMENTATION OF STUDY MATERIAL Receipt of the material used in this study was documented in a general logbook, which serves as a permanent record of the receipt, storage, and disposition of all study material received by TKL. On the basis of information provided by the sponsor, the study material was considered reasonably safe for evaluation on human subjects. A sample of the study material was reserved and will be stored for a period of 6 months. All study material is kept in a locked product storage room accessible to clinical staff members only. At the conclusion of the clinical study, the remaining study material was discarded and the disposition documented in the logbook. 4.3 APPLICATION OF STUDY MATERIAL Study material was applied to patch as instructed and patch was applied to the infrascapular area of the back, either to the right or left of the midline. 4.4 DESCRIPTION OF PATCH CONDITIONS Material evaluated under occlusive patch conditions is applied to a 2 cm x 2 cm Webril pad attached to a nonporous, plastic film adhesive bandage (3M medical tape). The patch is secured with hypoallergenic tape (Micropore), as needed. Material evaluated under semiocclusive patch conditions is applied to a 2 cm x 2 cm Webril pad. pad is affixed to the skin with hypoallergenic tape (Micropore). The 5.0 INTERPRETATION Sensitization is characterized by an acute allergic contact dermatitis. Typical sensitization reactions begin with an immunologic response in the dermis resulting in erythema, edema formation, and secondary epidermal damage (vesiculation), sometimes extending beyond the patch site and often accompanied by itching. Sensitization reactions tend to be delayed. The reaction typically becomes evident between 24 and 48 hours, peaks at 4872 hours and subsequently subsides. The reaction is often greater at Panel Book Page 75

78 TKL Study No. DS hours than at 48 hours. The severity of the reaction is generally greater during the challenge phase of an RIPT than that seen during induction. Irritant reactions are characterized as a nonimmunologic, localized, superficial, exudative, inflammatory response of the skin due to an externally applied material. The typical initial reaction does not develop much edema or vesiculation but results in scaling, drying, cracking, oozing, crusting, and erosions. The reaction is usually sharply delineated, not spreading beyond the patch site. Irritant reactions are typically evident by 24 hours and diminish over the next 4872 hours. Removal of the offending agent results in gradual improvement of the epidermal damage. The reaction seen at 72 hours is, therefore, less severe than that seen at 48 hours. Finally, the severity of the reaction experienced in the challenge phase is generally similar to that seen during induction. If the results of the study indicate the likelihood of sensitization, the recommended practice is to rechallenge the subjects who have demonstrated sensitizationlike reactions to confirm that these reactions are, indeed, associated with the product. Our preferred rechallenge procedure involves the application of the product to naïve sites, under both occlusive and semiocclusive patch conditions. Use of the semiocclusive patch condition helps to differentiate irritant and sensitization reactions. Generally speaking, if a product is a sensitizer it will produce a similar reaction under both occlusion and semiocclusion. Whereas, if the product has caused an irritant reaction, the reactions will be less pronounced under the semiocclusive condition. 6.0 DOCUMENTATION AND RETENTION OF DATA The case report forms (CRFs) were designed to identif, each subject by subject number and initials and to record demographics, examination results, adverse events, and end of study status. Originals or copies of all case report forms, correspondence, study reports, and all source data will be kept on hardcopy file for a minimum of 5 years from completion of the study. Storage was maintained either at a TKL facility in a secured room accessible only to TKL employees, or at an offsite location that provided a secure environment with burglar/fire alarm systems, camera detection, and controlled temperature and humidity. Documentation will be available for the sponsor s review on the premises of TKL Research, Inc. 7.0 RESULTS & DISCUSSION One hundred fourteen (114) subjects between the ages of 18 and 70 were enrolled and 102 subjects completed the study. See Tables 1 and 2 in Appendix I and Data Listings 1 and 2 in Appendix II. The following table summarizes subject enrollment and disposition. Number enrolled: 1 14 Number discontinued: 12 Lost to followup: 10 Voluntary withdrawal: 2 Number completed: 102 Source: Table 1, Appendix I Panel Book Page 76

79 TKL Study No. DS There were no adverse events reported. A summary of response data is provided in Table 3, Appendix I. grades are provided in Data Listing 3, Appendix II. Individual dermatological response 8.0 CONCLUSION Under the conditions employed in this study, there was no evidence of sensitization to Test Material REFERENCES Schwartz L, Peck SM. The patch test in contact dermatitis. Pubi Health Pep 1944; 59:2. Draize JH, Woodward G, Calvary HO. Methods for the study of irritation and toxicology of substances applied topically to the skin and mucous membranes. J Pharmacol Exp Ther 1944; 82: Lanman BM, Elvers WB, Howard CS. The role of human patch testing in a product development program. Joint Conf Cosmet Sci Toilet Goods Assoc 1968; Marzulli FN, Maibach HI. Contact allergy: predictive testing in man. Contact Dermatitis 1976; 2:1. Zhai H, Maibach HI. Dermatotoxicology. 6t ed. New York:Hemisphere, Stotts J. Planning, conduct and interpretation of human predictive sensitization patch tests. In:Drill VA, Lazar P, eds. Current Concepts in Cutaneous Toxicity. New York: Academic Press, 1980: Griffith JF. Predictive and diagnostic testing for contact sensitization. Toxicol Appl Pharmacol, Suppl 1969; 3:90. Gerberick GF, Robinson MK, Stotts J. An approach to allergic contact sensitization risk assessment of new chemicals and product ingredients. American Journal of Contact Dermatitis 1993; 4(1):14. Panel Book Page 77

80 APPENDIX I SUMMARY TABLES Panel Book Page 78

81 TKLStudyNo.DS Page lof I Table 1: Summary of Subject Enrollment and Disposition N (%) Subjects enrolled 114 Subjects completed induction phase 105 (92.1) Subjects completed all phases 102 (89.5) Total subjects discontinued 12 (10.5) Lost to followup 10 (8.8) Voluntary withdrawal 2 (1.8) Note: All percentages are relative to total subjects enrolled. See data listing 1 for further detail. Generated on 06/25/08:15:53 by DISPSMY.SAS /Uses: FINAL Panel Book Page 79

82 TKL Study No. DS Page 1 of 1 Table 2; Summary of Subject Demographics All Enrolled Subjects Age N (%) 18 to 53 (46.5) N (%) 45 to (50.0) N(%)65andup 4(3.5) Mean (SD) 44.9 (12.8) Median 45.7 Range 18.2 to 70.9 Gender N (%) Male 39 (34.2) N (%) Female 75 (65.8) Race Black 3 (2.6) Caucasian 64(56.1) Hispanic 47 (41.2) See data listing 2 for further detail. Generated on 06/25/08:15:53 by DEMOSMY.SAS fuses: DEMOGS Panel Book Page 80

83 108 TKL Study No. DS Table 3: Summary of Dermatologic Response Grades Number of Subjects by Product Product = Induction Reading Challenge Phase Make Response Up 4Xhr 72hr 96hr(*) ? Totalevaluable Number absent Number discontinued (*) when required Maximum Eli cited Response During Induction All Subjects Completing Induction (N1 05) Response n(%) Subjects 103 (98.1%)? 2(1.9%) Key to Symbols: = No reaction 7 = Minimal or doubtful response, slightly / different from surrounding normal skin + = Definite erythema, no edema ++ = Definite erythema, definite edema HH = Definite erythema, definite edema and vesiculation D = Damage to epidermis: oozing, crusting and/or superficial erosions p = Papular response >50% Generated on 06/25/08:15:53 by SUMMARY.SAS/USES: RESPONSE, PRODLIST, FINAL Panel Book Page 81

84 APPENDIX II DATA LISTINGS Panel Book Page 82

85 05/16/08 05/27/08 05/19/08 TKL STUDY NO. DS Page 1 of 4 Data Listing 1: Subject Enrollment and Disposition Study Dates Last Reading Completion Days in Subject No. Screened 1st Applic Chall Applic Ended # Status Study /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 10 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/ /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 II L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C 39 Key: Last Reading # (I=Induction Phase. C=Challenge Phase) Completion Status (CComp1eted, L=Lost to followup, S=Voluntary withdrawal, VProtocol violation, AE=Adverse event, O=Other) Generated on 06/24/08:14:58 by DISPLIST.SAS / Uses: DEMOGS, RESPONSE. FINAL Panel Book Page 83

86 TKL STUDY NO. DS Page 2 of 4 Data Listing 1: Subject Enrollment and Disposition Study Dates Subject No. Screened 1st Applic Chall Applic Ended /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/13/ /12/08 05/12/09 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 06/16/08 06/19/ /12/08 05/12/08 05/27/ /12/08 05/12/08 06/16/08 06/19/08 Last Reading # C C C C C C C C C C 9 C C C C C C C C C C C C C C C C C C 14 C Completion Status C C C C C C C C C C S C C C C C C C C C C C C C C C C C C L C Days in Study Key: Last Reading # (Ilnduction Phase, C=Challenge Phase) Completion Status (CCornpleted, L=Lost to followup. S=Voluntary withdrawal, VProtoco1 violation. AE=Adverse event, O=Other) Generated on 06/24/08:14:58 bydispljst.sas /Uses: DEMOGS, RESPONSE, FII 4AL Panel Book Page 84

87 05/16/08 05/23/08 05/21/08 05/23/08 TKL STUDY NO. DS Page 3 of 4 Data Listing I Subject Enrollment and Disposition Study Dates Last Reading Completion Days in Subject No. Screened 1st Applic Chall Applic Ended # Status Study /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 10 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 13 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/02/08 18 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 12 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 13 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C 39 Key: Last Reading # (l=induction Phase, C=Challenge Phase) Completion Status (C=Completed, L=Lost to followup. S=Voluntary withdrawal, V=Protocol violation, AEAdverse event, OOther) Generated on 06/24/08:14:58 by DISPLTST.SAS / Uses: DEMOGS, RESPONSE, F11 JAL Panel Book Page 85

88 06/16/08 TKL STUDY NO. DS Page 4 of 4 Data Listing 1: Subject Enrollment and Disposition Study Dates Last Reading Completion Days in Subject No. Screened 1st Applic Chall Applic Ended # Status Study /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 19 L /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /12/08 05/12/08 06/16/08 06/19/08 C C /16/08 05/16/08 06/16/08 06/19/08 C C /16/08 05/16/08 06/16/08 06/19/08 C C /16/08 05/16/08 06/16/08 06/16/08 19 L /16/08 05/16/08 06/16/08 06/19/08 C C 35 Key: Last Reading # (J1nduction Phase, C=Challenge Phase) Completion Status (C=Completed, L=Lost to followup, S=Voluntary withdrawal, V=Protocol violation, AE=Adverse event, OOther) Generated on 06/24/08:14:58 by DISPLIST.SAS /Uses: DEMOGS, RESPONSE, FINAL Panel Book Page 86

89 TKL STUDY NO. DS Page I of 3 Data Listing 2: Subject Demographics Subject No. Age Gender Race Female Hispanic Female Caucasian Female Caucasian Male Caucasian Female Caucasian Male Black Male Caucasian Male Caucasian Female Hispanic Male Hispanic Female Hispanic Female Caucasian Female Caucasian Female Caucasian Female Caucasian Female Hispanic Male Caucasian Male Hispanic Female Caucasian Female Caucasian Male Caucasian Female Hispanic Male Caucasian Female Hispanic Female Caucasian Female Caucasian Male Caucasian Female Caucasian Female Hispanic Female Hispanic Female Caucasian Male Caucasian Female Hispanic Female Black Male Hispanic Female Caucasian Female Hispanic Generated on 06/24/08:14:58 by DEMOLIST.SAS / Uses: DEMOGS Panel Book Page 87

90 TKL STUDY NO. DS Page 2 of 3 Data Listing 2: Subject Demographics Subject No. Age Gender Race Female Hispanic Female Hispanic Female Caucasian Female Caucasian Female Hispanic Male Hispanic Female Caucasian Female Hispanic Female Hispanic Female Caucasian Female Caucasian Male Caucasian Male Caucasian Female Caucasian Male Caucasian Male Hispanic Female Hispanic Female Caucasian Female Caucasian Female Caucasian Male Caucasian Female Caucasian Male Caucasian Female Hispanic Female Caucasian Male Caucasian Male Caucasian Female Caucasian Male Black Male Hispanic Female Hispanic Male Caucasian Male Hispanic Female Caucasian Female Caucasian Female Caucasian Female Hispanic Generated on 06/24/08:14:58 by DEMOLIST.SAS / Uses: DEMOGS Panel Book Page 88

91 TKLSTUDYNO.DS Page 3of 3 Data Listing 2: Subject Demographics Subject No. Age Gender Race Female Hispanic Male Hispanic Female Hispanic Female Caucasian Female Caucasian Female Caucasian Female Hispanic Male Hispanic Female Hispanic Male Hispanic Male Hispanic Male Hispanic Male Caucasian Female Caucasian Female Caucasian Female Caucasian Female Hispanic Male Hispanic Female Hispanic Female Hispanic Male Hispanic Female Caucasian Female Hispanic Female Hispanic Female Hispanic Female Caucasian Female Caucasian Male Caucasian Female Caucasian Female Caucasian Female Caucasian Female Caucasian Female Hispanic Male Hispanic Male Hispanic Female Caucasian Female Caucasian Male Caucasian Male Hispanic Male Caucasian Generated on 06/24/08:14:58 by DEMOLIST.SAS / Uses: DEMOGS Panel Book Page 89

92 X X 017 X X 022 X TKL Study No. DS1032U Page 1 of 5 Data Listing 3: Dermatologic Response Grades By Product and Subject Product = Induction Reading Challenge Phase Subject No MU 4Xhr 72hr 96hr(*) X X x 016 X X X X X X X X X X X Key to Symbols: X X X X X X = No reaction 7 = Minimal or doubtful response, slightly different from surrounding normal skin + Definite erythema, no edema ++ = Definite erythema, definite edema +H Definite erythema, definite edema and vesiculation N9G = No ninth grading NA Not applied NP = Not patched due to reaction achieved X = Reading not performed due to missed visit or subject discontinuation D = Damage to epidermis: oozing, crusting and/or superficial erosions p = Papular response >50% NR=Data not recorded MU = Makeup reading for missed induction visit (*) When required Generated on 06/24/08:14:58 by DETAIL.SAS/USES: RESPONSE, PRODLIST Panel Book Page 90

93 x x x TKL Study No. DS Page 2 of 5 Data Listing 3: Dermatologic Response Grades By Product and Subject Product = Subject No Induction Reading Challenge Phase MU 4Xhr 72hr 96hr(*) x x x x x x x x x x x x x x x x x (*) When required Generated on 06/24/08:14:58 by DETAIL.SAS/USES: RESPONSE, PRODLIST Panel Book Page 91

94 7 X 062 TKL Study No. DS Page 3 of 5 Data Listing 3: Deniiatologic Response Grades By Product and Subject Product = Induction Reading Challenge Phase Subject No MU 48hr 72hr 96hr(*) X X X X X X X X X X X X X X X X X X X X X X X X X (*) When required Generated on 06/24/08:14:58 by DETAIL.SAS/USES: RESPONSE, PRODLIST Panel Book Page 92

95 083 X 086 x 7 x TKLStudyNo.DS Page 4of 5 Data Listing 3: Dermatologic Response Grades By Product and Subject Product Induction Reading Challenge Phase Subject No MU 48hr 72hr 96hr(*) X X 085 X X X X x 087 X X X X x x x x x x x x x x x (*) When required Generated on 06/24/08:14:58 by DETAILSAS/USES: RESPONSE, PRODLIST Panel Book Page 93

96 X X X X X N9G TKi Study No. DS Page 5 of 5 Data Listing 3: Dermatologic Response Grades By Product and Subject Product = Induction Reading Challenge Phase Subject No MU 4Xhr 72hr 96hr(*) X X x X N9G X (*) When required Generated on 06/24/08:14:58 by DETAIL.SAS/USES: RESPONSE, PRODLIST Panel Book Page 94

97 APPENDIX III INFORMED CONSENT DOCUMENT Panel Book Page 95

98 TKL RESEARCH, INC. 4 FOREST AVENUE PARAMUS, NJ STUDY NO.: DS INFORMED CONSENT REPEATED INSULT PATCH STUDY PURPOSE You are invited to participate in this research study. The purpose of this research is to determine if the study materials can be applied to human skin without causing an allergic reaction. The study will involve a _minimum:of.ioflparticipants... STUDY MATERIALS The study materials include or may be components of cosmetics, moisturizers, lipsticks, skin care products, shampoos, shower gel/body wash, antiperspirants/deodorants, disinfectants, antibacterial, topical drugs, fragrances, soaps, sunscreens, fibers, adhesives, medications, insect repellents, antimicrobial (an ingredient used as a preservative), and/or any other materials which are intended for and/or may come into contact with human skin. Included is SLS which is a soap solution used as a control for comparison. STUDY DURATION This study consists of 13 visits (14 visits, if required) over 6 weeks, most visits lasting approximately 10 minutes. You will receive a schedule of visit dates and instructions. PROCEDURE You will be required to have completed a medical screening procedure. If you agree to be in this study and meet all the study entrance requirements, a small amount of the study material will be put on your back with a patch (i.e., a small adhesive square with a cotton pad). The patches will remain on your back for 24 hours and/or 48 hours and, in some cases, patches applied on Friday will remain on your back for 72 hours. You will return to TKL Research, Inc., at specified times to have the patches removed and/or the materials reapplied. At each of these visits, a clinical evaluator will examine your back to see if you are reacting. If you have a strong reaction at the study site, the study material will not be applied to that site but may be applied to another site(s). If you are a female of childbearing potential (i.e., not surgically sterile or have not experienced menopause), you must agree to prevent pregnancy throughout this study by using an accepted form of birth control [e.g., oral/ injectable/transtlermal contraceptive pill, IUD, condom/diaphragm with spermicide, abstinence (no sexual intercourse)]. If you are breastfeeding a child, you will not be permitted to participate in this study. Pregnancy and breastfeeding are prohibited to prevent any unforeseen risk to an unborn child or breastfeeding child. POTENTIAL RISKS Some of the study materials may be irritating under certain conditions but the degree of irritation is not expected to be greater than that described below. Individuals participating in this study may experience side effects such as redness, swelling, itching, cracking, peeling, or in rare cases, small blisters or sores. Reactions usually occur only where the study materials or patch materials (such as the patch tape adhesive) touch the skin. On rare occasions, the reactions may spread beyond the patch. A reaction may result in localized lightening or darkening of the skin, which may persist iii an occasional individual. Reactions may be due to either skin irritation or allergy to either study materials or patch materials (e.g., patch tape adhesive). It may be necessary to do additional application (rechallenge) to determine if an allergic reaction has occurred. If you should prove to be allergic, you can expect to react to this material if you encounter it at a later date. Whenever possible, you will be informed as to the identity of the material in order that you may avoid contact with it in the future. For any significant reactions that may occur as a direct result of your participation in this study, appropriate and reasonable medical treatment will be provided by TKL Research, Inc. at no cost to you to resolve the immediate problem. Provision of such medical care is not an admission of legal liability or responsibility Panel Book Page 96

99 TKL RESEARCH, INC. 4 FOREST AVENUE PARAMUS, NJ INFORMED CONSENT REPEATED INSULT PATCH STUDY STUDY NO.: DS for the condition being treated. If Such reactions occur, T personnel should be contacted immediately at (201) day, night or weekends. Extended medical care will not be provided.. _POTENTIAEEENEFITS:... zzz:z.: There is no personal benefit other than the satisfaction of participation in a clinical research study. SIGNIFICANT NEW FINDINGS You will be informed of any significant new findings that may affect your willingness to continue your participation. ALTERNATiVE TREATMENT Since this study is for research only, the only alternative is for you not to participate. WITHDRAWAL FROM STUDY Participation in the study is voluntary and you may refuse to participate or may withdraw at any time without penalty or loss of bencfits, other than financial, to which you are otherwise entitled. Your participation may also be discontinued at any time without your consent by the study doctor, or the study sponsor(s) (the company(ies) that makes the product(s) being evaluated). If you fail to comply with study procedures, your participation may be terminated. COST Your participation in the study will not incur any cost to you. FINANCIAL INCENTIVE You will be paid a sum of $ I 10_Go upon completion of this study. If in the judgment of the investigating personnel, it is best to discontinue your participation in this study due to an adverse experience or severe reaction you will be paid in full for your participation. If you are dismissed for refusal to obey rules or follow instructions you will not be paid. CONFIDENTIALITY AND AUTHORIZATION Reports prepared by TKL Research will utilize statistical information only and at no time will your name be used. A new federal regulation called the Health Insurance Portability and Accountability Act (HIPAA), which went into effect on April 14, 2003, describes how your personal health information may be used, disclosed and made accessible to you. This privacy rule is designed to protect the confidentiality of your personal health information. The following information describes how the HIPAA rule applies to you and your rights. This study can be performed only by collecting and using your personal health information. Your study records will be kept as confidential as possible under local, state and federal laws. Personnel from the following organizations may examine your study records: the sponsor, personnel associated with this study, regulatory agencies, such as the Food and Drug Administration (FDA) or Environmental Protection Agency (EPA), and the Institutional Review Board (IRB), a committee that has reviewed this study to help ensure that your rights and welfare as a research participant are protected and that the study is carried out in an ethical manner. Because of the number of individuals who may see your records, absolute confidentiality cannot be guaranteed. Personal health information that may be used and disclosed includes that which is obtained to determine your eligibility to participate and that which is collected from the procedures that are carried out It may identify you by name, address, telephone number, Social Security Number, study number, date of birth or other identifiers. Once the information is disclosed, it is possible that it may be redisclosed, at which time it is no longer protected by federal regulations, but may be by state laws. Panel Book Page 97

100 TKL RESEARCH, INC. 4 FOREST AVENUE PARAMUS, NJ STUDY NO.: DS INFORMED CONSENT REPEATED INSULT PATCH STUDY If the final study data are prepared for 5ubIication and other reports, your identity will not be revealed. Under these federal privacy regulations, you have the right to see and copy any of the information gathered z z_aboutyou,until:.yourstudy:records:arejioiongetkept by:thtstudy:.doctthowver,itm4yxip(be._ available until the study has been completed. There is no expiration date for this authorization. You may, by written notice to the study doctor, cancel your authorization to use or disclose your personal information at any time. If you withdraw your authorization, the information collected to that time may still be used to preserve the scientific integrity of the study. By signing this consent form, you authorize these uses and disclosures of your personal information. If you do not authorize these uses and disclosures, you will not be able to participate in the study. WHO TO CALL Additional information regarding this research is available either before or during the course of this study. If you have any questions or research related side effect or injury, you may contact Kathy (Ieorgeian, Director, Dermatologic Safety Testing at (201) ext The emergency 24 hr. phone number is A signed copy of this consent form will be given to you. I have read and understand the information given in this consent form. I have had an opportunity to ask questions and my questions have been answered. I voluntarily consent to participate. By signing this form I have not given up any of my legal rights which I would otherwise have as a research subject. ENTRY NO. PRINT NAME SIGNATURE DATE SIGNATURE OF WITNESS DATE Panel Book Page 98

101 Personal Care Products Council Committed to Safety, Quality & Innovation Memorandum TO: FROM: F. Alan Andersen, Ph.D. Director COSMETIC INGREDIENT REVIEW (CLR) John Bailey, Ph.D. Industry Liaison to the CIR Expert Panel DATE: January 4, 2011 SUBJECT: Toxicology Summary: Trimethylsiloxysilicate Momentive Toxicology summary for SR 1000 (Trimethylsiloxysilicate) th Street, N.W., Suite 3O0 Washington, D.C (fax) Panel Book Page 99

102 MOMENTIVE performance materia s The ccience hehtid the outions. Toxicology Summary for SRi 000 (r.., 12/3/20 10., i. c c e Acute Ocular Irritation Final report is dated April 23, Study was conducted at Toxikon Corporation; 15 Wiggins Ave. Bedford, MA Undiluted, SR 1000 was evaluated for its potential to produce an irritating/corrosive effect on the ocular tissue of male and female New Zealand White rabbits. The left eye of each animal was treated with 0.1 ml of material as a finely ground powder placed into the inferior conjunctival sac of the eye. The right eye remained untreated and served as the control. Eyes were examined at 1, 24, 48, and 72 hours after treatment and responses graded for ocular lesions according to the study guidelines. All of the animals exhibited an increase in body weight during the course of the study. No overt signs of clinical toxicity were evident during the course of the study in any of the animals. No fluorescein staining was evident in any of the test animals prior to treatment or at 24, 48, or 72 hours following treatment. No lesions were noted in the treated eyes of any animals at any of the observation points. No positive signs of irritation were noted in the treated or control eyes at any of the observation points. Based on these results, SR 1000 has been classified a nonirritant to the ocular tissue of New Zealand White rabbits. Acute Dermal Irritation Final report is dated April 30, Study was conducted at Toxikon Corporation; 15 Wiggins Ave. Bedford, MA Undiluted SR 1000 was evaluated for the potential to produce primary dermal irritation after a single topical 4hour application to the intact skin of New Zealand White rabbits. A dose of.5 gm was applied to the skin under gauze patches, and then kept in contact with the skin for 4 hours by wrapping with an impervious, nonirritating bandage. Following the exposure period the animals were observed for signs of redness and swelling at minutes, and then at 24, 48, and 72 hours. All six test animals exhibited a gain in body weight during the observation period. No overt signs of toxicity were evident in any of the animals during the course of the Product Stewardship, Comp[ince and Standards 260 Hudson River Road, VVaterford, NY USA Panel Book Page 100

103 e MOMENT WE performance materais The science behind the solutions. study. All animals survived the test. No signs of erythema and edema were noted at any of the observation periods. The Primary Irritation Index (P11) was 0 (zero). Based on these results and the evaluation criteria, SR1000 is considered a nonirritant to skin. Acute Oral Toxicity Final report is dated May 22, Study was conducted at Toxikon Corporation; 15 Wiggins Ave. Bedford, MA SR 1000, was evaluated for its potential to produce death following oral administration at a dose level of 5 g/kg in male and female Sprague Dawley rats. SR1000, suspended in corn oil, was tested to 10 rats (5 male and 5 females) in a limit test. No adverse clinical signs were present at any of the observation periods. All animals gained weight during the observation period. No animals died at this does level during the observation period. No lesions were noted in any of the animals at necropsy. Acute Dermal Toxicity Final report is dated April 30, Study was conducted at Toxikon Corporation; 15 Wiggins Ave. Bedford, MA Undiluted SR1000 was evaluated for its potential to produce systemic toxicity or death following a single 24hour topical application to the shaven trunks of male and female New Zealand White rabbits. The test substance was applied to the skin under gauze patches at a dose of 2 g/kg, and kept in contact with the skin for 24 hours by wrapping with an impervious, nonirritating bandage. The wrapping was then removed and the skin wiped and rinsed with corn oil to remove any remaining test substance. Following the treatment period, the animals were observed for signs of redness and swelling. Animals were observed daily for 14 days for clinical manifestations of systemic toxicity. All animals gained weight during the post treatment period. No overt signs of systemic toxicity were evident during the course of the study, and all animals survived the duration of the study. Slight signs of irritation (Grade 1) were noted at the application sites of 4/10 of the animals. There reactions reversed by day 2 of the observation period. No abnormalities were found during necropsy. Based on the absence of mortality and according to the criteria of the study protocol, SR 1000 is considered nontoxic by dermal administration at the limit dose of 2000 mg/kg. Product Stewardship 7 Compliance and Standards 260 Hudson River Road, Waterford, NY USA Panel Book Page 101

104 MOMENTIVE performance materiats The science behind the solutions. Local Lymph Node Assay (LLNA) Final report is dated March 6, Study was conducted at BSL BIOSERVICE Scientific Laboratories GmbH, Behringstralje 6, Planegg Germany. Based on the preliminary test SR1000 was assayed at three concnetrations: 15%, 30%, and 60% (in AOO 1+3 (vlv), respectively. The vechicle was Acetone! Olive Oil. Five mice per test group were treated by topical appication of the prepared test item to the entire dorsal surface of each ear once daily over three consencutive days. Five days after the first topical application all mice were injected intravenously with 3Hmethy tlhymidine. Approximately 5 hours after 3Hmethy tlhymidineinjection all mice were sacrificed and the draining auricular lymph nodes t were excised. A single cell suspension of the lymph node cells for each animal was prepared. The 3Hmethy tlhymidine incorporation was measured in a f3 counter and expressed as the number of disintegrations per minute (DPM). Determination of radioactivity was performed individually for each animal. The proliferative response of lymph node cells was calculated as the ratio of 3Hmethy tlhymidine incorporation into lymph node cells of test group animals relative to that recorded for control group animals. A stimulation index, ratio of test item! negative control, was calculated for each concentration. The stimulation index at concentration of 15% was 1.0, at 30% was 1.1, and at 60% was 0.8. All animals showed the expected weight development. At the daily clinical observation the animals did not show any visible clinical symptoms. No EC3 value (derived by linear interpolation) could be calculated as all concentrations tested showed a stimulation index below 3. Considering the reported data of this sensitization test and due to OECD 429 it can be stated that the test item SR1000 causes no reactions identified as sensitization. Bacterial Reverse Mutation Assay Final report is dated December 28, Study was conducted at BSL BIOSERVICE Scientific Laboratories GmbH, Behringstralje 6, Planegg Germany. Product Stewardship, Cornpiance and Standards 260 Hudson River Road, Waterford, NY USA Panel Book Page 102

105 MOMENT WE performance materials The science behind the solutions. SR1000 was evaluated in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 and Escherichia coli strain WP2uvrA, both in the absence and in the presence of an S9 exogenous metabolic activation system. A range finding study indicated that treatment with SR 1000 at doses up to 5000 rig/plate in the presence and in the absence of S9 resulted in normal growth in tester strains. Doses tested in triplicate in Salmonella strains were 30, 80, 250, 700, 2000 and 5000 rig/plate in both the presence and absence of S9. Revertant frequencies in SR1000 treated plates were similar to vehicle control values. No toxic effects of the test item were noted in any of the five tester strains used up the highest dose group evaluated (with and without metabolic activation) in experiment I and II. No biologically relevant increases in revertant colony numbers of any of the five tester strains were observed following treatment with SR1000 at any concentration level, neither in the presence or absence of metabolic activation in experiment I and II. All positive and vehicle control data were within acceptable ranges. Results were confirmed in an independent assay. SR 1000 was nonmutagenic in a SalmonellaEscherichia coli /Mammalian Microsome Reverse Mutation Assay. Product Stewardship, Compliance and Standards 260 Hudson River Road, Waterford, NY USA Panel Book Page 103

106 Personal Care Products Council Committed to Safety, Quality & Innovation Memorandum TO: FROM: DATE: SUBJECT: F. Alan Andersen, Ph.D. Director COSMETIC INGREDIENT REVIEW (Cifi) JohnBailey,Ph.D. if Industry Liaison to the CIR Expert Panel January 12, 2011 Safety Studies on Products Containing Silica Dimethyl Silylate Consumer Product Testing Co Exclusive repeated insult patch test on a lipstick containing 7% Silica Dimethyl Silylate. Experiment Reference Number: C Institute for In Vitro Sciences, Inc Topical application ocular irritation screening assay using the Epiocular TM human cell construct (eyeliner containing 2% Silica Dimethyl Silylate). Study number 1OAF14AF TKL Research Repeated insult patch test of an eyeliner containing 2% Silica Dimethyl Silylate. TKL Study No.: DS1O6O1O14. Consumer Product Testing Co Ophthalmological inuse safety evaluation of an eyeliner containing 2% Silica Dimethyl Silylate. Experiment Reference Number: C th Street, N.W., Suite 300 Washington, D.C (fax) Panel Book Page 104

107 )67 Consumer Product Testing Co. FINAL REPORT CLIENT: ATTENTION: TEST: Exclusive Repeated Insult Patch l cst Protocol No.: 1.D TEST MATERIAL: EXPERIMENT REFERENCE NUMBER: C O Richard R. Eisenbcrg, MI). Board Certitied Dcrrna!.ologist / FrKnk, RN. xecutive Vice President, Clinical Evaluations This report is submitted fr the exousive use of the person, pertrrerstp corporation to ni it is addressed, and neither the report nor tre name of these Laboratories nor any member of its slalt. may,e Ja.c.5 Corinec:don will hn advertising or sale of any procuct or procoss witnout Written authorjzation 70 New Dutch Lane Fairfield, New Jersey 4)7it a l 4 (( I I Rt tj)7: )8744 Panel Book Page 105

108 Consumer Product Testing Co. IU. 975 QUALITY ASSURANCE UNiT STATEMENT Study No.: C Q The objective of the Quality Assurance Unit (QAU) is to monitor the conduct and reporting of clinical laboratory studies. These studies have been performed with adherence to the applicable ICH Guideline E6 for Good Clinical Practice and requirements nrovided fti di. 21 CFR parts 50 and 56 and in accordance to standard operating procedures and applicable protocols. The QAU maintains copies of study protocols and standard operating procedure.. and has insroc1ed this study. All data pertinent to this study will be stored in the Consumer Product festing Company archive, unless specified otherwise. in writing by the Sponsor. Quality Assurance personnel involved: uality Assurance Date The representative signature of the Quality Assurroco kit sigdfis that this study has been performed in accordance with standard operating procedures am. study protocol as well as government regulations regarding such procedures and protocols. 7() New t)itch Lane f:articld, New Jersey U7OO2n 14 $73}O87 I I I Iax e)73) 8()a7234 Clinical ToxicoIoy Analvucal (1ei slr Mwrnbiologv Panel Book Page 106

109 C,çpiction C Page 3 Objective: To determine by repetitive epidermal contact the potential of a test material to induce primary or cumulative irritation and/or allergic contact sensitization. Participants: One hundred nine (109) ua1ified subjects, male and female, ranging in age from 16 to 79 years, were selected for this evaluation. One hundred (100) subjects completed this study. The remaining subjects discontinued their participation for various reasons, none of which were related to the application of the test material. Inclusion Criteria: a. Male and female subjeis, ae 6a and over. b. Absence of any visbie sk:n disease which might be confused with a skin reaction from the test material. c. Prohibition of use of topical or systemic steroids and/or antihistamines for at least seven days prior to study initiation. d. Completion of a Medical Hister: form and the understanding and signing of an lnformec Consent form, e. Considered reliable and caable of following directions. Exclusion Criteria: a. 111 health. b. Under a doctor s mr r taking medication(s) which could influence the outcome of the stucy c. Females who are pregnant or nursing. d. A history of adverse reactions to cosmetics or other personal care products. Test Material: L;, L c Co. 7 S (c D: / Study Schedule: Panel # lnitij.e. Date January 21, 2008 February 29, 2008 arith parental or guardian consent Panel Book Page 107

110 C080294, 10 Page 4 Methodology: The upper back between the scapulae served as the treatment area. Approximately 0,2 g of the test material, or an amount sufficient to cover the contact surface, was applied to the 3/4 x 3/411 absorbent pad portion of an adhesive dressing*. This was then app!ied to the appropriate treatment site to form an occlusive patch. Induction Phase: Patches were applied throc (3) times ocr week (e.g., Monday, Wednesday, and Friday) for a total c nine (9) aplications. The site was marked to ensure the continuity o patch application. Following supervised removal and scoring of the first Induction patch, participants were instructed to remove all subsequent Induction patches at home, twentyfour hours after application. The evaluation of this site was made again just prior to re application. If a participant was unable to report for an assigned test day, one (1) makeup day was permitted. This day was added to the Induction period. With the exception of the frst upcrve induction Patch reading, if any test site exhibited a moderate (2level :e ction during the Induction Phase, application was moved o an adjacent area. Applications were discontinued for the remainder of this test phase, if a moderate (2level) reaction was observed on this new test site. Applications would also be discontinued if marked (3level) or severe (4level) reactivity was noted. Rest periods consisted o twentyfbo hours following each Tuesday and l hursday removal, and ortye:ht hours following each Saturday removal. Challenge Phase: Approximately two (2) eeks after te final Induction patch application, a Challenge patch was appled to a virgin test site adjacent to the original lnduction patch site, following the same procedure described fbr Induction. The patch was removed and the site scored at the clinic twentyfour and seventytwo hours postapplication. *Manufactured by TruMed Technologies, Inc., Bumr ille. vfn Panel Book Page 108

111 C O Page5 Evaluation Criteria (Erythema and additional Dermal Seguebe): 0 0.5/ = No visibi skn reaction = Barely peceotib!e = Mild Moderate = Marked = Severe E Edema D Dryness S Staining P = Papules V Vesicles 13 = Bullae U Ulceration Sp = Spreading Erythema was scored numercally according to this key. if present, additional Dermal Seqw lac wern indicated by the appropriate letter code and a numerical value for seventy. Results: The results of each participant are appended (Table 1). Observations remained within normal limits throughout the test interval. Subject demographics are presented in Fable 2. Summary: Under the conditions of ti tidy, material, did not indicate a potential for drmal lrritafrjn cr allergic contact sensitization. Panel Book Page 109

112 0 CO Page 6 Table I Panel # Individuai Results Virgin Challenge Subject Induction Phase Site Number 24*hr *hr 72hr () ( Ii DID.c)F COMPLETE STUDY DII) NOT COMPLETE STUDY ) ( ( C * Supervised removal of E1 Induction and Challcie :uch = Subject not present for supervised removal Panel Book Page 110

113 C Page 7 1 able I (continued) Panel # lndividua Resehs Virgin Challenge Subject 1nductio. 1ase Site Number 24*hr hr 72 hr o o J:)!fl OT COMPLETE STUDY o ljo NOT COMPLETE STUDY o or Ct o o o ( ( ( ) ) 1) 1) ) ) ) 0 24* Supervised removal of l Induction and Challenge Pitch m Additional makeup day granted at th discretion fti:e cliiic suprvisor Panel Book Page 111

114 DID C l0 Page 8 Table I (continued) Panel # Individual Results Virgin Challenge Subject Induction Phase Site Number 24*hr *hr 72 hr DID NOT COMPLETE STUDY NOT COMPLETE STUDY * Supervised removal of l Induction and ChaJ1eng Patch Panel Book Page 112

115 CO Page 9 Table 1 (continced) Panel # Individual Results Virgin Challenge Subject Inductior Phase Site Number 24*hr *hr 72 hr o o 0 0 o ] DID NOT COMPLETE STUDY ( DID NOT COMPLETE STUDY Jo o o ( o o DID N(.Yi COMPLETE STUDY 24* Supervised removal of 1 Induction and Challenge Patch Panel Book Page 113

