Conclusion on the peer review of the pesticide risk assessment of the active substance isoproturon 1

Transcription

1 EFSA Journal 2015;13(8):4206 CONCLUSION ON PESTICIDE PEER REVIEW Conclusion on the peer review of the pesticide risk assessment of the active substance isoproturon 1 ABSTRACT European Food Safety Authority 2 European Food Safety Authority (EFSA), Parma, Italy The conclusions of the European Food Safety Authority (EFSA) following the peer review of the initial risk assessments carried out by the competent authority of the rapporteur Member State, Germany, for the pesticide active substance isoproturon are reported. The context of the peer review was that required by Commission Regulation (EU) No 1141/2010 as amended by Commission Implementing Regulation (EU) No 380/2013. The conclusions were reached on the basis of the evaluation of the representative uses of isoproturon as a herbicide on winter cereals (wheat, barley, rye, triticale) and spring cereals (wheat, barley). The reliable endpoints concluded as being appropriate for use in regulatory risk assessment, derived from the available studies and literature in the dossier peer reviewed, are presented. Missing information identified as being required by the regulatory framework is listed. Concerns are identified. European Food Safety Authority, 2015 KEY WORDS isoproturon, peer review, risk assessment, pesticide, herbicide 1 On request from the European Commission, Question No EFSA-Q , approved on 28 July Correspondence: pesticides.peerreview@efsa.europa.eu Suggested citation: EFSA (European Food Safety Authority), Conclusion on the peer review of the pesticide risk assessment of the active substance isoproturon. EFSA Journal 2015;13(8):4206, 99 pp. doi: /j.efsa Available online: European Food Safety Authority, 2015

2 SUMMARY Commission Regulation (EU) No 1141/2010 (hereinafter referred to as the Regulation ), as amended by Commission Implementing Regulation (EU) No 380/2013, lays down the procedure for the renewal of the approval of a second group of active substances and establishes the list of those substances. Isoproturon is one of the active substances listed in the Regulation. The rapporteur Member State (RMS) provided its initial evaluation of the dossier on isoproturon in the Renewal Assessment Report (RAR), which was received by the EFSA on 28 February The peer review was initiated on 10 March 2014 by dispatching the RAR for consultation of the Member States and the applicants Nufarm UK Ltd. (on behalf of the BCS-CGNS Isoproturon Task Force) and Makhteshim Agan Holding B.V. Following consideration of the comments received on the RAR, it was concluded that EFSA should conduct an expert consultation in the areas of mammalian toxicology and ecotoxicology and EFSA should adopt a conclusion on whether isoproturon can be expected to meet the conditions provided for in Article 4 of Regulation (EC) No 1107/2009 of the European Parliament and the Council. The conclusions laid down in this report were reached on the basis of the evaluation of the representative uses of isoproturon as a herbicide on winter cereals and spring cereals as proposed by the applicant. Full details of the representative uses can be found in Appendix A to this report. A data gap was identified in the section on analytical methods for an appropriate residue monitoring method for ground water. In the area of mammalian toxicology and non-dietary exposure, further data on the toxicological profile of the metabolites 2-OH-isoproturon and OH-monodesmethyl-isoproturon, present in significant proportions in straw (and rotational crops) are needed and a data gap was identified. The interim provisions of Annex II, Point of Regulation (EC) No 1107/2009 concerning human health for the consideration of endocrine disrupting properties are met leading to a critical area of concern. Regarding the scientific risk assessment a data gap was identified to investigate the potential endocrine-mediated effects of isoproturon. Groundwater metabolites are considered relevant based on the classification of the parent compound leading to a critical area of concern. Based on the new metabolism studies submitted in the framework of the renewal procedure, the residue definition for monitoring was confirmed as isoproturon (restricted to the uses on the cereal group only). Storage stability data for the metabolite RPA are required. No acute or chronic consumer intake concerns were identified. The data available on environmental fate and behaviour are sufficient to carry out the required environmental exposure assessments at EU level for the representative uses, with the notable exception that information is missing regarding the potential for aquatic organisms to be exposed to aqueous photolysis transformation products. The available groundwater exposure assessments result in the identification of the critical area of concern that there is a high potential for the representative uses assessed to result in groundwater exposure above the parametric drinking water limit of 0.1µg/L by the relevant metabolites of isoproturon in situations represented by all 9 pertinent FOCUS groundwater scenarios. In the ecotoxicology section data gaps and critical areas of concern were identified as regards the long term risk of isoproturon to birds and wild mammals. Data gaps and critical areas of concern regarding the risk to aquatic organisms were identified for isoproturon and the metabolite desmethylisoproturon. A data gap was identified for the metabolite desmethyl-isoproturon regarding the chronic risk to earthworms. EFSA Journal 2015;13(8):4206 2

3 TABLE OF CONTENTS Abstract... 1 Summary... 2 Background... 4 The active substance and the formulated product... 6 Conclusions of the evaluation Identity, physical/chemical/technical properties and methods of analysis Mammalian toxicity Residues Environmental fate and behaviour Ecotoxicology Overview of the risk assessment of compounds listed in residue definitions triggering assessment of effects data for the environmental compartments Soil Ground water Surface water and sediment Air List of studies to be generated, still ongoing or available but not peer reviewed Particular conditions proposed to be taken into account to manage the risk(s) identified Concerns Issues that could not be finalised Critical areas of concern Overview of the concerns identified for each representative use considered References Appendices Abbreviations EFSA Journal 2015;13(8):4206 3

