Regulation (EU) No 528/2012 concerning the making available on the market and use of biocidal products. Assessment Report

Transcription

1 Regulation (EU) No 528/2012 concerning the making available on the market and use of biocidal products Evaluation of active substances Assessment Report Margosa extract, cold-pressed oil of azadirachta indica seeds without shells extracted with Product-type 19 (Repellents and attractants) January 2017 eca: Germany

2 Product-type 19 April 2017 CONTENTS 1. STATEMENT OF SUBJECT MATTER AND PURPOSE Procedure followed Purpose of the assessment report OVERALL SUMMARY AND CONCLUSIONS Presentation of the Active Substance Identity, Physico-Chemical Properties & Methods of Analysis Intended Uses and Efficacy Classification and Labelling Summary of the Risk Assessment Human Health Risk Assessment Hazard identification and effects assessment Exposure assessment and risk characterisation Risk Characterisation Environmental Risk Assessment Fate and distribution in the environment Hazard identification and effects assessment PBT Assessment and Exclusion Criteria Exposure assessment and risk characterisation Overall conclusions Requirement for further information related to the reference biocidal product List of endpoints...24 APPENDIX I: LIST OF ENDPOINTS Chapter 1: Identity, Physical and Chemical Properties, Classification and Labelling...25 Chapter 2: Methods of Analysis...30 Chapter 3: Impact on Human Health...31 Chapter 4: Fate and Behaviour in the Environment...33 Chapter 5: Effects on Non-target Species...36 Chapter 6: Other End Points...37 APPENDIX II: LIST OF INTENDED USES APPENDIX III: LIST OF STUDIES

3 Product-type 19 April

4 Product-type 19 April STATEMENT OF SUBJECT MATTER AND PURPOSE 1.1. Procedure followed This assessment report has been established as a result of the evaluation of the active substance margosa extract, cold-pressed oil of azadirachta indica seeds without shells extracted with as product-type 19 (Repellents and attractants), carried out in the context of the work programme for the review of existing active substances provided for in Article 89 of Regulation (EU) No 528/2012, with a view to the possible approval of this substance. Margosa extract, cold-pressed oil of azadirachta indica seeds (CAS no ) was notified as an existing active substance, by, hereafter referred to as the applicant, in product-type 19. Commission Regulation (EC) No 1062/2014 of 4 August lays down the detailed rules for the evaluation of dossiers and for the decision-making process. On 26th of April 2006, the German Competent Authority received a dossier from. The evaluating Competent Authority (eca) refused the dossier as incomplete for the purpose of the evaluation on 31st of October After that Registration Service Srl, hereafter referred to as applicant, informed the Commission of their intention to take over the role of participant as regards the active substance/product type combination. On 31st of May 2010, the German Competent Authority received a dossier from the applicant. The eca accepted the dossier as complete for the purpose of the evaluation on 31st of August On 3 rd December 2015, the eca submitted to the Agency (ECHA) and the applicant a copy of the evaluation report, hereafter referred to as the competent authority report. In order to review the competent authority report and the comments received on it, consultations of technical experts from all Member States (peer review) were organised by the the "Agency (ECHA ). Revisions agreed upon were presented at the Biocidal Products Committee and its Working Groups meetings and the competent authority report was amended accordingly Purpose of the assessment report The aim of the assessment report is to support the opinion of the Biocidal Products Committee and a decision on the approval of margosa extract, cold-pressed oil of azadirachta indica seeds for product-type19, and, should it be approved, to facilitate the authorisation of individual biocidal products. In the evaluation of applications for product-authorisation, the provisions of Regulation (EU) No 528/2012 shall be applied, in particular the provisions of Chapter IV, as well as the common principles laid down in Annex VI. For the implementation of the common principles of Annex VI, the content and conclusions of this assessment report, which is available from the Agency web-site shall be taken into account. 1 COMMISSION DELEGATED REGULATION (EU) No 1062/2014 of 4 August 2014 on the work programme for the systematic examination of all existing active substances contained in biocidal products referred to in Regulation (EU) No 528/2012 of the European Parliament and of the Council. OJ L 294, , p. 1 4

5 However, where conclusions of this assessment report are based on data protected under the provisions of Regulation (EU) No 528/2012, such conclusions may not be used to the benefit of another applicant, unless access to these data for that purpose has been granted to that applicant. 2. OVERALL SUMMARY AND CONCLUSIONS 2.1. Presentation of the Active Substance Identity, Physico-Chemical Properties & Methods of Analysis The active substance margosa extract is a CO 2 -extract derived from cold-pressed Neem Seed Oil without shells (Azadirachta indica) using the manufacturing method developed by the applicant. It was clarified that the extraction including shells influences significantly the composition of the final extract. Furthermore, the shell is the nutrient medium for Aspergillus, which produced aflatoxins. Therefore, the extraction without shell is regarded an important criteria of the manufacture and it included in the description name of the substance. Margosa extract is a plant extract and consists mainly of the limonoids azadirachtin A, azadirachtin B, nimbin, salannin, together with co-extracted fatty acids (bound in glycerides) and a small amount of water. Margosa extract may contain traces of aflatoxins. Based on the rules set out for the identification and naming of substances the name of UVCB substances should include the manufacturing process and the species from which it is extracted. Therefore, the extract is named margosa extract, cold-pressed oil of azadirachta indica seeds. However, within this report the short form margosa extract will be used for convenience. Since margosa extract (UVCB) as such is regarded as the active substance, the physicochemical parameters are in general determined on the whole extract and not on the individual constituents. Margosa extract is thermally stable up to 340 C. The vapour pressure should be 3.8 x 10-7 hpa at 20 C. Furthermore, calculated values of the vapour pressure for limonoids and azadirachtins are given in the LoEP: The water solubility was calculated for some significant constituents. The partition coefficient was determined for some significant constituents. Sufficiently validated analytical methods for the determination of azadirachtin A, the other limonoids and other constituents in the technical material are available. The content of aflatoxins is determined An acceptable residue analytical method is available for detecting the analytical lead compounds nimbin and salannin in air. Sufficiently validated primary and confirmatory methods for detecting residues of the analytical lead compounds nimbin and salannin in soil, drinking water and surface water are missing and should be provided. Other analytical methods are not required, because no relevant residues are expected in food and feeding stuffs. Because the active substance is not classified as toxic or very toxic, residue analytical methods in body fluids and tissues are also not required. Identity, Physico-chemical Properties and Method of Analysis of Margosa extract (REPRESENTATIVE PRODUCT) Since the representative product is equal to the active substance, the information on the active substance applies. Missing information regarding: Acidity/alkalinity and if necessary ph value (1% in water) Storage stability stability and shelf-life. Effects of light, temperature and humidity on technical characteristics of the biocidal product; reactivity towards container material 5

