Antimicrobial AlphaSan Test Report Summary Table 08/21/01

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1 AlphaSan RC 5000 Physical Chemical Properties Acute Oral Toxicity, Rat EPA FIFRA 81-1 Acute Dermal Toxicity, Rat EPA FIFRA 81-2 Primary Dermal Irritation Rabbit EPA FIFRA 81-5 Primary Eye Irritation Rabbit EPA FIFRA 81-4 Skin Sensitization, Guinea Pig EPA FIFRA 81-6 Two-week Palatability Study in 90-Day Feeding Study in EPA FIFRA 82-1 Study was conducted in compliance with EEC Methods, Directive 92/69/EEC (OJ No. L383A, ) Part A, Methods A3, A6, A10, A16, OECD fasted rats 95 male + 5 female) were given a single dose of 5.0 g/kg body weight by gavage; the animals were sacrificed at 15 days for examination. 10 rats (5 male + 5 female) were given a single dermal application of 2.0 g/kg body weight; the animals were sacrificed at 15 days for examination Six rabbits were administered a single dermal dose of 0.5 g and observed for 4 days Six rabbits were administered a single ocular dose of 40 mg and observed for 7 days Guinea-pig maximization test by Magnusson and Kligman, 10 test and 5 control animals. 22 male and female rats were dosed from 0 to 1,000 mg/kg/day in the diet in 4 groups for 14 days. Groups of 10 male and 10 female rats were administered the test substance in the diet for 13 weeks at concentrations of 0, 30, 300, and 1,000 mg/kg/day. White Odorless Powder M.P.>1,000ºC D 4 20= Solubility: Ksp value less than Not Flammable Particle Size 93.47% < 4.022µm No deaths. Clinical signs of reaction to treatment were piloerecion and abnormal body carriage, with complete recovery by day 4. Oral LD 50 >5.0 g/kg body weight No deaths nor signs of systemic reaction. The sights of application showed no irritation or other dermal changes. No abnormalities observed. Dermal LD 50 >2.0 g/kg body weight No reactions were observed. Slight response in 4 rabbits, resolved in 2 days. No evidence of skin sensitization was found for any test animal. No deaths nor clinical signs of abnormality were observed. There were no significant differences in body weights. Target intakes of test substance were achieved. No deaths nor clinical signs of abnormality were observed related to treatment. Cholesterol levels were slightly elevated at the 300 and 1,000 mg/kg/day feeding level. No other microscopic changes were noted and all test animals were in general good health. For the study: NOAEL=1,000 mg/kg/day NOEL=30 mg/kg/day

2 AlphaSan RC 5000 Bacterial Mutation Assay EPA FIFRA 84-2 Mammalian Cell Mutation Assay OECD 476 Mouse Micronucleus Test EPA FIFRA 84-2 Acute Toxicity, Rainbow Trout EPA FIFRA 72-1 OECD 203 Acute Toxicity, Blue gill Sunfish EPA FIFRA 72-1 OECD 203 Acute Toxicity, Daphnia magna EPA FIFRA 72-2 OECD 202 Algal Growth Inhibition EPA FIFRA and OECD 201 in vitro assessment using Salmonella typhimurium with and without S-9 activation at 500, 150, 50, 15, 5, 1.5µg/plate in vitro using mouse lymphoma L5178Y cells with and without S-9 activation in vivo assay in mouse bone marrow cells with mice treated by single dose gavage at 1,250, 2,500, and 5,000 mg/kg Groups of 20 juvenile fish were exposed for a period of 96 hours in duplicate to 5 test concentrations from 1.0 to 10 mg/l with two controls. Groups of 20 juvenile fish were exposed for a period of 96 hours in duplicate to 5 test concentrations from 1.0 to 10 mg/l with two controls. Groups of 20 1 st instar Daphnia were exposed for a period of 48 hours in duplicate to 5 test concentrations from 1.0 to 10 mg/l with two controls. Selenastrum capricornutum cultures were exposed to seven test concentrations at 24ºC for 120 hours with continuous illumination. No evidence of mutagenic activity. Did not demonstrate mutagenic potential. No evidence of chromosome-damaging activity LC 50 =1.2 mg/l at 96 hours; NOEL=1.0 mg/l at 96 hours. LC 50 =1.7 mg/l at 96 hours; NOEL=1.0 mg/l at 96 hours. EC 50 =0.13 mg/l at 48 hours; NOEL=0.10 mg/l at 48 hours. E b C 50 = mg/l at 120 hours; NOEL=0.156 mg/l at 120 hours