116 C Page 10 Table 2 Panel # jct Data Number Initials Sex MC 63 F 2 KT 16 F 3 M13 76 F 4 SB 70 F 5 WB 73 M 6 EW 75 M 7 BR 76 F 8 JW 71 M 9 HL 74 F 10 MO 44 F DS 47 F 12 KK 47 F 13 LE 72 F 14 CC 71 F 15 ND 72 F 16 DO 74 F 17 HG 75 M 18 WP 78 M 19 MM 41 F 20 KV 32 F 21 TJ 35 F 22 HC 59 F 23 JO 56 F 24 CD 45 F 25 U 43 F 26 MB 77 F 27 J13 76 NI 28 DC 28 F Panel Book Page 114

117 C Page 11 Table 2 (continued) Panel # ] jecti)ata Subject Number Initials Age Sex 29 HF 36 F 30 AQ 60 F 31 JQ 55 F 32 MF 37 F 33 GL 78 F 34 WT 69 F 35 PT 71 M 36 MM 66 F 37 VM 62 F 38 FR 79 F 39 SV 37 F 40 FP 65 F 41 MI 66 F 42 LF 48 F 43 DG 69 F 44 JD 61 F 45 RP 57 M 46 JC 46 F 47 MD 40 F 48 MG 42 F 49 lb 19 F 50 HR 72 F 51 PR 60 M 52 MR 72 F 53 TR 20 F 54 LR 32 F 55 AA 47 F 56 AS 64 F Panel Book Page 115

118 Initials C Page 12 I able 2 (continued) Panel # Subject I)ata Subject Number Age Sex 57 AL 51 M 58 DL 24 F 59 EC 21 M 60 AL 61 F 61 GB 43 F 62 MR 24 F 63 in 64 LL 41 F 65 PR 43 F 66 3W 45 F 67 MP 55 M 68 SA 31 F 69 EB 61 NI 70 JES 66 F 71 BJS 38 F 72 MV 53 F 73 KR 38 F 74 DM 5] F 75 IT 59 F 76 EF 36 F 77 JT 58 M 78 CC 70 M 79 MR 53 F so AR 36 F 81 ET 48 F 82 CR 49 F 83 QM 30 F 84 ND 42 F 59 F Panel Book Page 116

119 C Page 13 Fable 2 (continued) Panel # $ject Data Subject Number Initials Sex 85 KC 43 F 86 MC 31 F 87 MB 55 F 88 SC 47 F 89 W13 35 M 90 JB 30 M 91 SB 57 F 92 JI) 38 M 93 RO 27 F 94 TE 29 F 95 C P 47 F 96 DP 25 F 97 CK 57 F 98 RV 69 F 99 PK 47 M 100 MD 47 F 101 GO 60 F 102 CA 40 F 103 PG 47 M 104 MC 35 F 105 EP 38 F 106 CB 39 F 107 DB 59 F 108 JA 44 M 109 MD 50 F Panel Book Page 117

120 FINAL REPORT udy Title TOPICAL APPLICATION OCULAR I RRITATION SCREENING ASSAY USING THE EPIOCULARTM HUMAN CELL CONSTRUCT Test Articles c o ic q r / Q Authors Greg Mon. BA. Monica Krcha, B.S. Study Compjetion Date 26 July 2010 Pert rming Jboratojy Institute for In Vitro Sciences, Inc. 30 W. Watkins Mill Road, Suite 100 Gaithersbure, MD Study Number I OAFI 4AF Laboratory Proycet Number 594. Page of 11 Panel Book Page 118

121 TABLE OF CONTENTS TABLE OF CONTENTS 2 SIGNATURE PAGE 3 TEST ARTICLE RECHP1 4 TOPICAL APPLICATION OCULAR IRRITATION SCREENING ASSAY USING THE EP1OCULAR HUMAN CELL CONSTRUCT INTRODUCTiON 6 MATERIALS AND METIIODS 7 RESULTS AND DISCUSSION 11 APPENDIX A SF (PROTOCOL) 19 PROTOCOL ATTACHMENT I APPENDIX B (ANALYZED DMA) BIB3 I Project 5942, Final Report 2 Panel Book Page 119

122 , 4 1 Ck SIGNATURE PAGE TOPICAL APPLICATION OCULAR IRRITATiON SCREENING ASSAY USING THE EPIOCULAR HUMAN CELL CONSTRUCT Initiation Date: 4 Jime 2( 0 Completion Date: 26 July 20 0 Sponsor: Sponsors Representative: Testing Facility: Institute Ibr In Vitr Sinces, Inc. 30 W. Watkins MiH Road, Suite 100 Gailhersburg. MD Archive I ocation: Institute f3r In Vitro Sciences. Inc. Gaithersburg. MD Study Director: Iy 2o to Greg Mutt. B.A, Date Laboratoiy Manager: Nathan R. Wilt, BS. Project 5942, Final Report 3 Panel Book Page 120

123 Protected TEST ARTICLE RECEIPT IIVS Test Article Sponsor s Ph sicai Storage Receipt Date Number Designation Description Conditions IOAF14 black solid(28 May OAF I biack solid i.jun 2010 temperature. room temperature * from exposure to light Project 5942 Final Report 4 Panel Book Page 121

124 TOPICALAPPLICATION OCULAR IRRITATION SCREENING ASSAY USING THE EPIOCULARTM HUMAN CELL CONSTRUCT Project 5942, FInal Report 5 Panel Book Page 122

125 INTRODUCTION The EpiOcular TM 1luman Cell Construct was used to assess the potential ocular irritation of the lest materials. The MTT (3 [4,5 dimethylthiazol.2 yl] 2.5 dipheny!tctrazolium bromide) conversion assay, which measures the NAD(P)Il.dependent microsomal enzyme reduction of MTT (and to a lesser extent. the succinate dehydroenase reduction of MTT) to a blue tbrmazau precipitate, was used to assess cellular metabolism after exposure to each test article for various exposure times!. The duration of exposure resulting in a 50% decrease in MTT conversion in test articletreated EpiOcIllarTM human ccli constructs, relative to control cultures was determined (ET 50 ). The tjurpose of this study was to evaluate the potentiel toxicity ofihe test articles. supplied by constructs after exposure to each test article for various exposure times. The laboratory phase of as measured by the conversion of MTT by EpiOcular tm1 human ccli the study was conducted From 7 June 2010 to 10.lune 2010 at the Institute for In Vitro Sciences, Inc. The test articles were tested in a screening assay to delennine the duration of exposure that resulted in an ELo endpoint. Berridge, MV., Ian, A.S.. McCoy, K.D., Wang. R. 1996) The t3in:hernical and Cellular Basis Of Cell Proliferation Assays That Use Tetrazolium Salts. Bioclieniica 4:1419. Project 5942, Final Report 6 Panel Book Page 123

126 MATERIALS ANI) METHODS Receipt of the EpiOcularThl 1lunian Cell Consti t Model Upon receipt of the EpiOcular Human Cell Construct Kit (MatTek Corporation), the solutions were stored as indicated by the manufacturer. The EpiOcular tm human cell constructs were stored at 28 C until used. On the day ol dosing. B iocnimtm Assay Medium was warmed to approximately 37 C. Nine tenths ml of Assay Medium were aliquoted into the appropriate wells of 6well plates. The sixwell plates were labeled to indicate test article and exposure time. The constructs were inspected for air bubbles between the agarose gel and Millicell insert prior to opening the sealed package. Cultures with air hubbies covering greater than 50% of the Millicell area were not used, The 24 well shipping containers were removed from the plastic hag and their surfaces were disinfected with 70% ethanol. The EpiOcular TM human cell constructs were transferred aseptically into the 6w l1 plates. The constructs were then incubated at 37±1 C in a hurnidilied atmosphere of 5±1% CC2 inair (standard culture conditions) for at least one hour. The medium was then aspirated and 0.9 ml. or fresh Assay Medium were added to each assay well below the EpiOcuiar incubator until treatment was initiated. TM human cll construct. The plates were returned to the Test Article Preparation dilution. As instructed by the Sponsor, each test article was administered to the test system without Assessment_of Direct Test Article Reduction of MTT Each lest article was added to a 1.0 mg/nil vitt (Sinna) solution in warm Dulbecco s Modi lied Eagle s Medium (DMEM) containing 2 mm Lgluinmiue (MTT Addition Medium) to assess its ability to directly reduce MTT. Approximately 100 iloithe test article. and 30mg of the test article,,were added to I ml of the MTT solution and the mixtures were incubated in the dark at standard culture conditions for approximately one hour. A negative control. 100 pl of sterile. deionized water (Quality Biological), was tested concurrently. If the MTT solution color turned blue/purple, the test article was presumed to have reduced the MTT. Water insoluble, test materials may show direct reduction (darkening) only at the interface between the test article and the medium. In cases where the test article was shown to reduce MTT, only those test articles that remained bound to the tissue after rinsing, resulting in a false MTT reduction signal. could present a problem. To evaluate whether residuai!cst article was binding to the tissue and leading to a false MTT ieduction signal, a functional check (using freezekilled control tissue) was performed as described in the section labeled Killed Controls (KC). The test articles were dark colored and their ability to directly reduce MTT could not be determined. A killed control experiment was performed concurrently in the screening assay to determine the extent of the direct MTT reduction (if any) by the test articles alone. Project 5942, Final Report 7 Panel Book Page 124

127 pu_determination The ph of the test article, was measured using ph paper (EM1) Chemicals Inc.). The test article was added to pil paper with a 0 14 p1 1 range in.0 ph unit increments to approximate a narrow p1i range. The pli could not he determined because the test article discolored the ph paper. The test article,, was a solid and therefore, the p1i was not determined. MTT Assay The EpiOcular TM cultures were treated in duplicate with the test articles at four exposure times of 240, 480, 960 and 1440 minutes. One hundred niicroliters of the test article, and thirty milligrams of the test article. were applied to each EpiOculaurM human cell construct. Due to its viscous rniture. an aliquot of the test article, was placed into a 3 ml syringe to help rid in loading the pipet tip. The pipet tip of the positive displacement pipet was inserted into the dispensing tip of the syringe so that the material could be loaded into the displacement tip under pressure. Simultaneously, the syringe plunger was depressed as the pipet piston was drawn upwards. If air hubbies appeared in the pipet tip, the test article was removed (expelled) and the process repeated until the tip was filled without air bubbles. A dosing device (fiatheaded cylinder ofshuhtly less diameter than the inner diameter of the tissue insert) was placed over the test article to assure even spreading over the surface of the tissues. Due to its viscous natem 30 mg samole of the test article. was measured and placed on the tip of a dosing device. The test article was then Ilatiened and the test article/dosing device was then placed on each tissue. Duplicate cultures of the negative control (exposure time control), I 00 ul. of sterile, deionized water (Quality Biological), were exposed for ; 480, and c,0 minutes. Duplicate cultures of the positive control, 100 1iL of 0.3% TritonX100 (Fisher), were exposed for 15 and 45 minutes. The exposed cultures were then incubated for the appropriate amount of time at standard culture conditions. After the appropriate exposure time, the EpiOcular cultures were extensively rinsed with Calcium and MagnesiumFree Dulbecco s Phosphate 13u Efered Sal inc (Ca MgFreeDPBS) and the wash medium wasdecanted. After rinsing, the tissue was transferred to 5 ml of Assay Medium for a 10 to 20 minute soak at room temperature to remove any test article absorbed into the tissue. A 1.0 mg/ml solution of MTT in warm MTT Addition Medium was prepared rio more than 2 hours before use. Thréeteiiths ml of MTT solution were added to designated wells in a prelaheled 24well plate. The EpiOcular TM constructs were transferred to the appropriate wells afler rinsing with CaMgFreeDPBS, The trays were incubated for approximately three hours at standard culture conditions. After the incubation period with MTT soiution. the EpiOcular 1 cultures were blotted on absorbent paper, cleared of excess liquid, and trcusterrcdto a prelabeled 24well plate containing 2.0 ml ofisopropanol in each designated well. rhe niales weic sealed with parafilm and stored in the refrigerator (2 8 C) until the last exposure time was harvested. The plates were then shaken Fir at least two hours at room teniperature At the end of the extraction period, the iquid within the Millicell 5 inserts was decanted into the well From which the Milliccll insert was taken. The extract solution was mixed and 200 tl were transferred to the appropriate wells of a 96well plate. Two hundred pl of Project 5942, Final Report 8 Panel Book Page 125

128 isopropanol were addcd to the two wells designated as the blanks. The absorbance at 550 mu (0D ) of each vell was measured with a Molecular Devices Vmax plate reader. 550 Killed Controls (K To evaluate whether residual test article was binding to the tissue and leading to a false MIT reduction signal, a functional check (using freezekilled control tissue) was perftrmed. Freeze killed tissues were received already prepared from Matlek, and were stored in the freezer until use. For the test articles, and single killed tissues were treated with the test article in the normal fashion for 240 and 1440 minutes (the shortest and longest test article exposure time). All assay procedures wer a aerformed exactly as described for the viable tissues. Single killedcontrol tissues were treated with the negative control (100 gl of sterile. deionized water) for 0.25 and 24 hours (the shortest and longest negative control exposure time). A small amount of MIT reduction is expected from the residual NADH and associated enzymes within the killed tissue. This background reduction of MTT was compared to the MTT reduction observed in the test articletreated killedcontrol tissues. the raw OD 0D The raw 0D 550 value of the negative conro1 treated killed control was subtracted from 50 values for each of the test articleireated killed controls, to determine the net 550 values of the test articletreated killed controis. The n C)D 0 values represent the amount of reduced MTT due to direct reduction by :est article residues. For the test articles. and, there was little or no direct MIT reduction in the test articletreated killed controls compared to the negative controltreated killed controls and the MTT reduction observed in the test articletreated viable tissue was ascribed to the viable cells. Presentation of Data The raw absorbance values were captured he mean 0D calculated. The corrected mean CD subtracting the mean 0D CD 550 value of the blank wells was 550 values of the negative controls were determined by 550 value oi the blank wells from therr mean 0D 550 values. The corrected 550 values of the individual test article exposure limes and the positive control exposure times 550 val ic of the blank wells from their 0D 550 values. were determined by subtracting the mean 0D All calculations were performed using an Excel sprcadsheel The following percent of control calculations were made: corrected 0D 550 of Test Article or Positive Control Exposure lime % of Control X 100 appropriate corrected mean OD 55 Negative Control Exposure time response curves were ploted with The % of Control on the ordinate and the test article or positive control exposure time on the abscissa. The EL from each plot, To determine the ET 50 0 value was interpolated, two consecutive points were selected, where one exposure time resulted in a relative survival greater than 50%, and one exposure time resulted in less than 50% survival. Two select points were used to determine the slope and the yintercept fur the equation ym(x) ± h. Finally. to determine lie ET 59. inc edluation was solved for y50. When all of the exposure time points showed grcate than 50% survival, the El presented as greater than the longest test article exposure time. 50 value was Project 5942, Final ReporE 9 Panel Book Page 126

129 Criteria for a Valid Test The assay results were accepted when: 1) The ET 50 value of the positive control fell within two standard deviations of the historical mean (updated every three months) and 2) The corrected mean 0D the corrected mean 0D 240 minutes). 550 value for the minimum negative control exposure time was within 20% of 550 value for the rnaximrm negative ontro exposure time (up to Project 5942, Final Report 10 Panel Book Page 127

130 Discolored RESULTS AND DISCUSSION MIT Assay The EpiOcularm cultures were treated in duplicate with the test articles, and, at thur exposure times of 240, 480, 960 and 1440 minutes. The negative control was exposed in duplicate for 15, 240, 480, and 1440 minutes. Table 1 summarizes the ET 50 results of the Topical Application Ocular Irritation Screening Assay Using the EpiOcuiar 1 1luman Cell Construct for the test articles and the positive control. The exposure time response curves are included in Appendix B. Since the positive control fell within two standard deviations of the historical mean ( minutes), and the corrected mean OD 550 value for the minimum negative control exposure time (1.474) was within 20% of the corrected mean 0D value for the maximum negative control exposure time (up to 240 minutes) (1.286), the assay results were accepted. 550 The test articles were dark colored and their ability to directly reduce MIT could not be determined. Therefore, a killedcontrol experiment was perlbrmcd. The results of the killed control experiment showed that there was little or no direct MTT reduction in the test articletreated killed control compared to the negative control treated killed controls and the MTT reduction in the test articletreated viable tissue was ascribed to the viable cells. The test articles, and.., cot id not be completely removed from the exposed tissues following the rinsing and soaking process after all exposure times. The residual test article prolonged the exposure to the tissues, which may have intluenced the toxic effeci. However, the ET 50 results were not affected because the test articles did not cause> 50% relative toxicity to the tissues up to the longest prescribed exposure time of 1440 minutes (ET 1440 minutes). 50 > Table I Assay Date. IIVS Test Article Sponsor s FT. Conc., 50 Number Designation (minutes) p1. 1OAFI4 Neat > 1440 DpH 9 June 2010 IOAF15 * Neat > 1440 ND Positive Control 0.3% TritonXI00 NA 29.4 NA DpiI p11 paper, the ph could nt be determined because the lest article discolored the ph paper NI) Not Determined, the ph was not deter nined because the test article is a solid. NA Material Benchmark Not applicable Project Final Report II Panel Book Page 128

131 material. ( i r ) 1? D, fl: t k I RESEARCH v i SUMMARY REPORT TKL Study No.: DSIO6OIO14 Date of Report: July Study_Title: Repeated Insult Patch Test Study Sponsor: TKL Protocol No.: TKLl000 Sponsor Reference No.: Study Objective: To assos: the. scnshiation potential of topically applied s:ucly Study Design: Standard RIPT rne:hodology with nine 24hour induction appiicai rs and a single 24hour challenge applica.ion followiag 1015 day rest period. Principal Investigator: Jonathan S. Dosik, MI) Dermatologist Director, 1)ermatologic Safety Testing: Kathleen Georgeian Manager, Dermatologic Safety Testing: Michelle Medina Corporate Office: IKL Rcscirch, Inc. Study Site: 365 W. Passaic Stren. Rochdlle l*a.rk, NJ 0 j62 l KL Research Inc. 48 Franklin Turnpike Ramsey, J Study Dates: Date S1d inii;ilcd: June 2, 2010 Date Study Ccmpletei: July 9,2010 Study Material: SPL Code_and Category u )untappiid Patch Conditiop Eye Liner 0.2 g Occlusive TKL Clinical Trials Division TKL Clinical Site DL4sion N rkl Recruitment Management Division Corporate Office: 365W Passaic Street, Rochelle ParkNJ Phone: 2(i1 587O5OO Panel Book Page 129

132 SUMMARY FEP(W.T(Contnued) TKL Study No.: 1)S Of 3 Number of Subjects: Enrolled: I I I Completed: 107 Discontinued: 4 Lost to followup: 2 \ohnllr ( withdrewal 2 Listing of Attached Tables: Appendix 1 Summary fables Appendix II Data Listings SUMMARY ANI) CONCLUSION: There were no adverse events reported. A summary ol response data is provided in Ta) V 3 Appcnfc I. Individual dermatological response grades are provided in Data Listing 3, Appendix I. Under the conditions employed in this study. there was no evidence of sensitization to SPI. Code STATEMENT OF QUAlITY CONTROL: The Quality Control Unit of the DermatologicI Safety Deiv nnent conducted a 100% review of ai study related documents. The protocol was reviev ed rior n % start of the study, and the medical screening forms and informed consent documents were reviev.ed nprocess of the study. The regulatory binder and study data were reviewed poststudy to ensure accuracy. The study report was reviewed and accurately reflects the data for this study. Panel Book Page 130

133 SUMMARY REPOf (Codhiied) TKL Study No.: DS Page 1 of 3 SiC NATURES: I have read Ihis report arni confirm thu o the best of my knowledge ii ucc:uratel describes the conduct and results of (he Sluny. Jo than S. Dosik, MI) Derrnatologi stipri ncipal Invesligator Director, Dermatologic Safety Testing 1)ate Michelle Medina i Ye Manager, Dermatologic Safety Testing Date 1 i Panel Book Page 131

134 I XIUiddV Panel Book Page 132

135 TKI. Study No. DS Page of I Table I Summary of Suhect Onrollment and Disposition Subjects enrolled N (%) i Subjects completed induction phase 107 (96.4) Subjects completed all phases 107 (96.4) Iota) subjects discontinued 1 (3.6) Lost to follow up 2 (1.5) VolunLuy withdrawal 2 ( I.8) Note: All percentages are relative to total subjects curolicd. See data listing I for further detail. Generated on 07/13/10:13:28 by DISPSMY.SAS / Uses: lina1. Panel Book Page 133

136 TK I Sud No. DS Page 1 of Table 2: Summary of Subject Demooiiphics \ll l:nrnlied Suhccts N(%) 111 to44 35(31.5) N (%) (49.5) N (%) 65 and up 2 (18.9) Mean (SD) 52.2 (12.5) Median 51.6 Range 19.7 to 75. ) Gender N 1%) Male N (%) Female 00 (90.1) I (9.9) Race 13ack Caucasian Ilispanic ) 2 I 11)5 (IX) (0 9) (94.6) See data listing 2 kr urther dciail. Generated on 07/13.10: 13:28 by DFMOSMY.SAS / Ltscc )\1(.CS Panel Book Page 134

137 TK Study Nw I)S Payc I of (able 3 Scmimarv of ()ermatoloaic Rep nse Number of Subjects by Produet C; I soles Prodoct Response total eoaluahle Number absent Number discontinued Indurticn 1h Chaflenc Phase 6 7 S ) ( ( Make Up 04 9 (04 451w I I 1 1 1) 3 0 1) hr 96hr ) 107 (07 Maximum Elicic:) 1 esoetse 1)urinc lndtienon All Subjects (aro eros Inductt.o N 107) Response n(%) Sublects 11)7 (100.0 ) (*) when required Key I) p to Symbols No reaction Definite er themu. Dc lint Ic to I)amagc no Minimal or doubt edema Definite ervihemo, delintic edema erythema. definite edema epiderm Papular response >50% 5: 7 oozing, crusting aiid andicor fiti vesieu lation respon m. slitehtlv superficial erosions I di tfercnt Eom surrounding nonnal skin ( LflciaLd on s h S1 \lmr t S S I Si s Cl SPD\sl PRODI IS I l\ Si Panel Book Page 135

138 APPENDIX II I)ATA L S1NGS Panel Book Page 136

139 p Jo I 4uj (1 ON A(IflJ.S IlL U0!I!SodS!Ø Pun wwiioui 1* S : 8uqs naurj sarnq pn; tic a a otoo.to oi9oa.o oiito,90 1/tft Pr I scsu uopaidwoa Sujpsqg 4iis II.4 JIIdóV Ii IZ ajjddjp; XC IQp&tp 0lfi0fl0 OIitW9O SW XC / OI/t0190 0l/t XC W OflOILO O1. Wi90 011Z XC : O11t0190 0I/Z0190 tb XC 3 a 01i60/LO OISOiLO 01i10I90 01iZ XC.3 a 0I to.lo oisoito OiitW9O 011t XL : It0I90 01IW XC M0 olrovoo 0I/t XC.3 a LO 01:9Cc/sO OIIW/90 01IZ0190 *00 XC t0 01:90/1.0 01It0190 OIIW/90 00 XC.3 a 01/ to/90 01st0190 too XC 3 a oiioa to 01W/sO OIItW9O OI/Z lie 3 3 or6olo 0L901L0 OIitO/90 0I Z0190 LAO tic a a L IL0 011W190 01It xt a a Oi.i,WLO oisaq.o OI,t0I90 0I1z tic a a ouoww Ol90io oiaoso ouzwco rio XE.3 a 0l6êL(i 01 90I.O Ol t t XL 3.3 0I/6WL( Ouroqo OIiW/90 L1O RE IL0 011t /t0/90 RIO XE.3 3 0l.tG LU 0I9tZfLO OIltOi9O 011W190 1W XE a a 01,60L0 OI 1JWtO 011z0 9O 01ZW9O OtO tic a a oi:6aj,fjj oioio ocizwo 01/C090 czo tic a 3 OIiqOâ.u,.OL 901J 011t090 01Th0190 LW XE % t t0190 PZ0 tic a oi.svw 0l 90iL0 oucoi9o OI/t0i90 XC i601L0 0I 90 LO oiaoico 011t0190 9W tic a a 0116 Li) 0v9(40 O1IZO10 OllZl90 LW tic.3 3 O1,601L0 OIWLO 0IIZ /t0/90 6W tic i60 LO Ol Z Z0190 0(0 XE L0 0l 90 L0 OlitO 90 OI:tO/90 ItO (.mtjj U3lIS1t).3 ISC1M unpaupaf j) si Supu.i sri OWI.%PVjI%t UoIIIqotA 1flSoJ(UJ.A fr%iaptp!m dcituflioa.s Inaoiioj 01 W(flJ polajdwoj3) Sf1015 U0!W1dW0 a iua*o ivnii 3st4uds:1a stxwi!iu :s35fl svnsiiasici Sq wcl:01,cla,ij to pinauaj Panel Book Page 137

140 TKL STUDY NO. DS Page 2 ot 4 Data isting I Subject 1/or lii ent and Disposition Subject o. Screcned 1st Applic Study 1)atcs ChrIl Applic LoPed Last Reac1in (trnplction Status /02/1 0 o :;oo 10 0/ 00: 1) ( C /02/1 0 06/12/10 07/06! Q1 0 C C OM)2/l ,02/10 06/02/ ,02/ / :0 (t7/ /10 06/ /10 06/ /02! 0 06/02/ /02. lo 06,102/10 I)iivs in Study 07 09, i0 C C /10 C C 38 07:06! ) C C 38 06/ ! o. 10 (COO! 0 C C /02: 10 06/02/10 0//CO / / /02/ ! It) 07:00 (1 ( 3$ 11/ ) / (70910 u :10 C C 3$ 1 C 38 1 ( 38 I /02. ] 0 00/02 ] , 0 07 OO:l 0 ( C : /021! 0 Ii) ] 0 C ( /02!!) 0 C i O I)) io 1 C /02! ] ((7 09.! 0 1 C / C C /02, : 0 07/09/ / 0 (160)2: 1(1 10 (17.09/! 0 C :02: :09/ : C C 3$ 052 ( 0Y02/I0 1171)6 0 C ( C C 3$ ( l ]() C C /02!10 06/02:10 0 Oñ T i1) C ( 7$ : )0 06:02) :02:10 1) :10 06/02 1(1 0/ ),0o10 1)6.02: Itt 0602/tO 1) C ( 3$ 07,09:10 C C 3$ 06:02: Oo. l ) 0 C C / (1602! 10 06/02/11) 07:00 0 ( /02/10 06/02/1 0 07: / /02/10 0 /OO )9 11) C C /)0 0700:10 C C.30 C C 3$ C C 38 Key. Reading 0 11 Induction Phase, C Challenge Phase) Status 1. Lou to toilow up. S event, Other) last (omplet:on 0 (C Coinpleted..l,iniar sviil a), V Piutocol s :ulatio:i, \l Adserse Generated ott /I 0:13:28 1w DISP1lST.SAS / Uses: D1/130135, RESPONSE, FINAl. Panel Book Page 138

141 I Protoco( TKI, STUDY NO. OS Page 3 of 4 Daoi Lsting Subject l/nroilnicni and Disposoton Study l)azes Subject \o tm ecncd I i ppio. (Ii ill \pplic I Last Reading Completion l)ays in mtdid I St abs Studa / /02/10 07/ /10 C C 3$ / / /0 07:09/10 C C 3% /i 0 06/ / /10 C C / , J11) C C 3$ /02/ )709/10 C C 3% ;10 06,02/10 0 Ota IU C C 3$ /02/ / )0.1(1 C C 3% 1)7(1 Oo!02 lo 00: IC 1, 0 il C C 7% 07) 06.02/ /l :4:0 C C 3% /02/10 06:02/10 1) :10 C C 3$ C 073 0o02/I0 0602/ /IC) C 07/0910 3$ /02/10 06/02/10 07:06 l (1 07/09/I 0 C C 3% l :10 C ( 3% 1) i )210 07/) lC( /02/10 06/02/IC) 0 (to, 10 (1700 ] 07% 0602/10 06/02/10.. 1)6 74/I )662/10 06/0211) Ill 1)7 00/10 C C 1) (1 C C 38 C C 3% 1(1 S 3 08(1 0602:11, /)0 0T Q9 10 C ( /02/10 06/02)1) 1)71)6 10 /1 09.1)) C C 3% /02/1(1 0602/1(1 0 o6 It) Ii 1)0/10 C C 3% () io :00/10 C C 3% 084 WY02 H) 0602/(0 071)6 II) C 1 3% /02/10 06/02/10 071( /I)) /) 0 061)2: ) 07.00/(0 C C. C 3% /02/ /11) C C 3% C 38 08% ) 06.02: /10 C C 3% l 1) 06:02:1(1 07/0611(1 0709/1 0 C C 3% / /l /lU C C 3% 09! (602,1(1 06:02/10 0 Xi, (1 C C 3$ ;02 I ) (COlt 10 1)7 09 1(1 ( C 2$ ,02/10 0,Ci, ii) C C 3% Key: I,ast Reading II linduc) ion Phase, C COal lenge Phase) Comp)etion Status (CCoinpleted. LLost to ) ollowup, S V( Iuntarv r ithdrcwa1, V event, OOther) victlattn, Al: Adverse Generated on 07/1 3/1(1:13:2% by DISPI,IS l.sas / Uses, (, 7.IOCS, RIISP( 40SF, ) INAI, Panel Book Page 139

142 Protocol FKI. STUI)Y NO. DS1064J10 Pane 4 ol 4 Data Li sling Subject Lurol ment and Disposition Study Dates Last Reading Completion Das in Subjeci No. Screcned 1st Applic Chall Aopbc Ludcd Status Study /02/10 06:02(10 0) i0 C ( 3$ /02/10 06/02/10 07/06 10 (t C C /02(10 06/02/1(1 07/06:10 07,09,10 C C / /02/10 07/ /09(10 C C /10 06/02/10 07/ /09/10 C C 3$ /02/10 06/02/10 07/ :09110 C C 3$ /02/tO 06/02/ (09/10 C C /02/I 0 06/02/10 07/ /l 0 C C /07 IO 06(02/10 074)610 07i09/l () ( C j /02] C C /02/10 06/ , /02/Il) 06 02/10 1)706:10 07/09 0 C C / l0 071)610 07/0910 C C /l ty 06 It) C ( /1(1 06:04/10 ti (19:1(1 C C /04/10 06 O$/IO 0 ; ( C C / t(6(07 lo 10 S 4 III 06/04/10 06, 04:I0 07/06 ] C C 36 Key: Last Reading 11 (1lnduct,on Phase, C Challenge Phasc) Completion Status (C Completed. L:Lost to cl low up, S Vi Iuntarv withdraw, I V v:olation, A 1/ Adverse event, C) Other) Generated on 07.13:10:13:28 by DISPIIS l SAS / Uses: Dl: logs, RLS[ ONSI/, FINAl. Panel Book Page 140

143 1K]. STUDY NO. DSI Page Data l.isting 2: Sub]eet Demographics Subject No. Age Gender Race 00] 54.2 Female Caucasian Female Caucasian Female Caucasian Fu:e Caucasan Fenia Ic Caucasian Female Caucasian Female Caucasian Female Cai eusian Female Caucasian 0 I ci ii ale Ca uea sian OIl 42.8 cemale Caucasian 0 I I.8 Female Caucus an I Female Caucasian Female Caueasmn Female Caucasian Male Caucasian Female Caucasian I.6 Female Caucasian Female Caueasian 02]).0 Female Caucus it 1) Female Caucasian Female ( eucasman Female Caucasian ) Female Caucasian cemnal; C aieaman ( Female Caucasi:mn Female Caucasian Female CaLicas man Femlc Caucasian Male Caucasian ,3 Fenia Ic Ca ness 1) Femna Ic i mm C a uca sian cmaic Caucasian F6mmmle Caucasian Fcnnmm Ic Caucus an Male Caucasian Female Caucasian Generated un 07] 3/10:] 3:28 by DEMOIIST.SAS / Uses: Di/MOGS Panel Book Page 141

144 TKL STUDY NO. DS1O6OIO Page 2 of S J)ala l.:stmg 2: Subject Demographics Subject No. Age Gender Race 03$ 36.0 Female Caucasmn Male Black Fema]e Caucasiu Ferns Ic ( aucasjan Female Caucasian Female Caucasian Male Caucasian Female Caucasian Female Caucasian Female Caucasian 04$ 68.6 Female Caucasian Female Caucasian Male ( aucasian ,9 Femalc ( auce$:in Female ( aucasian Female ( aucasian Female Caucasian Cinale Caucasian erna1e Caucasian Female Caucasian Female Caucasian Female Caucasian Female Caucasian 1) Female Caucasian Female Asian Female 11 spanic ,9 Female ( aueasian Female Caucasian Female ( aueasian Female Caucasian 06% 58.2 Female ( imuesian Cma Ic (.aucasjan male Hispanic FOmnale Caucasian Female Caucasian Fcmale Caucasian Female Caucasian Generated on 07/13/10:13:28 by 1)1MOLIS I.SAS / Uses: )7MOGS Panel Book Page 142

145 K1. Study No. DSIO6OIO Page of 5 Data listing 3: bermatologie Response Grades 13v Product and Subject Subjcct No. 1 InducI.on :eadii Challenge Phase i 9 MV 4$hr72hr 96hr() X N l) ( X Key to Symbols: NI) reaction 7 M inmal or doubtful response, Definite erytliema. no edema Definite ery:ima. delinite cd_t ns i Definite ervthema, definite edema and vesiculation i Wily d ffereni oip surrounding normal skin N9G = No ninth grading?a Not apphed NP Not one ied cue to re:utioii achieved X Reading not performed due to missed visit or subject jisenncuation D Damage to epidermis: Doting. crusting and/or superficial rosions p l apular response >50% NRData not rcorded MU Makeup reading for missed induction visit (*) \Vhen required Generated on 07/13/10:13:28 by D[/ l A1LAS USI/S: RI/SI (tnsp. PRODIlSI Panel Book Page 143

146 024 X iki. Studs No, VS l aee 2 of 5 Data 1_isting 3 13v Product VLrnnnoogtc Response Grades Prodoct and Subject Subject No. Induction Reading ( liaflcngc Ptiatc I MU 48hr 72hr 96hr(*) S t) X. X X X X Y X < X X X X V t)46 ( ) When rejuii cd Gencratcd on :] 3:25 by DII AI1_.SAS USES: l ESFONSE. l RO!)i 1Sf Panel Book Page 144

147 047 X._. TKL Study No. DSIO6OJO Page 3 ui 5 Data listing 3: l)ermatolegic Response Grades 13y Product and Subject Produst Subject Inducto cadnig Challenge Phase s tis( 057 (> (160 06> ML 48hr 72hr 96hi() ( (*) When required Generated on 07/I 3/1 0:13:28 by DVlAILSAS/USI/S: RLSJ C)NSI/. PROI)LIS1 Panel Book Page 145

148 ..... IKI Study No. DS Pago 4 ol 5 Data listing 3: Dennatologic Rc sponsc Gradcs 13y Product and Subject ProdcI SLibject No. 1 Inductori Reading Challenge Phase MU 48hr 721ir 96hr(*) t X X X N N X N N N N N N 079 OttO (4( % ( N9G (*) \Vhcn required Generated on 07/13.10:13:28 by DETAII..SASUSIS. 1/t.FcNSt PROD1.JS f Panel Book Page 146

149 N9G.. x.. III TKL Study No. DSIO6OIO Page 5 of 5 l)ata 1sting 3:t)crnatologie Respcnsc Grades By Prod oct and S uhjc t Product Subject Induct ion Rca ding C hail engr Phase MU 4Xhr72hr 9hhr(*) X X X X X X X X X X X X X X X X X X X X X. (*) When required Generated on 07 I 3 10:1 3:28 by l)1fa1l.sas:lses: RESPONSI:. PROI)l 1SF Panel Book Page 147

150 H ( Consumer Product Testing Co FINAL REPORT CLIENT: ATTENTION: TEST: Ophthaimoioicl lnsc Safety Evaluation Protocol No.: BUDYOI.1 U. TEST MATERIAL: (Q eii Ijc4 /O EXP FRI MENT REFERENCE N UMBER: ClO2474O I Reviewed by: Approved by: Michael ;wcil. PhD.. C.C.R.C.. C.C.R.A. Director, Clinical Evalucons I 1 Approved by: Joy Exec#iv ic PresiJem, Clinical Evaluations This report is sobrritlwi for th oxciusive use ct the person, paitrww!.f 0 corporalor iron ii ;cdressed. anl nerrier :ne r orl nor the name of These Laborafores nor ens ni br of Is 50 without written as thorizatio n if mar be Jn r. connection iri the advert dor (>1 asic iii tiny 2r00iJ0 Ci FOODSS 70 N(yvv [)ttlclt Lane Fairticki, NL W jrrrsev 070c: )7Th 6087 I I IIt x (0Tt) Panel Book Page 148

151 Quality Consumer ProducfTesting Co. L 97 QUALITY ASSURANCE UNIT STATEMENT Study Number: C1Q The Consumer Test material Testing Company, Incorporated (CPTC) Quality Assurance Unit (QAU) is responsible for monitoring the conduct, content and reporting of all clinical laboratory studies that arc conducted at CPTC. This trial has been conducted in accordance with ICFI GuideJine E6 for Good Clinical Practice, th requirements of 21 CFR Parts 50 and 56, other applicable regulations, CPTC Standard Operatin. Procedures, and the approved Trial Protocol. The CPTC QAU has reviewed all data, records, and documents relating to this trial and also this Fin:1 Report. The following QAU representative signatwe certifies that all data, records, and docurnenis relating to this trial and also this Final Report havebcen revieved and are deemed to he acceptable, ani the trial confbrms to all of the requirements as indicatnd above. All records and documents pertaining to the conduct cf this study shall be retained in the CPTC archives fbr a minimum octen (10) years. At any time prior to the completion of the tenth archival year, a Sponso may submit a written request to the CPTC QA Drmartment to obtain custody of study records once thc CPTC archive period has been completed. This transfer shall be performed at the Sponsor s expense. In the absence of a written request, studyrelated records shall be destroyed at the end of the CPTC archiva period in a manner that renders them useless. 7IL1 Assurance Representative Date 70 rscv Dutch Lane Fairfield, New Jersey ) Fax 973) Clii iicai Toxicology Analytical Chemistry Microbiology Panel Book Page 149