4 BACKGROUND Commission Regulation (EU) No 1141/ (hereinafter referred to as the Regulation ) as amended by Commission Implementing Regulation (EU) No 380/ lays down the detailed rules for the procedure of the renewal of the approval of a second group of active substances. This regulates for the European Food Safety Authority (EFSA) the procedure for organising the consultation of Member States and the applicants for comments on the initial evaluation in the Renewal Assessment Report (RAR) provided by the rapporteur Member State (RMS), and the organisation of an expert consultation, where appropriate. In accordance with Article 16 of the Regulation, if mandated, EFSA is required to adopt a conclusion on whether the active substance is expected to meet the conditions provided for in Article 4 of Regulation (EC) No 1107/2009 within 6 months from the receipt of the mandate, subject to an extension of up to 9 months where additional information is required to be submitted by the applicant(s) in accordance with Article 16(3). In accordance with Article 4 of the Regulation Germany (hereinafter referred to as the RMS ) received an application from Nufarm UK ltd. (on behalf of the BCS-CGNS Isoproturon Task Force) and Makhteshim Agan Holding B.V. for the renewal of approval of the active substance isoproturon. Complying with Article 11 of the Regulation, the RMS checked the completeness of the dossier and informed the applicants, the Commission and EFSA about the admissibility. The RMS provided its initial evaluation of the dossier on isoproturon in the Renewal Assessment Report (RAR), which was received by EFSA on 28 February 2014 (Germany, 2014). The peer review was initiated on 10 March 2014 by dispatching the RAR to Member States, to the applicant Nufarm UK ltd. on behalf of the BCS-CGNS Isoproturon Task Force and to the applicant Makhteshim Agan Holding B.V. for consultation and comments. In addition, EFSA conducted a public consultation on the RAR. The comments received were collated by EFSA and forwarded to the RMS for compilation and evaluation in the format of a Reporting Table. The applicants were invited to respond to the comments in column 3 of the Reporting Table. The comments and the applicants response were evaluated by the RMS in column 3. The need for expert consultation and the necessity for additional information to be submitted by the applicants in accordance with Article 16(3) of the Regulation were considered in a telephone conference between EFSA, the RMS, and the European Commission on 7 October On the basis of the comments received, the applicant s response to the comments and the RMS s evaluation thereof it was concluded that additional information should be requested from the applicants and EFSA should organise an expert consultation in the areas of mammalian toxicology and ecotoxicology. In accordance with Article 16(2) of the Regulation, COM decided to consult EFSA. The mandate was received on 15 October The outcome of the telephone conference, together with EFSA s further consideration of the comments is reflected in the conclusions set out in column 4 of the Reporting Table. All points that were identified as unresolved at the end of the comment evaluation phase and which required further consideration, including those issues to be considered in an expert consultation and the additional information to be submitted by the applicants were compiled by EFSA in the format of an Evaluation Table. 3 Commission Regulation (EU) No 1141/2010 of 7 December 2010 laying down the procedure for the renewal of the inclusion of a second group of active substances in Annex I to Council Directive 91/414/EEC and establishing the list of those substances. OJ L 322, , p Commission Implementing Regulation (EU) No 380/2013 of 25 April 2013 amending Regulation (EU) No 1141/2010 as regards the submission of the supplementary complete dossier to the Authority, the other Member States and the Commission. OJ L 116, , p.4 EFSA Journal 2015;13(8):4206 4

5 The conclusions arising from the consideration by EFSA, and as appropriate by the RMS, of the points identified in the Evaluation Table, together with the outcome of the expert consultation, where this took place, were reported in the final column of the Evaluation Table. A final consultation on the conclusions arising from the peer review of the risk assessment took place with Member States via a written procedure in June This conclusion report summarises the outcome of the peer review of the risk assessment on the active substance and the representative formulation evaluated on the basis of the representative uses as a herbicide on winter cereals (wheat, barley, rye, triticale) and spring cereals (wheat, barley) as proposed by the applicants. A list of the relevant end points for the active substance as well as the formulation is provided in Appendix A. In addition, a key supporting document to this conclusion is the Peer Review Report, which is a compilation of the documentation developed to evaluate and address all issues raised in the peer review, from the initial commenting phase to the conclusion. The Peer Review Report (EFSA, 2015) comprises the following documents, in which all views expressed during the course of the peer review, including minority views, can be found: the comments received on the RAR, the Reporting Table (9 October 2014) the Evaluation Table (20 July 2015) the report(s) of the scientific consultation with Member State experts (where relevant), the comments received on the assessment of the additional information (where relevant), the comments received on the draft EFSA conclusion. Given the importance of the RAR including its revisions (Germany, 2015) and the Peer Review Report, both documents are considered respectively as background documents to this conclusion. It is recommended that this conclusion report and its background documents would not be accepted to support any registration outside the EU for which the applicant has not demonstrated to have regulatory access to the information on which this conclusion report is based. EFSA Journal 2015;13(8):4206 5