6 (the possible increase of the aflatoxins content during storage should be addressed in this context) Technical characteristics of the biocidal product, e.g. wettability, persistent foaming, flowability, pourability and dustability will have to be submitted within the process of biocidal product authorization (see section 2.4) Intended Uses and Efficacy The assessment of the biocidal activity of the active substance demonstrates that it has a sufficient level of efficacy against the target organism(s) and the evaluation of the summary data provided in support of the efficacy of the accompanying product, establishes that the product may be expected to be efficacious. Margosa extract is intended to be used as a repellent to deter ants (e.g. Lasius niger) from entering buildings. It is aimed at non-professional users to be used in private houses. The intrinsic repellent efficacy of the active substance was proven in a simulated use trial conducted with the black garden ant (L. niger). Margosa extract was applied in a 2cm wide line, and the ants were deterred from crossing that line, even though the only food and water sources were behind the treated area. The intended use is the application of a biocidal product containing margosa extract on doorsteps and windowsills in a 2cm wide line, serving as a barrier for ants trying to enter the house from the outside. The application rate was 4 g margosa extract per linear meter on open-pored beech plywood, and 1.6 g margosa extract per linear meter on acacia terrace board. More specific testing should be assessed during product authorization, as the substance used in the efficacy trials was 100% margosa extract. The mode of action responsible for the repellent activity of margosa extract is not known. As the extract contains a mixture of limonoids, it is probably a combined effect of these that leads to a repellent effect. Since margosa extract is a non-lethal repellent, it does not exert a selective pressure for developing resistance. Ants that are repelled from entering a premise are free to feed in different, unprotected areas. Development of resistance is not expected. In addition, in order to facilitate the work of Member States in granting or reviewing authorisations, the intended uses of the substance, as identified during the evaluation process, are listed in Appendix II Classification and Labelling Currently, a harmonised classification according to Regulation (EC) No 1272/2008 (CLP Regulation) is not available for margosa extact. The toxicological properties of margosa extract, cold-pressed oil of azadirachta indica seeds as evidenced in the submitted toxicity studies do not require classification and labelling. The applicant has proposed considering the safety waring Keep out of reach of children. In the light of the reported intoxications of children with neem oil, the RMS supports the use of this precautionary statement. 6

7 Table 2-1 Proposed classification of active substance based on Regulation (EC) No 1272/2008 Hazard classes, hazard categories, hazard statements Wording Classification - - Remark: None proposed Table 2-2 Proposed labelling of active substance based on Regulation (EC) No 1272/2008 Hazard pictograms, signal words, hazard statements precautionary statements Labelling P102 Wording Keep out of reach of children Classification and Labelling of the biocidal product Margosa Extract Table 2-3 Proposed classification of the biocidal product based on Regulation (EC) No 1272/2008 Hazard classes, hazard Wording categories, hazard statements Classification None None Remark: Classification of the biocidal product is not required. Table 2-4 Proposed labelling of the biocidal product based on Regulation (EC) No 1272/2008 Hazard pictograms, signal words, hazard statements precautionary statements Labelling None Wording None 7

8 Remark: Classification and labelling of the biocidal product is not required 2.2. Summary of the Risk Assessment Human Health Risk Assessment Hazard identification and effects assessment Margosa extract is a CO 2 -extract derived from cold-pressed neem seed oil without shells (Azadirachta indica) using the manufacturing method developed by the applicant and acts as a repellent against worker ants. As a botanical extract it belongs to the group of substances with unknown or variable composition, complex reaction products or biological material (UVCB) with unspecified molecular and structural formula. The total content of limonoids was determined to be 2.7 ± 0.4 % including azadirachtin A. While in other margosa extracts azadirachtin A is regarded as lead compound, margosa extract in this CAR is considered different in composition and properties from other margosa extracts (e.g. NeemAzal, Fortune Aza, ATI-720 = NPI 720). This also applies to the content of aflatoxins which is much lower in this margosa extract (aflatoxin B1: < 2 µg/kg ( % w/w; sum of aflatoxins B1, B2, G1, G2: < 4 µg/kg ( % w/w). is, therefore, considered another substance. Consequently, studies performed with one of the above-mentioned extracts are not considered in this CAR and read across to those extracts is not applicable. Likewise, toxicity studies with neem products found in the open literature were considered not relevant for margosa extract due to different starting material or extraction procedures. The specification of margosa extract is covered by the batches of margosa extract as tested in the toxicity studies. Waiving Concept Chronic and carcinogenicity studies with margosa extract have not been performed. The waiving of such studies is deemed acceptable in view of the lack of pertinent findings in genotoxicity tests and repeat dose studies. According to Guidance on information requirements Guidance on regulation (EU) No 528/2012 The long-term toxicity study ( 12 months) does not need to be conducted if: Long-term exposure can be excluded and no effects have been seen at the limit dose in the 90-day study, or A combined long-term repeated dose/carcinogenicity study (8.11.1) is undertaken As only mild adverse effects (absolute and relative liver weight increases) were observed in the 90-day study in rats up to approx mg/kg bw/day and long-term exposure is not expected according to the use scenarios submitted by the applicant, omission of carcinogenicity study is justified. Therefore, a carcinogenicity study is deemed not necessary because the overall conclusion from the genotoxicity tests is that margosa extract is not genotoxic. Furthermore, the subacute and subchronic feeding studies did not reveal any finding that could be correlated with a pre-neoplastic event or lead to the expectation of different or more severe effects after chronic exposure. The effects in both studies were limited to liver changes. The subchronic 8