3 AlphaSan RC 2000 Acute Oral Toxicity, Rat EPA FIFRA 81-1 Primary Dermal Irritation, Rabbit Bacterial Mutation Assay EPA OECD 471 and 472 Acute Inhalation Toxicity (Nose Only) Study in EPA FIFRA 81-3 OECD 403 Avian Acute Oral Toxicity in Bobwhite Quail with Experimental Additive BIO-LIFE Associates EPA FIFRA OPPTS Oral Gavage Teratology Study in EPA FIFRA OPPTS Dust Explosibility Classification 10 Fasted rats (5 male + 5 female) were given a single dose of 5.0 g/kg body weight by gavage; the animals were sacrificed at 15 days for examination. Six rabbits were administered a dermal dose of 0.5 to one intact and one abraded skin site and observed for 3 days in vitro assessment using Salmonella typhimurium with and without S-9 activation at 16 dose levels between 2,000 and µg/plate. five male and five female rats were held in a tapered, polycarbonate restraining tube fitting onto a single tier of the exposure chamber and sealed by means of a rubber 'O' ring. Only the nose of each animal was exposed to the test atmosphere. The dose for each individual test bird was gravimetrically measured and dispensed at the appropriate dosage rate into gelatin capsules. The test material was administered by oral insertion of the gelatin capsule into the esophagus, followed by a hand massage down the esophagus towards the proventriculus. Groups of twenty-five mated female Sprague- Dawley CD strain rats were dosed orally, by gavage, with the test material from Day 6 to 15 of gestation inclusive. The dose levels were 100, 300 and 1000 mg/kg/day. A concurrent control group was dosed with vehicle only. All reasonable measures are taken to ignite the dispersed dust sample in air under ambient temperature and pressure conditions. No deaths. Clinical signs of reaction to treatment were piloerecion with complete recovery by day 3. Oral LD 50 > 5.0 g/kg body weight Transient very slight irritation of observed at the abraded sites of three animals. Primary Irritation Index was calculated to be 0.1. No evidence of mutagenic activity was seen at any dose level. The LC50 of the test material has been determined to be greater than 5.18 mg/l (1/5 mortalities in the male group, 0/5 mortalities in the female group). The acute oral LD50 of Experimental Additive was determined to be greater than 2000 mg/kg body weight. With regard to sublethal effects (diarrhea was noted 7.5 hours post-dosing and on Test Day 3 of the study in the 2000 mg/kg group), a NOEL was not achieved in this study. With regard to mortality, the NOEL was 2000 mg/kg body weight. Females dosed with Experimental Additive number from Day 6 to 15 of gestation at dose levels up to and including 1000 mg/kg showed no evidence of maternal toxicity and no significant effects on the growth and development of offspring. Sample tested was classified as NOT EXPLOSIBLE.