6 THE ACTIVE SUBSTANCE AND THE FORMULATED PRODUCT Isoproturon is the ISO common name for 3-(4-isopropylphenyl)-1,1-dimethylurea or 3-p-cumenyl-1,1- dimethylurea (IUPAC). The representative formulated products for the evaluation were Isoproturon 500 SC and Protugan 50 SC ; both are suspension concentrates (SC) containing 500 g/l isoproturon. The representative uses evaluated are spraying applications against grass weeds and broadleaved weeds in winter and spring cereals. Full details of the GAPs can be found in the list of end points in Appendix A. It should be noted that the maximum application rate considered under the Art. 12 review of MRLs was higher, e.g. 2 kg a.s./ha instead of 1.5 kg a.s./ha. CONCLUSIONS OF THE EVALUATION 1. Identity, physical/chemical/technical properties and methods of analysis The following guidance documents were followed in the production of this conclusion: SANCO/3030/99 rev.4 (European Commission, 2000), SANCO/10597/2003 rev (European Commission, 2012) and SANCO/825/00 rev. 8.1 (European Commission, 2010) The minimum purity of the active substance as manufactured is 980 g/kg for BCS-CGNS Isoproturon Task Force and 970 g/kg for Makteshim-Agan Holding B.V. respectively, both meeting the requirements of the FAO specification 336/TC/S (l990) (AGP: CP/250). N-nitroso-dimethylamine is considered a potential relevant impurity at a maximum content of 0.2 mg/kg but only for the BCS-CGNS Isoproturon Task Force source (see Section 2). The proposed specifications are based on batch data from industrial scale production. The assessment of the data package revealed no issues that need to be included as critical areas of concern with respect to the identity, physical, chemical and technical properties of isoproturon or the representative formulations. The main data regarding the identity of isoproturon and its physical and chemical properties are given in Appendix A. Methods of analysis are available for the active substance in the technical material and formulations and also for the relevant impurity. Isoproturon residues can be monitored in food and feed of plant origin by LC-MS/MS methods with LOQs of 0.01 mg/kg in each commodity group. Monitoring methods for food and feed of animal origin are not required as no residue definition was proposed. The residue definition for monitoring in soil was defined as isoproturon and desmethyl-isoproturon (see Sections 4 and 5). Compounds of the residue definition in soil can be monitored by LC-MS/MS with LOQs of 0.01 mg/kg for both compounds. Appropriate LC-MS/MS method exists for monitoring residues of isoproturon and desmethyl-isoproturon in ground water and surface water with a LOQ of 0.05 µg/l. It should be noted however, that the residue definition for monitoring in ground water was defined as isoproturon and its metabolites 1-OH-isoproturon, 2-OH-isoproturon, didesmethylisoproturon and propanoic acid-isoproturon, as a consequence a data gap was identified for an appropriate method able to quantify all the compounds of the residue definition. Residues of isoproturon in air can be monitored by HPLC-UV with a LOQ of 0.9 µg/m 3 or by LC-MS/MS with a LOQ of 1 µg/m 3. The active substance is not classified for a health hazard triggering a method of analysis for body fluids and tissues (see Section 2). EFSA Journal 2015;13(8):4206 6