9 NOAEL/LOAEL for liver weight increase (450/960 mg/kg/day) is almost 500/1000 times higher than the highest predicted human exposure. Tumour promotion as result of adaptive liver enlargement can therefore be ruled out at doses that humans are likely to be exposed to as a result of use as repellent containing Margosa extract. Studies on the effects of Margosa extract on fertility and postnatal development of offspring have not been conducted. A two-generation reproduction study is not deemed necessary, since human exposure is low when compared to the subchronic oral NOAEL of 450 mg/kg bw/day. Reproductive organ weights were not altered in this study and histopathological investigation did not reveal any abnormalities of these or any other tissues that might affect reproduction. This suggests that Margosa extract has no potential for reproductive toxicity at doses approaching the limit dose of 1000 mg/kg bw/day. Considering that exposure during or after use of Margosa extract is only 1.28/1.19 mg/kg bw/day in adults and 1.37 mg/kg bw/day in toddlers, there is no concern for reproductive or developmental effects associated with the use of Margosa extract as repellent against worker ants. In addition, a prenatal toxicity study in a second species (rodent) is not considered necessary, based on the results of a dermal study in rabbits where no embryo- or foetotoxicity was revealed at any dose, even in the presence of local maternal toxicity. In general, the limitations of the submitted data package were considered by applying an additional assessment factor for uncertainties in the derivation of reference values for margosa extract. Absorption, Distribution, Excretion, and Metabolism No studies on absorption, distribution, metabolism and excretion were submitted. Such ADME studies are performed with radioactively labelled compounds. Margosa extract contains only small amounts of limonoids. As the active substance is a complex mixture of various compounds, margosa extract is regarded as active substance in accordance with the Guidance Document on Botanical Active Substances Used in Plant Protection Products (SANCO/11470/2012- rev.8, 20 March 2014). Hence, results from ADME studies with labelled azadirachtin A are not meaningful for margosa extract. For risk assessment purposes, absorption of the active substance margosa extract from oral and inhalation exposure is assumed to be 100 % since actual data are not available. For dermal absorption default values according to EFSA guidance (EFSA 2012) are applied. Acute Toxicity Margosa extract is not acutely toxic when administered orally, dermally or by inhalation. It is not irritating to the skin or eyes of rabbits following single application and did not sensitise the skin of guinea pigs. Since irritation of the skin was not observed in a primary dermal irritation study, the conditions for classifying the active substance as irritating to the skin are not met. Short-term Toxicity Slight, reversible increases in liver weight were the only finding in 28-day and 90-day feeding studies with margosa extract in rats. This effect is not considered in the 90-d study as liver weight increases were up to 18 % compared to controls. The NOAEL was 990 mg/kg bw/d in the 28-day study and 450 mg/kg bw/d in the 90-day study. In a 28-day dermal study with margosa extract in rats, no systemic effects were observed. Slight erythema with or without desquamation was noted in all males and females receiving 9

10 500 mg/kg/day towards the end of the first week of dosing. In the dose groups receiving 1000 mg/kg/day incidence and time of appearance were similar and the grading ranged from slight to severe. The skin irritation disappeared in both groups during the second week of dosing and no further changes became apparent after that timepoint. Thus, classification for skin irritation following repeated exposure is not considered required. Genotoxicity Margosa extract did not show a mutagenic potential in bacteria and in Chinese hamster lung fibroblasts (V79 cells). Slightly positive effects for clastogenicity were observed with multiple damaged metaphases at the highest tested concentration (5000 µg/ml). This effect was observed in the presence of metabolic activation in the in vitro chromosomal aberration test in V79 cells. However, margosa extract was not genotoxic in the in vivo micronucleus test in mice exposed at dose levels up to and including 2000 mg/kg bw. In conclusion, based on the results of in vitro and in vivo genotoxicity tests, including adequate positive and negative controls, margosa extract is unlikely to pose a genotoxic risk to humans. Chronic Toxicity/ Carcinogenicity No chronic or carcinogenicity study has been submitted for margosa extract. The waiving of such a study is deemed acceptable in view of the lack of pertinent findings in genotoxicity tests and repeat dose studies (see also waiving concept). Reproduction Toxicity After dermal application of margosa extract to pregnant rabbits, local skin irritation occurred in all treated groups. Severity of the observed erythema and oedema was high in the highest dose group. It was considered that classification for skin irritation following repeated exposure of margosa extract was clearly demonstrated but severity in the relevant doses was considered not sufficient for classification according to CLP. Consequently, STOT-RE for skin irritation was not proposed. A slight, dose-related tendency towards reduction of maternal body weight gain was observed. Net body weight loss was observed in mid and high dose females but did not attain statistical significance. Overall, reductions in body weight gain in the mid and high dose groups were not accompanied by any sign of toxicity and the differences in terminal body weights were only marginal. This effect is, therefore, considered not to be biologically relevant. No embryo- or foetotoxicity was apparent. Small foetuses in all groups, including the control, were found mostly in litters of larger size and it appears that the higher proportion of such litters, rather than the treatment, contributed to the slightly increased number of small foetuses in the high dose group. Thus the maternal and the developmental NOAEL is 800 mg/kg bw/d, corresponding to the highest dose tested. Neither a prenatal toxicity study in a rodent species nor a two-generation study have been conducted for margosa extract (see also waiving concept). Neurotoxicity No specific studies for neurotoxic properties have been submitted. Available repeat dose studies in rats did not show any neurotoxic effects. Further Studies None 10

11 Medical Data No reports on adverse effects of margosa extract are available. Medical surveillance of an unknown number of plant workers occupationally handling with margosa extract did not reveal any obvious toxicity in humans. Reviews of the open literature on various neem products with unknown composition demonstrates evidence of poisoning incidents and side-effects in the use of neem products but are considered not applicable to margosa extract, "Margosa Oil" or "Neem Oil" is used as a traditional medicine in Asia and Africa. In case reports severe intoxications are described in children predominantly following oral administration of Margosa Oil as a home remedy for the treatment of various diseases (e.g. common cold, deworming). Margosa extract exerts no acute toxicity up to the limit dose of 2000 mg/kg bw in rats. In addition, no signs of toxicity were observed in repeated dose studies in rats (up to 90 days) and rabbits (treatment day 6-28) following oral (rats) and dermal (rats and rabbits) exposure. Hence, poisoning from margosa extract up to the limit dose of 2000 mg/kg bw is not to be expected. Summary & Conclusion Margosa extract is of low toxicity after acute and short-term/subchronic exposure and requires no classification/labelling according to Regulation (EC) No 1272/2008. Dermal irritation was noted after repeated application of the extract to the skin in two species (rats, rabbits), but not in the rabbit following single application. Thus, the conditions for classifying the active substance as irritating to the skin are not met In addition, the pregnant rabbits exhibited a reduction in body weight gain that may be attributed to maternal local irritation as the conceptuses remained unaffected. The following systemic Acceptable Exposure Levels (AELs) are proposed: Acute exposure: No endpoints relevant for an AEL acute are seen in the single-dose toxicity studies. Due to the lack of acute toxicity, the AEL acute was derived from the 28-d study in rats following oral exposure as a conservative approach. The NOAEL of 990 mg Margosa extract/kg bw/d from the 28-day oral rat study is considered appropriate to derive an AEL acute. Slight increases of liver weights were observed which were not considered adverse. Applying an assessment factor of 200 and assuming 100 % oral absorption, a systemic acute Acceptable Exposure Level (AEL acute ) of 5 mg/kg bw/d is proposed for short-term exposure to margosa extract. Medium-term exposure: Repeated-dose studies up to 90-d and prenatal toxicity studies are taken into consideration to derive the AEL medium-term. The NOAEL of 450 mg Margosa extract/kg bw/d from the subchronic (90 day) oral rat study is considered appropriate to derive an AEL medium-term. Applying an assessment factor of 200 and assuming 100 % oral absorption, a systemic medium-term Acceptable Exposure Level (AEL medium-term ) of 2.30 mg/kg bw/d is proposed for medium-term exposure to margosa extract. Long-term exposure: Chronic toxicity studies were waived based on the absence of any treatment-related adverse effects in repeat dose studies in rats and the absence of genotoxic effects mutagenicity assays. The most relevant NOAEL was observed in the subchronic (90 day) oral toxicity study in rats and it appears reasonable to use this value for the derivation of the AEL long-term. Applying a combined assessment factor of 400 (2 x 2 x 100) and assuming 100 % oral absorption, a systemic long-term Acceptable Exposure Level (AEL long-term ) of 1.1 mg/kg bw/d is proposed for long-term exposure towards margosa extract. 11