4 AlphaSan RC 2000 Acute Eye Irritation Study in the Rabbit EPA FIFRA OPPTS OECD 405 Acute Dermal Toxicity Study in EPA FIFRA OPPTS OECD 402 Skin Sensitization, Guinea pig EPA FIFRA OPPTS OECD Day Dietary Toxicity Study in EPA FIFRA OPPTS OECD 408 Mouse Lymphoma Assay In- Vitro EPA FIFRA OPPTS OECD 476 Antimicrobial AlphaSan Test Report Summary Table 08/21/01 Three rabbits were treated with a 0.1ml ocular dose of the test material. Assessments were made at approximately 1 hour and 24, 48 and 72 hours following treatment. A group of ten animals ( five males and five females) were given a single 24- hour, semi - occluded dermal application to intact skin at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of treatment. Magnusson & Kligman maximization study in the Guinea pig. 10 test and 5 control animals. Thestudy was performed in compliance with OECD guideline 406, and OPPTS Groups of 10 male and 10 female rats were administered to the test substance in the diet for 13 weeks at a concentration of up to ppm. This test was performed to meet the requirements of OECD 408 and OPPTS L5178Y TK +/ c mouse lymphoma cells were treated with the test substance at up to six dose levels, in duplicate, together with solvent and positive controls. 3-Hour exposures were used both with and without activation in experiment 1. In experiment 2 the exposure time without activation was increased to 24 hours. A third experiment was performed using a 3-hour exposure without metabolic activation to confirm a result obtained in experiment 1. This test was performed to meet the requirements of OECD 476 and OPPTS The test material, Experimental Additive , produced a maximum group mean score of 12.0 and was classified as a Mild Irritant (Class 4 on a 1 to 8 Scale) to the rabbit eye according to a modified Kay and Calandra classification system. The acute dermal median lethal dose (LD 50 ) of the test material, Experimental Additive , in the Sprague-Dawley CD Strain rat was found to be greater than 2000 mg/kg bodyweight. The test material, Experimental Additive , produced a 0% (0/10) sensitisation rate and was classified as a non-sensitiser to guinea pig skin. Preliminary results available. No draft report as of yet Experimental Additive induced small but reproducible statistically significant increases in the mutant frequency at the TK +/- locus in L5178Y cells in the absence of metabolic activation. Experimental Additive was therefore considered to be mutagenic under the conditions of the test.

5 AlphaSan RC 2000 Mouse Micronucleus Study EPA FIFRA OPPTS OECD 474 Dietary Two Generation Reproduction Study in EPA FIFRA OPPTS OECD 416 Determination of the Solubility of Silver and Zirconium from Experimental Additive Antimicrobial AlphaSan Test Report Summary Table 08/21/01 This in vivo study was performed to assess the potential of the test material to produce damage to chromosomes or aneuploidy when administered to mice. The study was conducted using the intraperitoneal route in groups of seven mice (males) at the maximum recommended dose (2000 mg/kg) with 1000 and 500 mg/kg as the two lower dose levels. This test was performed to meet the requirements of OECD 474 and OPPTS Objective of this study is to investigate the effects of the test material on reproduction, prenatal development and post-natal development of the rat when administered to two successive generations. This report was conducted in accordance with the Environmental Protection Agency (EPA) Good Laboratory Practice Standards, 40 CFR 160. Experimental Additive was considered to be non-genotoxic under the conditions of the test. Study is in-process. Expest draft report April A solubility study was performed on Experimental Additive , an antimicrobial agent zirconium phosphate-silver complex. The concentration of zirconium and silver were determined for solvents saturated with the test substance at 25ºC. The concentrations of zirconium and silver were determined in the following solvents: Water buffered to ph 5 Water buffered to ph 7 Water buffered to ph 9 Pure Water (Processed through a Milli-Q purification system) The solvents were equilibrated with an excess of the test substance at 30ºC for 24, 48 and 72 hours. After equilibration the samples were reequilibrated at 25ºC for 24 hours. The samples were then filtered and an aliquot of each sample was sent to Galbraith Laboratories for analysis by Inductively Coupled Plasma Atomic Emmision Spectroscopy (ICP- AES). The results are summerized below. Solvent Zirconium Silver Average Solubility ppb Water No Recovery 146, 87.0 ph 5 Buffer No Recovery 93.5, 100 ph 7 Buffer 95.0, , 102 ph 9 Buffer 113, , 9.8

6 AlphaSan RC 2000 Fortified samples of zirconium and silver were prepared in each of the solvents. Duplicate samples were prepared for each analyte in each solvent. Zirconium was added to each of the solvents at a level of 0.01 µg/ml. Silver was added to the Milli-Q water at 0.01 µg/ml. Silver was added to the other solvents at a level of 20 µg/ml. The results of the analysis of these samples are summerized below. Solvent Recoveries (%) Zirconium Silver No Recovery 146, 87.0 No Recovery 93.5, , , , , 9.8

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