7 2. Mammalian toxicity The toxicological profile of the active substance isoproturon and its metabolites was discussed at the pesticides peer review experts meeting 125 and assessed based on the following guidance documents: SANCO/221/2000 rev. 10-final (European Commission, 2003), SANCO/10597/2003 rev (European Commission, 2012) and Guidance on Dermal Absorption (EFSA PPR 2012). To assess the toxicological profile of the active substance isoproturon a complete set of valid toxicity studies was available. EFSA considered these toxicity studies representative of the proposed technical specifications based on the fact that there was evidence that the current specifications of isoproturon were devoid of genotoxic potential and given the toxicological profile of the isoproturon (i.e. carcinogenic and reproductive toxicant) impurities other than N-nitroso-dimethylamine (maximum content 0.2 mg/kg) are not expected to alter the toxicological profile of isoproturon. In the toxicokinetics studies, isoproturon was extensively and rapidly absorbed. Oral absorption was estimated to be greater than 90%. There was no evidence for accumulation. Excretion of isoproturon was predominantly through urine. The main metabolic pathway identified was oxidation of the isopropyl group and N-demethylation. In the acute toxicity studies, isoproturon showed low acute toxicity when administered orally, dermally or by inhalation to rats. It is not a skin or eye irritant or a dermal sensitiser. In short-term oral toxicity studies with rats, mice, dogs and monkeys the critical effects were observed in the liver (degeneration of hepatocytes, all species) and blood (haemolytic anaemia, rats, dogs and monkeys). Non-specific critical effects as reduced body weight gain were also observed in rats, dogs and monkeys. The dog was the most sensitive species. The relevant overall short-term oral NOAEL is 3.2 mg/kg bw per day (dog studies). Based on available genotoxicity studies, isoproturon (at least of the current production) is unlikely to be genotoxic. In long-term toxicity and carcinogenicity studies with rats and mice the critical effects included liver toxicity in rats and reduced body weight gain in both rats and mice. The relevant long-term NOAELs are 3.1 mg/kg bw per day for the rat and 20 mg/kg bw per day for the mouse. No evidence of carcinogenicity was observed in mice but hepatic tumours were observed in rats at the critical LOAEL of 15.2 mg/kg bw per day and above. Further mechanistic data were submitted on the mode of action for hepatic tumours in order to exclude the human relevance. The carcinogenic potential was discussed during the pesticide peer review expert meeting: the experts agreed that the applicants did not clearly demonstrate the non-relevance for humans of the mode of action for the liver tumours in the rat on the basis of the new mechanistic data provided. The experts considered that the harmonised classification and labelling as Carc. Cat. 2 (H351) would still be appropriate for isoproturon. In reproductive toxicity studies, fertility and overall reproductive performance was impaired, i.e. reduced mating index, pregnancy rate, number of implantations, litter size, pup weight were observed at 40 mg/kg bw. Parental toxicity including reduced body weight gain and histopathological changes in the liver, spleen and testes (retarded spermatogenesis) were observed from dose levels of 20 mg/kg bw per day. The parental, reproductive and offspring NOAELs are 10 mg/kg bw per day. The reproductive toxicity potential was discussed during the experts meeting: the majority of the experts considered that reproductive toxicity seen in rats can be due to reduced male fertility suggesting that classification as Reproductive toxicity Cat. 2 (H361f: Suspected of damaging fertility) 5 would be 5 It should be noted that proposals for classification made in the context of the evaluation procedure under Regulation (EC) No 1107/2009 are not formal proposals. Classification is formally proposed and decided in accordance with Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of substances and mixtures, amending and repealing Directives 67/548/EEC and 1999/45/EC, and amending Regulation (EC) No 1907/2006. OJ L 353, , EFSA Journal 2015;13(8):4206 7

8 required for isoproturon, as proposed by the RMS. In the developmental toxicity studies, there was no evidence of teratogenicity, and the relevant maternal and developmental NOAELs are 80 mg/kg bw per day for the rat and 40 mg/kg bw per day for the rabbit. No signs of delayed neurotoxicity were observed in domestic hens. Isoproturon did not show a neurotoxic potential in the standard toxicity studies. Isoproturon is classified as carcinogenic category 2 (harmonised classification under Regulation (EC) No. 1272/2008) and proposed to be classified as toxic for reproduction category 2, in accordance with the provisions of Regulation (EC) No 1272/2008 and therefore, the conditions of the interim provisions of Annex II, Point of Regulation (EC) No 1107/2009 concerning human health for the consideration of endocrine disrupting properties are met. With regard to the scientific risk assessment, results from the reproductive toxicity studies indicated that isoproturon may be an endocrine disrupting compound in mammals. Effects on fertility and overall reproductive performance in the two-generation reproductive toxicity studies in rats might be endocrine-mediated. Scientific literature indicated that isoproturon might have mild anti-estrogenic and anti-androgenic activity. Available data are not sufficient to rule out an endocrine-mediated mode of action; in particular the Level 2 tests, which are currently indicated in the OECD Conceptual Framework (OECD, 2012) and analysed in the EFSA Scientific Opinion on the hazard assessment of endocrine disruptors (EFSA SC, 2013), are missing. EFSA identified a data gap. Concerning the setting of health-based references values the agreed acceptable daily intake (ADI) is mg/kg bw per day, on the basis of the relevant long-term NOAEL of 3.1 mg/kg bw in 2-year study in rats based on critical effect at 15.2 mg/kg bw per day. A standard uncertainty factor of 100 plus 2 was applied. The ADI provides a margin of safety of at least 1000 relative to the LOAEL for liver tumours in rats (i.e mg/kg bw per day). The need of acute reference dose (ARfD) was discussed and the experts agreed to set an ARfD based on haematological effects observed in short-term dog studies since the onset of these effects can be acute. The agreed ARfD is 0.1 mg/kg bw based on the NOAEL of 10 mg/kg bw per day for haematological effects observed at 38 mg/kg bw per day in the dog studies (i.e. increased Heinz body formation at 4 weeks in the first 90 day dog study and methaemoglobin formation at the end of the second 90 day dog study). An uncertainty factor of 100 was applied. The agreed systemic acceptable operator exposure level (AOEL) is 0.03 mg/kg bw per day on the basis of the relevant overall NOAEL of 3.2 mg/kg bw per day in the dog studies based on liver toxicity and haemolytic anaemia at mg/kg bw per day. A standard uncertainty factor of 100 was applied. No correction factor for oral absorption is needed to derive the AOEL. The RMS estimated dermal absorption values of isoproturon in Isoproturon 500 SC of 0.1 % for the concentrate and 4% for the dilution and in Protugan 50 SC of 0.2% for the concentrate and 14% for the dilution. The RMS considered the representative use in cereals for the formulation Protugan 50 SC as a worstcase scenario for non-dietary exposure (i.e. operator, worker, bystander and resident). The estimated operator exposure was below the AOEL (31% of the AOEL) with the use of personal protective equipment (PPE) during mixing and loading and application according to the German Model (i.e. gloves during mixing and loading and application and protective clothing and sturdy footwear during application). Worker exposure was below the AOEL without the use of PPE (35% of the AOEL). Bystander and resident exposure was below the AOEL (maximum 32% of the AOEL; adult bystander). Toxicological studies were provided on metabolites didesmethyl-isoproturon, 1-OH-isoproturon, 2- OH-isoproturon, propanoic acid-isoproturon and desmethyl-isoproturon. Didesmethyl-isoproturon, 1-OH-isoproturon, 2-OH-isoproturon, and propanoic acid-isoproturon can occur in groundwater EFSA Journal 2015;13(8):4206 8