12 ADI/ARfD: If margosa extract is applied in accordance with the intended use no residues of margosa extract are to be expected in food of plant or animal origin. Therefore, no ADI and ARfD are derived Exposure assessment and risk characterisation Exposure of Professionals The biocidal product is a consumer product. It is not expected that professionals with a profession that does not involve biocides (e.g. housekeeper) use the product in their work environment. In that case, the rapporteur assesses that the pattern of use is similar to the pattern of consumers. Therefore, exposure and risk of professionals are estimated to be in the same range as of nonprofessionals (see chapter: Exposure of Non-professionals and Risk Assessment for Non- Professionals ). Exposure of Non-Professionals and the General Public The biocidal product margosa extract (100% active substance) is applied on doorsteps and windowsills as a barrier against ants and brushing with an application rate from 20 mg/cm 2 in materials like open-pores beech plywood to 53 mg/cm 2 in smoother materials like pre-treated terrace acacia board. Exposure may occur directly during application (including brush cleaning) or indirectly by contact with treated surfaces. The most relevant secondary exposure scenarios are exposure of adults by contact with bare feet and exposure of toddlers by touching treated surfaces with hands and subsequent oral ingestion. Primary exposure Application by brushing (inc. brush cleaning) Systemic inhalation exposure: 5.31 x 10-5 mg/kg bw/d Systemic dermal exposure: 1.28 mg/kg bw/d Total systemic exposure: 1.28 mg/kg bw/d Secondary exposure Exposure of adults by contact with bare feet Systemic dermal exposure: 1.19 mg/kg bw/d Total systemic exposure: 1.19 mg/kg bw/d Exposure of toddlers by dermal and oral contact Systemic dermal exposure: 0.95 mg/kg bw/d Systemic oral exposure: 0.42 mg/kg bw/d Total systemic exposure: 1.37 mg/kg bw/d Risk Characterisation Risk Assessment for Professionals The biocidal product is a consumer product. It is not excluded that professionals with a profession that does not involve biocides (e.g. housekeeper) use the product in their work environment. In that case, the rapporteur assesses that the pattern of use is similar to the 12

13 pattern of use of consumers. Therefore, exposure and risk of professionals are estimated to be in the same range as of nonprofessionals (see chapters: Exposure of Non-professionals and Risk Assessment for Non- Professionals ). Safety Measures for Professionals See Chapter: Safety Measures for Non-Professionals Risk Assessment for Non-Professionals and the General Public Summary risk assessment for non-professional primary exposure to margosa extact Exposure scenario Exposure adults (mg/kg bw/d) AEL/NOAEL (mg/kg bw/d) Exposure (% of AEL) MoE Medium-term exposure - application of MARGOSA EXTRACT by brushing (incl. brush cleaning) Inhalation 5.31 x / Dermal / Total / Summary risk assessment for secondary exposure of the general public to margosa extact Exposure scenario Exposure AEL/NOAEL Exposure MoE (mg/kg bw/d) (mg/kg bw/d) (% of AEL) Adults Acute exposure - contact to treated surfaces with bare feet Dermal / Total / Toddlers Acute exposure - contact to treated surfaces with hands when crawling Dermal / Oral / Total 1,37 5.0/

14 Safety Measures for Non-Professionals Primary and secondary exposure of non-professional users and the general public (including pets and domestic animals) to margosa extract after regular treatment with the biocidal product containing 100% of the active substance is acceptable with respect to human health. Specific safety measures are not required. Based on information provided by the applicant the biocidal product dried on rough surfaces like plywood within 24 h due to absorption into the wood. However, on acacia wood it remains liquid. Consequently, labelling of the biocidal product with Keep children away from treated surfaces until dried is proposed. Residues in food are not expected from the intended use. To ensure that residues in food do not occur, the following labelling is required: Avoid direct contact or contamination of food and feedstuff Environmental Risk Assessment The technical a.s. margosa extract is gained by CO 2 -extraction from cold pressed oil from neem tree (Azadirachta indica A. Juss.) seed without shells. With 97% in total, the main components of margosa extract in Pt19 are fats and fatty acids, existent mainly as trigycerides of oleic acid, stearic acid, linoleic acid and palmitic acid. Limonoids (belonging to the chemical group of triterpenoids) are representing the second group of relevant constituents of margosa extract, with less than 2% in total. The representative product assessed within this report is identical to the a.s., therefore only the a.s. had to be evaluated within the environmental effects assessment and not a formulated b.p. While it is known which components mostly contribute to the intended efficacy in PT19, it can also be deduced that mainly the limonoids should be regarded as relevant for (potential) adverse effects on non-target organisms in the environment. The limonoids from Neem tree are known to act as antifeedant and growth disruptor towards insects. Therefore the accompanying chemical analysis of the effect studies is based on salannin as the limonoid with the highest proportion in margosa extract. This also applies to recalculations to mean measured concentrations, if required. In addition, the effect assessment was supported by the physico-chemical properties of salannin for applying the equilibrium partitioning method. The use of a non-labelled test substance and the non-feasibility of synthesising reference compounds result in an unavoidable limitation of findings concerning the fate and behaviour of margosa extract i.e., the impossibility to determine the pathway of degradation of the test substance and the quality and quantity of its degradation products in a number of environmental fate studies (e.g. soil and water/sediment degradation, hydrolysis, aqueous photolysis) Fate and distribution in the environment Biodegradation Based on the results of a closed bottle test (OECD 301D) using margosa extract (identical with the product) the active substance was proved to be readily biodegradable fulfilling the 10-day window criterion (>60% degradation within 10 days). However, the study does not allow to draw conclusions if and to which extent the relevant compounds degrade. Based on the result of QSAR analysis none of the components azadirachtin, nimbin, salannin and stigmasterol can be considered as readily biodegradable. Considering the results of QSAR analysis and according to a decision of BPC WG ENV (III-2016), a degradation rate of k = 0 h -1 for biodegradation in sewage treatment plants is applied in the environmental exposure assessment of the active substance. 14