152 Cl Page 3 of 23 Objective: To evaluate the safety and ocular irritation potential of an eye liner following repetitive, daily use conditions. Participants: Thirtytwo female subjects, ranging in age from 1 9 to 63 years, were recruited and qualified for this trial. Thirtyone subjects completed this trial. Subject #1 6 did not complete th.s trial due to personai reasons unrelated o test material usage. Inclusion Criteria: a. Thirty healthy frnale subjects, ages 18 to 65. b. Agreed to discontinue the use rd their current eye liner and only use the test material. c. Agreed to arrive at the Testing Facility with no eye makeup on. d. Absence of any visible skin or eye disease that might have been confused with a reaction to the test material. e. Soft and hard contact lens wearers were empanelled, with tl:e remainder nonnon.act lens wearers. f. Agreed not to introduce the use of any new cosmetic, toiletry personal care test materials during the course of the trial. g. Regular user of an eye liner. h. Quali ing ophthalmic examination of subjects to ensure eye health and, if appropriate, the correct fit and condition of their contact lenses i. Completion of a Medical liistory form and the understanding and signing of an Informed Consent Form that includes a HIPPA statement. j. Considered deptnahle and ablate follow directions as outlined in the protocol. Exclusion Criteria: a. Ill health or taking medication, other than birth control, which couth have influencedthr purpose, ihtegritv or outcome of the trial. b. Females who were pregnant:noring or planning on becoming pregnart during the course o the trial. c. A history of adverse reactions to cosmetics or other personal care products. d. Currently on any systemic or topical corticosteroid. antiinflammatory, antihistamine therapy or any other medication which could have interfered with the ourpose or integrity of the trial. Test Material: Panel Book Page 150

153 Cl Page 4 ot 23 Trial Schedule: Initiation Date Completion Date June4,2010 June 18,2010 Methodology: Prior to acceptance, each subject received a qualifying ophthalmic examination by a Board Certified Ophthalmologist to ensure eye health and if appropriate, the correct lit of their contact lenses. The Ophthalmologist evaluated (by gross and slit lamp examination) the subjects eyelids, corneas, conjmictivae, irises, lenses, anterior chambers aid pupillary reactions, as well as visual acuity. The parameters for the examination were Visual acuity Pupillary Response External Slit Lamp: Pilpebral conunctivae Bulbar conjunctivac Fornices Cornea abnormalities Iris Lids enscs Findings were noted on Individual Ocular Examination Record Forms, The panel was comprised of 20 soft and 2 hard Contact lens wearers. The remaining 10 subjects were noncontact lens wearers. After completion of the ophthalmic examination, acceptable candidatcs received the test material and were instructed to use the test material at lcat once daily for 2 weeks. ccerdin& to the following directions: Instructions: I)iscontinuc the ue of your current eye liner and use only the te:t material provided for the duration of this trial. You are permitted to wear your regular cosmetic products for the duration of the trial. Do not introduce ay new cleansing products, moisturizers, toiletry, cosmetics or othcr nersonal care products during the trial interval. Do not wear eye makeup to the examinations. Panel Book Page 151

154 C Page 5 of 23 Methodology (continued): Usage Directions: At least once daily, use the applicator provided or your eydlining brush and apply the eye liner to your lash line as you normally would. You may reapply as often as desired. The time of application must be recorded on the daily diary. Keep out of reach of children. Do not let anyone else use the test material. Report any adve sc reactions or problems immediately to the laboratory staff. To document compliance, subjects were required to maintain a daily diary to record each use. A comprehensive ocuier examinaton was repeated for each subject after 2 weeks of test material usage, as previously described. A Sponsorsupplied questionnaire was issued at this time. All unused test material and daily diaries were returned at the final visit. Daily diaries and questionnaires were reviewed for completeness, prior o dismissal of the subjects.. Adverse Reactions: There were no adverse reactions. 1)eviations: Per Sponsor s direcivc, subjects were given permission to use their iwn brushes/applicators during the course of the trial due to the applicator brush that accompanied the test material being ineffective. Subject # 21 used the test material for 13 days, instead of 14, du w inadvertently missing application on the day of enrohmcnt. Results: All ophthalmologic& examination remained within normal limits throug oet the test interval, The results of all ophihalmological examinations are presented in Tables 12. Questionnaire tallies ure presented in Table 3. Panel Book Page 152

155 C Page 6 of 23 Results (continucd): Statistical analyses (ztest) on questionnaire responses are presented in Tible 4. A significant number of subjects answered with successful respoases (subjects who agreed with the positive statement or liked the particular test material attribute) for the following statements only: I liked the shade of the pro Juct The product was gentle to my eyes The product provided a smooth finish The product was easy to blend The product stayed true to its color The product was easy 1:o remove with the face cleanser A significant number of subjects answered with failed responses for the following statement: I like the application of the nroduct Subject demographics are listed in Table 5. A review of completed daily diaries revealed the following signiflcani. comments: Positive No. of Sublects Comments 2 Product was smooth on skin Color and texture good No problems Liked the product Stayed on nice once applied Product was great 1 Stayed on well with minimal smudging No. of Subjects Comments 8 Difficult/hard to apply/usc 4 Brush was a little awkward/too soft 4 Smeared/smudged easily 3 Did not like product Had to reapply Brush had to he wet before application I Felt liner would he better in a tube with applicator than a pod with a brush 1 Brush made it hard to do a fine line Brush could have been a little thicker 1 Prefer liquid eyeliner Tee i;2,ht, barely showed Panel Book Page 153

156 Cl O2474.OI Page 7 of 23 Summary: Under the conditions of this trial, test material, did riot indicate a potentiai Ibr ophthnlmological irritation or hypersensitivity and can be considered sale for use by both contact lens and noncontact lens wearers. Panel Book Page 154

157 uses Table 1 C Page 8 of23 Initial Ocular Examination Evaluation Date: June 4, 2010 Treatment Site: Eyes lest Iviaterial; HISTORY Glasses Worn: Past Eve Problems: Last Eye Bifocals! Subject # Initials Exam No Yes Dis Near Only Progressive No Yes I DF 4/2009 X X X 2 RC 12/2009 X X X 3 NR 6/2009 X x 4 MP 1/2010 X X X 5 GM 12/2009 X X 6 SP 2008 X X X 7 [3V 5/2010 X X X 8 JV 5/2009 X X 9 VP 10/ RK 1/2010 X I X..I X 11 MP 3/2010 X X 12 TC 6/2009 X T x ) 8/2008 X X X 14 VA 8/2009 X K X 15 YC 10/2009 X X X 6 LC 10/2009 X X 17 A13 9/2009 X X 18 RC 1/2010 X X 19 AM 10/2009 X X X 20 LK 2/2010 X X X 21 CS 7/2009 X X N 22 EA 2009 X. 23 HS 11/2009 X X X 24 CL 5/2009 X N 25 HG 8/2009 X X N 26 SV 2008 X X X 27 CG 5/2010 X X N 28 SV 2005 X N 29 LD 2009 X N X 30 BF 1/2010 X X N 3] RI 2009 X N N L 32 KC 10/2009 X N Comments: Subject # 4 has essential blepharospasm botox N Panel Book Page 155

158 Table I (co ntfn u ed) Initial Ocular Examination Cl02474.Ol Page 9 of 23 Eva]uation Date: June 4,2010 Test Material: Treatment Site: Eyes CONTACT LENS WEARER l ype: Schedule: V X Soil Gas txtended Subject # Initials No Yes SDFt Dispeable Permeable Daily Wear Wear I DP X X X 2 RC X X X 3 NR x 4 MP X 5 DM X 6 SP X 7 13V X 8 JV X X X 9 DP X X X 10 RK X 11 MP X X X 12 TC X 13 LD X X X 14 GA X X X 15 YC X X X 16 LC X X X 17 AB X X X IS RC X X X 19 AM X X X 20 LK X X X 21 CS X XV. X 22 EA X 23 HS X X X 24 CL X X X 25 HG X X 26 SV X V 27 CG X X 28 SV X 29 LD X X X 30 13F X X X 31 RI X X X 32 KC X X X Subject #19 has monovision Panel Book Page 156

159 20/ Table I (continued) Initial Ocular Gtvnination Cl02474.OI Page 10 of23 Evaluation Date: Treatment Site: June 4,2010 Eyes VIS(JAL ACUITY Were Glasses Worn During Visual Acuity F.xarn? Test Material: 01ST 01) OS V * Subject # Initials Yes No 20/ 20 20, 20 I OF X 20/ 20 20/ 20 2 RC X 20/ 20 20/ 20I 3 NR X 20/30 20/30 4 MP X 20/ 25 20/ 70 5 DM X 20/ 20 20/ 20 6 SP X 20/ 40 20/ 40 7 BV X 20/20 20/20 8 JV X * 2u/ 20 20/ 20 9 OP X 20/ 30 20/ RK X 20/25 20/25 11 NIP X 20/20 20/ TC X 20/ 30 20/ LD X 20/20 20/20 14 DA X 20/ 20i 15 YC X 20/ 25I 20/25 16 LC X 20/ 25 20/ A13 X 20/ /25 18 RC X 20/25 20/20 19 AM X 20/ / LK X 20! 20 20! CS X 20/ / EA X 20/ 30 20/ IS X 20/ 25 20/ CL X 20/ 30 20/ HG X 20/ 20 20/ SV X 20/ 25 20/ CG X 20/ 30 20/ SV X 20/ 20 20/ LD X 20/ 50I OF X 20! 25 20/ RI X 20/30 20/ KC X 20/30 20/30 Subject #19 has monovision Subject 4 4 has essential blepharospasm uses botox Panel Book Page 157

160 lable 1 (continued) Initial Ocular Examination C l age I of23 Oval ustlorl Date: June 4, 20 0 Treatment Site: Eyes 1 nt Mulcri a[ lye EXAMINATION LIDS CONJUNCTIVA COPNEA FORNICIIS IRIS Subtct# Initials Normal Abnormal Normal Abnormal Ncrna1 Abncimnl Nonnal Abnormal Normal Abnormal OF X X K X X RC X X K X X NR X X X X X 4_ MP X X X X X OM X X X x x ( SF X X A X X 7 L3V X X X X X z_ x A X X 9 DP X X X X A 10 RK X X A X X 11 MP X X A X X 12 TC X X A X A 13 LD X X A X X 14 DA X X X X 15 YE X X X X A 16 LC X X X A 17 AB X X X X X 18 RE X X A X X 19 AM A X X X X 20 LK X X A X X 21 CS X X A X A 22 EA X A A A X 23 1IS X X A I x x 24 Cl. X X A X X X X A X X 26 SV X X A A A 27 CO X X A X 28 SV X X A X X 29 ID X X x x 30 1W X X A X X 31 RI X X X X X 32 KC X X X X K Explain any abnormalities: None Panel Book Page 158

161 0 z L : : 7 fl rr z fl <<>>n xxx xxxxxxxxxx > xxxx><xxxxxxxx><xx><xx><xxx,><x z > cr Z1 xxxxxxxxxxxxxxxxxxxxxxxxx z z 0 xxx>xxxxxxx)x xxxxxxxxxxx xxx xxxxx C > 0 > 0 z C) 0 Panel Book Page 159

162 RC X X Norste! Table I (continued> lnoial Ocular Exainjuutan Cl Paste 3 o23 Evoluotton Date.lime 4, 2010 Tc.st Maturutl: Treatment Site: Eyes CONrACT LENS EVALuATION SCRATCHES DEPIT Subject 0 Initials N/A Absent Prasttt Absent Present Dacto?s Overall Comments 1_ OF X X NorttmiFotStssdy X X Norcss Fot Study NR X Nom,& For Study 4 MP X NointaiForS4 DM X For Study SP X Nuiral toe Study liv X Normal For Study JV IC Normal For Study OP IC IC Normal For Stud 0 RE X Normal tsr Study II Ml X X Noresal Fr Study 12 TC X Notu,siFsrStttdv 13 LD X Nonna: For Study 14 DA X X I NormaL Far 15 ic IC IC Norn:siE:r Study 16 LC IC X Nonna: For Study 17 AD X X Non si For Study Ill P.C X X Normal For Study TO AM X X Normal For Study 20 LK x X Normal Fr Study 21 CS X X Normal trr,r Only 22 NA X Nonual For Study 23 1 IS x IC Nonnal Pot Study 24 CL X IC Normat For Study 25 HG X IC i4oznnal For Study 26 SV X Normal For Study 27 CE X Normal For Study 20 sd x Normal For Study 29 LI) X IC Normal For Study 30 lip X IC Normal For Stu4y 31 Rl IC IC Normal For Study 32 KC IC IC Normal For Study Euptaitt arty scrateltru ur deposits: Subject #17 will wear crew tenses for study. Panel Book Page 160

163 Table 2 C10247&01 Page 14 of 23 Final Ocular Examination Evaluation Date: Treatment Site: June Eyes Test Product: VISUAL ACuITY Were Glasses Worn 1)1ST During Vsua.cuity Exam? OD OS Subject 11 initials Yes No 20/ 20 20/ 20 1 DF X 20/ 20 20/ 20 2 RC X 20/20 20/20 3 NR X 20/30 20/30 4 MP X 20/ 25 20/ 50 5 DM X 20/20 20/20 6 SP X 20/ BV X 20/ 20 20/ 20 8 iv X 20/ 20 20/ 20 9 DP X 20/ / RK 20/25 20/25 I I MP X 20/ 20 20/ TC X 20/ / LD X 20/ 20 20/ )A X 20/ 25 LP 15 YC [ X 20/ 25 20/ I.C 17 AB X 20/25 20/25 18 RC X 70/25 20/20 19 AM 20/200 20/40 20 LK X 20/ 20 20/ CS x 20/ 25 20/25 22 EA X 20/ 30 20/ IIS I X 20/ 25 20/ CL X 20/ 30 20/ HG X 20/ 20 20/ SV X 20/ 25 20/ CG X 20/ / SV. X 20/ 20 20/ LD X 20/ 50 20/ BF X 20/ 25 20/ Ri X 20/30 32 KC j / 30 LX Lx :X Subject 1119 has rnonovision Subject 11 4 has essential blepharospasm uses botox j20/30 Panel Book Page 161

164 X X X X. X Table 2 (continued) Fina) Ocular Examination Cl Page 15 of23 Evaluation Date: June 18, 2010 Treatment Site: Eyes Test Product: EYE EXAMINATION LIDS CONJUNCTIVA CORNEA FORNICES Subject # Initials Normal Abnormal Normal Abnormal Normal Abnormal Norma] Abnormal Normal Abnorrai DJ X x x X x 2 RC X X X X X 3 NR X X x 4 MP X X X X X 5 DM X X X X X 6 SP X X X X X 7 BV X X X X 8 JV X X X X 9 DP X X X X X 10 RX X X X X X 11 MP X X X X X 12 TC X X X X X 13 JJ) X X X X X 14 DA X X X X X IS YC X X X X 16 LC 17 Al3 X X X X X IS RC X X X X X 19 AM X X X x x 20 LK X X X X X 21 CS X X X X 22 EA X X X X 23 US X X X x 24 CL X X X X X 25 FIG X X X X X 26 SV X X X X X 27 CG X X X X sv x x 29 LD X X X X X 30 1W X X X X X 31 RI X X X X X 32 KC X X X X Explain any abnormalities: None x IRI S Panel Book Page 162

165 Yes Table 2 (continued) Final Ocular Examination ClO24740l Page l6of23 Evaluation Date: June 18, 2010 Treatment Site: Eyes Pest Product EYE EXAMINATION ANTERIOR CHAMBER LENSES Post Cataract Surgery Direct PUPILLARY REFLEX Conseritual Acc/Cnv No Yes No Yes Xo Subject II Initials Normal Abnormal Normal Abnormal OD OS I X X X X X 2 X X X X 3 X X X X 4 X X X X X 5 X X X X X 6 X X X X X 7 X X X X X 8 X X X X X 9 X X X X X 10 X X X X X 11 X X X X X 12 X X X X 13 X N X X N 14 X X X X X1 15 X X X X X X N X X X 18 X X X N 19 X X X X X 20 X X X X X 21 X X X N X 22 X X X X X 23 X X X X X 24 N X N X 25 X X N X X 26 X X X X N 27 X X N X X 28 X X X X 29 X X N X X 30 X X X N X 31 N N 32 X X X X Explain any abnormalities: None Panel Book Page 163

166 Normal Table 2 (continued) Final Ocular Examination C i Page 17 of23 Evaluation Date: June Treatment Site:_y Test Product: CONTACT LENS EVtLUATTON SCRATCHES I)FPOSITS Subject # Initials N/A Absent Present Aaett Present Doctor s Overall Comments I DF X X Normal Post Study 2 RC X X Normal Post Study 3 NR X Normal Post Study MP X Normal Post Study 5 DM X Normal Post Study 6 SP X Normal Post Sidy 7 BV X Normal Post Study 8 JV X X Normal Post Study 9 DP X X Normal Post Study 10 RK X Normal Post Study I I MP X X Normal Post Study 12 IC X Normal Post Study 13 LD X X Normal Post Study 14 DA X X Normal Post Study IS YC X X Normal Post Study 16 LC 17 A13 X X Normal Post Study 18 RC X X Normal Post Study 19 AM X Normal Post Study 20 LK X X Normal Post Study 21 CS X X Normal l ost Study 22 EA X Normal Post Study 23 HS X N Normal Post Study 24 CL X X Normal Post Study 25 HG X Normal Post Study 26 SV X Normal Post Study 27 CG X X Normal Post Study 28 SV X Normal Post Study 29 LD X X Normal Post Study 30 BF X X Normal Post Study 31 RI X X Post Study 32 KC X N Normal Post Study Explain any scratches or deposits: None Panel Book Page 164

167 Cl Page 18 of 23 Table 3 Questionnaire Tallies 1. 1 liked the appearance of the product. No. of subjects Liked a lot 6 Liked somewhat 9 Neither liked nor disliked 6 Did not like somewhat Did not like at all liked the shade of the product. No. of subjects Likedalot 17 Liked somewhat 6 Neither liked nor disliked 4 Did not like somewhat 4 Did not like at all liked the application of the product. No. of subjects Liked alot 6 Liked somewhat 4 Neither liked nor disliked Did not like somewhat 3 Did not like at all The product was gentle to my eyes. No. of subject Agree strongly 16 Agree somewhat 7 Neither agree nor disagree 6 Disagree somewhat 1 Disagree strongly Panel Book Page 165

168 C1O2474.O1 Page 19 of 23 Table 3 (continued) Questionnirc Tallies 5. The product was easy to apply to my eyes. No. of subecs Agree strongly 8 Agree somewhat 5 Neither agree nor disagree 2 Disagree somewhat 4 Disagree strongly The product provided an even application. No. of subjects Agree strongly 6 Agree somewhat 8 Neither agree nor disagree 3 Disagree somewhat 9 Disagree strongly 5 7. The product provided a smooth finish. No. of subects Agree strongly 8 Agree somewhat 9 Neither agree nor disagree 5 Disagree somewhat 3 Disagree strongly 6 8. The product was easy to blend. No. of subjects Agree strongly 6 Agree somewhat 12 Neither agree nor disagree 4 Disagree somewhat 5 Disagree strongly 4 Panel Book Page 166

169 Cl Page 20 of 23 Table 3 (continued) Questionnaire Tallies 9. The product glided on effortlessly. No. of subjects Agree strongly 7 Agree somewhat 7 Neither agree nor disagree 4 Disagree somewhat 4 Disagree strongly The product provided acceptable coverage. No. of subjec,s Agree strongly 6 Agree somewhat Neither agree nor disagree 4 Disagree somewhat 8 Disagree strongly 11. The product provided intense color with ease. No. of subjects Agree strongly 7 Agree somewhat 7 Neither agree nor disagree 4 Disagree somewhat 9 T)isagrce strongly The product stayed true to its color. No. of subiects Agree strongly 11 Agree somewhat 11 Neither agree nor disagree 4 Disagree somewhat Disagree strongly 3 Panel Book Page 167

170 Cl Page 21 of23 Table 3 (continued) Questionnaire Tallies 13. The product did NOT smudge / crease throughout the day. No. of subjects Agree strongly 3 Agree somewhat 9 Neither agree nor disagree Disagree somewhat 7 Strongly disagree The product stayed in place throughout the day. No. of subjects Agree strongly 3 Agree somewhat 13 Neither agree nor disagree 3 Disagree somewhat 6 Strongly disagree The product was easy to remove with a face cleanser. No. of subjects Agree strongly 13 Agree somewhat 9 Neither agree nor disagree 3 Disagree somewhat 6 Strongly disagree 0 Panel Book Page 168

171 15 effortlessly,.ll_. 3 Table 4 Cl Page 22 of 23 ztest on Questionnaire Responses (Successes I Failures) Question I Question 2 1 Question 3 1 Question 4 Question S I liked the I liked the shade I like the The product was The product was appearance of the of the product. application of the gentle to my eyes. easy to apply to product. product. my eyes. Successes Failures Total Trials P Value < < gniflcance n.s. Sig, j Sig. Sig. n.s.. Question 6 Qon7 Question8l Question 9 Question 10 The product The product The product was The product The product provided an even provided a easy to blend, glided on provided acceptable applicatioit smooth finish. coverage. Successes Failures Total Trials P Value j I_Significance n.s. Sig. Sig. n.jn.s Question 11 Question 12 )i2stion 13 Question [4 Question 15 The product The product The [ roduct did The product The product was provided stayed true to its NOT stayed in place easy to remove with intense color color, smudge/crease throughout the a face cleanser. with ease. throughout the day. day. Successes Failures Total I rials I p < < r gnificance,. n.s. Sig n.s. Significance observed at P0.05 ztest performed using SigmaStat lbr Windows Version I he responses were pooled into two categories: 1) Successes, subjects who agreed or liked the prodict attribute. 2) Failures, subjects who disagreed or did not like tte product attribute. Subjects that were undecided (neither agreed nor disagreed) were divided evenly between the two categories (half in the successes and half in the failures). Ifthcre was an uneven distribution of undecided responses, the successes category received the extraundecided response. Panel Book Page 169

172 TC C Page 23 of 23 Table 5 Subject Demographics Subject Number Initials Age Gender F I DF 54 F 2 RC 48 F 3 NR 1 4 MP 62 F 5 i)m 45 F 6 SP 63 F 7 BV 47 F 8 JV 20 F 9 LW 52 F 10 RK 46 F ii MP 44 F F 13 LD 60 F 14 DA YC 45 F 16 LC 63 F 17 AB 48 F 18 RC 32 F F F 21 Cs 34 F 22 EA 49 F 23 1IS 45 F 24 CL 44 F 25 HG 20 F 26 SV 33 F 27 CG 19 F 28 SV 45 F 29 LD 46 F 30 BF RI 42 F 32 KC 46 F Did not complete: Subject #16 Panel Book Page 170

173 CODING FORM FOR SRC INDEXING 6/10/8=" <:i/08/<:l2; Submilling Organization RESE.ARl:H & l:unsul T 1 NG CU Al~ ACUTe: Dt:::Rtr1AL,IJX II~I TY (LD::.IiI) in RAT~ WIIH CUVE:::I< u::r,<:::r OnTED 0e30':l~ ; DIMETHYL SlLIGUN~S & SILOXAN~b RXN PHUD WI 81 (67762<:l07) Panel Book Page 171

174 Degussa+ Degussa Corporation August 30, 1993 Office of Pollution Prevention and Toxies U.S. Environmental Protection Agency (TS790) 401 M Street, SW Room G099 East Tower Tunnel Washington, D.C Certified Mail Attention: 8(d) Reporting Re: Health and Safety Study Rule Section 8(d) [OPPTsB02040; FRL41821] Federal Register Notice May 14, 1993 Dear Sir: On August 26, 1993, we notified you that we became aware of the existence of some reportable health and safety studies that are not in our possession, but held by our parent corporation. We requested copies and have received them. The substance is , Dimethyl silicones and siloxane, reaction products with silica, sold under the trade name, Aerosil R202 and Aerosil R204. The studies were performed with Aerosil R202. The studies are:! Acute Dermal Toxicity (LP~ (in Ra.~"3t:X:'Gb3~ Degussa AGUSITNR, DG~ 2 Primary Eye Irritation (in Rabbits) Sc~~ 5 Degussa AGUSITNR, DGT..3 Acute Oral Toxicity (LDSO) (in Rats) 0CJ1"3CCOO 3,,(0 Degussa AGUSITNR, DGT 0 Thank you. r Primary Skin Irritation (in Rabbits) Degussa AGUS:TNR, DGT os The Ames Test on a Toluene Extract Degussa AGUSITNR, DKM oa ~~?'c:o:.::xjs<o'1 &" OJ~ "3 CoB o Sincerely, 9~~ John Lewinson, Ph.D. Manager, Product Regulatory Compliance JL93376 cc: H. Hasl, DCRP R. Marion, DCA 65 Challenger Road Ridgefield Park NJ JJ Panel Book Page 172

175 R C C RES EAR C H & CON S U L TIN G COM PAN V A G PROJECT _ACUTE DERMAL TOXICITY <LDsO) (STIJDY WITH AEROSIL R 202) IN RATS i:_cgu~;::~ ~ '"'J_ "".,,r\ ~S".. 001\2.... AG USIT NR. ' ::D.~T REPORT DATE: JUNE 10, 1985 o :;, po. Box CH 4452 Itingen Switzerland Phone 061 / 98 S2 S2 Tel~x RCC CH :2 Panel Book Page 173

176 ',K.:.J_::',, 0"';0. :,8,;E:\OSIL. R ~02 PREFACE GENERAL TITLE SPONSOR STUDY MONITOR TESTING FACILITY RCC PROJECT NUMBER TEST ARTICLE TEST SYSTEM Ac~te dermal toxiclty <L050) st~dy with AEROSIL R ZOZ In rats. OEGUSSA AG INOUSTRIELLE TOXIKCLOGIE POSTFACH 1345 ; HANAU 1 FEDERAL REPUBLIC OF ~ERMANY DR. O. WEIBERG Research & Cons~l~ing Company AG CH 4452 Itlngen / Switzerland AEROSIL R Z02 Rat PROJECT STAFF Study director, L. Ullmann Technical coordinators, R. Sacher H. Mohler Pathology: SCHEDULE Accll.mati.on April 9, 1985 Treatment / Observation April 16 to 30, 985 April 30, 1985 ARCHIVING Research & Cons~ltlng Company AG, CH 4452 Itingen / Switzerland, Raw data, protocol and report, d~pllcate of report. 4 Panel Book Page 174

177 RC:?ROJECT ~EROSIL R :C2 SUMMARY AND CONCLUSION GENERAL Th2 t2~t article AEROSIL R 202 was applled to ~he skln of rats of both sexes for 24 hours at a dose of 2000 mg{kg The following death rate was obser'led: o Z at 2000 mg/kg Based on these o!::lservations. the LOGIT model could not bioi to thi2 obsi2rvi2d rate of death applied tha.n 2000 rng/kg SYMPTOMS No signs of local or systemlc symptoms were observed l~ during the observatlon period. any rat NECROPSY No macroscopic organ change~ were observed. See App;mdix B1 ~ Panel Book Page 175

178 ~:: ~~O~~:T J46798 AERGS::'" R :C2 OB.JECTIVE" Th~ purpo~~ of ~h~s d~rmal (LDSO) study ~as to assess the tox~colog~cal profile of AEROSIL R ZOZ when admin~s~ered to rats by a s~ngle occl~si~e dermal appl~cat~on, follow~d by an observat~on period of 15 days Th~s study should prov~de a rational bas~s for a rlsk assessment in man. MATERIALS AND METHODS TEST SYSTEM Test system Rat, KFMHan. ~istar (outbred, SPFQuality) Rat~onal~ Recognized by the international guidelin~~ ~~ the recommended test system. So~rce Kleintierfarm Madoerin AG, CH 4414 Fuellinsdorf/Switze~land Number of animals per group 5 males 5 females Total number 0 f ani.mals 5 males 5 females Age at start of treatmqnt 8 10 I.lJQek s Body we~ght at start of treat.ment males: females: 234 ZS9 S g Idllntificat~on By ~nlque cag~ number and corresponding colorcoded ~pots. Randomlzation Randomly selected at time of delivery. Accllmatlon One week under laboratory conditions aft.er veteri.nary exam~natlon. q Panel Book Page 176

179 ~CC PRO~ECT O~6798 AEROSIL R 20: HUSBANDRY Room No. 136 Condl.tl.ons Standard Laboratory Conctitl.ons Al.rconditl.oned wl.th 1015 alr changes per hour, and hourly monl.tored envlronment wlth temperature 22+2 desr~es centl. grade, relatl.ve huml.dl.ty 'l., 12 hours artlfl.clal fluorescent 1l.ght/12 hours dark, musl.c/ll.ght period Accommodation Individually In MaKrolon type3 cages with standard softwood beddl.ng (Llgnocel, SChill AG, SWl.tzerland). Pelleted standard Kll.ba 343, Batch 18/85 rat maintenance dl.et (Kll.ba, Kll.ngentalmuehll2 AG, SWl.tzerland) ad ll.bltum Results of analysl.s for contamlnants are included in this report.!.later Community tap water from 1tlngen, ad llbitum. Results of analyses for contaml.nants are included In this report iest ARTICLE Identiflcat.:.on AEROS1L R 202 soll.d Batch Number unknown Purlty formulatlon; ~ngredil2nt5 aval.labl~ 1n the sponsor's files Stabllity of test artlcle Stabilitv of test article in vehlcle Routlne hyglenlc procedur~s were suffl Clent to assure p~rsonnel ~~alth and sa fety!() Panel Book Page 177

180 ~cc ~ROJECT ;EROSIL. R :02 10 T~st Artlcl~ Pr~paratlon Th~ test artlcl~ was plac~d lnto a glass beaker on a tared Mettler PK 4800 balance and the venlcle (Polyethylene glycol, PEG 400 Fluka AG 9470 Buchs/Switzerland) was added. A welght/ volume dllutlon was prepared US1~g a homogenizer Homogenelty of the test artlcle 1n th~ vehlcle was malntained dur1ng treatment using a magnetic stirrer. The preparation was mace 1mmediately prlor t~ doslng TREATMENT ApproXlmately 24 hours b~forq treatment, the backs of the animals were shaved wlth an electrlc clipper, ~xposlng an ar~a of approximately 20 square centlm~ters. Dn test day 1 the test article was applied evenly on the skin and cov~r~d wlth an OCC1US1VQ dr~sslng. The drqsslng was wrapped around thq abdomqn and fix~d wlth an ~lastic adheslvq bandage. Twentyfour hours aft~r the application, thq dressing was removed and thq skin rqactlon was assessed accordlng to the mqthod of Noaks and Sanderson (Noaks, D.N. and Sanderson, D.M. A MQthod for Determinlng th~ D~rmal TOX1Clty of P~StlCldQs. Brit. J. Industr. MG!d., ,1969). RatlonalQ: Dcclusive dqrmal application was provqn to be an efflcaclous mqthod to assess dqrmal tcxlclty. Applicatlon VolumQ/ kg body WQlght: Group 1: 20 ml at 2000 mg/kg ~ 11 Panel Book Page 178

181 RC:?ROJEC7 O~67?8 ':'EROSI~ ~ 20: 1.1 OBSERVA nons Mor~al1ty / V1abil1ty Four t.. m2s dur 1ng t.es:: day 1, and. dally durlng days 2 15 T~st day 1 (pr~ad.minlstratlon), 8 and 15. :. Symptoms Each animal had an examlnatlon for changes 1n app~arance and b2hav1or four tlmes dur1ng day 1, and dally dur1ng days 215 All abnormallties were recorded. The an1mals wer2 check2d :or th2 symptoms 11St2d below. PosltivQ flnaings are.. ndlcated under Appendlx A GENERAL BEHAVIOR NOSE aggressiveness crying restlessness / excltement nervousn2ss, fl2ar sedat.lon somnol12nc2 sleep C::lma RESPIRATION EYE chromodacryorrhea ekophthalmos mios1s mydrlasls whitish dlscharge lid adheslon rhlnorrhea epistax;:,s MOTILITY akinc!sla ataxla droppecl heacl hypericl.nesla hypoklnesia paralysis, flaccld paralysls, spastlc paddllng movements stlff movl2ments rolling movements hunched postur'i! BODY POSITION v2nt.ral body posit.ion lat.l2roabdomlnal posltlon curved bocty po~ltion /:1.. ~ Panel Book Page 179

182 RCC PROJECT AEROSIL R MOTOR SUSCEPTIBILITY spasms tonlc muscle spasms clonlc muscle ~pasms 0plst.hot.onus saltatory spasms trismus retchlng Straub phenomenon tremor musclet.wit.ching muscletwitching. generalized SKIN erythema edema necrosl5 VARIOUS loss of wel9.ht emaciatlon negat.ive co~neal reflex diarrhea ruffled fur necrosis of tlssue of application area salivat.lon pallor cyanosis PATHOLOGY Necropsy Necrops~es were performed by experienced prosectors. All animals were necropsied. All animals were anesthetized by int.raperitoneal inject.ion of sodium pentobarbital and killed by exsanguination. STATISTICAL ANALYSIS The LOGITModel could not be applied to the observed ra~es death. The LDSO was estimated wlthout use of a stat.istical mode 1. DATA COMPILATION of The following data were recorded on data sheets and transcribed for compilation ana analysis: behavior and appearance. mortality. body w"ights. macroscopic findings. Panel Book Page 180

183 ~~= ~RG~~C; C467 Q 8 ~EROSI:'" R :02 APPF;:NDIX A OBSERVATIONS AND RECORDS Panel Book Page 181

184 Degussa+ Degussa Corporation. August 30, 1993 Office of Pollution Prevention and Toxies U.S. Environmental Protection Agency (T5790) 401 M Street, SW Room G099 East Tower Tunnel Washington, D.C Certified Mail Attention: 8(d} Reporting Re: Health and Safety Study Rule Section 8(d) [OPPTS802040; FRL41821] Federal Register Notice May 14, 1993 Dear Sir: On August 26, 1993, we notified you that we became aware of the existence of some reportable health and safety studies that are not in our possession, but held by our parent corporation. We requested copies and have received them. The substance is ene2907, Dimethyl silicones and siloxane, reaction products with silica, sold under the trade name, Aerosil R202 and Aerosil R204. The studies were performed with Aerosil R202. The studies are: I Acute Qennal Toxicjty (I 050) (in Rats) /p'''r3co<x>'3g,;l.f Degussa AGU5ITNR, 8~12DGT z Primary Eye Irritation Qn Rabbits) ~'3&b ~ Degussa AGU5ITNR, 8~10DGlt.~ n ~..z Acute Oral Toxicity (LOSO) (in Rats) OfL)3CCOO3"" Degussa AGU5ITNR, DGT Q'C""I. u Thank you. f Primary Skin Irritation (in Rabbits) Qpo,s~~<'J Degussa AGU5ITNR, DGT 0 o ~ Th9 Ames Test on a IQluene Exb'ad 8"0~"3foB Degussa AGU5rrNR, ojijjJ.. Sincerely, 9~~ John Lewinson, Ph.D. Manager, Product Regulatory Comp6ance JL93376 cc: H. HasI, DCRP R.Marion, DCA 66 Cballenger lbld Rid{jBfieJd ParkNJ : Panel Book Page 182

185 R C C RES EAR C H & CON 5 U L TIN G COM PAN Y A G PROJECT 0468.:1...:1.. PRIMARY EYE IRRITATION STUDY UITH AEROSIL R 202 IN RABBITS REPORT.:::l.,..:>,. ~,., :.' ~ I ~. 0 0, DATE, JUNE S i DEGtJSSAAG _ USIT tm;..\. K~ P.O. Box CH 4452 Itin9~n Swit~~rland PnonQ 061 I Tel~x RCC CH Panel Book Page 183

186 RCC PROJECT AEROSIL R PREFACE GENERAL TITLE SPONSOR STUDY MONITOR TESTING FACILITY Primary eyq irritation study with AERQSIL R 202 in rabbits OEGUSSA AG INOUSTRIELLE TOXIKOLOGIE POSTFACH HANAU 1 FEDERAL REPUBLIC OF GERMANY DR. O. \JEIBERG Research & Consulting Company AG CH 4452 Itingen I Switzerland RCC PROJECT NUMBER TEST ARTICLE AEROSIL R 202 TEST SYSTEM Rabbit PROJECT STAFF Study director: L. Ullmann Technical coordinators: R. T. Sacher Porricello SCHEDULE Acclimation Treatment. ObsliiIrvation TQrmination ARCHIVING April April April 30 to May 3, 1985 May 3, 1985 Research and Consulting Company AG, CH 4452 Itingen / Switzerland, raw data. protocol and report, duplicate of report, test article reference sample 1 Panel Book Page 184

187 RC~ PROJECT 0468:1 AEROS!L R SUMMARV Under the condlt~on5 of ~h~s experlment. AEROSIL R 202 was found to cau5e a primary lrritation score of 0.1 when applied to the rabbit eye mucosa. In the area of application no discoloration of the cornea and conjunct~vae was observed which could be r~lat2d to effects of the test art~cle. No corrosion was observed at each of the measur~ng lntervals. According ~o EEC Council Directive, 67/548/EEC. April 1983 Brussels. Belgium, the mean value of the scores for each type of lesion, calculated for each animal separately. is the following: Animal No. Sex Mean hours Cornea opacity Iris ConJunct~vae redness chemos~s 31 male 32 male 33 female o 0.3 o oo o o o OBJECTIVE The purpose of this primary eye ~rritat~on study was to assess possible irritation potential when Slngle doses of AEROSIL R 202 'are placed in the conjunctival sac of rabbit eyes, This study should provide a rational basis for a risk assessment 1n man. Panel Book Page 185

188 RCC PROJECT D46a~: AC:ROSIL R MATERIALS AND METHODS TEST SYSTEM Species Rabbits, New Zealand ~h~te, KFM Rationale Source Total number of animals Age at start of treatment Body welght at start of treatment Identi fication Acclimat~on Recognized by the international guidel~nes as the recommended test system. Kleintierfarm Madoeri~ AG CH 4414 Fuellinsdorf 1 Sw~tzerland 2 males 1 female weeks kg By un~que cage number and corresponding ear tags. Four days under after veterinary examination. laboratory condit~ons HUSBANDRY Room No.: 129 Conditions Standard Laboratory Conditions. Airconditioned witn 1015 air changes per hour and hourly monitored environment with temperature 22+2 degrees cent~grade. relative humidity I., 12 hours artificial fluorescent light/12 hours dark, music/l~ght period. Accommodation Individually in stainless steel cages equipped with an automatic cleaning and drinking system (Dipl. Ing. U. Ehret GmbH, Versuchstiertechnik, D 7830 Emmendingen 1 FRG) Diet Pellet2d standard Kliba 341, Batch 6/85 rabbit maintenance diet (~lioa. Klingentalmuehle AG. Switzerland) ad libitum. Results of analysis for contaminants are included in this report. Uatli!r Community tap water from Itingen, ad libitum. Results of analysis for contaminants are included in this report. 9 Panel Book Page 186