9 above 0.1 µg/l (see Section 3). Genotoxicity studies provided for didesmethyl-isoproturon, 1-OHisoproturon, 2-OH-isoproturon, and propanoic acid-isoproturon indicated that they are not genotoxic. However, mechanistic data were not sufficient to demonstrate that these metabolites did not share the carcinogenic potential of isoproturon. In addition, no data on the reproductive toxicity potential of the metabolites are available. The metabolites are considered relevant groundwater metabolites because it could not be excluded that they share the carcinogenic and reproductive toxicity potential of isoproturon, which leads to a critical area of concern. Further toxicological data on metabolites 2-OHisoproturon and OH-monodesmethyl-isoproturon (present in significant proportions in straw (and rotational crops), see Section 3) are needed to conclude on whether they are of higher, equal or less toxic than the parent (see Section 3). Toxicity studies on metabolite desmethyl-isoproturon indicated that is of moderate acute oral toxicity to rats and showed no mutagenic potential in the Ames test. 3. Residues The assessment in the residue section is based on the guidance documents listed in the guideline 1607/VI/97 rev.2 and the guideline on extrapolation SANCO 7525/VI/95 rev. 9 (European Commission, 1999, 2011), the recommendations on livestock burden calculations stated in the JMPR reports (JMPR, 2004, 2007) and the OECD publication on MRL calculations (OECD, 2011). New metabolism studies on cereals (primary and rotational crops) using 14 C-isoproturon labelled on the phenyl ring were submitted and assessed in the framework of the renewal procedure. On primary crop, metabolism was investigated in wheat following post-emergence application at a rate of 1875 g/ha (1.25N). In addition, it was agreed that the rotational crop study conducted on wheat at 1566 g/ha (1N) with a plant back interval (PBI) of 30 days, was appropriate to support the preemergence uses on cereals. The assessment of the active substance was based mainly on these new studies conducted according to the current requested standards. The former studies, performed in the 1970s and 1980s and reported in the DAR for the initial approval of the active substance, were considered as informative only. Following post-emergence application, isoproturon accounted for 8% TRR only in immature plants, the main metabolite being identified as 4-isopropylaniline (28% TRR). At harvest, isoproturon was detected 18% TRR in straw and 3% TRR only in grain, where the main metabolite was identified as RPA , accounting for 19% TRR. Following pre-emergence application, at 30 day PBI, isoproturon was almost not detected, except in forage (11 % TRR) and residues in forage, hay and straw mostly composed of the hydroxy metabolites 2-OH-isoproturon (24 to 30% TRR), 1-OHisoproturon (13% TRR forage) and OH-monodesmethyl-isoproturon (15 % TRR in hay and straw). For the longer PBI of 213 and 324 days, OH-monodesmethyl-isoproturon and 2-OH-isoproturon were also identified as the major metabolites in rotational crops, especially in cereal straw (14% TRR), Swiss chard (15 to 25% TRR) and turnip leaves (16 to 29% TRR). The metabolism of isoproturon in wheat proceeds mainly via hydroxylation of the isopropyl group in combination with the demethylation of the dimethylurea group or the cleavage of this group leading to the aniline moiety. In addition, an uptake via roots also contributes to the presence of the major soil metabolites in plant (1- OH- and 2-OH-isoproturon). Based on these studies, the residue definition for risk assessment was initially proposed by the RMS as the sum of isoproturon and RPA , since RPA was observed in significant levels and proportions in cereal grains in the primary crop metabolism study. However, this metabolite was not detected in the additional field trials provided by the applicant and therefore, EFSA agrees with the RMS proposal, not to include RPA in the residue definition. In contrast, since the hydroxy metabolites (2-OH-isoproturon and OH-monodesmethyl-isoproturon) were concluded as the relevant markers of the residues in rotational crops and observed in significant levels in straw (0.32 and 0.51 mg eq/kg) in the rotational crop study simulating the pre-emergence application, their possible contribution to the animal intakes should not be disregarded. Therefore, EFSA is of the opinion that the residue definition for risk assessment should provisionally be proposed as the sum of isoproturon, 2-OH-isoproturon and OH-monodesmethyl-isoproturon, pending the submission of residues trials EFSA Journal 2015;13(8):4206 9