15 Regarding the freshwater compartment, the rate constant for biodegradation in surface water (kbio water ) is set to zero as well due to the lack of information on the biodegradation half-lives of the relevant components. A study about the degradation behaviour of azadirachtin A and B in an outdoor mesocosm experiment, provided by the applicant, was considered only as supportive information. Both isomers showed only low tendency to partition into bottom sediments accompanied by the dissipation of azadirachtin A and B from the water phase (DT 50 of 25 and 45 days, respectively). Also for the soil compartment, only supportive information is available from a published study, in which the degradation of azadirachtin A and B was investigated in a sandy loam at temperatures of 25 C and 15 C using a light regime for each environmental chamber of 16 h light and 8 h dark. Converting the DT 50 values gained from this study according to an average EU outdoor temperature of 12 C results in a maximum DT 50 of azadirachtin A of 56 days and for azadirachtin B for 75.3 days. Based on this, it can be assumed that azadirachtin A and B if potentially entering the soil would dissipate/be degradable. Since the tests were conducted with non-labelled material, neither mineralisation nor the formations of bound residues were quantifiable. Again, in the environmental exposure assessment of the active substance for the soil compartment no degradation was assumed due to the lack of knowledge about specific degradation rates of the relevant components. Abiotic Degradation All three constituent limonoids of margosa extract are susceptible to hydrolysis, though the determined half-lives vary greatly. The hydrolysis rate was highest for azadirachtin (DT50 = h at standard conditions of ph 7 and 12 C), whereas the mean hydrolysis half-life of nimbin was h (ph 7 and 12 C). The lowest hydrolysis rate was determined for salannin, corresponding to a DT50 of h (ph 7 and 12 C). Hydrolysis of the three limonoids is ph-dependent and in most cases an alkaline ph increases the hydrolysis rate, even though some results deviate from this observation. In Summary, hydrolysis might contribute to the degradation of azadirachtin and nimbin under environmental conditions, whereas it is negligible for salannin dissipation. Photolysis in water is irrelevant for the dissipation of the three limonoids, as they do not exhibit significant absorption in the visual light spectrum. However, the limonoids are susceptible to indirect photolysis in the atmosphere with calculated half-lives of 1.3 h up to 1.7 h. But as the volatilisation of the three limonoids is negligible due to the very low vapour pressure and Henry s Law coefficient, these processes are of minor importance for the environmental fate. Distribution and Mobility The adsorption behaviour to soil or sewage sludge of the constituent limonoids in Margosa extract was investigated using the HPLC method procedure according to the OECD Guidline 121. The determined arithmetic mean values for the Koc are 144 L/kg, 809 L/kg and 1766 L/kg for azadirachtin, nimbin and salannin, respectively. Bioaccumation A bioconcentration factor for the aquatic compartment was estimated on basis of salannin, which provides the highest log K ow of 3.5 out of the limonoids. Based on this, BCF for fish were estimated and below 100 L/kg wet weight for the limonoids (salannin: BCF = 95 L/kg ww). The calculated BCF value, BCF fish = 95 L/kg wet weight, indicated that limonoids have a low potential to bioaccumulate in aquatic organisms and would pose no unacceptable risk of biomagnification in the food chain. 15

16 Hazard identification and effects assessment Aquatic compartment A base data set of effect studies for the aquatic compartment is available for the environmental effect assessment. All tests were conducted with the pure a.s. Based on the complex composition and low solubility in water of margosa extract, the definition of a difficult substance in accordance to OECD Guidance Document No. 23 on Aquatic Toxicity Testing of Difficult Substances and Mixtures was met and due to the low solubility of the test substance, acetone has been used as a solvent. The stability of the test substance within the tests was monitored by analysing the concentration of salannin and, if required, recalculating from nominal to mean measured concentration based on the concentration of salannin. One acute study with fish was provided for the test substance margosa extract. The study is considered as key study for fish. After 96 h and based on mean measured concentrations, a LC 50 of 11.2 mg/l has been calculated (95 % c.i.: mg/l). One acute study with Daphnia magna was performed with the test substance margosa extract. The study is considered as key study for invertebrates. For 48 h, an EC 50 > 128 mg/l has been calculated based on mean measured concentrations. One 72 h growth study with the green algae Desmodesmus subspicatus was performed with the test substance margosa extract. The study covers both acute and long-term endpoints and is considered as key study for algae. After 72 h, a NOEC of 1.05 mg/l and an EC 50 > 237 mg/l has been calculated based on growth rate and mean measured concentrations. Fish are representing the most sensitive species in the aquatic compartment for margosa extract. Since short-term data from each of the three trophic levels are available, an assessment factor (AF) of 1000 should be applied. A PNEC water of mg/l has been derived based on the available data. Sediment No data for effects on sediment organisms are available. According to BPR guidance Vol. IV Part B Section an effect assessment for sediment organisms should be considered in cases where log K OW 3 applies. Out of the limonoids only salannin exceeds this trigger value (log K OW = 3.5 at ph 7). Therefore the equilibrium partition method (EPM) has been applied to identify a potential risk to sediment organisms as a screening approach. The input values for EPM calculation, Koc and Henry s Law constant, are based on salannin which also shows the highest Koc of the limonoids. The calculated PNEC sediment,epm for margosa extract is mg/kg wet weight sediment. Inhibition of microbial activity In a standard activated sludge respiration inhibition test with sludge from domestic sewage treatment plant no inhibitory effect of the test item up to and including a concentration of 100 mg a.s./l (nominal) was observed. Therefore the NOEC was 100 mg a.s./l and the EC 50 > 100 mg a.s./l. The risk to the microorganism population of a sewage treatment plant can be characterised to be low regarding the results of these studies. From this data a PNECmicroorganisms of 10 mg a.s./l (nominal) was derived. Terrestrial Compartment Effect data for the terrestrial compartment were not provided and are only required if on basis of EPM an unacceptable risk for the terrestrial compartment has been indicated, or if direct or 16