189 RC: p~aject 0468:: AEROSIL R Z02 9 TEST ARTICLE IdentJ.f1catJ.cn AEROSIL R 202 DescriptJ.cn So11d Ba~ch Number 1.mknown Pur1ty formulatj.on; J.ngrgdien~s avaj.lable J.n the sponsor's f11es. Stabi11ty of test artj.cle stable Safety precaut10ns Rou~ine hygien1c procedures were suff1 c1ent to assure personnel health and safety. Test Article PreparatJ.on: The ~es~ article was app11ed undiluted. TREATMENT A single dose was administered to the left eye of each animal. The right eye remained untreated and was used as the reference control. The application volume was per animal. Ra~1onale: Ocular contact is one of the probable routes for human exposure. /0 Panel Book Page 187

190 RCC PROJECT AEROSIL R RESULTS The results of the primary eye on the following pages. irritation test are reported TOXIC SYMPTOMS / MORTALITY No acute toxic symptoms were observed in the animals during the test period, and no mortality occurred. IRRITATION AEROSIL R 202 showed a primary irritation score of 0.1 when applied to the rabbit eye mucosa. COLORATION In the area of application no discoloration of the cornea and conjunctivae which could be related to effects of the test article was observed. CORROSION No corrosion of the cornea was observed at each of the measuring intervals. BODY ~EIGHTS The body weight gain of all rabbits was similar. NECROPSy Due to the results obtained, no macroscopic organ examination was indicated. /1 Panel Book Page 188

191 CODING FORM FOR SRC INDEXING ~CUTE OkRL TDXICITY (LD~0) STUDY WITH A~ROSIL R 2~2 IN RATS W~TH COVER LETTED 0830~3 DIMETHYL ~ILILONES AND SILOXANE (& > Panel Book Page 189

192 Degussa+ Degussa Corporation August Office of Ponution Prevention and Toxies U.S. Environmental Protection Agency (TS790) 401 M Street, SoN Room G099 East Tower Tunnel Washington, D.C Certified Mail Attention: 8(d) Reporting Re: Health and Safety Study Rule Section 8(d) [OPPTSS02040; FRL41821] Federal Register Notice May Dear Sir: On August 26, 1993, we notified you that we became aware of the existence of some reportable health and safety studies that are not in our possession, but held by our parent corporation. We requested copies and have received them. The substance is Sn Dimethyl siiicones and siloxane, reaction products with silica, sold under the trade name, AerosiJ R202 and Aerosil R204. The studies were performed with Aerosil R202. The studies are: I Ag.rte Dermal Toxicity (1050) (in Rats) gr,~'3ccxx>'3g:lf Degussa A<7U5rrNR DGT 2 Prj"'!!)' Eye Irritation (in Rabbits) a ~~?::&5 Degussa AGU5ITNR, S5001~DGT ODI~ ~3=3,,0.3 Acute Oral Toxic!!Y (1.050) On Rats) OegussaAGU5rrNR, DG~~ '.~ :J<J." "' ' Thank you. f Primary Skin Irritation (in Rabbits) Degusca AGU5ITNR, SfHlO13DGT ~C)acXCC>'?Jc~ ~ The Ames Test on a ToJuene Extr'ad &' 0)'3OQ:)C"3 Co9 Oegussa AGU5ITNR, ol(;;t.CD <=) o Sincerely, g~~ John Lewinson. Ph.D. Manager, Product Regulatory Compliance JL93376 cc: H. HasI, DCRP R. Marion. DCA Panel Book Page 190

193 R C C RES EAR C ~ & CON S U L T r ~ G COM PAN Y A G PROJECT ACUTE ORAL TOXICITY <LoSO) STUDY UITH AEROSIL R 202 IN RATS i DEGUSSA AG USIT NR. g~.. REPORT / DATE; JUNE l.:) ;...) :..,.~! C, ~,.. 0 0, '"" c.. P.O. Box CH 44S2 ItingQn SwitzQrland PnonQ 061 I 98 S2 S2 TQ1QX RCC CH Panel Book Page 191

194 RCC?ROJECT :'ERO=:~ Fl =:02 :3 PR~FACE GENERAL TITLE Acut2 o~al toxlclty <LD50) 5tuCY wlth AEROSIL R 202 In ~at~ SPONSOR STUDY MONITOR TESTING FACILITY DEGUSSA AG INDUSTRIELLE TOXIKOLQGIE POSTFACH HANAU 1 FEDERAL REPUBLIC OF GERMANY CR. O. lojeiberg RQ5Qa~ch & Con5ultlng Company AG CH 4452 ItingQn I SWltzQ~land RCC PROJECT NUMBER TEST ARTICLE TEST SYSTEM AEROSIL R 202 Rat PROJECT STAFF Study dl~qctor: L. Ullmann TQchniCal coordlnator5: R. SachQr H. MohlQr Pathology: SCHEDULE Acclimation TrQatm12nt / Ob512rvation ARCHIVING April la, 1985 April 17 to May May 1, 1985 RQ512arch & Con5ulting Company AG. CH 4452 Itlng12n I SWltZQrland Raw data, protocol and ~Qport, duplicatq of ~Qport, tq5t articll2 rqfl2rl2nc12 ~ample 4 Panel Book Page 192

195 SUMMARV ::::~NCLus:::aN ;c.. 0..:, _.._._... _.,",,: ~ : c 5 ': ~ ~ t ~ :: ~,=. c.~,~s : :.. : : c=...5 =.,~ ~.i", "" n :. ~ :. _ ~ oi.= ;.: ;! ~! :~~~ ~~~;! ~ ~r~: S~,~~;.?: ~ ~C5~ O~ 180C ~g~'~~ l~~ fc:~=~~~; aqat~ ~~~~ ~a~ ots~~~~~ ~,""'., '..J. 7~~ ac~:~ ~ra: ~=50 ~: oo~~~~~c ov~ a p~r~c~ than J..OOO rng/kg S't~?TOr:S ~~spnea was OC5~~~~~ l~ ~~i ~nlmal ~~thln : tc 5 ~o~r~ aftqr ~~~~ a~tlcl. ~=p~:~3t~on.:,. (. :.::::C20?S'( ~ ~y~ to t~~~"lc~l ~~fflc.~~~l;s ~d~5~ng~ t~~~ ~~~~~~~:. ~~ ~~;~~~ ~~~Q g~oup~ ~~~~ ~~~54~1~ p. Panel Book Page 193

196 ~c: '~a~e:7 ~~6/87 ~ERC::::_ ~ :O~ OBJFCTIVE' Th~ purpo~~ of t~l~ ~tud~ wa~ to a~s~~~ th~ tox~colog1cal profllg of AEROSIL R 202 wh~n adm1n1~t~r12d t :] rat5 by ~:lngl~ ol'"al 9a"ag~, follo~d by an ob~'2i'"vatlon p12l'"lod of 15 day5. rh1~ 5tudy ~hould providg a rat10nal ba515 fol'" ~ rl5k a~5~~mqnt 1n man MATERIALS TEST SYSTEM AND METHODS Rat. KFMHan Ui~tar (outbl'"l2d, SPFQuality) RatLonalQ RQcogniZQd by thq Lntgrnat10nal guldqlinq~ 85 thg I'"QcommQndgd tg~t 5y~tgm. SOUI'"CQ K1Q1ntiQrfarm MadOQrln AG, CH 4414 FUQllln5dorf / SWLtZQrland NumbQr of anima15 pqr group Total numcqr an lmal~ of AgQ at ~tart of trqatmqnt 9 to 11. w~qk~ Booy WQight at 5tart of trqatmqnt ~01 9 By uniquq cagq numbqr and corl'"q~ponding colorcooqd 5POt~ Rancloml.Zatlon Randomly ~Q1Qctgd at tlmq of d21lvqry 1n gl'"oup~ of fivq OnQ WQQk undqr laboratory condit1on5, aftgr v~tgrinal'"y Qxamlnat~on Panel Book Page 194

197 ~c: ~RC~E:~ J~6787 ":'E~OS:~ R :0: HUSBANDRY Room No 136 Condltlon5 Standa~d Labo~ato~y Cond1t1on5 Al~condltlonQd wlth 1015 al~ c~angq5 PQ~ hou~. and hou~ly monlto~qd Qnvl~OnmQnt wlth tqmpq~atu~q 22 2 dqg~qq5 CQntlg~adQ. ~QlatlvQ hum1dlty I., 12 hou~5 a~tlflclal fluo~ Q5CQnt 11ght/12 hou~5 da~k. mu51c/llght pq~fod Accommodatlon G~OUp5 of flvq 1n Mak~olon typq3 cagq5 wlth 5tandard 50ftwood oqd:llng C LlgnOCQl, Schill AG, SWl tzq~ land) 01Qt PQllQtQd 5tandard Kllba 343, Batch 18/95 rat malntqnancq dlqt (Kllba, KllngQntalmuQhlQ AG, SWltzQ~land) avallablq ad Ilbltum RQ5ult5 of analy5l5 :or contamlnant5 arq lncludqd 1n th15 ~Qport UatQr Comm~nlty tap water from ItlngQn, avallablq ad Ilbltum. RQ5ult5 of analy5q5fo~ contaminant5 a~q lncludqd 1n th15 ~Qpo~t iest ARTICLE AEROSIL R 202 Batch NumbC2~ unk nown fo~mulatlon; lng~qdhi!nt5 avallablq In thq 5pon50~'5 fllq5 Stability of tq5t artielq Stability of tq5t a~tielc2 dilution 5tablQ fo~ at lqa5t 2 hou~5 RoutlnQ hygiqnic P~OCQdU~Q5 WQ~e 5uffl C1Qnt to a55u~q pq~50nnql health and 5ahty 10 Panel Book Page 195

198 ~cc?rg~~cr AE~OS:~ =1 ::0: loo ThQ ~QS~ ~rtlclq was placqd ln~o a glass bqakq~ on a ta~qd MQt~lQ~ PK 4800 balancq and thq vq"lclq <PolyQthylQnQ glycol PEG 400, Fluka AG. CH 9470 Buch5 / S~lt=Q~land) was addqd A WQlght by volumq dllutlon was p~~parqd USlng a homogqnl=q~ (Ult~aTur~al(, JankQ and Kunk~l, StaufQn, FRG) HomogQnQlty of thq tqst a~tlclq l~ thq vqhiclq was malntalnqd du~lng t~qat~qnt uslng a magnqtlc 5tl~~Q~ (AUQ~alttmann, SWltZQrland). TREATMENT ThQ anlmals ~QCQivQd thq tqst articlq on a mg/kg body ~lght basq by oral gavagq aftqr bqlng fastqd for 12 to 18 hour5 (accq5s to watq~ was not lntqr~uptqd) Food was agaln prqsqntqd approximatqly onq hour aftqr doslng. Rat.lonalQ: Oral adm.lnlstratlon has bqen provqn to bq an QfficaCious mqthod to assqss tqst articlq absorpt.lon. Appllcatlon VolumQ/ kg body WQ.lght: Grcup 1 20 ml at 1000 mg/kg Panel Book Page 196

199 CODING FORM FOR SRC INDEXING LJTSIZI Submitting Organization DC:GUSSA CORP RES 8. CCJNS CCJ SKIN IRRITATICJN STUDY WITH AcROSIL R 202 IN RABBITS OCCLUSIVE APPLICATION) WITH COVER LETTER DATED METHYL SILICONES AND SILOXANc (677&291217) Panel Book Page 197

200 Degussa+ Degussa Corporation August 30, g rd) Office of Pollution Prevention and Toxies U.S. Environmental Protection Agency (TS'r790) 401 M Street. SW Room ~ East Tower Tunnel Washington, D.C Certified Mail Attention: S{d) Reporting Re: Health and Safety Study Rule Section S(d) [OPPTS802040; FRL41821] Federal Register Notice May 14,1993 Dear Sir: On August 26, 1993, we notified you that we became aware of the existence of some reportable health and safety studies that are not in our possession, but held by our parent corporation. We requested copies and have received them. The substance is , Dimethyl silicones and siloxane, reaction products with silica, sold under the trade name, Aerosil R202 and Aerosil R204. The studies were performed with Aerosal R202. The studies are: f AaJte Dermal ToxicitY (LOSQ) (in Rats) B'O,'3fXXX>3G:Lf Degussa AGUS'rITNR DGT Thank you. 2 Prim!!>' Exe Irritation (in Rabbits) ~~ 5 Degussa AGUsITNR DGT..3 Acute Oral Toxicity (LOSO) (in Rats) S~3cxx)o '3"" Degussa AGUS'rITNR DGT. r Primary Skin I~n (in Rabbits) ~o,sa:::cc@ DegussaAGUS'rITNR ~ 5 The Ames Test on a Toluene extrad Dr Pl3b00C> "3 Co > Degussa AGUS'rITNR, DKM. OD..0 :..l ~/).:! I == c ;... Sincerely. 9~~ John Lewinson. Ph.D. Manager, Product Regulatory Compliance JL':'93376 cc: H. HasI, DCRP. R. Marion. DCA.. ~ 65 Cb8llenger Road ~.PaIkNJ b'1s6il:. 2bi064i06100,.. ~ ':. :",. ~:'...,. ~": :. ",' Panel Book Page 198

201 RES E A R : H R C C C 0 :J 5 U L N G ~ G PROJECT PRIMARY SKIN IRRITATION STUDY ~ITH AEROSIL R 202 IN RABBITS (4HOUR OCCLUSIVE APPLIC~TION) o REP 0 R T DATE: MAY 30, ~985 r OEGUSSA AG USIT NR. OQ1~... :Db I P.O. Box CH 4452 It1nge~ Switzerland Phone 06~ I 98 S2 52 T~lex RCC CH Panel Book Page 199

202 RCC PROJECT AEROSIL. R :0: 7 SUMMARV UndQr thq condit~ons of th~s QXPQr~mQnt, AEROSIL R :02 was found to causq a pr~mary ~rr~tat~on scorq of 0.2 whqn appl~qd to intact rabb~t skin. :. In thq arli!a of application no discoloratl.on c"f tho;! sk 1.n was obsqrvli!d whl.ch could bli! rli!latqd to li!ffects of thq test art~c12. No corrosion effect had occurrqd on thq"skin at each mqasuring intci!rllal. According to EEC Council DirCi!ctillCi!, 67/S48/EEC, April 1983 Bru5sCi!ls, BCi!lgium, thq mqan valuci! of thci! scorci!s for Qach typq of lci!sion, calculatqd for Qach animal sqparatli!ly, 1.5 thci! following: Animal No. SQX 34 mah 3S fqmalq 36 hmalq Mli!an 1, 48, 72 hours ErythQma 0.7 o Edema o OBJECTIVE ThQ purp05q of this primary skin irritation study was to assess possiblr irritation potrntial whrn 5in91R dosrs of AEROSIL R 202 ar'2 placrd on thq skin of rabbits. ThiS study should prollidq a rat~onal in man. basis for a risk assqssmqnt Panel Book Page 200

203 ~:..: :~;: :.~:: ~~.;.~ ::;G:~.:::?:s: ~ :~: MATERIALS AND METHODS TEST SY57Ei': Rabbl.1'.;s ~~W Z~alan= ~hl.:e. KF~ :SPFayal~ty) Rat~onala To tal ~ '.imb.r 0 f arnmais Recognl=ed by tr.~ lnternatlon~~ su~delines as the rec~m~e~dac :~st system :. Kle;~tierfarm Mado.ri~ AG CH 4414 Fuall~~sderf! Sw~t=.r:and 1 malas 2 females AS. at start of treatr.~~t ~eks Body ~ight at start of ti'"ea::':nqn~ <;1 Ide;'! tl hca ticn By unl~ue ear tass. cage number and correspcndi;'!g Acclimation Feur days u~der tast co~dltlons aft~r veterlnary ~xaminatlon. HUSBANDRY Room No : 129 Cond~tlons Stan~ard Laboratory Condltlons Alrcondltioned with 1015 alr c~anges p~r ho~r and hourly monitored envlronment with temperature 22 2 aegrees centigr~de. relatlve humidity l. 1: hoyrs artif1c1al fluor.scent light/12 hours dark, music/light per~od Accommodation Individually in stainless steel cages eculcped wlth an automatlc cleaning and drinking system (Dipl. I;'!g. ~. Ehret GmbH, Versuchst1ert.chnlk. D 7830 Emmendingen I FRG). Diet P.lleted standard Kliba 341. Batch 6/85 rabbit malntenance diet (Kllba, Kl1ngentalmuehle AG, SWit=erland) ad libltum. Results of anaiysls for contamina~ts are i;'!cluaed in this report. Ua~er Cor;;mun:.~y tap wat'ir frcm rtinge~. ad!loitij;.i ~e'5;jlts of '!'.nalysls for ~ontami~ants are lncluded 1n th~5 reccrt 9. Panel Book Page 201

204 R:: QROJECT AEf;CS~L R 20: TEST ARTICLE ':'EROSIL R 202 '.5oll.d unk nown formulation; l.ngr~di~nt5 availabl~ l.n th~ 5ponsor'5 fil~5 Stability of tq5t csrtl.cli! Stabl.lity of t~5t articl~ in v~hicl~ 5tabl~ for at l~a5t 2 hour5 Routl.n~ hygl.~nic proc~dur~5 WQri! sufficient to CS55UrQ per50nnql h~alth and 5afety. Test Articl~ Prepcsratl.On: Th~ te5t article '.IIa5 l!ppli~d mol.5t~n~d 'llith tap water. TREATMENT Approximately 24 hour5 before treatment, the dorsal fur wa5 shaved with an electric clipper, exp05ing an area of approximately 100 squ;srq centimeter5 (10 cm 10 cm). On test dolly 1, O.S 9 of the test article was ;spplied to the intact 5kin of the shaved area. It was covqred with a 3.0 cm x 3.0 cm patch of surgical gau4e. ThQ gauze W85 cov~rqd with aluminl.um foil and then cov~red with an OCClU5iv~ dre5sing. The drr551ng was wrapped around the abdomen and anchored with an ela5tl.c bandage. Ratl.onalQ Skin contact i5 one of the probable routes for human exposur~. 10 Panel Book Page 202

205 ~c: =~8;EC7 J=~aGC ~ERCS!~ R ::J: :3 RESULTS ;h~ ~~sul~s of the pr~mary sk~~ ~rrltatlc~ t~s: ar2 ~~ported on ~h~ followir.g pages. IRRI7ATICN ~EROSIL R 202 sl".owed c. primar~' lrrltatlor: scq"re of 0.2 when appllec tc l~tac~ rabbit sk~~ ;" COLORATION In the area of appllca~lon no discoloration of ~he skin was ooserved WhlCh could be related to effects of the test article. CORROSION No destructlons or irreversible alterations of the t.~ated skin wer'2 observed. No corrosion effect 'JJas eviden~ on ths: sic In. BODY ~EIGHTS The body welght gain of all rabblts 'llas similar. TOXIC SYMPTOMS / MORTALITY Ne acute tox1c symptoms were observed i~ test period, and no ~ortality occurred. the anlmals during the NECROPSY Due to the results obtained. no macroscopic organ '2Xamlna~lon IJJ2.S 1~(llcat2d Panel Book Page 203

206 CODING FORM FOR SRC INDEXING MUTAG~NILITY T~ST (~ R TULUbN~ EXTkRLl ~ROM 2~2 WlfH co~~~ LETIE~ OArED ~83~~3 IMcTHYL SILICUN~S AND SILuXANc (~77o~907) Panel Book Page 204

207 D6gussa+ Degussa Corporation August 30, 1993 Office of Pollution Prevention and ToxiCS U.S. EnvironlT.ental Protection Agency (T8790) 401 M Street. SW Room G099 East Tower Tunnel Washillgton, D.C C.rtiiied Mail Attention: 8(d) Reporting R.: Health and Safety Study Rule Section 8(d) (OPPTS802040; FRL41821) F~ Register Notice May 14, 1993 o.arsir: On August 26, 1993, we notified you that we became 8W8At of the existence of some reportable health and safety studies that are not in our possession, but MId by our parent corporation. W. requested copies and have received them. The substance ii , Dimethyl silicones and SiIoxane, reaction products with silica, sokt under the trade name, A8IOsiI R202 and A8IOsii R204. The studiel were performed with AefOIiI R202. The stuc:tiei...: I Acu!t [)tnnii Tpvirjly (I P5!) (in Rau) Sf,'3 X)CXf3.~ [)egi" AGUSITNR,850012DGT %. b::i Eye '~..:' RebitI) ~~5 18 AOUsrr DGT, ~ " o Thriyou. ~, 9~~ John lhinion, Ph.D. tmniigir, PnMtuct RegI..y CompIIIa Jl83378 cc: H... OCRP R. M8rtan, DCA 0IiIIIIiCiii Eil... 0 '1 hii NJ 0&1 iali'4id) Panel Book Page 205

208 r DEGUSSA AG USIT NR.I ~lf: QQ ~tt' :]2 ~t:tl ~".~ l~: :... :. :'1Y. C ',_, ~:: I,' '.' _ <:f T,,"coiogy L' \/ "'~'_ :/ fjf 1'''~:''.:" Ci~~~,' Z.l!.,..(~.,:r Str~'3a Mainz BACIER!AI. MUTAGENICITY TEST ON A TOLUENE EXTRACT FROM... : AEROSIL R 202 c '. Report No. SP 598 (12 pages) Report date: January 10, 1984 Sponsor: Oegussa AG, Zweigniederlassung ~olfgan9, Hanau 1, Federal Republic of Gennany Panel Book Page 206

209 (, R~SCLTS A~D DISCUSSIO~ Controls Control plates without mutagen showed numbers of spontaneous revertant colonies per plate similar to those described by Ames' group (2). As positive controls NmethylN'nitroNnitrosoguanidine. NethylN'nitroNnitrosoguanidine anu benzo(a)pyrene 4,Soxide were used in the experiments for direct mutagenicity and 3methylcholanthrene, benzo(a)pyrene, 2aminoanthracene and benzo(e)pyrene in the experiments with an activating system. Benzo(a)pyrene is activated well by liver preparations from 3methylcholanthrenetreated rats. but only poorly by preparations froc phenobarbitaltreated rats. The reverse is true for 3methylcholanthrene (2). Aroclor 1254 induces monooxygenases which are induced by phenobarbital as well as monooxyger.ases which are induced by 3methylcholanthrene (12). Both benzo(a)pyrene and 3methylcholanthrene were activated in our experi~ents. This indicates that both metabolizing systems were functioning. Also 2aminoanthracene. which requires Noxidation for activation. was strongly mutagenic. Inhibition of epoxide hydrolase by l,l,ltrichloropropene oxide was controlled by studying its effect on the mutagenicity of benzo(e)pyrene. l,l,ltrichloropropene oxide increased the mutaenlclcy of benzd(e)pyr~ne severalfold. the sutaaenicitie. of various positive controls, such as those of benzo (a)pyr.n., 3..thylcholanthr.ne and benzo(a)pyrene 4,5ox1de 1n TA 100. and tho of 3..thylcholanthrene and NechylN'nitroNnitrosoguanidine in TA 98, are aood indications for the presence of the Ifactor. p~~ 101, which.. y be lo.t. Various compounds. ineludina these controls, show sutagenicity in bacterla with the ~factor, but are inactive or much less active in respective bacteria without t~is Ifactor. In addition to this qualitative assay for the pre.ence of pkm 101 by direct d.teraination of the relevant property (increased mutagenicity), a quantitative as.ay with esti.. tion of the fraction of bacteria which Brow on ampicillincontainina plates va. performed. These experiments.howed that virtually 100 : a! the bact.ria of the used cultures of TA 100 or TA 98 po d the Rfactor.. ~.. Panel Book Page 207

210 7 TOLUESE EXTRACT FR0!1 AEROSIL R ZOZ The extract was prepared by Degussa AG. The dry weight of the extract from 50 gram Aerosil R 202 was 2.1 gram. This extract was tested for mutagenicity with Salnonella typhimurium TA 100. TA 1537 and TA 98 as well as Escherichia coli WP2 uvra. Six different concentrations from 15.8 to 5000 ~g per plate were used in the experiments for direct mutagenicit.y. In the experiments with 59 Mix. a concentration of 5 ~& per plate vae tested additionally. The extract appeared dissolved (or was possibly 'inely dispersed) up to the highest concentration used. For estimation of toxicity. his+ mutants were added as an internal standard to some plates and their survival was determined. A significant toxicity occurred only at the highest concentration of extract and then only in the direct test. Hence. toxicity and solubility allowed mutagenicity testing with an excellent sensitivity. The results of the mutagenicity tests are sht"jti on the table on page 8. No mutagenicity was observed under any experimental conditions. This included the case where the epoxide hydrolase inhtbitor and glutathione depletor trichloropropene 2.Joxide. was added to the 59 Mix in order to increase the sensitivity of the test towards coopounds which are activated to mutagenic epoxldes (this experiment was performed with TA 98). Kainz. January ~~.... Dr Glatt. ~... Prof. Dr. F. Oesch '... q. Panel Book Page 208

211 BACTERIAl. MUTACENICITY TF.5T ON A TOLUENE EXTRACT FROM AEROSIL R 202. I Rt:SUl.TS: The extract WclS not mllt.1,~, nic andcr any ekpertment~l conditions. J Panel Book Page ~

212 CODING FORr.1 FOR SRC INDEXING M.::r;:)ll~he No. r.:.:::;:;':'::':;':';';':"'~,jots ~4 t.:..::.:...;...::..=~ HOECHST CELANESE CORP Do~um6nl TIII& INITIAL SUBMISSION: 28DAY DUST INHALATION TOXICITY STUDY OF CABOSIL N70TS IN ALBINO RATS WITH COVER LETTER DATED ~~0692 Cn&mical Calegory "CABOSIL N70TS Panel Book Page 210

213 t...g... _, ~. ~E\\Q Hoechst Celanese \\q 2 <?51 D Department of Environmental. Health & Safety Affairs (DEHSA) November 6, 1992 RAJ16992 Document Processing Center (fs790) U.S. Environmental Protection Agency 401 M St., S.W. Washington, D.C A.ttn: TSCA Section 8(e) Coordhator,,, _. f"", i Hoechst Celanese Corporation,, : _,_:,~ Route PO Box 2500 Somerville, NJ Telex Fax I~~IIIIIIII II! 8EHQ9Z8~78 HUT 1V17/92 _ Dear Sir or Madam: In accordance with the requirements of TSCA Section 8(e), Hoechst Celanese Corporation hereby submits a report of 3 28 day dust inhalation smdy in rats of CABOSIL~ TS720. treated fumed silica (CAS No ~7). The chemical identity given in the report CABQSIL N7QTS, according to Cabot Corporation, is identical to CABQSIL~TS720. The toxicity report was recently obtained from Cabot Corporation. In this smdy. 40 male and 40 female rats were exposed to CABQSIL~ TS720, Treated Fumed Silica at a concentration of 60 mglm 3 for 6 h1day, 5 days/week for up to 4 weeks. During the first day of treatment. signs of excessive toxicity were observed and a decision was made to reduce the exposure to 30 mglm 3 for the remainder of the experiment. Despite this adjustment, by the end of treatment day 2, nir.e male rats had died. Necropsy of these animals revealed evidence of a lethal, acute pulmonary hemorrhage accompanied by bronchiolar plugs with emphysema in five out of five rats examined. The acute toxicity exhibited by this surfacealtered material is much greater than would be expected for untreated amorphous silica. CABOSIL~ TS720. Treated Fumed Silica is being investigated by the Hoechst Celanese Corporation for use as an additive in polymeric materials. Once the polymers are manufactured, the treated fumed silica is incorporated into the polymer matrix presenting no opportunity for consumer exposure. Hoechst Celanese Corporation will take adequate measures to protect its workers from excessive exposure to this material. This submission contains no confidential business information. Ifany fulther information is required, do not hesitate to contact Dr. Michele R. Sullivan. Director. Product Safety at (908) III :::z (:? SusanEng~ Vice President. Environmel!tal, Health & Safety Affairs CERIIFLED MAll1 RETURN RECEIYf REQUESTED File: Log No. 154 Panel Book Page 211

214 TOXIGENICS' STUDY DAY COST INHALATION TOXICIT7 S'!'UDY OF CABOSIL ~73TS IN ALBINO RATS SUBMITTED TO: CABOT CORPORATION CABOSIL DIVISION P.O. BOX 188 TUSCOLA, IL SOBMITTED BY: TOXIGENICS, INC. 183G EAST PERSHING ROAD DECATUR, IL JlJNE 29, 1983 ToxiGenics A Subsldiarv of Whittaker CorcorafJon1/, Panel Book Page 212

215 I. Summary_ Stl:dy No. Page = 42 Pages A 28day dust inhalation toxicity study using albino rats ~as conducted with CABOSIL N79TS. Exposures were for 5 days per week fer 4 consecutive weeks. Four designated groups, TI, TII, TII! and TIV, each consisting of 19 male and 19 female rats, were exposed to an atmosphere of the test article. After weeks 1, 2 and 4, groups TI, TII and TIV, respectively, were sacrificed and examined. The ti~eweighted average (cumulated) gravimetric concentrations for weeks 1, 2, 3 and 4 of the study were; ~.037, 0.031, and ~g/l ai~, respectively. Two Vehicle Control groups, VCI and VCII, were exposed to air only and were sacrificed and examined after 2 and 4 weeks, respectively, of exposure. Each Vehicle Control group consisted of 19 males and 10 females. Test group TIII was designated a recovery group with onehalf (5,I, I, I r I.: :.j : ii! I I: Ii males and 5 :emales) sacrificed after 6 and 12 weeks, respectively, of recovery. (Following 4 weeks of exposu:e.) The 6 and l2week recovery groups will henceforth be referred to as TiIIA and TiIIB, respectively. i :i During ~he first 2 days of the study, 9 male rats died..~ This was apparently due to test article at a level of 0.06 mg/l air during the firt six hours of the study (day 1). Subsequently, test article target concentration was set at 3.03 mg/l air. Animal observations noted included; crusty eye, crusty muzzle, crusty nose, crusty substance around ear tag, eye closed, ToxiGenics A SubSidiarY 01 Whittaker C.j~=;:;rauon I Panel Book Page 213

216 I I I I I I I I I I I I I J ~ I level of ~g/l Study No. Page 2 air for various times up to 4 weeks was inflammation was more localized and less fulminant. collagen proliferation. of 42 irregular breathing, irritable, lacrimation, salivation, sca~, red stained fur and yellow/brown stained fur. tically from corresponding controls during the course of the Pages study. Recovery group animals appeared to gain weight at a rate cocparable to untreated animals of the same species and strain. Hematology data indicated a decrease in the relative number of lymphocytes and an increase in the number of neutrophils after 2 and 4 weeks cf exposure. This finding is consistent with the chronicactive pu~onary Mean bocy weights of test anlmals did not differ statishistologically. leukocyte couct. Serum chemistry data were not remarkably different from comparable cortrois.... inflammatory process seen At week four there was an increase in total Histological examinations of rats exposed to CABOSIL,.".VoJ _...,. I N73TS at 0.06 ~g/l air for six hours followed by an exposure associated with a chronicac~ive pulmonary inflammatory process. Six weeks after the four weeks of exposure the pulmonary Twelve weeks after the four weeks of exposure, the pulmonary inflammation was e»en more localized, however histologic markers of chronicity were evident. namely interstitial fibrosis and interstitial Granulomatous responses were not seen. What proportions of toe chronic pulmonary lesions were due to the initial severe injury relative to the subsequent lower Tod3enics 1 A Subsi<liarv of Whittaker Corporation. Panel Book Page 214

217 Study No. Page of 42 Pages dose level (9.669 mg/1 air compared to G.931 mg/l air) cannot be determined from tbis datat:t ~.~D~ S. R1chard Dudek, Ph.D. St~rector Stephen V. Becker, D.V.M. Pathologist All work relating to this study was done in conformity wit~ the FDA Good Laboratory Practice Regulations (21 CFR 58). The study was inspected by a Quality Assurance Specialist on the dates shown below. Management, including the Study Director, was informed of the results of these inspections/audits on the dates shown. The data in the report ~ere compared with tbe raw data and are in agreement. The report and study file were examined to assure tttat any p~oblems found during Quality Assurance inspections were corrected, and if necessary, their effect on the study documented. Phase inspections conducted January 21 and 24, Final Data Inspection and Report Audit conducted 17, and 29, 1983; reported to Management, including the Director, June 21, l ~~Z~~""T""'. Supervisor. Quality Assurance June 15, 16. Study 1Jz.o,.I '03 ~ The raw data relating to this study, as well as specimens, and the final report are stored at ToxiGenics, Inc. Storage is as per FDA GLP's and may include volume ~eduction by conversion to certified microform. Date of Report: June 29., ~9.83 II. Introduction A 28day inhalation study was conducted in rats to determine the subchronic toxicity of CABQSIL N70TS. The exposure regimen for tbis study was 6 hours per day, 5 days per week for a total of 28 days. During the 28day exposure period, groups of 20 animals (10 males and 10 females) were scheduled for sacrifice and evaluation I after and 20 exposures. These groups were designated TI, TII I: and TIV, respectively. One group. TIII, of 20 rats (10 males and 10 I: ToxiGenics iii A SubSidialv of Whittaker CoroorabOn ~ ~I I Panel Book Page 215

218 I Study No. Page of 42 Pages \ females) was designated as a recovery group with one half of its ani~als (S males and 5 females) sacrificed after 6 and 12 weeks of recovery. These groups were designated TIIIA and.tiiib, respectively. scheduled evaluations. Given below are the study groups and their Clinical Rats Pathology Group M/F Weeks.Body Weight Weeks Necropsy Weeks Initial Histology Baseline 13/13 3 VCI 13/U 2 VCII 19/13 4 TI 19/HJ 1 TII 19/13 2 TIIIA SIS 13 TIIIB SIS 16 TIV 13/13 4 3,1,2 2 3,1,2,3,4 4 9,1 1 9,1,2 2 3,1,2,3, ,1,2,3, ,1,2,3,4 4 x x ve= vehlcie control, a1r only T,. Test group = Not determined The study was performed at ToxiGenics, Inc., 1833 East Pershing Road, Decatur, IL 62S26. The study was initiated on 3anuary 23, 1983 and completed on May 13, III. Personnel The principal tech~ical staff involved in this study is listed below. B. Richard Dudek, Ph.D., Study Director Peter V. Churukian, B.A., Section Bead, Chronic Inhalation Toxicology Panel Book Page 216 ToxiGenics,:.. A SubsidiarY 01 Whitlaker CorporatiOn OJ

219 I, I I I I I I I I I I I I 1 I I I I I IV. I barcode of the number were affixed to each animal's individual then sacrificed. and glass. Study No. Page of 42 Larry L. Morath, B.S., Section Bead, Acute and Subacute Inhalation Toxicology william J. Koretke, B.S., Inhalation Toxicology B. Chris Pegram, B.A., Inhalation Toxicology Patrick M. McKeown, B.S., Inhalation Toxicology Deborah A. unique within ToxiGenics. Reimer, B.S., Inhalation Toxicology K. Mickle Hayward, Inhalation Toxicology Dr. Stephen Becker, D.V.M., Pathologist Procedures A. Test system Onehundred forty (79 male and 79 female) rats l were used for the study. The animals were identified with an ear tag baseline clinical chemistry and hematology determinations and 1 Rattus norvegicus (CD rat), Charles River Breeding Laboratories, Inc., port~qe, MI Pages Only healthy animals were used for the study as determined by body weight and clinical observations during quarantine. groups of lb male and 19 female rats each were randomly selected by computer and designated as either VCI, VCII Control), TI, TII, TIII and TIV. The number and a computer readable cage compartment. At the end of the 19day quarantine period, 19 males and 19 females were selected for blood collection for (Vehicle Animals were housed and exposed in 5 cubic meter chambers constructed of stainless steel Six ToxiGenics A Subsidiarv ofwhi llaker Corcorali~ 0 Panel Book Page 217 I~

220 Panel Book Page 218

221 Panel Book Page 219

222 Study No. Page 13 42G1171 of 42 Pages Week Mean 1 G G S.D. = Standard Oevlatl0n Mass Median Diameter (microns) Geometric S.D. Cumulated Mean ~2766 Cumulated Geometric S.D "35 B. Mortality and Detailed Observations During the 28day exposure period 9 animals from the various designated groups were found dead on the days and times listed below. Time Study Found Day Dead VCI VCII TI TII 1 AM PM AM 2 3 PM NOTE: All anlmals found dead were males. AM prio~ to exposure initiation. PM = After exposure te~ination. = No deaths occurred. TIII 1 TIV 1 These deaths apparently resulted from a tuneweighted average concentration of test material equal to G.G6 mg/l of air during day 1. Subsequently, the target exposure level was lowered to G.S3 mg/l of air for the remainder of the study. A summary of a1l observations noted during the 28day study is presented in Table 1. All individual animal observations are I presented in Appendix B. Panel Book Page 220 ToxiGenics \'l. A SubSidiarv 01 ~t\lnaker Corporation

223 Study No. Page of 42 Pages TABLE 1: SUMMARY OF ANIMAL OBSERVATIONS 28DAY DOST INHALATION TOXICITY STUDY IN ALBINO RATS TEST ARTICLE: CABOSIL N7GJTS Number of Occurrences Observation 1/CI (No. Males/No. Females) VCII TI TII TIII TIV Crusty eye GJ/13 13/G 13/1 G/9 III 1/2 Crusty muzzle lij/g GIG G/2 2/6 5/9 6/7 Crusty nose GIG 2/13 4/8 6/1G 9/9 9/9 Crusty substance around ear tag llg 1/2 I:J/G a/3. a/3.2./2 Eye closed a/3 G/GJ 3/3 3/" I:J/liJ 2/1 Irregular breathing I:J/" "I" 7/9 6/lliJ 9/lllJ 9/HJ Irritable 1/3 lij/lij 3/liJ 3/G 3/3 3/1:J Lacrimation a/" "/G "/3 "II "I" "/G Red stained fur ala GIG III G/3 3/9 1/8 Salivation G/3 GIG 4/1:J 5/2 2/1 6/" Scab I:J/I:J l/a G/a G/liJ "/2 "/G Yellowlbrown stained fur a/l:j ala GIG 3/" 3/1 III Ị. ToxiGenics \~ A SubSidiary of WMlaker Corccratiol" Panel Book Page 221