10 where these metabolites are analysed for and pending the submission of additional information to conclude on the carcinogenic and reprotoxic properties of these hydroxy metabolites. For monitoring, the residue definition was limited to isoproturon only. Theses residue definitions are restricted to the cereal group. A large number of residue trials conducted in the 1970s, 1980s and to a small extent in the early 1990s were submitted. Samples were analysed using methods including a hydrolysis step and therefore, assumed to analyse all component containing the 4-isopropyl-aniline moiety. Considering that residue levels were measured below or close to the limit of quantifications, it can be concluded that the metabolites containing the 4-isopropyl-aniline moiety (mono and didesmethyl-isoproturon, 4- isopropyl-aniline) are not expected to be present in significant levels in cereal grain and straw at harvest. Six additional trials conducted in 2013 were submitted where samples were analysed for isoproturon and RPA While accounting for 19 % TRR in cereal grains in the metabolism studies (0.017 mg/kg), RPA was not detected in in grain and straw in these field trials (<0.01 mg/kg). Based on this residue dataset, a MRL of 0.05 mg/kg was proposed for cereal grains. Since any individual component was detected above 0.01 mg eq/kg in the different plant commodities investigated at the latest plant back intervals (213 and 314 days) representative of typical crop rotations for cereals, it was concluded that significant residues are not expected in rotational crops, provided that isoproturon is applied according to the supported uses. Residue trial data are supported by storage stability studies, showing isoproturon to be stable for at least 14 and 24 months in cereal straw and grain respectively, when stored at <-18 C. Processing studies on cereals were not provided and are not requested since residues in grains were mostly below the LOQs. Metabolism and feeding studies in livestock were not submitted while, using the highest residue levels observed in cereal straw (0.53 mg/kg) the trigger value of 0.1 mg/kg dry matter (DM) for the submission of these data is exceeded for beef and dairy cattle (0.13 and 0.31 mg/kg DM). However, having regard to the large dataset provided for cereals (95 trials), the intakes by livestock are all calculated to be below 0.1 mg/kg DM when the second highest value of 0.15 mg/kg in straw is considered (corresponding to the 99 th percentile value). Therefore EFSA agrees whit the conclusion of the RMS that, based on the available information, a ruminant metabolism study should not be required. However, this assessment should be considered provisional, pending the submission on the information requested on the hydroxy metabolites. The consumer risk assessment was conducted with the EFSA PRIMo model. No chronic or acute consumer intake concerns were identified. Using the MRL values proposed for cereal grains, the highest chronic intake was calculated to be 4% of the ADI (WHO, cluster B) and the highest acute intake 0.7% of the ARfD for wheat (DE, Child). 4. Environmental fate and behaviour The rates of dissipation and degradation in the environmental matrices investigated were estimated using FOCUS (2006) kinetics guidance. In soil laboratory incubations under aerobic conditions in the dark, isoproturon exhibited low to moderate persistence, forming the major (>10% applied radioactivity (AR)) metabolite desmethyl-isoproturon (max. 14 % AR) which exhibited moderate to medium persistence. 1-OH-isoproturon (max. 9 % AR), 2-OH-isoproturon (max. 9 % AR), didesmethyl-isoproturon (max. 7.5 % AR) and propanoic acid-isoproturon (max. 9 % AR) reached levels triggering assessment for groundwater exposure. A data gap has been identified for one further reliable aerobic soil DegT50 for didesmethyl-isoproturon as only two reliable values were available (see Section 7). Mineralisation of the phenyl ring 14 C radiolabel to carbon dioxide accounted for % AR after days. The formation of unextractable residues (not extracted by acetonitrile / water or methanol / water and similar extraction systems) for this radiolabel accounted for % AR after days. In anaerobic soil incubations isoproturon was essentially stable. Isoproturon exhibited high mobility in soil. Desmethyl-isoproturon exhibited high to medium soil mobility and 1- OH-isoproturon and 2-OH-isoproturon exhibited very high soil mobility. It was concluded that the adsorption of these compounds was not ph dependent. There is a data gap for a batch soil adsorption EFSA Journal 2015;13(8):