17 long-term exposure represents a relevant exposure pathway. All of these conditions do not apply; therefore no effect data is required. Similar to sediment, the EPM has been applied as a screening approach, resulting in a PNEC soil,epm of mg/kg wet weight soil. Atmosphere Due to the very low vapour pressure of margosa extract ( hpa at 20 C) and the small Henry s Law constants of the constituent limonoids ( Pa m3/mol, Pa m3/mol and Pa m3/mol for azadirachtin, nimbin and salannin, respectively) only negligible volatilisation and transfer to the atmosphere is expected. Additionally, the tropospheric half-lives of the three limonoids were calculated to be in the range of 1.3 h to 1.7 h. According to these findings, accumulation and long-range transport of margosa extract or the constituent limonoids in the atmosphere followed by wet or dry deposition is not expected PBT Assessment and Exclusion Criteria The PBT assessment was performed in line with the REACH legislation (Guidance on Information Requirements and Chemical Safety Assessment Part C: PBT/vPvB assessment, and Chapter R.11: PBT/vPvB assessment, Version 2.0), following the PBT and vpvb criteria laid down in Annex XIII of the REACH Regulation (EC) No 1907/2006. In addition, exclusion criteria according to Article 5(1) of the BPR were assessed according to ECHA Guidance on BPR Vol. IV Part B. The PBT assessment presented in Doc II-C 13.5 covers all constituents 1% (w/w), i.e. the lipid fraction, stigmasterol as well as individual limonoids salannin, nimbin and azadirachtin (A+B). v(p) Assessment P criterion: Half life vp criterion: Half life > 40 d freshwater or >120 d in freshwater sediment or > 120 d in soil > 60 d water or > 180 d in freshwater sediment or >180 d in soil According to the Closed Bottle test (OECD 301D), margosa extract is considered to be readily biodegradable fulfilling the 10-day window criterion. Generally, it is assumed that a chemical giving a positive result in a test of this type will rapidly biodegrade in the environment. However, it has to be noted that if a substance consists of more than one constituent PBT/vPvB assessment needs to be performed for all individual amounts 0.1 % (w/w). In case of margosa extract (CO 2 extract from cold pressed Neem seed oil), this involves the individual limonoids (i.e. salannin, nimbin, and azadirachtin), stigmasterol as well as lipids (diand triglycerides, free fatty acids). In case of di- and triglycerides as well as free fatty acids it was considered scientifically unjustified to perform a PBT assessment, since they are ubiquitous and essential components of all living organisms. In contrast to lipids, the substance class of triterpenoids, comprising substances such as the limonoids, (i.e. azadirachtin, nimbin, and salannin) is of higher complexity and structural diversity, although being of ubiquitous distribution, too. Triterpenoids are produced by many plants and animals as well. This supports the assumption that common pathways for the breakdown of limonoids exist in nature. Due to the lack of screening tests conducted with the individual limonoids, biodegradability was assessed using QSAR calculations in the first step. 17

18 Furthermore, available information regarding abiotic degradation (i.e. hydrolysis) as well as the degradation of azadirachtin A and B in soil was taken into account. Based on this and due to the lack of further information about the degradation of azadirachtin, nimbin, and salannin in the aquatic and/or soil compartment as well as about the formation and degradation behaviour of relevant degradation products, it is concluded that all limonoids are considered potentially fulfilling the (v)p criterion. Based on QSAR results and due to the lack of further information, it has to be concluded as well that stigmasterol potentially fulfils the (v)p-criterion. B-Assessment a-criterion: vb-criterion: BCF > 2000 L/kg wet weight BCF > 5000 L/kg wet weight The bioaccumulation for fish was calculated for all constituents exceeding 0.1 % (w/w) of Margosa extract. Based on the lack of measured bioaccumulation data, the assessment was performed following a screening approach on basis of the respective log Kow values. An octanol-water partitioning coefficient log Kow > 4.5 indicates that the substance is potentially (v)b. Out of the limonoids, sa lannin provided the highest octanol-water partitioning coefficient, but does not exceed the screening trigger value (salannin : Kow = 3.5; nimbin: Kow = 3.0; azadirachtin: Kow = 1.3), indicating a low potential for bioaccumulation for this group of constituents. Therefore the bioaccumulation ("B") criterion for the limonoid fraction of Margosa extract is not fulfilled. Based on physiological considerations, the bioaccumulation of glycerides and fatty acids as well as for stigmasterol contained in the a.s. can be considered as not relevant for the assessment of the B criterion. In conclusion the B criterion is not fulfilled for Margosa extract. T-Assessment T-criterion: Chronic NOEC < 0.01 mg/lor CMR or endocrine disrupting effects No chronic effects data is available for the Margosa extract, therefore screening on basis of acute effects has been performed. Rainbow trout (0. mykiss) represents the most sensitive aquatic organism in an acute test, with LC 50 = 11.2 mg/l. Margosa extract cannot be considered as potentially toxic because all L(E)C 50 to aquatic organisms are higher than 0.1 mg/l. Also considering that neither CMR properties are reported nor criteria for endocrinedisrupting effects (see next section) are met for the a.s., it can be concluded on the basis of the provided effect data for Margosa extract that the T criteria are not fulfilled. However, based on available data for the limonoids as relevant constituents of Margosa extract the T criteria should be considered as potentially fulfilled. Conclusion of the PBT assessment The PST assessment for Margosa extract as a.s. and for the individual components ;;::: 0.1 % (w/w), i.e. the lipid fraction, stigmasterol, as well as the limonoids azadirachtin, nimbin and sa lannin, is summarised in the following table. Constituent (v)p (v)b T Conclusion Lipids assessment scientifically unjustified not PST scientifically Stigmasterol pot. vp not T pot. vp unjustified 18