224 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /19/65 04/28/94 8D DOW CORNING CORP FOOD & DRUG RESEARCH LABS INC CHRONIC (8MONfH) FEEDING snmies WITH ANTIFOAM A IN RABBITS.WI1H. COVER LETTER DATED ' Chemical Category SILOXANES AND srtlcones~dime ( :9) Panel Book Page 222

225 : & &4 April 20, 1994 _, 1 I.'..~~.~.~...~~:~~S ~'E~j ~~6~: TSCA Document Processing Center (TS790) Office of Prevention, Pollution and Toxic SUbstances U S. Environmental Protection Agency 401 M street, S.W.. washington, D.C Attn: 8(d) Health and. Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41821] Health and Safety Data Reporting; 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow Corning Corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Cbemica1 Substance(s): siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Dimethyl silicones and siloxane, reaction products with silica 7% in Antifoam A Title of Submitted StUdy: CHRONIC (8Month) FEEDING STUDIES WITH ANTIFOAM A IN RABBI~S Dow Corning corporation November 19, 1965 Manufacturer: Dow corning corporation 2200 West salzburg Road Midland, HI Panel Book Page 223

226 2. I you require further general information concerning ~is submission,p1.ease contact Dr. Rhys G. Danie1.s at For further informatio~ concerning the submitted study or the toxico1.ogical properties of the 1.isted chemical. substance, p1.ease contact Dr.Robert G. Meeks at sincere1y, ~E.B Alvin E. Bey.~ u.s. Area VicePresident Corporate Director HES Panel Book Page 224

227 A sc::ies of extensive investigations of a num.ber of polyslloxane:,,cc.mpounds is in progress in these Laboratories. The: studies were auth,:)rized,:)n September by 0,:),,' Coi"ning Corporation and are being carried out in accordance with protocol.:; developed, u1 con1'ercnce.. with these Laboratories, dated Sept.::mbe:" The present report covers the results of that part of the studies identified in the protocols, as "Phase 0 Groups 3 and ~1l. which was designed to investigatc' the. efiects of feeding Antiioa= A to rabbits in rations containing both. normal and elevated le...els of cholesterol. For the purposes oi comparison. groups w.:rc included which receivec. only ::ations containing equivalent aitlounts o tlus s~erol. Antiioam. A is a,iscous milky white fluid. The sample io~ these studi..s,,..as recei...ed on S~ptember ';. labele<i: "Dow Corning. Medical _1.ntiioam A Com?ound. 1 gal. Lot :No. H 0019:' ":J;he cholesterol sam.ple was labeled:' looe G~1; Cholesterol USP E'or Investigational Usc Only: Cont:o'ol No. ";'071; ~utritional Bioche=icals Corpo:o'atio~. Ohio.,t Cleveland. Panel Book Page 225

228 Procedure Thirty young adult albino rabbits weighing be~ween Z.5 and 4.5 kg were acclimatized t" laboratory collchticns and distributed as follows: Group Addition to Basal Diet (Purina Rabbit Chow) Males Females 3 1"0 Antiioam A 'f~ Antiioa:n A o. S% Choi~sterol 18 None (control) o. S% Cholesterol 3 3 Purina Rabbit Cno,... served as the basal ration. The animals,,,erc housed indi...iduall) in raisedbottom cages. ~atio:ls and fr~sh watcr were a...ailab:!.~ ad libitu:1.l. Appearance. behavior, and sur'\rival were observed daily and body wdghts r.:corded weekly. Attcr.1.?"ts to rccorc. food intake proved futile and were discontim:ec aiter the first fe''' '... eeks. At 0, i, and S :r.onths "t:"", following cetcrminations we=c made i::. au rabbits: hcn~oglobin conce:::t=ation, hematocrit, red cell fr.agility. totai. and differential!eu~ocyte counts,. crill" ph, specific gravity. albutnin;... glucose, acetone, bile salts and pigments, and the microscopic appeara::.ce of th~ centrifuged sedim.cr.ts. In adcetion, at 8 n10nths blood urea Panel Book Page 226

229 ni~rogen and serum, cholesterol levels. and serum alkaline phosphata sc and glutamic pyru'i.ic tr:j.nsaminasc activities were evaluated.. At the. time of the final examinations. specimens of blood and urine wcre also collected and submitted to the Do..." Corning orga:tization for silicon analyses. All rabbits ""'ere then sacrificed by e.'csanguination under Nembutal anesthesia. The viscera a:ld carco.::;scs werc examined for gross abnormalitics and 10 critical organs were weighed (liver. spleen. kiclnc) s. heart. lungs. brain. gonads. and adn~.:.a. adrcnals, and thyroids) and conl.puted as percenta :cs of body weight. Represen"tativ..: sections of approximatc.ly 25 organs (livcr. gall bladder, spleen. kidneys, esophagus, stolnach, large and small intcsti.."1.cs. pancreas. urinary bladd~=, adrcnals. gonads and'adnexa, thyroid. pit:.:.itary. thymus, rned:i.astina.l and mesenteric lym.ph nodcs. hc<,.rt. aort.."\.. l~gs. marrow. skeletal muscle, peripheral nerve. skin. spinal cord. and o::"ain) wc!"c ~reser,,cd i per cent formalin. All of these ''"cr ;: processed. sectioj:.~d, stained with hematoxylin and eosin and cxaminc:.d :1l.icroscopically. Panel Book Page 227

230 , Results the Antifoam A group receiving the unsupplemented basal ration (No. 9609lvL) died after 21 weeks. Dea'th appeared to be du to pulmonar}~ irritationa.~d.';:':;c:.. ". congestion. signs of "'hich had been seen while the rabbit '\Va,s alive.. ' This':... may represent an i':'lcidental finding, since respiratory diesease is common' in rabbits, or it.nay have resulted from inhalation oi the po,,,dered diet~', Nos. 96l0~ t a::td 96lZl\. 1 in the group receiving b.jth the Antiioam A plus the cholesterol supplement died during the 3.!:1d and loth werk, respectively. Extensive hepatic injuries were seen in both animals. No. 9719F of the group receiving 'the cholcsterolsuppleme:1ted ration died during the 28th week "..ith si:nilar h.::patic findings. All rabbits in the group receiving the unsupplemented basal ration survived to the terlnination of the study. No significant beha,oio:::al changes ''1:ere noted at any tl..ne. The charact~r 0::: i1 equency of excreta appeared normal, exceptior occasional episodes oi diarrhea. which was seen in all groups regardle~s oi th~ prc~ence Or abscnc..~ oi either c~olestcrol '.:>r Antiio~l1n A in the diets. Food intake records were discontin..:.cd because rabbits tend to scatter large am.ounts oi iood rendering euor:s of acc~:ate recording useless. The rations '\.vere preparec. in po,vcered form. in order to permit complete hor."log~nous distribution of the silicone sa,:nples anc. cholesterol in the diet, and this agg::a, at.:d th.: problc~.. In all' g=oups' including the controls.,.. ".:.4:~": Panel Book Page 228

231 nasal and ocular irritation. evidencc by nasal exudate, sneezing, and iridal inflanunation were not~d curi::.g the one to ~yo hours after ea.ting. These bouts of respiratory irritation nlay have led to the pulmonary inflammation and permitted the iniection ~"hich :lpparcnuy icd to the early death of No. 9609).1. Finding~ at oth =~ 'l.utopsics also ren~cted this irritation. Inasmuch ~s tho? rabbits ",'crc rnatur~ at the initia~ion of the study; body weight changes throug.hout the study wcr~ rninim:ll (Table 1). Several rabbits in the control ~roup (c. g., ~os. '1652M and 96561\'1) showed small net wdght losse$ as did one rabbit (~o. 9659:1\.1) in the cholesterol group a.nd another (No. 967ZF) in the Anti{oam A + cholcsl;crol g:roup. Over the total 36weck pc:iod net weight cha.::.g~s '\'aried irorn losses kg to gains of 0.9 kg with no signiiicant ciiifcrcnces noted ::lmong the groups or anlong tbe sexes. The l.;:c.kocyte data. (Table 2.)...al:iec widely am.ong the groups includin~ the control>' bu~ g.enerally rcn:.ained within the normal 1intits for those paran"lct<.'r.::; obser'\ cd for raobits in thcse' Laboratories. Deviations from no:mal wc'.o:: sec':;. ~!":.fuc' diffe=c~tia.l CO\l.."'lts of Nos. 9607M {AntifoamA}, 9612:1'\"1 (.'\o;,::lloa::.':. A.. chol.:.stcrol). <!:'1d 9659F (cholesterol). These clw.nges.inay havc o~c':'1 i:'...::idcr.tal t" th~ t!:"a::lsient respiratory iniections and acute parasitic inic'~utions com:"1"lol1 i..;., rabbits. One rabbit (:No. 9672F, :~'tifoa.m..:j,. + chol,:~st~rol) showed ::!.:l. c."trcme leukocytosis (> cells per m.m 3 ) e...idently re~atcc. to e)':tens~""c he?a~ic injury discussed belo'\v. Panel Book Page 229

232 The hematologic determination::; (Table l) revealed c_ num.ber of ab:l.orn1.~itics in the ;;roups rc:ceiving cr.olestcrol. either with or without Antifc:l.m A. Specifically, anemia.... as evident in the terminal examination:; of. Nos_ 9610M. 9611::"1. and 9672F of the A!)tifo..rn A + choicsterol g::ou? and Noo 96:>9::vt of the cholcste:ol grol:p. These hem.oslobin values iell below 10 S per 100 Inl 'which is considered the lower limit of nonna!. Hcznatocri'ts...':erc cons:a:1t with the expected cell voluincs a.t these hcmcgloblj1 levels with the exceptio~ oi those sc~n in the terminal ~"":'ll~i=1ation of No. 967:?F where n1icroscop~c < X:llnination of the cells rev... a),~c. hypochromia. The erythrocyte!rag~lity varied widely. but these "\,,,"crc within the no:::nai : an~... and it was c,,idcnt that :1.0 changes in blood suriac~ t<'n~ion oecur::cd with chronic adrni:listration of either ::hc AntifoaIn A alo:lc or in con1bination with the st.~rolo The biochemical dat01 (Table 3) detc:::ninc<.l only at tcnnination rcvealcc. cxtrelncly high sc:..m cholesterol values for all c::'olestcrolfed animals regarc.lcss of t!~c presence or abse::cc of AntH'oam A in the diets avct;tgi:lg 6 to 10 tirn~~s :10rI1.1a'._ Slight clc... ;.tions we:'c noted among Nos. 9655::'\., 9715F, and 9716= of the co:otrols. ::>~t these wcre not of the order of t~;lg.nitlld(' of tho,;:c 5':C;1 in the steroi~::=eatcdrabbits. In several ir. sbnc.,; an1on~ t!lo;.o,;c r:'l::>b~rs thc serul"!":. d:.oh~stcrol lc"\'cl was be~ond the range 0; mcasnrclncnt. The.h'..."a!ucs a?pca.: in the table as 600 mg: per cent. thc Up?.:"::" lln"'l1t. 0: :'l1ea.~u::"cmcnt with ~~e method used. Panel Book Page 230

233 Other biochemic.'ll determinations sho,""'cd that there were no effects of the silicone or saicon~~tcroid adm.i:listration OD normal liver arod biliary function. These did not apparently renect the rnorphological status of these organs as detern"llned in the subsequent microsc"pic c..~n"lin..'\tions of the tissues taken at autopsy. Urine!indings (Table 4) were characteristic of.'this species where lipoida~ urines Olre common. There were several instances of albuminuria in all g"c'ups. No ::iij::niiicant abnormalities in glucose or other parameters were noted 1:1.he urine c:;.ar:.1ir..ations oi th~ ~"'o tes".: and the 1:'... 0 control groups. Th~ orgar weight data (Trtble 5) exp:.esscd on an absolute basis and... :; p... r cent 01 body wc:ight wc:.e ~pproxi:natel~' ~qui...alcnt \...hen the Antifoarn. A group was compared to the basal ration group or when the cholesterol groups,",..ith a.nd without the silicone were compared. ':i:'he addit:ion of cholc:stcrol to th.;: dict, h::>wever, ob"'iously caused increased live)." weights (as well as scrurn levels) and caused sigr.iiicant ele... ations in the weights of othcr organs. A:r..ong the...ari.:us groups it appeared that the s..erol was depositec. in a =:'l~:;::bc:" 0: :is:;;ues. This deposition caus~d chans.:~ in total organ...\,'<~ight anc..j.':'; "'.:ill be seen bc~ow, in tissue morphology. No di ::ect ('ffeet of 3c.:=:..inis:ratior..:>f. the silicone itseli was noted inasmuch as tho:" responses in ea.ch of th...~ siliconctrca.:eci. groups did not differ from that of the '.:orresponding controls. Panel Book Page 231

234 Gross findings at autopsy (Table 6) 'were particula:!:"ly num.e;ous in the two groups which had 0.8 per cent cholesterol in the diet', but 'there appeared to be no difference with the addition of the Antifoarn A.,TyPic.al cholesterolinduced changes occurred in me liver, heart and aorta:; ~,.' addition, changes in the adrenals and spl~en were noted. Renal and pul!nonary changes '\'ere scattered. although they appeared to be.more numerous in the basal control group. It should he noted that the population of that grol~p was 't"wice that of the other g:oups. Histopathological e..a=.inations coniirmed 'the gross findings (Table 7). All anil:"l.als which received O. S per cent chol~stcrol in their diet sho,...cd fibrotic changes and cirrhosis of the liver. SO!"1'le of those,...hich also had recei... ed t.~e A:::ltiioam A ~ cholestt"::"ol in their diet showed fooi::'ly material in hepatic cells, "ioam cells" (histiocrtcs containing lipidlike material) in t..l:!.e brar.ches of the hepatic artery. foa..ny material. in the sinusoids of the splcen, and scattered "foam cells" in the adrenals and kidneys. Fatty :":laterial w~s seen L.""l coronaries, somcti!nes oblitcrating tht> lun1cn in hearts of some of these cholesterol treated animals. The occur:::cncc of "foar:: cells", however. was no"t licited to the c~pc'rimcn~land,"o:1t:"ol gro..:.ps which received additional 0.8 per cen1: cholcst~roi. The:;;e o.::curred in the 1:"l.u.::o:::a and muscle of. "the stomach, 'the intes"tiz:al mucosa, in th~ re;"lc:.l pel,is, adrcr.al medulla, in occasional heart vessel,;, inti::::a oi the aorta, and,;kin of the basal ration group. To sor:.1e extent, tne p~esencc of t~esc hist~cc~;'"tes containing lipoidal?'la1:e_riai. Panel Book Page 232

235 may be conside::ocd as nonspccific. ex..:cpt whcn 'they occur in large ownbers and in :nany animals of a given groljp. e. g.. cholesterol control gr~up. Summary and Conclusions A "tudy corn.paring the cffcc'ts of die:.ary administration to rabbits of I per cent Antiioam A. 1 per cent :~tiioam A i 0.8?er cent cholesterol; and 0.8 per cent cholesterol alone with those of unsupplern.ented diet has. been co!nplcted. No signiiic3:lt dii"c:ences wc::c noted in tr.e physiological status '01 the animals betwcen the group that received only Anrifoam A and compared to the basal co!ltrol group. Sigiriiica::J.t dific:t"~ncesin,a nwnbe:: of p3.ra:"t~et~rs were rioted "'hen comparisons.'\'ere n"lade betwecl': the <:holesterolied groups (both with and without _"\nd::{r.u A~ ",.~:.'1 the b~sa:l centre!s. \.:1'\o11.g the latter abno::=alities occ:.:.=rcd in the he=ogram.s (... here anc:r.:ia "''o"'::'s seen i..." a nu:nber of cholest",::vlt:::ciltcdrabbits ai',the" termination of ClC st:.:.cy aiter S months of feeding); in ~he bib'od che~ical",,. ','. values ("...'here cholesterc'lemia '\v~s in all choles"tcrol:"':trca:tcd r,abb~t~); arid'. il"tthc size and >~"lorphology of a n'.l.rri.ber of organs (whe~e~~ke~:(iipid. :,;;" "';~~;:>~'~~~l_'".. ~ ::C~i:::<::c:~::::::::::':c:::r:2i:~~%;t~~1:i~r~;i' + choles"te."!"ol1;rou?:s.. Panel Book Page 233

236 It w~s concluded t!lat '\\rhcrcas a marked number of. tissue, abnormalities have been induced by the inclusion. i., a complete,rabbit ration, of cxcess cholesterol a: a lc'\'cl of 0.8 per cent, no sigriificant adverse effects of th~ ingcstion of I per CC:1t: Antifoam A have been noted in this chronic (SlT1onth) feeding study in 'ra.bbits. Panel Book Page 234

237 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /13/66 04/28/94 8D DOW CORNING CORP FOOD & DRUG RESEARCH LABS INC CHRONIC (IYEAR) FEEDING STUDIES WITH ANTIFOAM A IN RATS WITH COVER LEITER DATED Chemical Category SILOXANES AND SILICONES, DIME ( ) Panel Book Page 235

238 DOW CORNING 8» April 20, ~994 TSCA Document Processing Center (TS790) Office of Prevention, Pollution and Toxic Substances u.s. Environmental Protection Agency 40~ M street, s.w. Washington, D.C Attn: Sed) Health and Safety Data Reporting Rule Re: 58 FR 285~~ (May ~4, ~993) [OPPTS82040; FRL41821] Health and Safety Data Reporting; 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow Corning Corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical Substance(s): siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Dimethyl silicones and siloxane, reaction products with silica 7% in Antifoam A Title of Submitted study: CHRONIC (lyear) FEEDING STUDIES WITH ANTI FOAM A IN RATS Dow Corning Corporation May 13, 1966 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland, MI Contains No C8' DOW CORNING CORPORATION, MIDLAND, MICHIGAN TELEPHONE: (517) Panel Book Page 236

239 2 If you require further general information concerning this submission, please contact Dr. Rhys G. Daniels at Sl For further information concerning the submitted study or the toxicological properties of the listed chemical substance, please contact Dr. Robert G. Meeks at Sl sincerely, ~ E. 'B~ Alvin E. Bey 0 u.s. Area VicePresident Corporate Director HES Panel Book Page 237

240 I ~ ~_:..:.._:.. ~fj~:. ~ A series oi toxicologi<:al investigations of pol)'siloxanc compounds is in progress in these Laboratories. The studiel> were authorized on September by thp. Dow Corning Corporation and are being carried Ollt in accol"dancc with p::,otocols developed in conference with these Laboratories da.ted December The present report covers expcrimel'ts, idcn;;iiied in the protocols as Phase D, Groups 3 and 4. which were designed to detcrmm<: r.he effects of feeding Antifoam A or a mixture o Antifoam A and cholesterol to ra:'~. For purposes of cornpa.:dson, two control groups '''"Cre included, one receiving i.~(' basal ration ollly al1d the other the basal ration supplemented with cholesterol lor one year. The test sample was subtnittc'd on Septctnber and was identified as o11o\'\,'s: "Do\o: Corning. l\1cciic<..1.<\ntifoam A Compound; i gal. Lot No. HOOI9". Tho:: cholest':tol used was purchasea from Nutritional Biochemicals Corpo:ation. Cr.", clajld, Ohio and was labeled: "Cholestc )l usp Fo!" Ll\,"Cstiga.tional Usc O:lly: Contl'ol l\:o. 1,071: 1000 Gm." The effects of thc ~oly:siloxancadministratior. under condition::; of normal <::.nci high ste:01 intake we::c determined by comparing a s~'des of pararnct'"': s of physioio~ic<ll dc\ clopm.cnt and status (growth,efficiency of food utiliz~hicln; blo"ci ch(,rnical. hematological. and urine analytical dctcrnlinali"nl<; Z':10 h~",ot11o:rpholvf:.y)arno:lg groups.cceiving. thc basal ration alone or :;~~ ':"'~',<,nt.,a with Antifoarn A, Antifoam A in conlbination with cholestcrol. '. ',\'ith cholestcrol alone, o\"c:: a Iyear period. 1 Panel Book Page 238

241 'EI'~' :~ :~f,... '. "',:s.3'1 ~r' Procedure Fifty albino weanling rats of the FDRLstrain. bred in these Laboratories. 'were.andornly distributed into test and control groups Group Addition to Basal Diet 1'viales Females 3 1'ro Anti:foam A :; 5 4 1'0 Antifoam A + :; 5 0.8% Cholcstc;'ol IS None (cont:ols) O. 8% Cholc:Herol :; :; All diets as '\... cll as ample s:.li='plics of f::csh tap '\... ah~r we:... ;:J.vailablc ad lioituln to the rats. They we:e housed indi'\" ually in :'ais~,dbottoln. wl"ca:csh cag~'s,.\ppearan,:c. behavicr, survival, and ~ncral physlcal condition wcn~ obs.:"l'vcd daily. Body weights and fooci consumption,\,.. ere determined weekly fo, the initial 12 weeks of the study. From these data the effi.::iency of fooe utilization (:::FU,' g,a.ms gain in body weight pc'r 100 grams food COnSUlll';c.) w;).s C<l1C'\11at~ cl. :30<:Y '\\ ::ip,hts wc:c recorded bi'\'licckly through thi.' rest v~ the :> ~w.;:ek?e.ioci. 2 Panel Book Page 239

242 . rfjr:.. Prior to the feeding ph.:l.ses of the study, base line values for the following clinical laboratory parameters ".ere de~eritlincd: Blood Urine hemoglobin, hematocrit, total and differential leukocyte COU:l.:s. erythrocyte fragility, seruin cholesterolle...els, alakaline phosphatase and glutainic pyrm:i.c t.ansot':1.inase activities: specific gravity, ph, albuinin, glucose, ketone bodies, occul~ blood, bilc salt and piglnents, and n.:'::roscopic,'xamination of thc centrifuged sediment. These tests were c..rricc1 out using sar:nplcs of c<l.rdiac blood and l:l:dcler urine obtained dir~ctly fro:n 15 weanling rats of each sex, selccted at randon"l [rain the FDRL stock colony. Thl~ Incnns and ranges (= 1 stanclard deviations) '...ere calculi.l.ted for cach of thcsc?al'ain<~rs. These ranges werc accepted <1:; :11<.' initial values ugain:h which a111abol'atory da;:a derived throllghou~ the s:ud)' were compar...d. Aftcr 3, 6, and 12 :":':.or.ths of administ::<.l.t:'on of the test dic~s, hcxnatological ex<llninations we::e made on samplo.:::s of blood taken froin th~ ret':'oorbital sinus of each rat, a:~d u:ine exanli:~ationswere Inade on pooled sal"':'l?les colh'ctccl 0'",': 2';'hour pc:iods from g,roups ;:,r 2. 01" 3 ::ats of the SalTI<;' s,"x in each clos3f,c' group. D\:!"ing tl.e 1!lTIonth exaxninations. addition21 seru\1"'\ sa:t'lplcs W,'l"e collected and analy7.cd for cholesterol. alkalin,~ ph05pnata~c, :'..I':!d glut:l!ni.:?yruvic ~r3ns<:..rni=1ase. Sam?les of blooe!. an~ urine were subn'litted t\l :)0'... Corning for silicon an...lysis. 3 Panel Book Page 240

243 ,. _,.. ~~. ~:_""":' '':_'!~.;; :!..;...~.~.y' During the week fvllowing the final series of clinical cxamilkltions, the animals "\'e:e sacrificed by cxsanguinatjon while under pentobarbitol sodium anesthesia and the iatcrna1 o:"~ilns WlOre examined grossly at necropsy. Th~ li...er. kidneys.,;plct'n. hea:t, gonads.and adnexa. adrenals. th~ roids. brain. pituitary. and lungs of each uf th,:, :at", in the 4 groups... ere weighed and organ:bodr weight rali.:>!' calculated. Represent.:l.tive samples of all tissues and organ:; werc fix.:d in 10 pc: cent formalin. Hcmatoxylincosinstaineci ::;cctions of the following tissues fl'om all rats werc examined mic:oscopically: liver (2 sections), splcen, sto:nach (2 levcls). slnall intcstine (3 lcv('ls), large i"tcst:'ne (2. 1('vc1s), panl'cas, kidneys. bladder. adrcn~. gonads. thyroids. pituitary. thy:~us, csophilgus. mcdiastinal and nh'"ocnteric lyrn})h nodes, hea:. and ao:ta. lungs. b..'nc rn~.rrow (sn1car frorn the ternur), skin, "keh~tal muscle...ith attached?cripheral r.c:"l. C, spinai,ord (3 lev,~ls). b::ain (4 le"l.'clsl, uterus 01' sclni"al vesicles, prostat<:, anc. all tisst..:.0 n1asscs Seen grossly. 4 Panel Book Page 241

244 ~_.:_~.::~...,.'' ~'r Results There were no <..bnonnalities in the gcneral appearance or behavior oi the rats throughout the course of the 52wcek study. The appcarance and [I'cqucncy of excretion \,I:{"~'e nonna!. One or two rats in each group!ail("d to sur...h c. The' mortality figures!ollow:!l.1atct"ial 1% Antifoa m A 1'fa Antifoan:. A + O. S~~ Chal('st~:"~"l Group (Si7.c) 3 (5M.Sn.; (S~!, S Fj Male FClnale no. deaths o z o None (conl~jl) O.S'fc Chole::Herol 18 (1 0 ~1, 1OF) 19 (SM, SF) o 1 ),.~ost of these d(. aths occnr:ed durin~ the :il"lal! 0 weeks of the ;;tudy, the a??2.rcllt ca~sc of <k<lth ilj...vl... ing acut.' pu.!.:":"lon.j.ry pathology. It is cvident fro:.. t::, dist:ib~l~oll oi these aca.hs that thc:e were no c.ficc!s on s..:.:vi...al 0: A::1tifo;"I"':"l A, cholcstc!"ol 0: their con"lbination. Tr.:= :"::.:l1..',,; of the A::ti:oa:n A g:ou? showec. ~ignii"icantlygrcate: );:"0,"t1l (1' <G. OS) lh;:w basal <:o::.trol (Ta.~i", I ) lhl"o~ghout the study. There w<' :~. no olh~ : signi:ic2,r:t di:::,' :cnccs i:1 t!1..' rate of growth among the two te::'! groups. the b;;'''.3.l <ll:':' :~... cholc;;tcrol co::1trol gl'"o,,:,ps. \,... h ~n averages LO= th~ soon,,' 5("X wer,' (". :"\1iJa:cC.. Howe...":. a general trend was evident. i. c. chole:"~.."rolf~d :'n~dc": ::'h"::wcc ;.::r"atcl'" growth that other males whereas cholcstc:vliecl i,'males ::'hl.jwed 10...cr net gain::' than comparable females receiving c:t~cr the test or o3.~al ri:l.t:'otl. ThllS o...erdosage oi this steroid p:ccursor <i??ea!"t'ci. to aifect each sex diiierently. S Panel Book Page 242

245 ..,~..;.~:,:.:' ~~~. "!Zl.( ~ \1'..,.,.....,:... ~,~ Food intakc was c')lupar<lblc in all groups. Comparison of E!'"'U values bct... ccn groups paralleled the gro... U1 respo:"ses. It is evident that there wcr.c no dclctc"rious cffccts on grewth!lg[cd on comparison oi the iou:: g.roups. Initially, the normal :angcs for hem...tological param.eters in weanling rats (mean ± 1 standard deviation) were as follows: Determination Male Female Hem.oglobin, g/100 =1 Hcm,atocrit. per cent Leukocyte count, x 10 3 / min 3 Di ferentia1 count, ocr cent Neutrophil:;;. Lymphocyte s Eo::> invphils ~10nocyt<~s Basophils E:ythrocyt~ Fr<l ility, 2 initial. pc" r cent x l(~ final. per <"cr.t X 1(j"2 9.S S 38.':., :;)., 1';. ; 29. '7 67. S o 1. 1 o 2.7 o 50 5'; & l ~acrificcc. at approximately 0:lC month 01 age. The normal values u..""ldergo changes with the O:l:;('t <'>: :::c:"l.:.a1 In<lturit~, a:1c subsequently with the physical <l.nd :;ystc:nic e:lect::: v1 a;:il~;:. Thc a...era.ge data ior hematological paran"lcters after 3, 6. :I:ld I;: :1h)l'1ths of test n"latc:i..! administration arc shown i:l Table 2. The ;)'o:i~1.al,;hiit ~11 JWl:trophil:lyrnphocytc ratio which occurs witr. age :s seen i::: this 'S?edeS il~ all group, as is the generalized decrease 6 Panel Book Page 243

246 in packed cell volunle. There were no shifts in h~mo lobin content of the blood of the males but there ",vas a notablc continuous fall in that of the females, which appcars to be a function of the c:.dmillistration of the dietary cholc'stcrol. Statistic"l colllparison of all thp. parameters of hem.atopoictic fw1.ction showcd no significant diffcre~\ces (1' >0. 05) among the groups at any period. Blood chemical analyses (madt: tcrmir.a.uy (lnly) revealed no signi Hcant diiiercnces among groups of the sanlc sex in urea nitrogc::l, serum alkaline pnosphataso:! activity, or serum glutamic pyruvic transaminase activity (Table 3). ~[a.les recehing a coitlbi."'lation of Antifoam A + cholesterol showed a sigllifica:1t elevation in serum cholesterol content on comparison ".vith all othe 1" groups of the same sex. No such trend was notcd among the females but ill g,chcral the values for the fernalcs were higher than in the mall::::. Lar<::<:::...ariations in Serll:11 cholcstc:ol levels among indivici.uals of all mammalian species is well recognizcd and most often associated with dietary i::ltake. Since no...alues in exccss of 110 mg choiestl~rol pci' 100 ml seruln WC1"e noted. n.:> biological sig:::.ificance is attached to this int.cq;rou? variation. In ocreral there appeared to be no cifec;;s of acministrat o::l of.tunifoa:n A, chol~stcrol or their com.bination on any of th~ I::~tabolic, hepatic. or rz~lal fu.."1ztions rei1ected in the various ser;,\:~';. cons:.itl1~ni.s :'C'lo which the l:lood ",vas ar:alyzec. 7 Panel Book Page 244

247 'fi'~j I~._,.~.:,.,...,.. ~,...j The urinc analy:;es (Table 1) showed no significant intcrgroup variations. reflecting lack of eifect on renal function of Antifoam A, cholestet'ol or the combinatiol:. Minimal traces of glucose were noted in the urine of the rnales of L'1e Antifoanl A + cholestcr01 group at the terminal period and among the fen"lales of the Antifoam A group at 6 months. These findings often occur at isolated periods in untreated ra.ts. Traces of occult blood in the urine of. males in the cholesterol control group may be significant '...hen considered in the light of thc histopathological findinj;s discussed bdow. A decrease in ovarian and acircnal weights was not<:'d among the fcmal':$ recciving chole::..croicontaining dicts with anc. ""'ithout added polysiloxanc:s. This i~ support for the concept t.hat excessive leve1s of administration of the steroid affect hormenc produ;:tion. All ch<l.ngcs appear to be cl1oie~terolinduccd;no c! <>cts of ~ilhonl' a.dn~l..istration werc seen (Tabh 5). There were no ::oigniiicant findings at nee ropsy of the rats in the four groups indicative of the effcct~ of adnlinistratio:l of either t~st material or the combi::'l.:ltion thereof (Tabic 6). The apparent preponderance of anolnali.:s in tl1c bas::l.i control grou? is. in actu~l.ity, i:lcicativc of its double siz.e. It is \Ji int":'cst tv note th:it there were neither gross nor m.icr("~copic changes i:1.cicativc of the c:fects of cholesterol described above (Table 7). Thcr~ "... ~~rc' very few in~:i:'lgs i:1 the Antifoan"l A group as 8 Panel Book Page 245

248 .:...I~: compared ~o all othc'r group,;. but there was no significa.nt incidence of a.ny type of lc:.sion in the.:holestcrol control group when compared to the basal control <;roup. Interstitial alld peribronchial inflammation was observed in all of the test gro~ps and ~be controls and is not considered related to the treatment..all Obscl vatiolls reported were not associ'lted with the incidence of this condition. There were '10 indications tint administration of Antifoam A, cholesterol, or the combi:ilatior. thereof, caused abnormal growths or neoplas=s. In general. the changes seen in the tissue and cellular architecture reilected the incidence of lesions characteristic of normal 13 to 14month old rats in these Laboratories. Sllmnlary and Conclusions A study colnparing the cffects of dietary ::..dministratio11 to rats of 1% Antiioanl A. 1% A:l.tifoa:n A + 0.8% cholesterol, and O. S% cholcs terol alone 'with those of an unsupplclll.c:1[cd diet has been co=plctcd. No significant dhferences in a. I1\.:.n:.ber of pa.::amcters 'were noted when con"lparisons WerFmade bet'l.vecn the cholcstc:olfcd groups (both with and 'l.vithout Antifoam A) and the basal cont:ol. The 2'.ailablc cvidenc,, points to. e::rfcrcilce in nornl"l.fcn"lale en~~c'::l'ine pr.:>duction ;;;.nd lor utilization when the cholesterol level in th ~ diet is c:..:c."~s;,;i'l. e. The cffec~s i.n the male appeared to be the rcvc:sc of thl')sc in the: fcln.?~e. indicating sex steroid l:"::ll<:.tionships. There were no significant ci.ihe::ences obser'l."cd between the cholcstercl and Antif :lan"l A + cholcs.crul groups. It was conch.:ded t11.:'.t.'1.d:ninistration of 10/0 Antiio;:;.m A in the diet of rats for more tha:t one year did not have any aevers e effects. 9 Panel Book Page 246

249 CODING FORMS FOR SRC INDEXING Microfiche No. OTS / /28/94 8D DOW CORNING CORP ATLAS CHEMICAL INDUS INC DOW CORNING A.'l>.ITIFOAM A (MEDICAL GRADE): A TERATOGENIC POTENTIAL STUDY IN RATS WITH COVER LETTER DATED Chemical Category SILOXANES AND SILICONES, DIME ( ) Panel Book Page 247

250 DOW CORN!~G April 20, 1994 (;fie2fains No C81 TSCA Document processing Center (TS790) Office of Prevention, Pollution and Toxic Substances U.S. Environmental Protection Agency 401 M Street, S.W. Washington, D.C Attn: 8(d) Health and safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41821] Health and Safety Data Reporting; 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow corning corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical Substance(s): Siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Di~ethy~ cilicones and siloxane, reaction products with silica 7% in Dow Corning Antifoam A Title of Submitted Study: DOW CORNING ANTIFOAM A (MEDICAL GRADE): A TERATOGENIC POTENTIAL STUDY IN RATS Dow Corning corporation May 4, 1970 Manufacturer: Dow corning corporation 2200 West Salzburg Road Midland, MI DOW CORNING CORPORATION, MIDLAND, MICHIGAN TELEPHONE: (517) Panel Book Page 248

251 2 If you require further general information conc~rning this submission, please contact Dr. Rhys G. Daniels at For further information concerning the submitted study or the toxicological properties of the listed chemical substance, please contact Dr. Robert G. Meeks at Sincerely, ~e.b~ Alvin E. Bey 0 u.s. Area VicePresident Corporate Director RES Panel Book Page 249

252 DA1'EMA~ 4, 19'70 REPORT NQ BMPD7, ATLAS CHEMICAL 'J. / 1;(1 (l / (/7) r'f' 'I. '7 ((, \ /. 1, XN~USTR:IES'l I.NC. BICMEDICAL RES:EARCa: DEPA!<.TMl:.~' TITLE DOW CORNING' ANTU'01:.M A (MF.~T~& GRADE) : A ~ERATOGENIC POTEN~IA~ ~~~DY IN RATS PROJECT 125(17 ; :EQUESTEO BY: J. F. ~REON, Ph.D.*.COPIES. TO : REPORT OF' MR. L..E. GONGWER MR. k. A. SAATM..~ K. HeBBEN, V.M.D. {See Inside Cover For Distribution. List).. *See note on signature page. K. K. KIMURA, M.D. DIRECTOR, aiomedical RESEARCH DEPARTMENT ~~~fj~1 I. ':"~.E : ~:';ic~a. t i~ fiie.. 1 Panel Book Page 250

253 1 Panel Book Page 251

254 :~,r'c'\.."1.d ~ c~t..;:. to a~d ir: t:.he ev.=1.uat.l.o:l c i; 'the saf~t:.y vi' t.::~s cn~?c~!~~ _~ ~:~e for~ul~tion o~ cert~i~ dr~g products. 2 Panel Book Page 252

255 111_ CQ~CLUS!O~S D~~ Corr.ing An~~fo~~ A (M~dical Grade), g~ven orally at dosage levels ~? to 3.80 g/kg/day frv~ day 6 through day 15 of gestation, caused no dete~table ernbryo~oxic or teratogenic effect ~n f~~uses c:?rcgna~t rats. This con~luslon is ba5~d. upon the abscr.ce of sign~fica~t dif=erences bet~een 0= ~~ong the four l~vcls tes~ed =egard~ng l> viability of fetus~s at l~paro~~my, 2) resor?tions, 3) g=os5 external anomalies. 4) soft ~~S~;UE anorn~lies, 5) skel~ta: anomalies, and 6) avera~c fct~l ~elghl. 3 Panel Book Page 253

256 ~13~erial : Dow Corning rl~~ifoam A {DC nntife~~ A, ~edlc~l Grad~l, Stuart Division dd5~~~atio~ Lot Xl~29063, was recc~ved ~ro~ th= S~uart Div~5ion o~ F~bruary 14, Th~s ~~ter~al ~et ~he specif~=at~ons and was a?proved tor usc 1,n prcc1..lct::.o:r.. {See Tabl..:: 1 :or specifications a~d Table 2 for a~alysis of Lot M l An~rnals: \ :.::.star d02:rp;c.:} rats, designated.j.s S~r, MW3, ~~rc procured from ~aror Rcs~a=ch South, Puerto R~co. The rats, 240 fer.:alc and 60.nale, were appro~irnately 75 to SO days of.j.gc upon rece~pt and were all~wcd a period of t~n days fer.j.ccl~matization prio~ to us~. The female rats were housed ~n pairs ~n w~rc hanging cages: t~~ male rats were placed ~r. ~nt!.l.v':'duill cages. Al~ rat.s were r.:a.::.n~ained on Purill';" 1..:1:) oratory Chew, ad l~bitum, throughou~ th~ s~udy.! 1.:>t. ~ nq a;jc Assl.::mment to ::;o~aqe Levels: I...hen the l<l~~ WC1C ~P?rox~ffiat.ely as to 103 cays of ~ge, one male c.:>t ~as P::C.5~:1t.:d t.o t.\,'0 pa~re 1. fernal~ r,,,ts for a period of fj..\i~ d~y5. or unt:l viable ~Ferma~ozoa were dernons~rated 2n a vcg~nal :im~cl.l. Vagl.llal sme..us \,'ere t::l~en da::..ly from all female ~a~s io~lowl.ng ~~cscn~at20n to the ~ale. If, after a pe~~od of 5 d~ys, no cv~d~ncc of copulatlon w~th a male rat was seen 2~ clth~r f~md1e r~t, the male was discarded as no~virile ~nd a m~l~ ~c~ons~rated to be v.::.rile ~as p~esentec to the fc~ale ra~s_.; Panel Book Page 254