11 study for the metabolite didesmethyl-isoproturon ensuring an appropriate range of environmentally relevant soil ph are investigated, (considering the different tautomers and possible zwitterions of didesmethyl-isoproturon, see Section 7). In two lysimeter studies (4, 1.2 m depth sand monoliths investigated at a single site in Germany, 2 having autumn applications with the other two receiving spring applications) parent isoproturon was present in leachate with annual average concentrations being above 0.1µg/L in one spring and one autumn treated monolith (at and up to µg/l respectively). In each year in the other monolith of each replicate, leachate concentrations were < 0.1µg/L. Radiolabeled resolved transformation products had annual average leachate concentrations < 0.1µg/L. In laboratory incubations in dark aerobic natural sediment water systems (8 investigated), isoproturon exhibited moderate to high persistence, forming the major metabolites desmethyl-isoproturon (max. 19 % AR water and 13 % AR in sediment) and didesmethyl-isoproturon (max. 14 % AR in water). The unextractable sediment fraction (not extracted by acetone / water or acetone) was a sink for the phenyl ring 14 C radiolabel, accounting for % AR at study end ( days). Mineralisation of this radiolabel accounted for % AR at the end of the study. The rate of decline of isoproturon in laboratory sterile aqueous photolysis experiments was slow relative to that occurred in the aerobic sediment water incubations in 2 investigations. In a third investigation photolysis was more significant, resulting in 2 major but unidentified transformation products (ascribed as B and C ) being formed in a ph 7 buffer. Therefore data gaps have been identified for a description of what was done in relation to attempting to identify these two major photodegradates and addressing the potential for aquatic exposure and risk from these photodegradates (see Section 7). This results in an assessment being not finalised in relation to there not being an aquatic risk assessment for potential major phototransformation products. The necessary surface water and sediment exposure assessments (Predicted environmental concentrations (PEC) calculations) were carried out for the metabolites desmethyl-isoproturon and didesmethyl-isoproturon, using the FOCUS (FOCUS, 2001) step 1 and step 2 approach (version 2.1 of the Steps 1-2 in FOCUS calculator). For the active substance isoproturon, appropriate step 3 (FOCUS, 2001) and step 4 calculations were available 6. The step 4 calculations appropriately followed the FOCUS (FOCUS, 2007) guidance, with no-spray drift buffer zones of up to 5 m being implemented (representing a % spray drift reduction) and vegetative buffer strips of up to 20 m (reducing solute flux in run-off by 80 % and particulate erosion runoff by 95%) being implemented for the run-off scenarios. The SWAN tool (version 3.0.0) was appropriately used to implement these mitigation measures in the simulations. However, risk managers and others may wish to note that whilst run-off mitigation is included in the step 4 calculations available, the FOCUS (FOCUS, 2007) report acknowledges that for substances with K Foc < 2000 ml/g (i.e. isoproturon), the general applicability and effectiveness of run-off mitigation measures had been less clearly demonstrated in the available scientific literature, than for more strongly adsorbed compounds. The necessary groundwater exposure assessments were appropriately carried out using FOCUS (FOCUS, 2009) scenarios and the model PELMO for the active substance isoproturon and its soil transformation products desmethyl-isoproturon, 1-OH-isoproturon, 2-OH-isoproturon, propanoic acid-isoproturon and didesmethyl-isoproturon. For didesmethyl-isoproturon as soil adsorption data were not available a K Foc of 1 ml/g (to reflect minimal soil adsorption) was used in simulations. The potential for groundwater exposure from the representative uses by isoproturon and desmethylisoproturon above the parametric drinking water limit of 0.1 µg/l was concluded to be low in geoclimatic situations that are represented by all 9 FOCUS groundwater scenarios. For the relevant metabolites 1-OH-isoproturon, 2-OH-isoproturon, propanoic acid-isoproturon and didesmethylisoproturon, annual average concentrations moving below 1m were predicted to be above this parametric limit for 3/6, 0/6, 2/6 and 6/6 scenarios respectively in spring planted cereals and 9/9, 7/9, 9/9 and 9/9 scenarios respectively in autumn planted cereals. 6 Simulations correctly utilised the agreed Q10 of 2.58 (following EFSA, 2007) and Walker equation coefficient of Simulations correctly utilised the agreed Q10 of 2.58 (following EFSA, 2007) and Walker equation coefficient of 0.7 EFSA Journal 2015;13(8):

12 A satisfactory literature search on published peer reviewed scientific literature for the metabolites of isoproturon that reach levels that trigger assessment in environmental compartments was not available (see Section 7). The PEC in soil, surface water, sediment, and groundwater covering the representative uses assessed can be found in Appendix A of this conclusion. 5. Ecotoxicology The risk assessment was based on the following documents: European Commission (2002a, 2002b), SETAC (2001), and EFSA (2009). On the basis of the available data and assessments, a low acute dietary risk to birds and wild mammals was concluded for the representative use of isoproturon. A low risk was also concluded from secondary poisoning and exposure via consumption of contaminated water. During the Pesticides Peer Review Expert Meeting 128, the experts discussed the dietary long-term risk assessment for birds and wild mammals. As a high long-term risk to both birds and wild mammals was identified at tier one, several refinement options were discussed. Some refinement options were considered as acceptable, but still a high long term risk was identified for all considered focal species; therefore a data gap for a further refinement of the risk assessment was identified. A large number of aquatic ecotoxicological data was available for isoproturon. The risk assessment showed that the risk to aquatic organisms was driven by the toxicity of isoproturon to algae and aquatic plants. During the Pesticides Peer Review Expert Meeting 128, the experts agreed on a NOAEC, derived from one of the available mesocosm studies, in which several species of algae and aquatic plants were tested. An assessment factor of four was agreed upon. EFSA considered that the applicability of this NOAEC for FOCUS pond scenarios presented some uncertainties, as the comparison between the mesocosm exposure profile and those predicted by FOCUS models was not straightforward. A high risk was identified for all scenarios comparing the NOAEC with the PEC values calculated at step 3. With 5 m no-spray buffer strip + 20 m vegetated field strip used as mitigation measures, the risk was still high in the majority of the scenarios (high risk in 7 out of 9 scenarios). It is also noted that, using PEC sw at step 3 and 4, high acute and chronic risk was identified, in at least one FOCUS scenario, for all aquatic organisms which are not covered by the mesocosm NOAEC (fish, Daphnia, Chironomus). The risk assessment for the metabolite desmethyl-isoproturon was driven by the toxicity to algae and aquatic plants. A high risk was identified when comparing these toxicity values to PEC calculated at the step 2. No step 3 PEC calculations were provided. A data gap for further refining the risk assessment for aquatic organisms was therefore identified for isoproturon and the metabolite desmethyl-isoproturon. Toxicity of the metabolite didesmethyl-isoproturon to algae and aquatic plants was several orders of magnitude lower than the toxicity of isoproturon and a low risk to aquatic organisms was identified. A low risk for all groundwater metabolites was concluded by comparing PEC gw to the respective ecotoxicological endpoints. The available data and risk assessments were suitable to conclude a low risk to honey bees and nontarget arthropods. A low risk to earthworms was concluded for isoproturon, however a data gap was identified for addressing the chronic risk of the metabolite desmethyl-isoproturon to earthworms, as this metabolite was found to be more toxic than isoproturon in acute testing, but no information was provided for chronic toxicity. Low risk to soil macroorganisms and soil microorganisms was concluded for isoproturon. A low risk to non-target terrestrial plants was concluded when mitigation measures comparable to a 5 m no-spray buffer strip are applied. A low risk to sewage treatment organisms was also concluded. EFSA Journal 2015;13(8):