19 Product-type 19 March 2017 Salannin pot. (v)p not (v)b pot T Nimbin pot. (v)p not (v)b pot T pot. vp (aquatic) Azadirachtin A + B pot. P (soil) not (v)b pot T pot vp, pot T pot vp, pot T pot vp, pot T Assessment of PBT criteria was performed on basis of data for the whole plant extract as well as for the relevant components of Margosa extract, consisting mainly of lipids and limonoids. For the group of lipids, including triglycerides, diglycerides - mainly based on stearic, oleic, palmitic and linoleic acid - as well as the corresponding free fatty acids, neither P, B nor T criteria are fulfilled. Therefore, this group of constituents neither meets PBT nor vpvb criteria. Stigmasterol potentially fulfils the (v)p-criterion but neither the B criterion nor the T-criterion. Therefore, t his constituent neither meets PBT nor vpvb criteria as well. The limonoids salannin, nimbin and azadirachtin had to be assessed separately. Azadirachtin consists of azadirachtin A and B, but no specific data was available for further PBT assessment. Azadirachtin potentially fulfils the (v)p criterion, does not fulfil the B criterion and potentially does fulfil the T criterion. Salannin and nimbin were potentially fulfilling (v)p as well, both were not fulfilling the B criterion and both were potentially fulfilling the T criterion based on a read-across approach. In conclusion, none of the relevant compounds of Margosa extract fulfils the criteria of a PBT or a vpvb substance. Therefore, the whole Margosa extract does not represent a PBT/vPvB substance. Based on the P- and T-criterion, the limonoids are potentially fulfilling two of the three PBT criteria. If PBT guidance and criteria are strictly applied, this conclusion has to be transferred to the active substance: Margosa extract potentially fulfils two out of three PBT criteria. Based on the contained limonoids and both on the lack of information on breakdown/transformation products from hydrolysis and aerobic biodegradation in soil and - with regard to the T-criterion - on read-across with margosa extract PT18, the following studies for the limonoids and their transformation products could allow achieving a definite conclusion for the PBT assessment: OECD 309 test (Aerobic Mineralisation in Surface Water - Simulation Biodegradation Test) On the basis of the results of the OECD 309, if needed : a test OECD 307 (Aerobic and anaerobic transformation in soil including identification of transformation products) or a OECD 308 (water/sediment degradation study including identification of transformation products) shall be performed Long-term effect test with aquatic insects The data are needed for the renewal process. A stepwise approach is conceivable, starting with investigating the T criterion. However, the applicant is advised to contact the eca before performing the studies. It should be considered that the limonoids represent only a small fraction of the whole extract ( < 2%), whereas azadirachtin A and B make up < 0.2%. In contrast, margosa extract approved in PT18 consists to a large share only of limonoids, mostly azadirachtin A/B. Therefore, a read-across from PT18 regarding the T criterion could be regarded as a worstcase approach (furthermore azadirachtin was regarded as being 'not P' for PT18). In addition, based on the data requirements applying during submission of the dossier the submitted data set should be regarded as complete ("How to deal with extracts and oils of plant or animal origin?", endorsed at the 23rct CA-Meeting, and "Guidance to Member States and industry on the data requirements for naturally occurring substances used as attractants/repellents", endorsed at the 18th CA-Meeting). Due to this, further information should only be considered at the renewal stage of margosa extract in PT19. If P and T status could be confirmed by further 19

20 data, this could render the active substance margosa extract as a candidate for substitution, eventually triggering comparative assessment for biocidal products containing the active substance. Endocrine-Disrupting Properties ED-criteria: endocrine-disrupting properties that may cause adverse effects in humans, or endocrine-disrupting properties identified in accordance with Articles 57 (f) and 59(1) of Regulation (EC) No 1907/2006. Until the Commission has specified scientific criteria, also transitional criteria apply: carcinogen category 2 and toxic for reproduction category 2, or toxic for reproduction and having toxic effects on endocrine organs. Margosa extract contains around 2% limonoids. These limonoids salannin, nimbin and azadirachtin are known to exhibit biological activity by inhibiting insect growth and development. Biological effects of azadirachtin include antifeedant activity, insecticidal activity and inhibition of hormone and enzyme activity, and similar effects are described in open literature for salannin and nimbin. The limonoids azadirachtin, salannin and nimbin are known to similar inhibit ecdysone 20-monoxygenase. This enzyme is responsible in insects for the conversion of the molting hormone ecdysone to its more physiologically active metabolite 20- hydroxyecdysone. Extracts from Neem tree with higher content of limonoids, especially of azadirachtin, are used as insecticides (product type 18) and the effects on insects described above represent intended effects. Besides insects, no information is available on endocrine disrupting effects on further non-target organisms, especially considering vertebrates (including mammals and humans). Studies available in the human health risk assessment for margosa extract did not mention any indications for endocrine disrupting effects. The BPR and supporting guidance does not suggest that endocrine effects on arthropod should be considered during assessment of endocrine-disrupting properties. Article 5(d) of the BPR only refers to endocrine-disrupting properties that may cause adverse effects in humans or which are identified in accordance with Articles 57(f) and 59(1) of Regulation (EC) No 1907/2006 as having endocrine disrupting properties. The active substance was not identified as SVHC under REACH and adverse effects in humans are presently not described in literature. In addition, the transitional criteria provided in the BPR are not met; the a.s. has neither been classified as carcinogen category 2 and toxic for reproduction category 2 nor as toxic for reproduction and have toxic effects on endocrine organs. The Commission still has to adopted definitive criteria for endocrine disruption. It should be noted that a neem extract with higher content of limonoids is authorised and used in line with the BPR as intended endocrine acting active substances. Therefore it can be concluded that in agreement with the BPR the present criteria for endocrine-disrupting properties are not fulfilled Exposure assessment and risk characterisation Exposure assessment For environmental exposure estimation data about one representative biocidal representative product is provided by the applicant. For the life cycle stage production, no exposure assessment has been performed as the production process takes places in a closed facility. As the representative product is technical identical to the a.s., no exposure assessment for the formulation has been performed. 20

21 According to the intended use emission to the environment are expected only for the life-cycle step non-professional use. Two different models were applied to estimate the emissions to the environment. First, a consumption based assessment was conductedincorporating specific information about the product and the intended use. The second model applied is a tonnage based model, which is more generic approach estimating emission from the produced and/or imported tonnage of the assessed active substance. Consumption Scenario The consumption based environmental exposure was assessed for the application and the cleaning step. The environmental exposure has been assessed applying the EU Technical Guidance Document (TGD) on Risk Assessment (2003), the OECD Emission Scenario Document Number 18 for Insecticides, Acaricides and Products to Control Other Arthropods (PT18) for Household and Professional Uses (July 2008) and the OECD Emission Scenario Document Number 22 (2009) for Coating Industry (Paints, Lacquers and Varnishes). The exposure of a.s. in the life cycle stage non-professional use of the biocidal representative product Margosa extract barrier repellent is estimated considering the application steps by non-professionals indoor and subsequent cleaning steps of the product. The application of the b.p. as a repellent barrier is envisaged to prevent garden ants Lasius niger from entering into domestic premises. Due to the proposed non-professional indoor use of the ready-to-use product the application mode can be described as: Targeted barrier surface application of a product in gel form with a brush A detailed description of emission scenarios for gel application including the input and output values is given in chapter II-8. Regarding the cleaning step, two general cleaning methods (wet and dry cleaning with emission to waste water or solid waste) are described in the OECD ESD No.18. Since the product is an oily liquid, it is not expected that residues of margosa extract can be removed by dry cleaning methods. Thus, the exposure pathway of solid waste to municipal landfill is negligible. However, the wet cleaning process is relevant for the environmental risk assessment of the margosa extract product. In addition, releases of margosa extract to the sewer due to wet cleaning of the brush were included in the exposure assessment. Assuming that residues of a.s. removed through wet cleaning may be emitted to waste water, the STP is considered as the primary receiving compartment for margosa extract. Subsequently, surface water, sediment, soil and groundwater represent secondary receiving compartments. Tonnage Scenario The tonnage based environmental exposure has been assessed applying the EU Technical Guidance Document (TGD) on Risk Assessment (2003) and the ECHA ESD PT19 for repellents and attractants (2015). The applicant provided the production tonnage of the active substance margosa extract PT 19 for the years 2012 to This tonnage data represents a complete overview as the applicant is the only company producing and/or importing this active substance in/into the EU. The emission modelling contains a safety margin in order to cover a potential future increase of active substance production in the European Union. As the dummy product is very specific with regard to the target organism, unequal spatial and temporal distribution of product use in the EU is predicted. In conclusion, the product is applied only in areas where the target organism Lasius niger is occurring in detrimental amounts during the seasons in which the occurrence of Lasius niger is so high to cause adverse effects. These temporal and spatial variations of product use were integrated into the parameterisation of the model. 21