257 ~~n~mum of 5C :cm~lc =ats. Th~ ~~ccess~o~ of co?ula~lon Th.:.s proccdu:c cont..:.r.ucd u~t.ll d group of 25. phlsl:mably!?rcgn.:l.n~, fcr.a.l~ ~os~~~ L~v~ls 3~~ Adm:nlSt~5~lon: Three cos~y~,cvcl~ of UC A~:~ oam A (Medlcal G~a~e). en a log in~crv~l ~.;'VC.!. 1: ). 3.80g DC Ant.:.!oarn ~ '~edical Grad~)/kg/day icr Days 5 through 15, 20g DC A~t.~foarn A.~ed~cal Grade)/ks!d~y ~r Da~3 6 ch=~ugh 15, O.3Sa :lc A!':tlfoa."'Tl A i~'ed.:.ca.l Grade\/kg/d'lY tor Days6 through S ~ap wa~erikg/ddy for CKys 6 through!5, :Control level'. 5 Panel Book Page 255

258 V. P.ESUI.?S F~~ale ra~5 usee i~ ~h~s ex?eri~e~~ are identified te:t:_,le alse shews ~r.e fa~,,= of e';eh =a:. s~c::rrr.a~ozc~ or vagir..al plug \"er~ fo"..lnd, Of t::'e 99 rats 98 survived to " '. ~.. ','..:' in ~;rh.i'c..~>:: term. '>'~~':. clden~, 3 ra~s ~ere ~csed i~correc=ly and da:a from them wer~' excluded. Of t:.he 95 z:at:.s :s'\,;r'j:'v:,~c; to term and correctl~~.dcse,(j,;,:, 28...;ere ZO'..lnc :'0 te :lo",pregna t: upon laparo':om::'. T::'e number of female ra:.s <Jresumed pregnant, t~e., nur..ber>:. sulvivlng to tcrm id::j".l 20 of gest:.<l~io:l), anti the nu.rnber of ~~ablc fetuses a': Day ~0 are l~s~cd below according :0 dosage Dosage So. of No. Rat.s 1':0. Rat:.s Level!: f Fe:nale No. Dled Sur "iving with Vial':ll~ DC A:;~.:.. toa!:l Rat:.s Pre or :'lis te Terr.! Fe~uses at' A i11gl surncd d"sed Prler but:. Not Day 20, of i9/kg.day) _?rcc'lnar..t:.... 'Co Term Preqnant 11: * Gesta~ion ") _::l ~..;a.'t;er (q/kg/cay) (Cont.rol) TC'_<11 39 '4 28 ~13,.. Based on prese:1ce of sperm 1.n a va?inal' smear.. o :::::.a..,;gina~.~. ~~~~~~~Cd o?re5~~ed gesta~lon.. 6~. to be nor.pregn~nt uponla~,ar~~c?m~_~'?;'?~~: ~~~O:~k:?~~ by ass~gi.ed ~u~~er according 1;;0 dosage leval i:l Table 3. 9 Panel Book Page 256

259 ...,...,..,, :... _,.,.,:~. sig:l 't.ha't. OCCi.:.':'S aro~:::i D", ~... :J _...J' B. Embrvo~oX1citv resorbed. and d~ad fe't.uses at 't.he t~mc of Cacsa~ca~ dc~lvary ~D~\' 20 or. scsce.t.:.c:c) ar~ prescn:.ed i~ Table 4 fo~,=::ac::..l.r:.d,:,v.!.cllal Also presentee a~e the total ~~plan't.ation s~tcs and the O... Clr.l~s. ra't.s fro~ 't.~e overall popc~atic~ shows an avcr~gc of The number of?lacental $ltes r~ 10 Panel Book Page 257

260 ,.~,~ Panel Book Page 258.;, 'r, rj r' ;.'.." :. I;, I'!' () G 0,),. (, '_: l~)., ~, ~,) ~.', I,, ;t:" r:; I:""' ~) n. ~ '1 :1 CI f" ~,..) " 'r. :J t,.: " ",,; II \~ <; f" r;' r, 1) Ii I", ~ 'J.. c: "1 o.:. t...j '1',' (J c.... ~ w ;0;' ~~ ~ rj (,1.:. ('. i:' ":': ~ c f, r.,.' '..; oj :t :;, ~f4 ". " ::, (;i ~;, ~ ~ I~),e. " ", D (',',,.. l,~ 10" I~ ~!, C', ('. rj " 0:1 I... rj rc..., I' (J r ~~, iy., I () Ul,'.,J ' '< ~.. t"\ O:(~, w r:> 0 fj..,, 0 r " ' " i' ~~!'J \~I (J f. r) I (J tj th,~, l." I' " :1 C,\ U.,. I~l CI J' :;J n, I ' ',' " 0 I) '.,.. ", Il rj (J " In ~I f). 'II r, r.,!_: I': c, <, ;.' I' " PJ VI r. ~.l ~~, :1 'I, U. f:i 1" <: r. I' n, " :1 D, " I' " " 0 10 r.,,j '"~,.' It,,~ Ir :.: " II, C." " c' :,1 I, :, )~, Il' I', I~ D " ':: I~j ~: 0 /.'~. '\,: " 1;'., c ::1 0, :1 " II, I,' :,. t.,,":.,,1 I, fj I~ It':,~ ~, II. ;.; I' ::r '\,: C' ',' r.. rl rt,'i ~. (, (;l r:'.. (0 I~ 0 Ii> I~ '. 0," r;'1 1...,_. c; :1' ~ /), 0 1/1 () 1:, f';> :,. III I ' '" 11,,,, D " II.J ", \!I OJ... J v. I~ r t'. I~ :0: I' n, 0 f) 11 :r () 'r. 1"1 rj <. Co '1 ':::; 0 I" I. li1 0 H, <: l~ rj tr 'f; In I ' til ft no f' 0 r, :l Ij C') Ul n. I'... p. I" :.1 I: I h ti " ". '. :./ I~ I' I'.:.: f. (.,., t,, 0 o~ Ul,. l~ () In In f" f1 II 0 IJ :J (~I 11 :l tj to :J.., ll, '" t" :" :; :J r~ li1 (fi {li,:.. Ul :1 IJ " r,. t, (t. 1Il :1' r' :',. I: n. III 11, I~ fj '11 (J I) I', If, r' :1 " "... I,' I:, f~ r: CD I~, ~ ''1 O;tJ 1\ IS I~ r, :1 " 0 lii.. '/1,... ~ I, fli (1),.~ ~ r.> \lj t~, IJ '" I ' L10... IV 0 \li 0 " r~ '" I~ I'~ 1 ft rt Q,~ I e,,i., 'Il :1 I' Ul ;'j I' r: <. ' H ().\ rl lr n :J \li fj r l n I 'u. 0 ",. li1... p, 11' :j' I,' rt 1/1,ll :.,. ld I' It ::1' :J u UI I' ID ~J I!l ru ~: rj' Iii " ~, r., I' IJ l\i 01:, :1 li1 It r; r;..ci.",~ 1/1 0 r: III 0 I 0 I), W t1 I!l 1':1 I~ rj I) 0 " :J" :0: :,' :J rj 'UlOtr rj rl r>: :,' l~ ru :1' toj '" Ul '1. ~' fl 11 CD l') :1 ;T tj',., Q II I~' () 'll II> III l\i 11 '1 11> ",: ro, II. 0' rl,.,,... l~ I). rt n. oj D rj rt I' r IJ,., ti III t;' ::l III I)' :1" rl V' () Ul :j r1 't] :ti ~J :,;.: tf f) I; hl 0 fo () rr r ~rotjf:: (') 1\,J f, II co. :J I" (1) 11> ~I rtl/lrt:j.~ V' (1 r~ ()..: rl ',3 III t1 IV 0' O'Ioooto COUll,. () 0 (~ r... X fll, t,., lu 'IJ 0 (j) () 1/1 II fj. '11 f/j 'I '0 /, )." I~ 11 ", ;:r rd tr :0: I~ " Iv :J fj,", X \1l IL~ () " IV t1 0 VI 0 0 r).j 1.", :J et 11 (1 n, (1 0 n I~ th. (1 I' :1'.' rl,", '0 l{ ~l HI {J If. III r.,,., I~' " f', l/l 0 l\i 0 (' n 1\ n. 0 If>,1 f,,~ rl :1 l\i " I, '1 \lj :,. 0 :1 rj l~ ~U '0 ~U :11 n. tf rr rj,: r'..: V' ~, 0 :1 " (~ r) f.i..:,~,",". ii' '0 C) III fj :J '. ~ VI D UlliUlQ :1 :J '.I I,' r.. ',1 'n,'.. :J 0 O 'II.:l " r..: '" UJ 0 ij,; J 1' f" I~, :J),., I{ I ' " r. r, :J f. I'., "; fj tr ~ rl tt. rl 11 (J ~~,0. I) I' VI, V' 0\ lo',.i,'t... t' ~ 0 (J." :'J ".: 0 l3 (~J 0.. I v..:. l)t '40 :J :~ (I u, r, II rt tt, I:,~ ~',~ ~ o ~:...'1 ly r: rj II. " " 0 '.. til I~ f,' f,l :J, r~ v. f, 11 l) " III \lj 1'..,.. f/j rt I,) 11 rj.. fj ~1 :J'fj III rj ft ('10 r: tr. Ul f..!/l

261 A::. a':lalysis 0:: covariance fo=.,lacc~':.al s::. cos ar:c v~~~lc tc~u5c5 by dosage levels rtrca~~e~~s} sho~s ~~ for v~able =c:.uscs as tested by ':.~ F ~e5~. ~he F valu~, :or cl:c grou.ps sec::.... ':.::::'s ::.eas'.;rc~e::t a:=e ::o~ sig::.':'f':"cant. {Sec T~~ cistr~b~t~on of ~ale to c~ale fetuses apprc~ches D 1 '':0 1 ra':.~o. (Se:e ':'ablc 5.} C. Fetal Exa~inat':'cn fetus w~s Gross ano;:1al~es: A careiul cxamina':.ion of each mace at laparotomy for general co~for~3tion and any ex~ernal gross anomalies. The following table lists the nur.bcr of fetuses examined and the number of anomalies that ~c=e observed. Do::;ag~ Lev.::l of ~o. Fetuses with Exte;:nal Anomalies DC Ant.i fca;n Ku..'!lber 0::: Hematomas A " lgl Fet.uses on Hind (C':k.:'i/ d~\r) Exananed!\one Runt;.~!:'nenrlaq~S.r..caudacc ""3:~ 191 """T'88 O J la 3 a, c: L 1 0 I'Jater {q/kg/oav ~ ~ icor.:..::;cll 'i'c t.al 711 6'~9 3" ,' ~~l:.iplc findings 0::0 fe~u.s w~:.h multiple findings was a runt, with~uc ~ ~~~l, an~ hac s~all a~eas of h~atorr.as on corsal tr.o=acic 2.2 Panel Book Page 259

262 ar~a as ~cll as o~ both h~~d feet a~d the S~h l~f~ c~g~t. A l~~~~r~ute, also srr.all b~t not considered a runt, hac s~~~lar :t~e average nur.~e~ for ~he dosage level ~as 10_9 fetuses) a~d ;~ed ~o ~lac~~~al or ~~?la~t si~es ~~ t~e le=t ~t~~inc hor~. ~:~cc ~hc cl~~ ~as fa~rly a~y?ical, and s~ncc irnplantatl0~ o! e~b=yos occu~cd prior to :rca:~ent, these fincinss are?robabl~ ~~~r~b~~adl~ co gcne~~c factors ra~~er ~ha~ ~c t~c t~ea~~cn~ "ltn DC Ant~foa~ A. Vary~ng ccgrqes of ~ematorr.as ~ere scen :n cne to 3 fe:us~s from aach dos~ge level. :nclucing =~e ~a:er co~=rol l~vcl. ~nd arc considered inc~dental 0: may have b~cn ~c~~d~~=all~.:.ncuc:cd cur ~n~; r(?moval from the I.:.tert:.s or :::.ur':':l.g clea:ls.:.nq" Soft t.:.ssue anomal~es: i3ou~~' 5 f lx.:lt.:.vc ;,"ere reexamined for g::oss ehternal anc;:.allcs ot ~~Vcndagc~, Integu:nen: and fo:: conformation. Each fet~s ~as t.:ler. sl.:.c..::c.:.::. th.:.n cross sect.:.ons and exai:1inec for s:.z~, s::'a~)c, plac~~~n~, ~~~ Lcx~u=e of l~ternal organs. The 't.abl~ belo';... 1':'$t.$ t.~,~ n:':"~l::>cr of fetuse:s examined by this r:lethod, a.nd the n~~~0r o~ a~o~alics no~cd by dosage levels. 13 Panel Book Page 260

263 DOS.. l~;'''' L,~vcl of DC A:1l:fo~m ~umbe~ of,n. i:~:c;j F!:t.uscs ~:~l~:~:/d,:,,"l 3.': :::;.:a..~ir:~d '; :':\..:r:":~o~~,')= ~ct.uscs... ;.~l' Ano~3.1ic~.. Scft: T: l;;5uc Her ;~a~e~ Dila~~on 0: :~on(: Di, 2t:r~~q!i' Rena] P,: l,, ~s 1 e O 0 0 C I ":/~:G/':;il'l) 3.30 (Co:;t.rol) la 1a. Tc~al HernIated diaphragm a~d dilation o~ the renal pelv:s ~l~oui) o~ fetuses since i~ is generally at~:ibu~ablc to ~c~c~ic of ~ny a~orna.lous 1ndings 1n c~uscs prepared for ~one st.alnlnq :J~~::9 0 _"' Lcvc 1 ot ~1C i\ot:l.!oam i\ (!'!G 1 ('1 /k.:l.:dl.'t."v I 3.ti0 i.?o n. ~,:3 ",'.i l'_' r ':1/ L;l, 'd.'l.\ ).5. :3 (I (C,::l::tr:)ll ~urocr 0:: F~tu:::es!::xamlned 1.,, ~ Number of Fetuses with Skeletal Defects Ncne So11~ Centrum , ')..:> 14 o 1 (13th ~to=~clc) o c 1 Panel Book Page 261

264 ~x~~~~cc exccp~ for ~~e ~o~specific spli~ ce~~r~~.!'!:~owed good calci icatio:: of the c=a~iu:n, generally a well ccf~n~d hjo~=, 7 cervical, 13 ~~oracic, 6 l~~~ar, ~ sacral, a~d fro~ 3 to 6 cacdal vertebra. Thirtee~ pai~s of r~bs wc=e ~c~~crs of ~he manu~ri~~ and generally 3 to 4 stcrne~ra; ~hc clavlcl~s ~ere well de=ined as w~re the scapulae. The forcl~~bs wcr~ d~velo?ed without no~iceable variatio~; 4 h.~cacarpals =h~rd phalanges were observed. were 1d~Il~lfied as thre~ separa~e paired bones. The ie~u.::, t~bia, and f~bula were develop~d w~chout not~cca~le var~atlo?, ~nd g~n~rallyi metatarsals and phalanges were o~5~rvcd in numbers coinciding with the forcdigits. :c"t.uses...i thin a litter,.;ere remarkably sirnllar ~n sk~l~tal SL~uc~ure and observable foc~ of calcificat~o~. v~~;a=:on b~=wccn l~ttcrs may be attr~buted to dlf crencc5 in ~0~ WhlCh ~~~ have ~een as much as 20 hours. Averaq,; fetal '.;eight: &"'l an~l.~!sis of CO"Jar:..ancc ier a~orag~ l~~~e= Slze ar.d average fe~al b01y weigh~ by dosage l~v~l~ (~rca~m~nts, shows no s~gn~ficant difference betwc~~ ~~e ~2Ju~~~d t~~aule~~ ~ea~s for average fetal body weight a~ ~ested (See Exhibit 10.) There is no indicatio~ of any ~=c~~recnt ~r coc~o~nd relationship. 15 Panel Book Page 262

265 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /06/83 04/28/94 80 DOW CORNING CORP GENETIC EVALUATION OF DOW CORNING 1520 SILICONE ANTIFOAM IN THE AMES BACTERIAL ASSAY, WITH COVER LETTER DAJED 4/20/94 Chemical Category SILOXANES AND SILICONES, DIME ( , 18%); * Panel Book Page 263

266 DOW CORN I NG April 20, 1994 TSCA Document Processing center (TS790) Office of Prevention, Pollution and Toxic Substances U.S. Environmental Protection Agency 401 M street, s.w. Washington, D.C. 204~~ Attn: 8Cd} Health and Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41821] Health and Safety Data Reporting: 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow Corning Corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical Substance(s): Siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 18% and Dimethyl silicones and siloxane, reaction products with silica 2% identified as Dow Corning e 1520 Silicone Antifoam Title of Submitted study: GENETIC EVALUATION OF DOW CORNINGe 1520 SILICONE ANT I FOAM IN THE AMES BACTERIAL ASSAY Dow Corning Corporation June 6, 1983 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland,MI DOW CORNING CORPORATION, MIDlAND, MICHIGAN TB.Ei"HONE: (517) Panel Book Page 264

267 2 For purposes of health and safe'ty data reporting und,":r section 8ed) of TSCA, the general PROPRIETARY designation on the attached toxicological study has been waived by Dow Corning. If you require further general information concerning thjs submission, please contact Dr. Rhys G. Daniels at For further information concerning the submitted study or the toxicological properties of the listed chemical substance, please contact Dr. Robert G. MeeKS at Sincerely, ~ t:.. 'B Alvin E. Bey ~ 0.5. Area VicepVesident Corpcrate Director HES Panel Book Page 265

268 GENETI:C EVALtJATI:OR 01' OOW CORHDIG 1520 SI:LI:COm: ANT:IFOAJI m THE AHES BACTBRZAL ASSAY Dow corning corporation 1.983I June 6, ABSTRACT The test material was evaluated for genetic activity in a microbial assay (Ames, et al., MUTATION RESEARCH, 31.:347, 1.975) with and without the addition of mammalian metabolic activation. No evidence of genetic activity was observed. Panel Book Page 266

269 3 OBJEC'l'IVE The obj~etive of this study was to evaluate the test material ~or gen~tic activjty in a microbial assay (Ames, et al., MUTATION RESEAR~I, 31:347, 1975) with and without the addition of mammalian metabolic activation preparations. MATERIALS A. Test Material TX834l8l02 B. Indicator Microorganisms Salmonella typhimurium, str. TA1535 TA1537 TA1538 TA98 TAlOQ C. Activation System 1. Component Final Concentration/ml M9C12 KCl NADP Glucose6phosphate Sodium phosphate, ph 7.4 Homogenate fraction equivalent to 25 mg of wet tissue 8 ~ moles moles moles moles moles 2. S9 Homogenate A 9000 x~ supernatant prepared from SpragueDawley adult male rat liver induced by AROCLOR 1~54 five days prior to kill. Purchased from LittonBionetics, Inc., Kensington, Maryland. Stored until use at 76oC. D. Positive Control Chemicals Chemicals used for?ositive controls in the nonactivation ana activation assays. PROPPIfT AP''': r~;. COIRNNC ccpr:;r.: :,r;,,", ';',' ':::'..,,:' I Panel Book Page 267

270 't Nonactivation Activation Chemical" SodiWII Azide (AZ) 9Amino Acridine (AA) 2Nitrofluorene (NF) 2Anthramine (ARTH) Water or Saline Ethano' Dimethyl~ulfoxicie Dimethylsulfoxide E. Solvent Concen~rations given in Results section. ""Previously shown to be nonmutagenic. Deionized vater was used to prepare dilutions of the test material. The solvent employed and concentrations of chemicals are recorded in th(! Results section. EXPERIMEN'l'ALPROCEDURE A. Spot Plate Test Approximately 10 8 cells from a l6hour culture of each indicator strain were added to separate test tubes containing 2.0 ml of molten overlay agar supplemented with biotin and a trace of histidine. For nonactivation tests, O.S JIll. sterlle saline was added and the contents of the tube poured over the surface of a sterile petxi dish containing approximately )0 ml of hardened base agar (Minimal Agar Davis Difco Corp.) and allowed to solidify. In activation tests, just prior to pouring, an aliquot of the reaction mixture (0.5 ml containing the 9000 ~ ~ liver homogenate) was added to each of the overlay agar tubes which was then mixed and pcured over the base agar. When all plates are hardened, the test m3terial~ were spotted on individual plates of each bacterial strain, with and without activation, in 10 ul quantities. The plates were then coverc3 to prevent photoreactivity of chemicals and within one hour the plates were transferred to a darkened incubator and h~ld at 37 0 C for 72 hour~. B. OVerlay Plate Test Approximately 10 8 cells from a l6hour culture of each inoic~tnr,tr~'n was added to separate test tubes containing 2.0 ml of moltcn :l'.7ar supplemented with hiotin and a trace of histidine. For n~n~~~ivation tests. five dose levels of the test compound and 0.5 ml st~!'"; 1!'.11 i nr were added to the contents of the appropriate tubes and pollr.'.~ 'Wr:T I;hf' surfaces of selective agar plates. In actf.vation tests, fi w' rl i ff:,"nt conc.entrations of the test chemical were added to the appropr i.it: I;uhc':; with cells. Just prior to pouring, an aliquot of z:~action mixttl!'"f CO.5 rr.l containing the 9000 x ~ liver homogenate) was added to each of the activation overlay t:jbes, mixed and the contents poured ~ver thc ~urfac( of a minimal agar plate and allowcrl to soudify. The p'.atc~ were incubated for 72 hours at 3,oC and scored for the" numbcr of colc,"jr.~ Panel Book Page 268

271 5 growing on each plate. The concentrations of all chemicals are given in the Results section. Positive and solvent controls usinq ~th dir~ctly active positive chemicals and those that require metabolic activation v~re run vi tt each assay. c. Recordlra Tbe number of colonies an each plate were counted using a MOdel CIIl Autc.atfld Colony Counter (Mew Brunswick Scientific Co., Inc.) Each count listed in the Results section is the average of ten replicate counts on each plate. The ~esults are presented as revertants per plat~ for ~ach indicator strain eapla,ed in the assay. Positive and solvent controls are provided as referer.ce points. RESOLTS A. $POt Plate Test 'l'be test material was negative against all five strains employed, vi th and without metabolic activation. B. OVerlay Plate Test See Table I for results and concentruti~ns t~sted. IM'!'ERPRETM'Ial OF RESOL'l'S AND CONCLOSl:ORS The test material vas examined for mutagenic activity in a series of in vitro microbial assays employing Salmonella strains as indicator organis:ns..n;ecompound was tested directly and in the presenoe of liver microsomal enzyme preparations frolll Aroelorindueed rats. '!'he following results were obtained: A. NonActivation Test Results The results of the tests conducted on the mate~ial metabolic system were all negative. in the absence of a B. Activation Test Results The results of the t~st oonduct;d on the material in the presence of a metabolic system were all negative. c. Conclusion The test material,.~83418l02,failed to exhibit mutagenic activity in both th~ activation and nonactivation systems and is considered not t~ he mutagenic under the conditions employed. PROF~'ETAF.Y, Fe!! u:;t v."!'..., ;>r'., COINII'IG ceg'o~;r~." ':01<'1'. Panel Book Page 269

272 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /94 80 DOW CORNING CORP ANALYSIS OF DOW CORNING ANTIFOAM A COMPOUND IN THE FEED USED FOR DETERMINING THE TERATOGENIC POTENTIAL OF POL YDIMETHYL SILOXANE IN THE RABBIT, WITH COVER LETTER DATED 4/20/94 Chemical Category SILOXANES AND SILICONES, DIME ( ); * ( ) Panel Book Page 270

273 DOW CORNING April 20, 1994 TSCA Document processing center (TS790) Office of Prevention, Pollution and Toxic Substances u.s. Environmental Protection Agency 401 M street, S.W. Washington, D.C Attn: 8(d) Health and Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41821] Health and safety Data Reporting; 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow Corning corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical Substance(s): Siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Dimethyl silicones and siloxane, reaction products with silica 7% in Antifoam A compound Title of Submitted study: ANALYSIS OF DOW CORNING4!I ANTIFOAM A COMPOUND IN THE FEED USED FOR DETERMINING THE TERATOGENIC P0TENTIAL OF POLYDIMETHYLSILOXANE IN THE RABBIT Dow Corning Corporation October 11, 1984 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland, HI DOW CORNING CORPORATION, MIDlAND, MICHIGAN TREPHONE: (517) Panel Book Page 271

274 2 For purposes of health and safety data reporting under Section 8ed) of TSCA, the general PROPRIETARY designation on the attached toxicological study has been waived by Dow Corning. If you require further general information concerning this submission, please contact Dr. Rbys G. Daniels at For further information concerning the submitted study or the toxicological properties of ~e listed chemical substance, please contact Dr. Robe~ G. Meeks at Sincerely, ~e.b~ Alvin E. Bey 0 u.s. Area VicePresident corporate Director HES Panel Book Page 272

275 ANALYSiS OF DOW CORHDlG ANTiFOAK A COJlPOUlm Dr THE PEBD USED FOR DBTERHDITNG THE TERATOGENIC PO'l'BliTIAL OP POLYDDlBTlIYLS:ILODIiB Dr THE RABB:IT Dow Corning Corporation 1984i october 11, 1984 ABSTRACT Analysis of DOW CORNING8 Antifoam A Compound in feed was conducted on three rabbit feed mixtures that were formulated by Ralston Purina with 0.5%, 1.0% and 2.5% DOW CORNING8 Antifoam A compound, Food Grade. Four sets of samples were collected. The first set was on the first shipment, the second set was on the second shipment, the third set was at the end of the first shipment, and the fourth set was on feed that had been in the feeder for two days. Each sample was analyzed for composition and distribution of linears and cyclics of the test chemical in the feed. The test results indicate that no significant amounts of DOW CORNING8 Antifoam A compound was lost by evaporation during storage or while in the feeders. There was no significant difference in distribution of linears and cyclics between pure DOW CORNING8 Antifoam A Compound and the DOW CORNING8 Antifoam A compound that came from the refrigerated and unrefrigerated feed. consequently, the results of the study indicate the feed was properly prepared and that the integrity of the feed mixtures was maintained during the study. Panel Book Page 273

276 3 PURPOSE '!'be purpose of the i'lvestigation ",as to determine the ccmposition and distribution of linears aild c:ycllcs of DOW CO~ Antifoam A Cc:cpound content in the feed formulated by ita:lston Purina. '1'EST MATEIUALS A NJIIple of DOW COI\NING Antifoam A compound (Food Grade: Lot ) was submitted to Ralston PUrina CaDpany. Richmond. 'l'bey were requested to formulate three pelletized rabbit c:hoi.t mixtures of DOW CORNING Antifoam A Compound at dietary concentrations of 0.5'. 1.Ot. and 2.51 (".,,Iv). FOur mixtures were received fco!ii Ralston Purina. '%tley were labeled control. (Lot 36731), 0.5\ Antifoam A Cclllpou.""ld {Lot 36732). 1.0\ Antifoa: A Cclllpound (Lo~ 36743) and 2.5 I Antifoam A (Lot 36734). The feed vas received in bio sbipmedts. These bags of mixed and control. feeds were refrigerated at all times except dur1.ng f~ding of the animal.s. At au times the bags of feed were kept on pallets in the re!rigerator. Analysis of feed wag conducted on the first sbipment. the second shipment, feed that had been left for two days in t:be feeders. and the end of the first shipment. All sajiiples were grour.d with a mortar and pestle and placed in glass vial.s. The samples for checlc of total 51 content were ",ashed with three aliquots of methyl isobutyl ketone (Ml:BK). "l'be decanted a1iquot!l. of MIBr.: were stored in Cilpped glass vials in a freezer until ablpcaent to Anal.~"t!cal Department for analysis of tou! Si by atomic absorption. '!'be samples for cneck of distribution of lineara add cyeuca of the DOW CORNING Antifoa:ll A Compound in t!le feed V\!re washed in toluene overnigbt. 'l'be de~ted al.lquots of toluene ",ere stored in capped glllss vials in a freezer I1ntll sbipsteot to Analytical Department for analysis by 9aS partition cbrolm.tograpby (GPC;. RESULTS 'l'he composition test rc:lults are included in Table l.. 'l'bere were no apparent dlf erences between the reported and measured composition of ~ samples collected for the three feed ldixtures. Gas partition chromatography (GPC) analysis was c:c>nducted on two samples from each feed mixture for each set of samples. Listed in Figures 15 are GPC analyses of samples from the first shipment. Fi9ure 1 is tile conttol feed GPC. F:.gw:e 2 is the ~e DOWi CORNING Antifoam A Callpound GPC. Figures 3. 4, and 5 are GPC analyses of the 0.5\. 1.;\. and 2.51 DOW CORNING Antifo:llll A compound mixtures, respectively. There were sollie unidentified materials in the control. feed GPC. "l'be areas in the control feed GPC had to be subtracted rclll the DOW CORNING Antifoa:n A Compound/feed ldixtures GPC analyses to obtain the correct distribution of, DOW CORNING PROPRIErA11.Y Panel Book Page 274

277 4 llnears and cyellcs of IY.)W CORNING A.."1tlfoam A Compound In the s.u:ples. '!'able II is a summary of the data from the GPC aaal.yds of the eed aixtures. There vas no significant difference in distribution of llde~s and c:yclics betveer. pure Dow' Corning Antifoaa A CoIIIpound and the DOW CORNING Antifoam A CCIIlpound that came frem the refrigerated.nd unrefrigerated feed. CONCLOSIONS 'l'he test results indicate that no significant amounts of DOW CORNING Antifoam A Compound was lost ~. 6~~poration during storage or while in the feeders. There was no significano; o!1fference in distribution of Unears and c:yclics between pure DOW CORNING Antifoam A Compound and the opw CORNING Antifo.ul A Compound that came froll! the refrigerated and unrefrigerated feed. CoosC! quently, the resw.ts of the study indicate the feed was properly prepared and that the integrity of the feed liiixtures vas Dl41ntained during the study. DOlV corumm PROPi\:ETARY Panel Book Page 275

278 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /23/84 04/28/94 8D DOW CORNING CORP TERATOGENIC POTENTIAL OF DOW CORNING ANTI FOAM A COlv1POUND, FOOD GRADE IN RABBITS, WITH COVER LETTER DATED 4/20/94 Chemical Category SILOXANES AND SILICONES. DIME (631486~9); * ( ) Panel Book Page 276

279 DOW CORNING April 20, 1994 TSCA Document Processing Center (TS790) Office of Prevention, Pollution and Toxic Substances U.S. Environmental Protection Agency 401 M Street, S.W. Washington, D.C Attn: 8(d) Health and Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41821] Health and Safety Data Reporting; 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow corning Corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical substance(s): siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Dimethyl silicones and siloxane, reaction products ~Jith silica 7.0% in Dow Corning Antifoam A Compound Title of Submitted study: TERATOGENIC POTENTIAL OF DOW CORNING ANTI FOAM A COMPOUND, FOOD GRADE IN RABBITS Dow corning corpora~ion October 23, 1984 Manufacturer: Dow corning corporation 2200 west Salzburg Road Midland, MI , DOW CORNING CORPORATION, MIDLAND, MICHIGAN TELEPHONE: (517) Panel Book Page 277

280 2 For purposes of health and safety data reporting under Section 8ed} of TSCA, the general PROPRIE~ARY designation on the attached toxicological study has been waived by Dow corning. If you require further general information concerning this submission, please contact Dr. Rhys G. Daniels at For further information concerning the submitted study or the toxicological properties of the listed chemical substance, please contact Dr. Robert G. Meeks at Sincerely, ~E.B~ Alvin E. Bey 0 u.s. Area VicePresident Corporate Director HES Panel Book Page 278

281 2 File Nt:).: IlliO'} Authors: Waheed H. Siddiqui Series :,0.: 1(J()051:!'):! Barncstine Stanton Reference No. TXCl!.2:,rlO05 Gary B. Kolesar Location: Date: Midland, MI Claire R. Devrip<; Ti tle: Teratogenic ~ten~~dl of DOW CORNING z hntif02m A Compound, Food Grade irl Rabbits t'\.8stract The teratogenic potential of OOW CORNING e Antifoam A Compound, Food Grade (hereafter referred to as DOK CORNING e Antifoam A Compound) administered in the diet was studied in rabbits. Pregnant New ~aland white raboits were given food grade OOW CORNING Antifoam A Compound in the diet at concentrations of 0, 0.5, 1.0, ane 2.5% from the morning of day 6 through the morning of day 19 of gestation. There were no overt signs of toxicity in the dams, and no statistically significant differences in feed consumption at any level compared to the control group. There was no evidence of adverse effects in mean maternal body weight or liver weight of the treated females. The incidence of resorptions among the total fetal population was not altered by feeding DOW CORNING e Antifoam A Compound to pregnant rabbits at th~ three dietary levels. Term fetuses were examined by standard terato:ogical techniques. Male and female pup weights were unaffected by the matprnal trcat.:ncnt. DOW CORNING Anti foam.'\ CO::lpoLOnd had no statistically significant adverse effects on the incidence of external, visceral or skeletal abnormalities. On the basis of these results, it is concluded that COW CORNING Antifoam A Compound i~ not cmbryotoxic or teratogenic in rabbits at dietary level~ up to 2.Si. DOW CORNING PROPRIETARv Panel Book Page 279

282 ~ I N'I'RODUC7ION. DOW CORNING Antifoam A Compound (food grade) is co~nly used as a defoamer in the pr~~~s~ing of foods at levels up to 10 ppm and also is found in se~eral antifla~ulent formulations. This material consists of polydimethylsiloxan~ fluid, a chemically inert polymer (Rochow, 1951), about 10% or less conce~tration of cyclic species (cyclic tetramer through cyclic hexamers) and silica. This study was designed to assess the teratogenic potential of DVW CORNING k~tifoam A Compound in the ptegnant New zealand white rabbits. The test material was given in the rabbit feed and this route of administration is simulated the route of exposure to humans. II. MATERIALS AND METHODS A) Test Method Food grade DOW CORNING Antifoam A Compound (polydimethylsiloxane), Lot No , supplied by the Medical Products Plant, Hemlock, Michigan, was used in this stedy. The molecular distribution of linears and cyclics in the DOW CO~~ING Antifoam A Compound were analyzed by gas ~artition chromatography and founn within the specified limits for this material (Appendix 4). B) Experimental Animals Adult ~ale and female New Zealand white rabbits weighing a9proximately 3.5 kg, were p~=ch~sed from Langshaw Farms, ~u9usta, Michigan. upon arrival at the ~oxicology Department, all rabbits were thoroughly examined for their physical health by a qualified consulting veterinarian and by a qualified animal technician. The rabbits were housed individually in stainless steel ~ire mesh c~ges for an acclimation period of three weeks. During this period, rabbits DOW CORNING PRO?Hlt r o.\!\' Panel Book Page 280

283 6 exhibiting sickness, disease, infection or physical abnormalities were discarded. The animals were fe~ PURINA Laboratory Rabbit Chow (Ralston Purina Company, St. Louis, Missouri) and tap water ad libitum. Each rabbit was identified by a number on a metal ear tag. The animal number also appeared on the outside of each cage to insure that each animal was returned to its own cage after physical examination, sanitation or any study related reasons. All rabbits were housed in rooms maintain~d at 22 0 C ± 2 0 C temperature, 45% (3050%) relative humidity and a light cycle of 12 hours. C. EXperimental Design 1) Animal Groups A standard outline of the experimental design is shown in Table I. Ninety six (96) female rabbits anc 30 proven bucks oe the same line were utilized in this study. A female was introduced to an untreated proven buck twice with a four hour interval. The day of successful mlting determined by observing mounting and positive vaginal smears was counted as d,_ 0 of pregnancy. Just prior to first mating, 0.1 ml of FSH, chorionic gonadotrophin, was injected to each female rabbit through the marginal ear vein to simulate ovulation. Breeding continued until 1524 mated female rabbits per group were obtained. Animals were assigned to four grvups ~uch that their body weights on gestation day zero (GOO) were nearly equal across dose groups. Twentyfour (24) time mated female rabbits were assigned to four groups. Dams were rank ordered by body weight on GOO and assigned to control and treatment groups. One animal ~~z ~zzigned a~ random to each dose group from the four lightest animals, then from the next four lig~~est animals, etc., until six animals ~ere assig~ed to each group per day for four days. DOW CORNING PROPRIETARV Panel Book Page 281

284 7 2) RC'ltc and Dose Selection DOW CORNING Antifoan. A Compound is co~only used as a defoamer in the procl~sing of foods at levels up to 10 ppm. Thus, the ~est chemical at the dietary concentrations of 0.5, 1.0 and 2.5% is sao, 1,000, and 2,500 fold (respectively) higher than normal human consumption, assuming that the total food for human con;,umption contains the maximum allowed level of polydimethylsiloxane (PDMS). The route of administration in the present study simulates the route of exposure to humans. 3) Treatment The DOW CO~~ING Antifoam A Compound of welldefined purity (gel permeation c~romatography) was used for incorpo~ation in tre test diets at concentrations of 0, 0.5, 1.0, and 2.5%. The test ma~erial was mixed with rabbit chow by Ralston Purina Company (RichmC'.ld, Indiana). The test diets were stored in a reirigerated ~oom. The stability and homogenicity of the test chemical in diets were determined at the beginning and at the end of dietary feeding. The rabbit PURINA diets containing a (control), 0.5, 1.0 and 2.5% DOW CO&~ING Antifoam A Co~pound were fed ~o b~ed females on days 6 through 18 of gestation.!iater and feed were provided au libitum. D) Maternal Observations f.nimals were observed daily during the tt"eatment and posttr'eatmenr. perioos for physical appearance, abnormal behavior, signs of (loc?" and systemic) toxicity, and death. i'he ;;'Iu.tc:::,:~l body wei9ht~ were recorded on days 0, 6, S, 12, 15, 18, 24, and 29 of gestation. Food consuj:iptions of all the female nnw r.nrning PROPRIETtl.R"! )JUl' _... Panel Book Page 282

285 10 III. RES~LTS AND DISCUSSIONS No clinical symptoms or untoward behaviora~ reactions were observed among the test or control group animals. '1'\.'0 animal" in the c:mtrol S:::'>'';o ciied ouring t~e study. External examination did not reveal any a~~0~mal f!ndings. Autopsy of thes~ animals show~d the absence of normal feed in the go~trcintestinal tract of one rabbit. The left a~d right lobes of the same animal also showed consolidation and acute pneumon1a. The cause of death in this case was most likely due to the pneumonia. The gastrointestinal tract of the other female con~ainc2 normal fced, fecal material and an exl~cted amount ce fat. The fat was slight.ly jaundiced in appearance. The liver appc?cr.::o ~o show some degree of mottling and was also slightly jaundiced in color. No other gross changes were noted in this case and a cause of death wa~ not established u~n gross examination. The homogenicity and stability date of linear ar.d cyclic components of DOW CORNING Antiioam A Compound in the rabbit feed are summarized in Tables II and III. These results indicai:e that DON CORNING Antifoam A Cor::pound was uniformly distributed in the feed and no signi~icant arne.. It '.'as lost by evaporation during mixing, pellitizing, storage or ~hile in the feeders. There was no significant difference in distribution of linear and cyclic species between pure DOW CORNING Antifoam A Compound and that extra~ted from the refrigerated and nonrefrigerated feed. Consequently, the results of these analyses confirm that the diets were p:cperly 9repared and the integrity of the test material in the feed was main':ained during the study period. The mean maternal body Weight data on days 6, ~, 12, 15, 18, 24, and 29 of gestation are Fresented in Table IV. Individual values are reported in ~?Cndix 1. No st~tistical significant differences in mean body weights w~re observed between control and experimental animals. No signs of feed refusal were obs~rvcd in Qny of the rabbits in this study and no effects were noted on OOW p.,r'i CORNING?~O?R\tl Panel Book Page 283

286 11 average food consumption throughout the experimcntal period by feeding diets containing 0, 0.5, 1.0, and 2.5% DOW COWlING Antifoam A Compound (Table V). Thc absolute and relative liver weights were not affected by feeding DOH CORNING Antifoam A Compounrl at different concentration levels and all "alucs were comparable with the control (Table V). The effects of feeding diets treated with DOW CORNING Antifoam A Compcund ~~ the c~tcome of pregnancy and fetal development are shown in Table VI. The i~cidence of pregnancy w~s not altered ~y feedi~g diets containing DO~ CORNING Antifoam A Compound. Of the 24 animals bred in each group, 2:. females (including one dead rabbit) were found pregnant in the control and 22, 21, and 23 females in the diet groups containing 0.5, 1.0, and 2.5% DO, CORNING Antifoam.; Compound, respectively. No females had totally resorbed litters in this study except one dam was observeu to have only one early resorption site at 1.0% dose level. One rabbit fed the his~~st concer tratio~ of DOW CORNING Antifoam A Compound has Pasteurella mu!tocida infecticn in the uterine horns and a second female from the same group was obscrved to have twisted uterine horns with dead fetuses. The data of these t~~ fc~ale; were not included in this report. No statistically significant differer.~;es on the number of implantation sites per dam, mean preimplantation, or postillplantation losses, percentage ~esorptions and number of corpora lutea were observed among treatment groups. No adverse effects on the number of :.ive fetuses per liter, mean litter size, male or female pup weights, or crc wnrump length were observed among litters o[ rabbits treated with DO~ CO~~ING Antifoam A CompoOlnd. ThrE:e dead fetuses were observed, o..e each among litters of rabbits fed diets containing 0, 1.0, ~nd 2.5%. Thc external, ~isccral and skeletal alterations of fetuses are prescnte i in thc form of major and minor anomalj.cs in Tablc VII. No statistically significant effect of any treatment on major or minor anomalies could b" found. Ma~or anomalic$ were observed in three ietuses from the 0.5% do~e, two DOW CORNING PROPi~lt I.~ Panel Book Page 284

287 12 fetuses from 1% and one fetus feom the ~.5% dose gro~ps. The anomalies were spina bifida in one fetus and dilated renal pelvis in two fetuses at the 0.5% dose, misshapen scapula in two fetuses at the 1% dose, and multiple anomalies of fac~, heart and other tissues in cne fetus at the 2.5% dose. The occurrence of these anomalies m?y have been spontaneous rather than treabmentrelated for the following r~asons: (1) Neither the total number of malformed fetuses nor each individual maltorma~ion showed a coseresponse relationship, and (2) the type and incidence of these anomalies were comparable to those reported for the normal population of New zealand rabbits (Palme~, 1968; Hartman, 1974) and those observed in our historical control. The incidence of skeletal variants in the treated litters was not significantly increased over the control incidence. Aberrations in skeletal development included one or two extra ribs, rudimentary ribs and spurs, unossified fifth sternebra, and delayed ossification of skull. None of these aberrations were considered treatment related since their incidences at each dose was statistically not different from their respective control incidences. In a review, Khera (!98l) concluded that the fetal skeletal aberrations quite often are transitory in nature and tend to disappear during further. dev~lopment. In the absence of major malformations or when the fetal skeletal aberrations are similar and not significantly different from controls, they may not warrant serious attention. In conclusion, the results of this st:.jdy indicate that DOW CORNING Antifoam A Compound is not embryotoxic, fetotoxic. or teratogenic in New zealand white rabbits wher. fed in the diet up to 2.5% during the period of major organogenesis. DOW CORNING PROPRIETARY Panel Book Page 285

288 CODING FORMS FOR SRC INDEXING Microfiche No. OTS /15/74 04/28/94 8D DOW CORNING CORP PHARMACOKINETIC AND METABOUC STUDIES ON DOW CORNING ANTIFOAMS A AND M IN MICE, MONKEYS AND HUMANS WITH COVER LEITER DATED 04/20/94 Chemical Category DOW CORNING ANTIFOAMS A AND M (CAS # ) Panel Book Page 286

289 DOW CORNING April 20, 1994 TSCA Document Processing Center (TS790) Office of Prevention, Pollution and Toxic Substances u.s. Environmental Protection Agency 401 M Street, S.W. Washington, D.C Attn: 8(d} Health and Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040~ FRL41821) Health and Safety Data Reporting~ 40 CFR Part 716 ITC 30Listed Siloxanes and Silicones Dear Sir: The enclosed information is submitted on behalf of Dow Corning Corporation, Midland, Michigan, , in compliance with the referenced health and safety data reporting rule. Listed Chemical substance: Siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 91% and Sil~xanes and silicones, dime, hydroxyterminated 2.9% in Antifoam M compound siloxanes and silicones, dime (Dimethyl silicones and siloxanes) 93% and Dimethyl silicones and siloxane, reaction products with silica 7% in Antifoam A Compound Title of Submitted study: PHARMACOKINETIC AND METABOLIC STUDIES ON DOW CORNING* ANTIFOAMS A AND M IN MICE, MONKEYS AND HUMANS Dow Corning Corporation April 15, 1974 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland, MI DOW CORNING CORPORATION, MIDLAND, MICHIGAN 48686{J994 TELEPHONE: (51 7) Panel Book Page 287

290 2 For purposes of health and safety data reporting under Section 8(d) of TSCA, the general PROPRIETARY designation on the attached toxicological study has been waived by Dow Corning. If you require further general information concerning this submission, please contact Dr. Rhys G. Daniels at For further information concerning the submitted study or the toxicological properties of the listed chemical substance, please contact Dr. Robert G. Meeks at Sincerely, ~e.b~ Alvin E. Bey _0. U.S. Area Vicepresident Corporate Director HES Panel Book Page 288

291 2 Abstract This study was cond\:l.cte' to fulfill a request of the Joint FAO (Food Additives Orgcu~ization)/iiHO (World Health Organization) for metabolic studies in several species on DOW CORNING$ Antifoam A and M Compounds, i.e., polydimethylsiloxane fluids containing silica. DOW CORN:rNG<!l Antifoam A and DOW CORNING0 Antifoam M Compounds hereafter will be referred to as A and M Compounds. M Compound is prepared from po1ydimethy1si1oxane polymer which contains less than 0.022% concentration of low molecular weight species (cyclic trimers through cyclic hexamers) compared to an approximate 10% level present in A Compound. Initial studies in mice demonstrated that there was a significant increase in urinary and biliary silicon after oral administration of A, but not M Compound. The increase in urinary and biliary silicon appears to represent organosolub1e silicon rather than inorganic silicon from absorption of silica. Complete balance s't: i ies over a sevenday period were then conducted in unanesthel ~ed rhesus monkeys with 1 "Clabe11ed polydimethy1si10xane flt:...a.s identical to the polymer fluids in A and M Compounds except that these fluids contained no silica. There was no more than 0.25% of the dose as organoso1ub1e silicon detected in expired air and urine after oral administration of 21.8 or 41.8 mg/kg M fluid. There was less than 0.01% of the dose found in the sum of approximately forty different tissues 72 hours after dosing percent of the dose was recovered from the feces. Gas chromatography analysis of toluene "which had been used to extract organoso1ub1e silicon from the feces, revealed that no low molecular weight po1ydimethy1siloxane species had been produced during passage through the gastrointestinal tract. On the other hand, following oral administration of 1.15, 13.7 or 18.0 mg/kg A fluid, there was a statistically significant (p<.ol) amount of organosolub1e silicon present in expired air, urine and bile. The percent of the dose found at all dose levels in expired air, bile and urine was respectively (0.40.9%, tt 36 hr); (0.10.3%, tt 36 hr) and (2.12.5%, tt 24 hr). The remainder of the dose was recovered from the feces. Blood levels were below the limit of detection, i.e., <1 pg/m1. Studies were then conducted in humans under conditions of controlled dietary and fluid ;!\take. A and M Compounds were administered orally in a single dose of 100 mg/kg at different times to one group of L"ldividuals, while A and M Emulsions were administered orally in a single dose of 100 mg/kg emulsion (30 mg/kg solids) at different times to another group. Total and organoso1uble (methyl ethyl ketone) urinary silicon output as well as organosoluble (toluene) silicon output in expired air were the parameters used to assess absorption and elimination of silicon in these materials. 'There were no increased silicon levels found Panel Book Page 289

292 3 in any of the parameters measured after either oral M Compound or Emulsion. There were increased silicon levels in all three of the parameters measured after either oral A Compound or Emulsion. It is probable that the increased silicon levels in ~ose parameters investigated represent organosoluble silicon rather than inorganic silicon (silica). The average percent of dose found in urine over six days was 1.8% and 3.3% for A Compound and A Emulsion respectively with a halflife of 24 hours in both cases. At least 0.5% of the dose was found in expired air with a halflife of 8 hours, and has been identified as low molecular weight cyclosiloxanes (octamethylcyclotetrasiloxane and decamethylcyclotetrasiloxane). The organosoluble silicon in urine has not been identified.. Panel Book Page 290

293 7 3. Composition of A and M Emulsions GLC data were also obtained on Dow corning IS Antifoam Emulsions. I~ was of interest to see if other emulsion ingredients (water, s~earate emulsifiers and preservative) promote siloxane bond cleavage and cyclic formation. M Emulsion contains six GLC peaks in the low molecular weight cyclosiloxane region; however, none of the peaks appear to be low molecular weight dimethyl cyclics e (Me2SiO)x, dllnethyl linears [Me~SiO(~iO)~SiMe3]'or siloxanols [HO(~!O)XH]' as determined by spikin: the emulsion with appropriate standards. Mass spectrometer analysis of M Emulsion yielded a series of esters derived from phydroxybenzoic acid, i.e. OH~~OR, UI 6 where R = CH 3, C2H 3, C2H 7, C4H~. The alky1~hydroxybenzoates are present to the extent of 0.2%, and are included in the emulsion as a preservative. 4. Determination of Silicon in Biological Samples The atomic absorption spectrometer analysis of silicon method is included in detail in Appendix IV. This method was used for the determination of organosoluble silicon. Total silicon (including inorganic) was determined in an emission spectrometer as defined in Appendix V. B. Biological The mouse, rhesus monkey and man. were the experimental subjects used in this study. Experiments using the rhesus monkeys were delayed over a period of two years, , while adequate analytical and metabolic methodology was developed. A part of the method was presented at the Society of Toxicology Meeting at New York in March of The method bas been written for submission to the Journal of Toxicology and Applied Pharmacology. The manuscript will be submitted in April, 1974, along with the first in a series of papers dealing with the phannacokinetics and metabolism of organosi1icon compounds. The entire methodology manuscript, as it will be submitted, is included in this report as Appendix VI. For clarity, the methodology specifically used for the mouse and human studies is presented simultaneously with the results obtained in those species. III. Results A. Mouse Study Male Buckberg mice, about 2S go, were used for this study. The animals were kept in in4ividual cages, fasted 4 hours prior to oral dosing and after dosing given water ad lib during the 24hr experimental period. There were twelve miceper group: (1) The Panel Book Page 291

294 12 control group received 0.5 ml/kg sesame oil. All other groups also received 0.5 ml/kg sesame oil as vehicle for the test materials. (2) The silica group received 6 mg/kg (Si0 2 }x. This is the approximate amount of silica contained in A and M Compounds. (3) A Compound at 0.5 ml/kg. (4) A Emulsion. The animals received 1.67 ml/kg which represents the same amount of polydimethylsiloxane in group 3. (5) Antifoam 351 Compound. This antifoam was an experimental preparation similar to M Compound. It contained <.02% low molecular weight polymeric species, i.e., a "lowvolatility antifoam"; however, it did contain 350 cs polydfmethylsiloxane rather than 1000 cs similar to that found in Antifoam M. The dose was 0.5 mli:icg. (6) Antifoam 352 Emulsion. This 30% emulsion was made from 351 Compound and the dose administered was 1.67 ml/kg. (7) DOW CORNING 360 Medical Grade 350 cs polyd~ethylsiloxadefluid. This material contains no silica or emulsifiers and was given in a dose of 0.5 ml/kg. At 4, 12 and 24 hours after treatment, four mice from each group were anesthetized with methoxyflurane and urine was removed directly from the urinary bladder of each mouse and pooled for silicon analysis by emission spectroscopy. The bile was removed directly from the gall bladder and handled in the same way. Precautions were taken to avoid silicon contamination, i.e., acidhardened glass s}~inges and nonsiliconized needles were used. Table 1 shows there were no statistically significant differences in urinary or biliary silicon concentration at the time periods tested among the control, silica, 351 Compound, 352 Emulsion, or 360 Medical Grade silicone fluid groups. There was a statistically significant (p<.05) increase in urinary and biliary silicon concentration compared to the control groups at the time periods tested for A Compound and Emulsion. ~his increase in silicon represents only a small part of that potentially available for absorption from the gut. B. Monkey Study Rhesus monkeys were surgically prepared and were placed in an individual metabolism chamber for a period of at least 72 hours as described in Appendix VI. The I~Clabelled polydbnethylsiloxane fluid used in these studies was synthesized in the Dow Corning Radiochemistry Laboratory from randomly labelled dimethyl cyclic siloxanes. Both polymer components of the A Compound were labelled and the final viscosity of the A fluid was 335 cs, while that for M fluid was 932 cs. Specific activity of A fluid (335 cs) = 0.50 mci/g and M fluid (932 cs) = 0.91 mci/g. Neither A or M fluid contained silica in this portion of the study. The data are summarized in Table 2. Three monkeys received A fluid (without silica) at dose levels of 1.15, 13.7 and 18.0 mg/kg. These doses were calculated after determining the amount of radioactivity remaining in syringe, needle and gavage tube used to administer the aose~ Approximate to 0.9 percent of the dose Panel Book Page 292

295 15 was found in the expired air. The halflife for pulmonary elimination was 46 hrs. Less than 0.1 percent of the dose was lost as volatiles from urine, flatus and fecal material in the lower portion of the chamber. Approximately 2.0 to 2.5 percent of the dose was found in the urine with a halflife of 24 hrs. Only small amounts, percent of dose, were found in the bile in the first 24 hrs. after dosing. Fecal material collected over 92 hrs. contained 8090 percent of the dose. The time of elimination of the bulk of the fecal radioactivity varied between 18 and 72 hours. Blood levels were below the detection limit,!..,!:.., <1 llg/m1. M fluid was administered orally at 21.8 and 41.8 mg/kg. Only o. 01 O. 02 percent of the dose was recovered in expired air and only 0.22 and < percent of the dose was recovered in the url~e. Fecal el~ination accounted for 9397 oercent of the label. Monkey id30 who received M fluid was sacrificed at 72 hrs. after the oral dose. A complete autopsy was performed. Tissue samples were cut into sections of <5.0 roms, and they were placed in liquid nitrogen. The tissues were hammer milled to a fine powder and were placed in scintillation vials in duplicate weighings of 10, 50 and 100 mg. Three ml of cold distilled water was added to each Vial. The contents of each vial were dispersed by an ultrasonic cell disruptor until a uniform dispersion was obtained. Aquasol, 9.5 ml, was then added and vigorously shaken. The samples were cooled in the counter overnight and they were counted for 10 minutes. There was less than O. 01 % of the dose found in the sum of approximately forty different tissues 72 hours after dosing (Table 3). All fecal samples from monkey 413, who also received M fluid, were extracted with toluene and the extracts analyzed by GPC. The GPC fractions were collected and radiochemically assayed individually. The distribution of counts conformed to the GPC curves for DOW CORNING~ 360 Medical Grade fluid (Appendix VII). C. Human Study. Two individuals volunteered to take part in early 1973 pilot studies designed to determine the methodology and protocol guidelines for a later extensive study in eleven persons. None of the data obtained in the early study will be presented; however, the conclusions drawn were identical to the more definitive study. The eleven individuals were divided into two groups. The vital statistics of these individuals are listed in Table 4. Each individual selected his own diet. The diet was not siliconfree. Only beer was excluded from the diet because of its high and variable silicon content. There was no restriction on the quantity of food or Panel Book Page 293

296 18 liquid consumed, although a number of individuals elected to keep their caloric intake at a lower than normal level and lose weight over the thirteenday study. Absorption of A or M Compound or Emulsion was based on silicon elimination in expired air and urine. Thus, the same diet regiinen that each individual selected on day I was consumed every day throughout the experiment to stabilize silicon intake. Each day began at midnight just after the last urinesa~ple was passed prior to going to bed. Urine was collected and measured under clean conditions designed to minimize silicon contamination; ~.~., polypropylene collectors, storage beakers and dust covers were used, and all equipment was rinsed with deionized/distilled water. Urine collections were separated into three or four periods over the 24hour day depending on whether an individual slept eight or six hooj.rs; however, data presented are based on mg silicon per 24hour urine s~~ple. Urine was stored in the cold for not more than 24 hrs. A 2 ml sample of urine was pipetted into a polycarbonate vial and mixed with potassium hydroxide for emission spectroscopy analysis of total silicon. A 10 ml sample of urine was placed in a vial with 2.5 g sodium chloride and shaken for 1 hour against 10 ml of methylethyl ketone..the mixture was separated by centrifugation and the methylethyl ketone removed for atomic absorption analysis (organosoluble silicon). Table 5, and Figs. 5 and 6, give the individual and group mean values for total and organosoluble urinary silicon. Organo50luble silicon varied for 820% of total urinary silicon during the control period. Although there is a considerable variation in the range of either total or organosoluble silicon in urine among. individuals, we have not been able to ascribe these differences to age, sex, race, type of diet, fluid intake, urinary output,. or body weight, Table 6. It is of interest that the three individuals in the Compound Group who appear to be consistently high in urinary silicon, BBF, TWM, was, are the only individuals who smoke. We are continuing to explore this observation. At the start of the fifth day, M Compound or Emulsion was administered to the appropriate groups. Table 7 lists the formulation data on which all calculations were based to express percent of dose eliminated. There were no statistically significant increases in total or organasoluble urinary silicon eliminated on days 5, 6 or 7 compared to the selected day 4 control following administration of either M C.ompound or Emulsion, Table 8. These data are shown graphically in Figs. 7 and 8 for both total and organosoluble silicon. A Compound or Emulsion was administered to the respective groups at the beginning of day 8 (midnight of day 7)..This is the time in a 24hour period when the stomach is least filled with food Panel Book Page 294

297 27 for 78 hours. We were interested in comparing absorption among.individua1s under conditions where gastrointestinal activity is likely most similar in a group of individuals, i.e., the fasted state. Table 9 shows a statistically significant (p<0.05) increase abovethe day4 control in total urinary silicon on days 8, 9 and 10 for both A Compound and A Emulsion groups. Organoso1uble urinary silicon was statistically significantly increased (p<o.05) above the control level on days 8 through 13 in both groups. These findings were noted for A Compound as well as A Emulsion. Figs. 9 and 10 graphically depict these data. The daily urine output of all individuals in both groups on the ~~tifoam study is shown in Table 10. An analysis of varia~ce of the mean urinary output of either the Emulsion or Compound group reveals no statistically significant treatment differences at any time period when A or M Compound or Emulsion, were administered. Table 11 summarizes the individual total increase in urinary silicon over a sixday period following oral administration of A Compound and Emulsion. Every individual showed an increase in both total and organoso1uble urinary silicon, although there was a t,rofold variation in the absolute values in the case of A Emulsion, and as much as a five to sixfold variation in the absolute values in the case of A Compound. The degree of absorption and urinary elimination of Antifoam A does not correlate with sex, race, age, caloric intake/day, body weight, fluid intake, or control urinary silicon levels. The percent of dose of A Compound or Emulsion eliminated in the urine of man based on total silicon was 1.8 and 3.3% respectively (Table 12). These values compare favorably with the % of dose eliminated in the urine of monkeys based on a similar urine balance study using 14Clabelled A fluid (Table 2). About onequarter of the silicon appears to be based on organosoluble silicon. It is not likely that the increase in total silicon is referrab1e to the silica present; because, no increase was seen in total urinary silicon after oral M Compound or Emulsion, although these materials contain a slightly greater amount of silica than present in A Compound or Emulsion. The discrepancy between the percent of dose based on total versus organoso1ub1e silicon probably relates to a low extraction efficiency with the organic solvent. Preliminary experiments with various organic solvents and salting out procedures, and employing standards suggested indicated an extraction'efficiency of 2530%. The low molecular weight polymeric species present in Antifoam A should be practically insoluble in urine at the apparent level present. Thus, there may well have been metabolic alteration in these species to increase their water solubility. We are continuing in an attempt to characterize these apparent polar metabolites in urine. Panel Book Page 295

298 34 Four men receiving M Compound and A Compound were used for sampling expired air after dosing. The expired air was collected by fitting the supine individual with an anesthesia mask which was connected to a one way flow valve. The valve allowed the inspiration of fresh air but directed all expired air through a polyvinyl chloride tube to a collapseable polyethylene bag reservoir. The expired air...as drawn by partial vacuum from the reservoir through polyvinyl chloride tubing into a single air scrubber described in Appendix VI. The scrubber was charged only with 250 ml toluene. The air was not passed over hot copper oxide..two collection systems were available so that two individuals could be used at once. Expired air was scrubbed for twohour intervals alternately from each of two men because of tightfitting mask discomfort. In this way ~~pired air was sampled for a 6hr period after dosing with M Compound by alternating two groups of two volunteers each. The same procedure was used following dosing with A Compound; however, the total scrubbing periods were extended to 8 hrs with an additional one hour period for each volunteer at 12 and 36 hours after dosing. The toluene used to extract volatile compounds from expired air was sampled and changed at 'the end of each air scrubbing period. Samples of extract were submitted for atomic absorption analysis and gasliquid chromatography/mass spectroscopy analysis of organosilicon compounds. No organosilicon compounds were found in the expired air of four individuals who ingested M compound (Table 13). On the other hand, air extracts from four individuals receiving A Compound had detectable amounts of silicon containing material. An average of 7.58 mg of silicon or 21.6 mg polymer appeared in expired air compared to a mean intake of 6100 mg polymer which represents 0.35% of the dose in the first eight hours after the oral dose of Antifoam A. This is an underestimate in that the material continued to be excreted for at least thirteen hours. These data compare favorably with the range of % of the dose found in the expired air of three rhesus monkeys. A toluene extract of the 68 hr expired air sample of ~VE:S given A Compound was analyzed by gas chromatography/mass spectrometry using Multiple Peak Monitoring. The retention time and response for the ions at m/e 281 and 282 of (Me 2 SiO)4 was determined with standards varying from 1 to 32 ng/lll. The sample was found to contain principally octamethylcyclotetrasiloxane, (Me2SiO)~, at 26 ppm and decamethylcyclopentasiloxane, (Me 2 SiO)s, at 1 ppm. Panel Book Page 296

299 36 TV. Discussion After siloxane antifoam compounds are administered orally, only the low molecular weight cyclosiloxanes, if present, are absorbed from the entire gastrointestinal tract in man. The percent of dose absorbed after M Compound, if any, was below the limits of detection. The 24 percent of total dose absorbed after A Compound is relatively constant in the doses that were examined, i.e., 1, 10 and 100 mg/kg. The percent absorbed based on the avarlable low molecular weight cyclosiloxanes is ca. 2040% when pulmonary and urinary elimination are both considered. The emulsifiers present in A Emulsion may have enhanced the human absorption of cyclosiloxanes in this studyi however, there is simply insufficient data to verify that conclusion. On the other hand, it has been shown that the administration of similar low molecular weight siloxanes in lipid vehicles (e.g., sesame oil, corn oil, oleic acid) does enhance their absorption(l).. 1. Bennett, D.R., Gorzinski, S.J. and Le Beau, J.E. Structure Activity of Oral Organosiloxanes on the Male Reproductive System. Tox & Appl. Pharmacol. 21:5567, The inorganic silica p~esent in the Dow Corning Antifoam M formulations, Santocel C does not appear to be absorbed. The polymer components that are not absorbed are eliminated unchanged in the feces. Those that are absorbed are principally eliminated in the urine ("'75%) with a halflife of about 24 hrsi whereas, the remaining 25% are eliminated principally in expired air with a halflife of 36 hrs. A minor component is present in bile also with a halflife of 36 hrs. Thecyclosiloxanes eliminated in expired air are unchanged whereas those eliminated in urine may have been changed to more polar metabolites. The only human data that we are aware of which might contradict these findings is that of Frazer. He has stated that silicone Antifoam A.and M are absorbed in trace quantities or not at all. Frazer fed antifoam to human subjects and then measured colorimetrically urinary silicate by estimating the stannous chloride reduction of the silico molybdenum complex formed by the reaction between silicate and acid molybdate. The color from the interfering phosphate complex was eliminated by the addition of sulphuric acidoxalic acid reagent. This method will not detect organosiloxanes unless they are metabolized completely to inorganic silicic acid derivatives. Our study has shown that no silica is absorbed, and only low molecular weight cyclosiloxanes are absorbed. The latter are eliminated unchanged in expired air and althoughmore polar in urine, still can be extracted with organic solvents. Thus, Frazer did not utilize appropriate methodology to detect absorption. Furthermore, the great variation in just the control Panel Book Page 297

300 37 urinary silicon levels determined by Frazer compared to other data shown in Table 14 likely suggest that the interfering urinary phosphate was not eliminated completely. This suggestion ~s supported by the work of Jankowiak and Le Vier (2) of Dow Corning 2. Jankowiak, M.E. and Le Vier, R.R. Elimination of Phosphorous Interference in the Colorimetric Dete~ination of Silicon in Biological Material. Anal. Biochem. 44:462472, who have recently examined this problem of phosphorous interference in biological samples. On the basis of dat~ presented in this study', we conclude that at a single dose level ofo.25 mg/kg (equivalent to 10 ppm daily dietary intake) of A Compound, containing ca. 10% low molecular weight polymeric species, the potential exists for a maximum total absorption of 25 pg/kg organosiloxane depending on the character of the diet. However, at a dietary intake level of 4000 ppm, there is essentially no absorption of silicon in any form if M Compound or Emu1sion is administered which contains less than 0.022% low molecular weight species at or below dodecameth~lhexacyc10siloxane. V. New Compound :Index None VI. Information Retrieval Index DOW CORNrNG~ Antifoam A Fluid DOW CORNrNG~ Antifoam A Compound DOW CORNING~ Antifoam A Emulsion DOW CORNrNG~ Antifoam M Fluid DOW CORNING S Antifoam M Compound DOW CO~1ING~Antifoam M Emulsion Mouse, Buckbeig Monkey', Macaca mu1lata Human Silica, Santoce1 aerogel Po1ydimethylsi10xane Polyd.imethylcyc10si10xanes Pharmacokinetic Data on Organosilicon Compounds Metabolic Data on Organosi1icon Compounds Silicon Analysis VII. Documentation The info:rmation presented iii this report has been assembled from individual experiments contained in individual pocket files from 1968 to 1974, and are located for convenience in the general files under the title of Antifoam Study Submitted to FAO/WHO Panel Book Page 298

301 CODING FORMS FOR SRC INDEXING Microfiche No. OTS New Doc ID Old Doc ID Date Produced 10/12/98 Date Received 11/06/98 TSCA section 8D SUbmitting Organization DOW CORNING CORP Contractor HUNTINGDON LIFE SCIENCES LTD Document Title BACTERIAL REVERSE MUTATION ASSAY OF DOW CORNING AF EMULSION FOOD GRADE, WITH COVER LETTER DATED 10/30/1998 Chemical Category DIMETHYL SILICONES AND SILOXANES ( ) Panel Book Page 299

302 ~~~~~ ". DOW CORNING October 30,1998 CERT # Z <.0 co TSCA Document Processing Center (7407) Office ofpollution Prevention and Toxics U.S. Environmental Protection Agency 401 M Street S.W. Washington, D.C Attn: 8(d) Health and Safety Data Reporting Rule I 0... ~:::'"J '' r>'1.,ot.0:)! t"rt ~~~ ~.t:=; Re: Dear Sir: 58 FR28511 (May 14,1993) [OPPTS82040; FRL41721] Health and Safety Data Reporting Submission oflists and Copies ofhealth and Saf~ No CD' The enclosed information is submitted on behalfofdow Corning Corporation, Midland, Michigan, , in compliance with a health and safety data rule issued under Section 8(d) ofthe Toxic Substances Control Act (TSCA) and with an effective date ofjune 14, 1993 (Sunset Date June 30, 1998), and in accordance with 40 CFR 716 (Health and Safety Data Reporting). Listed Chemical Substance: C Dimethyl silicones and siloxanes, 25 wt.% Dimethyl silicones and siloxanes, reaction products with silica, 2 wt.% Remainder: water and organic nonionic surfactants and preservatives in DOW CORNING AF Emulsion, Food Grade '2.;', N W ~... " >~.. :., <'0 co ~,P~ Title ofrecently Completed Study: BACTERIAL REVERSE MUTATION ASSAY OF DOW CORNING AF ~ EMULSION FOOD GRADE 0 :.:.::~,.::: ::;;: (") 1'"'1 C==;O ry N I'v Dow C()rning Curporatlon Midland, Michigan 48G8()'099~1 Phone: (517) 1% 1000 Dow Corning Corporation October 12, ""'""'" Panel Book Page 300

303 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland, Michigan For purposes ofhealth and safety data reporting under TSCA Section 8(d), the general INTERNAL designation on the attached health and safety study is waived by Dow Corning. Ifyou require further information regarding this submission, please contact Dr. Rhys G. Daniels, Regulatory Compliance Specialist, Regulatory Compliance Group, HERA Americas, at or at the address provided herein. Sincerely,.OJ?//.qf,<API"" Michael P. Hill l Americas Vice President and Corporate Director Health and Environmental Sciences (517) RGD98183 Panel Book Page 301

304 DCC Study No h1emal No. DCN 088/ Report No Security Internal Bacterial mutation assay ofdow Corning"" AF Emulsion Food Grade ABSTRACT In this in vitro assessment ofthe mutagenic potential of Dow Coming AF Emulsion Food Grade, histidine dependent auxotrophic mutants ofsalmonella typhimurium (strains TA1535, TAI53?, TA98 and TAIOO) and tryptophan dependent mutants of Escherichia coli (strains WP2 and WP2 uvra) were exposed to the test substance diluted in water. which was also used as a negative control. A preliminary toxicity test and two independent mutation tests were performed in the presence and absence.of liver preparations from Aroclor l254induced rats (S9 mix). All the tests included a preincubation stage. Dose levels of up to 5000 Ilg/plate were tested in the preliminary toxicity test. This is the standard limit dose recommended in the regulatory guidelines this assay follows. No signs oftoxicity were observed towards the tester strains so 5000 Ilg/plate was again chosen as the highest concentration in the mail1 mutation test. Other dose levels used were a series of ca halflog Jo dilutions ofthe highest concentration ( , 150 and 50 Ilg/plate). No evidence of mutagenic activity was seen at any dose level of Dow Coming AF Emulsion Food Grade in either mutation test. The concurrent positive controls demonstrated the sensitivity of the assay and the metabolising activity of the liver preparations. It is concluded that, when tested as an emulsion in water, Do\y Corning"" AF Emulsion Foodgrade shows no evidence of mutagenic activity in this Bacterial Reverse Mutation Assay. Page 3 of25 Panel Book Page 302

305 DCC Study No Extemal No. DCN 088/ Report No Security Intemal Bacterial mutation assay ofdow Coming'" AF Emulsion Food Grade n. EXPERIMENTAL PROCEDURE A, Test Substance Dow Corning"" AF Emulsion Food Grade, lot number HH0868 1I, was received at Huntingdon Life Sciences Ltd. on January 29, The test substancewas an offwhite liquid ':ll1cl was stored at room temperature. The Huntingdon Test Substance Data Sheet indicated that the test substance was stable until August 30, The test substance as received is n.:garded as the "pure" material and is representativeof Do\\' Corning'" AF Emulsion Food Grade. Test substance characterisation was carried out at Huntingdon Life Sciences Ltd (Huntingdon Life Sciences Schedule Number: DCN/089, DCC Study Number: 8736). All remaining test substancewill be returned to the Sponsor after the completion ofall the relevant stu~lies. B. Bacterial Strains: The following strainswere used: S typhil111lri1l111 TAI535 htsg46ifa uvrb S'.typhil11llrilll11 TA1537 hisc3076 rja uvrb S. typhil11llrilll11 TA98 hisd3052 rja uvrb pkmlo 1 S. typhimllrilll11 TAIOO hisg46 rfa livrb pkmi0l E coli WP2 trp E coli WP2 trp livra The strains of S. lyphil1111rillm were obtained from Professor B.N. Ames, University of Califomia, Berkeley, California, USA. The strains ofe. coli were obtained from the National Collections oflndustrial and Marine Bacteria, Aberdeen, Scotland. Batches of the strains were obtained from master stocks held in liquid nitrogen. The test batches were aliquots of nutrient broth cultures and were stored at 80 C. The test batches used in this assay were less than 6 months old. Dimethyl sulphoxide (DMSO) was added to the cultures at 8% v/vas a cryopreservative. Each batch offrozen strain was tested, where applicable. for cell membrane penneability (ifa mutation), sensitivity to UV light and the pkm101 plasmid which confers resistance to ampicillin. The batches of strains used all responded as expected in these tests. The responses of the strains to a series of diagnostic mutagens were also assessed. The responses obtained for each strain were as expected. Histidine/tryptophan auxotrophy was demonstrated by recording the number of spontaneous revertant colonies that grew on the solvent control plates. For use in tests an aliquot offrozen culture, which had been thawed at room temperature, was added to 25 ml ofnutrient broth (Merck No.2) and incubated, with shaking, at 37±2 C for 10 hours. after which they were stored at 4 C until testing to prevent further growth. These cultun.:s provided at least 2 x 10 9 cells per ml which were measured photometrically. Page 9 of25 Panel Book Page 303

306 ". DCC Study No External No. DCN 088/ Report No Security Internal Bacterial mutation assay ofdow Corning"" AF Emulsion Food Grade C. Positive Control Compounds: The SUibility of the positive controls and their stock 'solutions, which were prepared every two weeks, were demonstrated by the ability to provoke a mutagenic response on the positive control plates. The dose levels selected for the positive controls were below toxic levels and were the lowest levels that consistently produce at least a doubling ofmean revertants over the concurrent mean solvent control. These were prepared by diluting from the stock solutions. The following records for each batch of the positive controls are kept to satisfy USEPA TSCA GLP Standards, 40CFR Part 792 Subpart F a&b: identity, supplier, grade, purity/composition, expiration date, storage and solubility. These records are kept in laboratory books and were obtained from the container labels. supplier catalogues and certificates ofanalysis when supplied. In the absence ofs9 mix Identity: Supplier: Batch number: Appearance: Grade: Purity: Storage: Expiration date: Solvent: Concentration: Identity: Supplier: Batch number: Appearance: Grade: Purity: Storage: Expiration date: Solvent: Concentration: NEthylN'nitroNnitrosoguanidine Sigma Chemical, Poole, Dorset, England. 67F3700 Pale yellow crystalline powder Standard Laboratory Reagent 98% 20 C None supplied DMSO (soluble at a minimum of I mg/ml) 5 /lg/plate for strain TAl535 3 /lg/plate for strain TAl00 2 /lg/plate for strain WP2 and WP2 uvra 9Aminoacridine Sigma Chemical, Poole, Dorset, England. 35HI408 Yellow powder Standard Laboratory Agent Minimum 98% 4 C None supplied DMSO (soluble at a minimum of8 mg/ml) 30 /lg/plate for strain TAl537 Page 10 of 25 Panel Book Page 304

307 DCC Study No External No. DCN 088/ Report No Security Internal Bacterial mutation assay ofdow Coming"" AF Emulsion Food Grade. Identity: Supplier: Batch number: Appearance: Grade: Purity: Storage: Expiration date: Solvent: Concentration: 2Nitrofluorene Aldrich Chemical Company, Gillingham, Dorset, England Beige powder Standard Laboratory Agent 98% 4 C None supplied DMSO (soluble at a minimum of I mg/ml) I flg/plate for strain T A9R In the presence ofs9 mix Identity: Supplier: Batch number: Appearance: Grade: Purity: Storage: Expiration date: Solvent: Concentration: 2Aminoallthracene Aldrich Chemical Company, Gillingham, Dorset, England Green powder Standard Laboratory Agent 96% 4 C None supplied DMSO(soluble at a minimum of 2 mg/ml) 2 flg/plate for strains TAI535 <md TAI flg/plate for strain TA98 I J.1g/plate for strain TAIOO 50 ~lg/plate for strain WP2 10 flg/plate for strain WP2 llvra The teclmical details ofthe solvent used for the positive controls were as follows: Identity: Supplier: Batch number: Appearance: Grade: Purity: Stability: Storage: Date of arrival: Expiration date: DMSO Fisher Scientific, Loughborough, Leicestershire, Engl<ll1d Clear colourless liquid AnalR 99.96% Stable at room temperature Room temperature April 1997 Not supplied The location of the methods of synthesis, fabrication and derivation of the positive control SubsWlces is llllkno\\11. Page II of25 Panel Book Page 305