22 Product-type 19 w ithout shells extracted w ith March 2017 Alike the Consumption Scenario, the STP is considered as the primary receiving compartment of margosa extract in the Tonnage Scenario. Subsequently, surface water, sediment, soil and groundwater represent secondary receiving compartments. Summary of PECs The PRCs for the Consumption Scenario and the Tonnage Scenario have been estimated for the aquat ic compartment including STP, surface water, and sediment, and for the terrestrial compartment including soil and groundwater. As the PEC 9 roundwater estimation according to the pore water calculation model of the TOG resulted in unacceptable risks for the groundwater after sewage sludge application, the groundwater risk assessment was refined by using FOCUS model PEARL (transport and fate simulation tool). Risk characterisation Stages of the product 's life-cycle considered as relevant for the Consumption Scenario are targeted barrier surface application (including brush residues) and wet cleaning steps for nonprofessional use. Residues of b.p. removed through wet cleaning can be emitted to waste wat er, therefore STP is considered as the primary receiving compartment for the b.p., which is identical with the active substance. Subsequently, surface water, sediment, soil and groundwater represent secondary receiving compartments. The same environmental emission pathway applies to the Tonnage Scenario. Aquatic Compartment STP represents the primary receiving compartment for the Consumption Scenario and the Tonnage Scenario. Secondarily affected compartments in the aquatic environment are surface water and sediment. PNECmicroorganism is based on the NOEC from an act ivated sludge respiration test and an AF of 10. PNECwater was derived on the basis of three acute effect studies with fish representing the most sensitive trophic level and an AF of For the sediment compartment no effect studies were available and the equilibrium partitioning method (EPM) was used therefore as a screening approach: Table 2-3 PEC/PNEC ratios for different exposure situations concerning the hydrosphere Exposure Environmental PEC PNEC PEC I PNEC scenario comoartment STP 299 ua/l ua/l 0.03 Consumption Surface water ua/l 11.2 µg/l 2.19 Scenario Sediment µg/kg 439 µg/kg WW 2.19 WW STP ua/l ua/l 0.00 Tonnage Surface water 1.80 µq/l 11.2 µq/l 0.16 Scenario Sediment µg/kg 439 µg/kg WW 0.16 WW Estimated PEC/PNEC ratios for surface water and sediment are exceeding the t rigger value of l, indicating an unacceptable risk if the exposure estimation is performed without further refinement and not considering degradation in STP. Based on a tonnage scenario an acceptable risk, i.e. PEC/PNEC < 1, was found for all scenarios relevant for the aquatic compartment. It should be noted that the t onnage-based approach was considered as appropriate for demonstrating a safe use only for a.s. evaluation and that, as a general requirement, also the 22

23 Product-type 19 w ithout shells extracted w ith March 2017 consumption- based approach should indicate a safe use at the stage of product authorisation. Terrestrial Compartment including Groundwater One emission pathway was identified for the exposition of the terrestrial compartment with margosa extract, which applies to the Consumption Scenario and the Tonnage Scenario: Emission via wastewater to STP leading to releases to soil via sewage sludge application and subsequently, leaching to groundwater Based on a lack of effect data for the terrestrial compartment, PNEC 50 ; 1 was calculated as screening approach by applying the EPM. Groundwater assessment is based on the limit of 0. 1 µg/l set for biocides (active substances) in Council Directives 2006/118/EC and 98/83/EC. As the exposure estimation according to the pore water calculation model of the TGD resulted in PEC 9 roundwater > 0.1 µg/l after sewage sludge application, the groundwater exposure assessment was refined by using FOCUS model PEARL (transport and fate simulation tool). The groundwater assessment was refined for the two different emission models (Consumption Scenario or Tonnage scenario) and types or sewage sludge application (on arable land or grassland) Table 2-4 PEC/PNEC ratios for different exposure situations terrestrial compartment concerning the Exposure scenario Environmental compartment µg/kg Soil Consumption WW Scenario Groundwater One safe (refined) scenario µg/kg Soil Tonnage WW Scenario Groundwater Six safe (refined) scenarios PEC PNEC PEC I PNEC 350 µg/kg WW µg/l acceptable 350 µg/kg WW µg/l acceptable The screening approach via EPM did not indicate an unacceptable risk for soil ( PEC/PNEC > 1) for the consumption -based scenario, which considers neither a refinement of the exposure estimation nor degradation in STP. The limit value for the groundwater of 0. 1 µg/l (Council Directives 2006/118/EC and 98/83/EC) is exceeded at none of the assessed locations (Tonnage Scenario, grassland) and at up to seven locations (Consumption Scenario, arable land). However, at the 47th CA-Meeting held in July 2012 it was agreed that at the level of active substance assessment at least one of the nine locations needs to show a safe scenario. The modelling results comply with this requirement, as the location Sevilla is safe for all assessed scenarios. Consequently, the risks for the groundwater of the Consumption Scenario and the Tonnage Scenario can be considered as acceptable. Considering tonnage data the risk assessment could demonstrate an acceptable risk for the terrestrial compartment. It should be noted that the tonnage-based approach was considered as appropriate for demonstrating a safe use only for a.s. evaluation and that, as a general requirement, also the consumption- based approach should indicates a safe use at the stage of product authorisation. Toxicity to honeybees and other beneficial arthropods 23