308 Dee Study No External No. DCN 088/ Report No Security Internal Bacterial mutation assay ofdow Coming~AF Emulsion Food Grade m. ASSESSMENT OF RESULTS For a tcst to be considered valid, the mean of the solvent/vehicle control revertant colony numbers for each strain must lie in the following ranges based on the laboratory's historical control values. TA1535 TA1537 TA98 TAlOO WP2 WP2uvrA l Also, the positive controls must cause at least a doubling of mean revertant colony numbers over the mean of the solvent/vehicle control. The mean number of revertant colonies for all treatment groups were compared with those obtained for solvent/vehicle control groups. Plates that were contaminated, had excess precipitation or exhibited excessive toxicity were rejected. However, there were be no fewer than two plates per condition. giving an acceptable test. The mutagenic activity of a test substance was assessed by applying the following criteria: (a) If treatment with a test substance produced an increase in the mean revertant colony numbers of at least twice the mean concurrent solvent/vehicle controls, with some evidence of a positive doserelationship, in two separate experiments, with any bacterial strain either in the presence or absence of S9 mix, it was considered to show evidence of mutagenic activity in this test system. No statistical analysis was performed. (b) Iftreatment with a test substance did not produce reproducible increases ofat least 1.5 times the mean concurrent solvent/vehicle controls, at any dose level with any bacterial strain, it was considered to show no evidence ofmutagenic activity in this test system. No statistical analysis was perfonned. IV. MAINTENANCE OF RECORDS The protocol and all amendments/deviations as _well as all raw data, a sample of the test substance and study related documents generated during the course ofthe study at Huntingdon Life Sciences Ltd.. together with the original final report will be lodged in the Huntingdon Life Sciences Archives, HlUltingdon. Huntingdon Life Sciences Ltd.. P.O. Box 2, Huntingdon, Cambridgeshire, PEI8 6ES, England. Page 15 of25 Panel Book Page 306

309 DCC Study No External No. DCN 088/ Report No Security Internal Bacterial mutation assay ofdow Corning"" AF Emulsion Food Grade Such records will be retnined for a minimum period of five years from the date of issue of the final report. At the end of the tivg yem retention period the client will be contacted and advice sought on the future requirements. Under no circumstances will any item be discarded without the sponsor's prior approval. V. RESULTS The individual plate counts for Dow Corning"" AF Emulsion Food Grade obtained in the preliminary toxicity test with the testt:r,trnins nre shown in Table I. No toxicity was observed towards the tester strains so 5000 J.lg/platc wns clloscn as the top dose level for the main test. The mean number of rew:rtnnt colonies, together with the individual plate counts for Dow Corning"" AF Emulsion Food Gmde obtnined in the first main test with the tester strains are shown in Table 2. Positive control mutability checks are sho\\~1 in Table 3. The mean number of revertant colonies, together with the individual plate counts for Do\\' Coming"" AF Emulsion Food Grade obtained in the second main test with the tester strains are sho\\11 in Table 4. Positive control mutnbility checks are sho\\~ in Table 5. No substantial increases in revertant colony numbers of any of the tester strains were observed following treatment with Dow Corning"" AF Emulsion Food Grade at any dose level, in the presence or absence ofs9 mix, in either mutation test. No plates were rejected due to excessive precipitate or toxicity. No contamination was observed on any of the plates reported. However, in the first test TAl535 was contaminated. so this part of the test was repeated and the data reported obtained from the repeat test. The mean revertant colony counts for the solvent controls were within the historical range. The concurrent positive controls demonstrated the sensitivity ofthe assay and the metabolising activity of the liver preparations by causing increases over double the concurrent solvent control. VI. CONCLUSION It is concluded thnt, when tested as an emulsion in water, Dow Corning"" AF Emulsion Food Grade shows no evidence ofmutagenic nctivity in tlus Bacterial Reverse Mutation Assay. Pngc 16 of25 Panel Book Page 307

310 CODING FORMS FOR SRC INDEXING Microfiche No. OTS New Doc ID Old Doc ID Date Produced 07/14/99 Date Received 08/10/99 TSCA section 8D Submitting organization DOW CORNING CORP Contractor HUNTINGDON LIFE SCIENCES LTD EYE IRRITATION STUDY OF DOW CORNING AF EMULSION FOOD GRADE (CONTAINS 25 WT.% OF DIMETHYL SILICONES AND SILOXANES) IN THE RABBIT, WITH COVER LETTER DATED 08/04/1999 Chemical Category DIMETHYL SILICONES AND SILOXANES ( ) Panel Book Page 308

311 DOWCORNING """""...,.:l"'",~.;;, > RECEIVED QPPT CBIG 1999 AUG 10 Mi [i; [; I August 4, 1999 CERT # TSCA Document Processing Center (7407) Office ofpollution Prevention and Toxics U.S. Environmental Protection Agency 401 M Street S.W. Washington, D.C Attn: 8(d) Health and Safety Data Reporting Rule Re: 58 FR (May 14, 1993) [OPPTS82040; FRL41721] Health and Safety Data Reporting Submission ofusts and Copies ofhealth and Safety Studies Dear Sir: The enclosed information is submitted on behalfofdow Corning Corporation, Midland, Michigan, , in compliance with a health and safety data rule issued under Section 8(d) ofthe Toxic Substances Control Act (TSCA) and with an effective date ofjune 14, 1993 (Sunset Date June 14,2003), and in accordance with 40 CFR 716 (Health and Safety Data Reporting). Listed Chemical Substances: Dimethyl silicones and siloxanes, 25 wt% Dimethyl silicones and siloxanes, reaction products with silica, 2 wt.% Remainder: water and organic nonionic surfactants and preservatives Title of Recently Completed Study: in DOW CORNING AF Emulsion, Food Grade EYE IRRlTATION STUDY OF DOW CORNING AF EMULSION FOOD GRADE IN THE RABBIT Dow Corning Corporation July 14, 1999 Contain NO cal r;; :"', ~'I,:l._~. I,..,., z< C)jTl DO Dow C)tnln~ l:orporatiol1 \Helland, \lilhig.1l1 IKGK(;"O\lM Phont.': CH 7) "l~h)"'1o()(} Panel Book Page 309

312 Manufacturer: Dow Corning Corporation 2200 West Salzburg Road Midland, Michigan For purposes ofhealth and safety data reporting under TSCA Section 8(d), the general INTERNAL designation on the attached health and safety report is waived by Dow Corning. Ifyou require further information regarding this submission, please contact Dr. Rhys G. Daniels, Senior Regulatory Compliance Specialist, Regulatory Compliance Group, HERA Americas, at or at the address provided herein. Sincerely, ~~ Director, Laboratory Heal Sciences Health and Environmental Sciences (517) RGD99I10 Panel Book Page 310

313 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Corning AF Emulsion Food Grade in the rabbit ABSTRACT The objectiveofthis study was to assess the eye irritation potential of Dow Corning AF Emulsion Food Grade, following a single instillation to the rabbit eye. The study was conducted using EPA Health Effects Testing Guidelines, Subpart E Specific OrganlTissue Toxicity Primary eye irritation, September 27, 1985 (described in Federal Register Vol. 50, No. 188) and subsequent revisions. Subpart E provides detailed information relating to data requirements of40 CFRPart798 and supports the Toxic Substances Control Act (TSCA). Dow Corning AF Emulsion Food Grade, was administered at a dose of0.1 ml into the lower everted lid ofone eye ofsix male rabbits. All rabbits were scored for irritation for three days (approximately 24, 48 and 72 hours) after test substance instillation. Additional observations were made four days after instillation in three rabbits and seven days after instillation in two rabbits. There was no evidence ofsystemic response to treatment. A single instillation ofdow Corning AF Emulsion Food Grade into the eye ofthe rabbit elicited a diffuse crimson colouration ofthe conjunctivae and slight swellingoftheeyelids in oneanimal. Slightredness ofthe conjunctivaealone was seen in the remaining five animals. Ocular reactions had resolved completely in all animals by one, three or seven days after instillation. All corneal and iridial scores for all animals at all observation times were zero. The mean conjunctival redness score (24, 48 and 72 hours) was 0.6. As a result ofthe ocular reactions observed, Dow Coming AF Emulsion Food Grade is classified as nonirritant in the European Union. Page 3 of22 Panel Book Page 311

314 DCC Study No External No. DCN 087/983467/SE Report No. 1998IO Security Internal 1. INTRODUCTION Eye Irritation Study ofdow Corning AF Emulsion Food Grade in the rabbit Eye Irritation Study ofdow Corning AF Emulsion Food Grade in the Rabbit The objective of this study was to assess the eye irritation potential of Dow Corning AF Emulsion Food Grade following a single instillation into the rabbit eye. II. MATERIALS AND METHODS A. Test Substance: Dow Corning AF Emulsion Food Grade, lot number HH086811, was received at Huntingdon Life Sciences Ltd. on January 29, The test substance was an offwhite liquid, and was stored at room temperature. The Huntingdon test substance safety data sheet indicated that the test substance was stable until August 30, The test substance, as received, is regarded as the "pure" material and is representative of Dow Corning AF Emulsion Food Grade. All the remaining test substance will be returned to the sponsor after the completion of all the relevant studies, with the exception of a 1 g sample which will be retained by Huntingdon Life Sciences Ltd. Test substance characterisation was carried out at Huntingdon Life Sciences Ltd. (Schedule number DCN/089, DCC Study number 8700). As the test substance is administered as supplied, assessment of solubility was not applicable in this study. The ph of the test substance was measured at Huntingdon Life Sciences Ltd. using a ph meter; an undiluted sample of the test substance gave a ph measurement of 2.9. This was confirmed in the characterisation test and reported in DCN/089. The absorption ofthe test substance was not quantitated. B. Dosage Formulation: The test substance was administered undiluted. C. Animals: New Zealand White rabbits weighing 1927 to 2663 g at receipt were obtained from Harlan UK Ltd, Shaw's Farm, Blackthorn, Bicester, Oxon, England on March 19, 1998, and kept in isolation. They Were observed daily for signs of illhealth and following a review of health monitoring procedures (absence of clinical observations and satisfactory body weight gain) by a veterinary officer, six healthy rabbits were randomly selected from the stock order after 28 days of acclimatisation. All six rabbits were male. The animals were identified by a numbered aluminium tag placed through the edge of one ear on arrival. These numbers were unique within the Huntingdon Life Sciences Ltd. Acute Toxicology Department throughout the duration ofthe study. The cage was identified by a coloured label displaying information relating to the study. Rabbits ofthe New Zealand White strain were chosen as the test species as they have been shown to be a suitable model for eye irritation studies and are the species recommended in the test guidelines. The rabbits were dosed by instillation into the eye as the test substance may come into contact with the eye during handling Or use. Page 8 of22 Panel Book Page 312

315 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Coming AF Emulsion Food Grade in the rabbit D. Food and Water: The rabbits were provided, ad libitum, with a standard laboratory diet, SDS Stanrab (P) SQC Rabbit Diet (supplier: Special Diet Services Ltd, Witham, Essex) and drinking water via an automatic watering system (supplier: Anglian Water). Autoclaved hay was supplied three times weekly. The batches ofdiet were analysed once, by the supplier, for nutrients, possible contaminants and microorganisms, likely to be present in the diet, and which, if in excess may have an undesirable effect on the test system. Results of routine physical and chemical analyses of drinking water performed by the supplier were made available to Huntingdon Life Sciences Ltd. as quarterly summaries. Water was supplied in conformity with EC Directive IEEC and UK Water Act 1989 and subsequent amendments. No contaminants capable ofadversely affecting the integrity or interpretation of the results from this study were known to be present in the basal diet or the drinking water during the conduct ofthis study. The Study Director reviewed the feed and water analyses. A certificate of analysis for each batch of diet used is appended to this report together with the most recent summary of results ofroutine physical and chemical analyses ofdrinking water performed by the supplier. The certificates of analyses will be lodged in Huntingdon Life Sciences Ltd. Archives. Samples ofwater were taken from the drinking water at source in the animal room prior to the study start The samples were analysed for microbial contaminants (total viable count, coliform count and E.Coli count) by Huntingdon Life Sciences Ltd. DepartmentofMicrobiology. A certificate ofanalysis is appended to this report E. Housing and Environment: The rabbits were housed individually in a suspended metal cage with perforated floor measuring 45.5 cm high, 76 cm wide and 60.5 cm deep (floor area 4598 cm2). The cage size is in compliance with UK animal welfare guidelines. Absorbent cage liners were placed in the pan below the metal mesh floor ofthe animal cage to absorb liquids. During the treatment phase of the study, animal room temperature and relative humidity were continuously recorded, using a seven day recorder. Minimum and maximum parameters were noted daily and ranged from 18 to 23 C and 40 to 62%, respectively. Air exchange was set to provide approximately 18 air changes per hour. Fluorescent lighting was controlled by means of a time switch and provided 12 hours of artificial light ( hours) which was followed by 12 hours ofdarkness in each 24 hour period. F. Methods: 1. Animals: The six male rabbits were allocated to the study using a random numbers table. Animals were in the bodyweight range 2593 g to 2984 g and were approximately 14 to 16 weeks ofage on Day 1 ofthe study. Page 9 of22 Panel Book Page 313

316 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Corning AF Emulsion Food Grade in the rabbit 2. Dosing: The New Zealand White rabbit has been shown to be a suitable model for eye irritation studies and is the species recommended in the guidelines. The rabbits were dosed by instillation into the eye as the test substance may come into contact with the eye during handling or use. The eyes of each animal were examined prior to instillation of the test substance to ensure that there was no preexisting corneal damage, iridial or conjunctival inflammation. One animal was treated in advance of the others, to ensure that if a severe response was produced, no further animals would be exposed (pilot animal see Table I). A single 0.1 ml dose ofthe test substance, was placed undiluted into the lower everted lid of one eye of each animal. The eyelids were gently held together for one second after instillation before releasing. The contralateral eye remained untreated. 3. Observations: The rabbits were observed daily for mortality and morbidity. 4. Body Weights: The rabbits were weighed on arrival and immediately prior to dosing. 5. Clinical signs: The rabbits were observed daily for any signs of ill health and toxicity. Observations were made at the cageside during the twice daily standard mortality and morbidity checks and when animals were removed from the cage to determine ocular responses. 6. Ocular Responses: The eye oftherabbits was examined 1 hour and 1,2 and 3 days after instillation (approximately 24, 48 and 72 hours). Additional observations were made four days after instillation in three rabbits and seven days after instillation in two rabbits. Observation ofthe eyes was aided by the use ofa handheld light. At each interval, ocular irritation was assessed according to the following prescribed arbitrary numerical system (based on Draize JH, Appraisal of the Safety ofchemicals in Foods, Drugs & Cosmetics, Assoc. Food & Drug Officials ofthe US, Austin, TX; 1959): (A) (B) Cornea Opacity: degree ofdensity (most dense area used) No ulceration or opacity 0 Scattered or diffuse areas, details ofiris clearly visible 1* Easily discernible translucent areas, detaijs ofiris slightly obscured 2* Opalescent areas, no details ofiris visible, sizeofpupil barejy discernible 3* Opaque, iris invisible 4* Areaofcornea involved One quarter (or less) but not zero Greater than one quarter but less than half Greater than halfbut less than three quarters Greater than three quarters up to whole area J The Total Score =(A x B) x 5, Maximum Total = 80 Page 10 of22 Panel Book Page 314

317 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Coming AF Emulsion Food Grade in the rabbit (C) Iris Normal 0 Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any ofthese or combination ofany thereof, iris still reacting to light (sluggish reaction is positive) 1* No reaction to light, haemorrhage, gross destruction (any or all ofthese) 2* Conjunctivae The Total Score = C x 5, Maximum Total = 10 (D) Redness (refers to the severe reading ofpalpebral and bulbar conjunctivae, as compared to the control eye) Blood vessels normal Some blood vessels definitely hyperaemic (injected) Diffuse, crimson colour, individual vessels not easily discernible Diffuse beefy red o 1 2* 3* (E) (F) Chemosis (lids and/or nictating membranes) No swelling Any swelling above normal (includes nictating membranes) Obvious swelling with partial eversion oflids Swelling with lids about halfclosed Swelling with lids more than halfclosed Discharge o 1 2* 3* 4* No discharge 0 Any amount different than normal (does not include small amounts observed in inner canthus ofnormal animals) 1 Discharge with moistening ofthe lids and hairs just adjacentto lids 2 Discharge with moistening ofthe lids and hairs and considerable area around the eye 3 The Total Score =(D + E + F) x 2, Maximum Total =20 The maximum total score is the sum ofall scores obtained from the cornea, iris and conjunctivae. Maximum total score possible = 110. * indicates a positive effect. Page 11 of22 Panel Book Page 315

318 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Coming AF Emulsion Food Grade in the rabbit For each animal, mean scores for corneal opacity, iris lesions, conjunctival erythema and conjunctival chemosis were calculated by adding the scores for the respective parameter at 24, 48 and 72 hours and dividing bysix. The irritation potential ofthe test substance was assessed according to the criteria described in Appendix 4 which are based upon the European Economic Community guidelines (93/2IIEEC): 8. Animal Disposition: After the final observation (April 20, I998 for pilot animal or April24 or 28, 1998 for remaining five animals) the rabbits were sacrificed by an intravenous overdose, into the marginal ear vein, of phenobarbitone sodium B.P. 200 mg/ml (Expiral manufactured by Sanofi Animal Health Ltd., Rhodes Way, Walford, Hertfordshire, England) and discarded without necropsy. G. Location of Study records: The protocol and all raw data as well as a sample of the test substance and study related documents generated during the course of the study at Huntingdon Life Sciences Ltd., together with a copy of the final report are lodged in the Huntingdon Life Sciences Ltd., Archive, Huntingdon, England. Such records will be retained for a minimum period offive years from the date of issue ofthe final report. At the end of the five year retention period the client will be contacted and advice sought on the future requirements. Under no circumstances will any item be discarded without the client's prior approval. H. Statistical Analyses: None conducted III. RESULTS A. Mortalitv: No deaths occurred during the study. B. Body Weights: The weight for the rabbits used in this study was in the range 2593 and 2984 g at treatment initiation. C. Clinical Signs: There were no signs ofsystemic reaction to treatment. D. Ocular Responses: A single instillation of Dow Corning AF Emulsion Food Grade into the eye ofthe rabbit elicited a diffuse crimson colouration ofthe conjunctivae and slight swelling of the eyelids in one animal. Slight redness of the conjunctivae alone was seen in the remaining five animals. The mean conjunctival redness (24, 48 and 72 hours) was 0.6. Ocular reactions had resolved completely in ali animals by one, three or seven days after instillation. All corneal and iridial scores for allanimalsatali observationtimes werezero. Page 12 of22 Panel Book Page 316

319 DCC Study No External No. DCN 087/983467/SE Report No Security Internal Eye Irritation Study ofdow Coming AF Emulsion Food Grade in the rabbit IV. CONCLUSION Under the conditions ofthis study, Dow Corning AF Emulsion Food Grade elicited very slight to welldefined conjunctival irritation (diffuse crimson colouration). Based on European Economic Community guidelines (93/21/EEC) this test substance is classified as a nonirritant in the European Union. Page 13 of22 Panel Book Page 317

320 CODING FORMS FOR SRC INDEXING Microfiche No. OTS New Doc ID Old Doc ID Date Produced 07/20/99 Date Received 08/10/99 TSCA section 8D Submitting organization DOW CORNING CORP Contractor HUNTINGDON LIFE SCIENCES LTD SKIN IRRITATION STUDY OF DOW CORNING AF EMULSION FOOD GRADE (CONTAINS 25 WT.% OF DIMETHYL SILICONES AND SILOXANES) IN THE RABBIT, WITH COVER LETTER DATED 08/04/1999 Chemical Category DIMETHYL SILICONES AND SILOXANES ( ) Panel Book Page 318

321 DOWCORNING August 4,1999 RECEIVED appt C8lC J999 AUG 10 Mill: 42 CERT # TSCA Document Processing Center (7407) Office ofpollution Prevention and Toxics U.S. Environmental Protection Agency 401 M Street S.W. Washington, D.C Attn: 8(d) Health and Safety Data Reporting Rule Re: 58 FR28511 (May 14,1993) [OPPTS82040; FRL41721] Health and Safety Data Reporting Submission oflists and Copies ofhealth and Safety Studies Dear Sir: The enclosed information is submitted on behalfofdow Corning Corporation, Midland, Michigan, , in compliance with a health and safety data rule issued under Section 8(d) ofthe Toxic Substances Control Act (TSCA) and with an effective date ofjune 14, 1993 (Sunset Date June 14, 2003), and in accordance with 40 CFR 716 (Bealth and Safety Data Reporting). Listed Chemical Substances: Dimethyl silicones and siloxanes, 25 wt.% Dimethyl silicones and siloxanes, reaction products with silica, 2 wt.% Remainder: water and organic nonionic surfactants and preservatives in DOW CORNING AF Emulsion, Food Grade N c.:>?': Title ofrecently Completed Study: SKIN IRRITATION STUDY OF DOW CORNING AF EMULSION FOOD GRADE IN THE RABBIT Dow Corning Corporation July 20, 1999 Contain NO CBI Dow Curnin~ Corpllratinn ~lidland. Mkhi~an ~~!ih(i()9~c! Phmw: 1:>1 7)!9f)IOOO I Panel Book Page 319

322 Manufacturer: Dow Coming Corporation 2200 West Salzburg Road Midland, Michigan For purposes ofhealth and safety data reporting under TSCA Section 8(d), the general INTERNAL designation on the attached health and safety report is waived by Dow Corning. Ifyou require further information regarding this submission, please contact Dr. Rhys G. Daniels, Senior Regulatory Compliance Specialist, Regulatory Compliance Group, HERA Americas, at or at the address provided herein. Sincerely, Patrick W. Langvardt Director, Laboratory Health Sciences Health and Environmental Sciences (517) RGD9911 I Panel Book Page 320

323 DOW CORNING CORPORATION HEALTH & ENVIRONMENTAL SCIENCES TECHNICAL REPORT Report No.: Title: Study No.: External Testing Facility No.: Test Substance: Study Director: Sponsor: Study Monitor: Testing Facility: Study Completion Date: Security Statement: HUNTINGDON LIFE SCIENCES LTD PO BOX2 HUNTINGDON CAMBRIDGESIDRE Skin Irritation Study ofdow Corning AF Emulsion Food Grade in the Rabbit 8734 DCN 086/983059/SE Dow Corning AF Emulsion, Food Grade Brenda I Parcell, M.LA.T. Dow Corning Corporation 2200 W Salzburg Road Midland Michigan USA C.D.Kary Huntingdon Life Sciences Ltd. PO Box 2 Huntingdon Cambridgeshire PE186ES ENGLAND 20 July 1999 PEl86ES ENGLAND This report may be reproduced and shared with any Dow Coming employee. Distribution outside the corporation must be approved by the Director of H.E.S. When this INTERNAL report is no longer needed, it may be placed in office waste baskets for destruction. Page I of 18 Panel Book Page 321

324 DCC Study No External No. DCN 086/ SE Report No Security Internal Skin Irritation Study ofdow Corning AF Emulsion, Food Grade in the Rabbit ABSTRACT The objective of this study was to assess the skin irritation potential of Dow Corning AF Emulsion, Food Grade following a single dermal application to the rabbit. The study was conducted using EPA Health Effects Testing Guidelines, Subpart E Specific Organ/Tissue Toxicity Primary dermal irritation, September 27, 1985 (described in Federal Register Vol. 50, No. 188) and subsequent revisions. Subpart E provides detailed information relating to data requirements of 40 CFRPart 798 and supports the Toxic Substances Control Act (TSCA). Dow Corning AF Emulsion, Food Grade was administered at a dose of 0.5 ml to one intact skin site on the clipped dorsoiumbar region of six male rabbits. Each treatment site was washed with warm water four hours after semioccluded application of the test substance. The rabbits were scored for irritation at 60 minutes and 24, 48 and 72 hours following removal ofthe dressings, following which the study was terminated. There was no evidence of systemic response to treatment. No dermal irritation was observed following a single semiocclusive application ofdow Corning AF Emulsion, Food Grade to intact rabbit skin for four hours. The Primary Irritation Index (PH) was calculated to be 0.0. Page 3 of 18 Panel Book Page 322

325 DCC Study No External No. DCN 086/983059/SE Report No. 1999IOOOO47071 Security Internal Skin Irritation Study ofdow Coming AF Emulsion, Food Grade in the Rabbit I. INTRODUCTION Skin Irritation ofdow Corning AF Emulsion, Food Grade in the Rabbit The objective of this study was to assess the skin irritation potential of Dow Corning AF Emulsion, Food Grade following a single dermal application to the rabbit. II. MATERIALS AND METHODS A. Test Substance: Dow Corning AF Emulsion, Food Grade, lot number HH086811, was received at Huntingdon Life Sciences Ltd. on January 29, The test substance was a offwhite liquid, and was stored at room temperature. The Huntingdon Test Substance Data Sheet indicated that the test substance was stable until 30 August The test substance, as received, is regarded as the "pure" material and is representative of Dow Corning AF Emulsion, Food Grade. All the remaining test substance will be returned to the sponsor after the completion of all the relevant studies, with the exception of a 1 g sample which will be retained by Huntingdon Life Sciences Ltd. Test substance characterisation was carried out at Huntingdon Life Sciences Ltd. (Schedule number DCN/089, DCC Study number 8736). As the test substance is administered as supplied, assessment ofsolubility was not applicable in this study. The ph of the test substance was measured at Huntingdon Life Sciences Ltd. using a ph meter; an undiluted sample of the test substance gave a ph measurement of 2.9. This was confirmed in the characterisation test and reported in DCN/089. The absorption of the test substance was not quantitated B. Dosage Formulation: The test substance was administered undiluted. C. Animals: New Zealand White rabbits weighing 1927 to 2663 g were obtained from Harlan UK Ltd, Shaw's Farm, Blackthorn, Bicester, Oxon, England on March 19, 1998, and kept in isolation. They were observed daily for signs of illhealth and following a review of health monitoring procedures (absence of clinical observations and satisfactory body weight gain) by a veterinary officer, six healthy rabbits were randomly selected from the stock order after 12 days of acclimatisation. All six rabbits were male. The animals were identified by a numbered aluminium tag placed through the edge of one ear on arrival. These numbers were unique within the Huntingdon Life Sciences Ltd. Acute Toxicology Department throughout the duration ofthe study. The cage was identified by a coloured label displaying information relating to the study. Rabbits ofthe New Zealand White strain were chosen as the test species as they have been shown to be a suitable model for skin irritation studies and are the species recommended in the test guideline. The rabbits were dosed by topical application as the test substance may come into contact with the skin during handling or use. Page 8 of 18 Panel Book Page 323

326 ~~~~~~ DCC Study No External No. DCN 086/983059/SE Report No Security Internal Skin Irritation Study ofdow Coming AF Emulsion, Food Grade in the Rabbit D. Food and Water: The rabbits were provided, ad libitum, with a standard laboratory diet, SDS Stanrab (P) SQC Rabbit Diet (supplier: Special Diet Services Ltd, Witham, Essex) and drinking water via an automatic watering system (supplier: Anglian Water). Autoclaved hay was supplied three times weekly prior to the study start.. The batch of diet was analysed once, by the supplier, for nutrients, possible contaminants and microorganisms, likely to be present in the diet, and which, if in excess may have an undesirable effect on the test system. Results of routine physical and chemical analyses of drinking water performed by the supplier were made available to Huntingdon Life Sciences Ltd. as quarterly summaries. Water was supplied in conformity with EC Directive 80/778/EEC and UK Water Act 1989 and subsequent amendments. No contaminants capable ofadversely affecting the integrity or interpretation of the results from this study were known to be present in the basal diet or the drinking water during the conduct ofthis study. The Study Director reviewed the feed and water analyses. A certificate of analysis for each batch of diet used is appended to this report together with the most recent summary of results of routine physical and chemical analyses of drinking water performed by the supplier. The certificates of analyses will be lodged in Huntingdon Life Sciences Ltd. Archives. Samples of water were taken from the drinking water at source in the animal room prior to the study start. The samples were analysed for microbial contaminants (total viable count, coliform count and E.Coli count) by Huntingdon Life Sciences Ltd. DepartmentofMicrobiology. A certificate ofanalysis is appended to this report. E. Housing and Environment: The rabbits were housed individually in a suspended metal cage with perforated floor measuring 45.5 cm high, 76 cm wide and 60.5 cm deep (floor area 4598 cm2). The cage size is in compliance with UK animal welfare guidelines. Absorbent cage liners were placed in the pan below the metal mesh floor ofthe animal cage to absorb liquids. During the treatment phase of the study, animal room temperature and relative humidity were continuously recorded, using a seven day recorder. Minimum and maximum parameters were noted daily and ranged from 20 to 22 C and 46 to 57%, respectively. Air exchange was set to provide approximately 18 air changes per hour. Fluorescent lighting was controlled by means of a time switch and provided 12 hours of artificial light ( hours) which was followed by 12 hours ofdarkness in each 24 hour period. F. Methods: 1. Animals: The six male rabbits were allocated to the study using a random numbers table. Animals were in the bodyweight range 2286 g to 2552 g and were approximately II to 14 weeks ofage on Day I ofthe study. Page 9 of 18 Panel Book Page 324

327 DCC Study No External No. DCN 086/983059/SE Report No Security Internal Skin Irritation Study ofdow Coming AF Emulsion, Food Grade in the Rabbit 2. Preparation: One day prior to application of the test substance (approximately 24 hours), hair was removed with electric clippers from the dorsoiumbar region of the rabbit taking care not to damage the skin, exposing an area of skin approximately 100 mm x 100 mm (approximately 10% of the total body surface area). The skin was not abraded. Clipping was repeated as necessary for evaluation of dermal responses, throughout the observation period. 3. Dosing: On the day oftreatment (April 1, 1998), a single 0.5 ml dose ofthe test substance, was applied using a 5 ml plastic syringe, under a 2ply 25 mm x 25 mm (6.25 cm2) porous gauze pad to the previously clipped intact dorsoiumbar skin site of each rabbit. The treatment area (approximately 100 mm x 100 mm) was covered by a semiocclusive dressing (Elastoplast elastic adhesive bandage 7.5cm) encircled firmly around the trunk of the animal, the end of which was secured with "Sleek. The animals were not restrained during the exposure period and were returned to their cages immediately after treatment. At the end ofthe 4 hours exposure period the dressings were carefully removed and the treated area of skin was washed with water (34 C) to remove any residual test substance. The treated area was blotted dry with absorbent paper. 4. Observations: The rabbits wereobserved daily for mortality and morbidity. 5. Body Weights: The rabbits were weighed on arrival and immediately prior to dosing. 6. Clinical signs: The rabbits were observed daily for any signs of ill health and toxicity. Observations were made at the cageside during the twice daily standard mortality and morbidity checks and when animals were removed from the cage to determine dermal responses. 7. Dermal Responses: The treated skin of the rabbits was examined on Day 1 (approximately 60 minutes after removal ofthe dressings) and on Days 2, 3 and 4 (24,48 and 72 hours after removal of the dressings). At each interval, dermal irritation was assessed according to the following prescribed arbitrary numerical system (based on Draize JR, Appraisal ofthe SafetyofChemicals in Foods, Drugs & Cosmetics, Assoc. Food & Drug Officials ofthe US, Austin, TX; 1959): Page 10 ofl8 Panel Book Page 325

328 DCC Study No External No. DCN 086/983059/SE Report No. 1999IOOOO47071 Security Internal Skin Irritation Study ofdow Corning AF Emulsion, Food Grade in the Rabbit G. Location of Study records: The protocol and all raw data as well as a sample of the test substance and study related documents generated during the course of the study at Huntingdon Life Sciences Ltd., together with a copy of the final report are lodged in the Huntingdon Life Sciences Ltd., Archive, Huntingdon, England. Such records will be retained for a minimum period of five years from the date of issue of the final report. At the end ofthe five year retention period the client will be contacted and advice sought on the future requirements. Under no circumstances will any item be discarded without the client's prior approval. H. Statistical Analvsis: None conducted. III. RESULTS A. Mortality: No deaths occurred during the study. B. Body Weights: The weight for the rabbits used in this study was in the range 2286 to 2552 g at treatment initiation. C. Clinical Signs: There were no signs ofsystemic reaction to treatment. D. Dermal Responses: No dermal irritation was observed following a single semiocclusive application ofdow Coming AF Emulsion, Food Grade to intact rabbit skin for four hours. The Primary Irritation Index (PH) was calculated to be 0.0. IV. CONCLUSION Under the conditions of this study, Dow Coming AF Emulsion, Food Grade elicited no dermal irritation. Page 12 of 18 Panel Book Page 326

329 21CFR Please In The In In The Personal Care Memorandum Products Council Committed to Safety, Quality & Innovation TO: FROM: F. Alan Andersen, Ph.D. Director COSMETIC NGREDIENT REVifiW (CIR) John Bailey, Ph.D. Industry Liaison to the CIR Expert Panel DATE: January 20, 2011 SUBJECT: Comments on the Scientific Literature Review of Silica Silylate, Silica Dimethyl Silylate, Trimethylsiloxysilicate and TrifluoropropyldimethyllTrimethylsiloxysilicate p.2, 7 concentration of use table provided for Silica Dimethyl Silylate, Trimethylsiloxysilicate and Trifluoropropyldimethyl/Trimethylsiloxysilicate did not include Silica Silylate in the title of the table. Therefore, Silica Silylate was not included in the concentration of use survey. Silica Silylate will be included in the next concentration of use survey. p.2 If particles of Silica Dimethyl Silylate used in perfumes are all >90 urn, why are effects on the lung of concern? p.2 (reference 12) states that Silicon Dioxide is an approved anticaking agent for use in food at up to 2%. 21CFR (reference 13) states that Silica aerogel is GRAS. Although these substances are related to Silica Dimethyl Silylate, it would be helpful if the actual substance listed in the CFR was stated. p.3 What was the radioactive material used to label Antifoam A and Antifoam M in the oral study in monkeys? Please provide the reference for the monkey and human studies. p.3 Please revise the following sentence. Increased silicon levels were not found in urine, feces, or expelled air after either oral Antifoam A. p.3 It is not clear what was being measured in the human study (reference 14?). Was a radiolabel used? Were they just measuring silicon? p.3 provide the particle size of the Silica Dimethyl Silylate used in the rat inhalation study (reference 15). p.3 describing the concentration that did not result in mortalities in rats, please add that this was a single four hour exposure, and include the reference. p.4 second sentence of the first paragraph uses basal diet and basic diet. Is there any difference between these two diets? p.4 the discussion of results of the oral rat study (reference 24), it is not clear what is meant by atrophic hepatocytes with decreased appearance. p.45 the description of the developmental study of Antifoam A (reference 27), it would be helpful to know how many rats per dose group were actually pregnant. It states that 28 rats were not pregnant, and 3 rats were excluded because of dosing errors, but it is not clear to which groups these rats belonged. The number of dose groups in this study is also not clear. p.6. Please revise the following incomplete sentence. An in vitro mammalian chromosome aberration test of silica dimethyl silylate (63, 125, 250, 500 ig/ml) using Chinese hamster ovary (CHO) cells with and without metabolic activation th Street, N.W., Suite 30O Washington, D.C (fax) Panel Book Page 327

330 Please What In It In Please CAS It In p.6 The p.7 p.7 p.7 In p.7 p.7 2 year rat study of Silica Dimethyl Silylate (reference 26) states: No carcinogenic effects (nature and incidence of tumors were comparable with the historical control data) and no other treatment related significant changes were observed. NOAEL is 100 mg/kg bw when mixed into the diet. The lack of carcinogenic effects should be stated in the Carcinogenicity section. include the route of exposure (oral) used in the study for which Silica Dirnethyl Silylate was found to be eliminated in the feces. Elimination primarily in the feces following oral exposure suggests Silica Dimethyl Silylate was not absorbed. is the source of the NOAEL of 500 mg/kg and LOAEL of 1000 mg/kg? These values are presented after a description of an inhalation study suggesting the route was inhalation, but the units for the inhalation studies are all concentrations, e.g., mg/rn 3, rather than doses. Please provide the route of exposure and the species for the NOAEL and LOAEL. the Summary, please provide the route of exposure and the maximum dose that did not result in reproductive and developmental toxicity. Please indicate that multiple species were studied in reproductive and developmental toxicity studies. the Summary, please mention the mammalian cell genotoxicity study. is not correct to state that There were not carcinogenicity studies discovered. As indicated above, the 2year rat study of Silica Dimethyl Silylate did not see increased tumor incidences compared to controls. p.7 the Summary, please provide the maximum concentrations of Silica Dimethyl Silylate that were not irritating to rabbit skin and eyes. p.9, Table 1 provide references for this table. The CAS numbers for the chlorinated compounds ( , , , ) and the other names including chlorine should not be listed under Silica Dimethyl Silylate. Although a supplier may consider the chlorinated compounds to belong under the name Silica Dimethyl Silylate, it is likely that the INCI committee would have given the chlorinated compounds a different name as the fluorine containing compound was given a different name. p.9, Table 1 reports for the CAS numbers listed as coming from trade name ingredients were checked (by Tara Gottschalck). The following CAS numbers were considered appropriate and added to the Dictionary database. Silica Silylate: , ; Silica Dimethyl Silylate: , ; Trimethylsiloxysilicate: p.10, Table 2 Please delete Silsoft 876 under Trimethylsiloxysilicate. This is an error in the Dictionary that has been corrected. The attached sheet shows the correct INCI name for this trade name. p.12, Table 4 is not clear why NS is included among the footnotes to Table 4 and Silica Silylate which has not yet been included in a concentration of use survey, is not included in this table. p.13, Table 5 Reference 3 (both the single and 3 day exposure studies) should be presented in the Absorption, Distribution, Metabolism and Excretion section under inhalation exposure. p.13, Table 5 the description of reference 53, it is not clear what is meant by controls showed signs of recovery at both time periods. What happened to the controls? The duration of the recovery period used in this study is not clear. 2 Panel Book Page 328