13 For the ecotoxicological assessments, one peer reviewed paper was available in RAR concerning the potential endocrine activity of isoproturon. The available study used a recombinant yeast screen to detect receptor mediated (anti-) estrogenic and (anti-) androgenic activity; cultured Xenopus oocytes were used to measure effects on the ovulatory response and ovarian steroidogenesis. Some antiestrogenic and antiandrogenic activities were reported in the study, along with inhibited ovulation without altering hormone levels. This in vitro study was considered as relevant supporting information, but the reported results are not considered strong evidence of endocrine disruption activity. Pending on the outcome of the data gap in Section 2, further ecotoxicological tests might be necessary to address the potential endocrine disrupting properties of isoproturon. EFSA Journal 2015;13(8):

14 6. Overview of the risk assessment of compounds listed in residue definitions triggering assessment of effects data for the environmental compartments 6.1. Soil Compound (name and/or code) isoproturon desmethyl-isoproturon Persistence low to moderate persistence Single first-order DT days (20ºC pf 2 soil moisture) moderate to medium persistence Single first-order DT days or biphasic kinetic DT days (DT days, 20ºC 40-45% MWHC or pf 2.5) Ecotoxicology Low risk to soil dwelling organisms Low acute risk to soil dwelling organisms. Data gap for chronic risk to soil dwelling organisms EFSA Journal 2015;13(8):

15 6.2. Ground water Compound (name and/or code) isoproturon Mobility in soil high mobility K Foc ml/g >0.1 μg/l 1m depth for the representative uses (at least one FOCUS scenario or relevant lysimeter) Pesticidal activity Toxicological relevance No Yes Yes Ecotoxicological activity Negligible exposure via groundwater becoming surface water, but see 6.3 desmethyl-isoproturon high to medium mobility K Foc ml/g No Assessment not triggered Assessment not triggered (Moderate acute oral toxicity in rats; Ames test: negative). Negligible exposure via groundwater becoming surface water, but see 6.3 didesmethyl-isoproturon data gap, conservative value of 1 ml/g used for modelling µg/l No Yes (it cannot be excluded that they share the carcinogenic and reproductive toxicity potential of the parent; not genotoxic) Low risk to aquatic organisms when groundwater becomes surface water 1-OH-isoproturon very high mobility, extrapolated from 2-OHisoproturon winter µg/l, 5 post emergence scenarios < 0.1µg/L spring µg/l, 3 pre & post emergence scenarios < 0.1µg/L No Yes (it cannot be excluded that they share the carcinogenic and reproductive toxicity potential of the parent; not genotoxic) Low risk to aquatic organisms when groundwater becomes surface water 2-OH-isoproturon very high mobility K Foc 9-13 ml/g winter µg/l, 2 pre & all post emergence scenarios < 0.1µg/L No Yes (it cannot be excluded that they share the carcinogenic and reproductive toxicity potential of the parent; not genotoxic) Low risk to aquatic organisms when groundwater becomes surface water propanoic acidisoproturon Very high mobility K Foc ml/g winter µg/l, 6 post emergence scenarios < 0.1µg/L spring µg/l, 4 pre & post emergence scenarios < 0.1µg/L No Yes (it cannot be excluded that they share the carcinogenic and reproductive toxicity potential of the parent; not genotoxic) Low risk to aquatic organisms when groundwater becomes surface water EFSA Journal 2015;13(8):

16 6.3. Surface water and sediment Compound (name and/or code) isoproturon (water and sediment) desmethyl-isoproturon (water and sediment) didesmethyl-isoproturon (water only) Unidentified photolysis metabolite B (water only) Unidentified photolysis metabolite C (water only) Ecotoxicology High risk to aquatic organisms/sediment dwelling organisms High risk to aquatic organisms Low risk to aquatic organisms Data gap Data gap 6.4. Air Compound(name and/or code) isoproturon Toxicology Low acute toxicity by inhalation to rats (LC 50 >1.95 mg/l (4 h, whole body exposure, aerosol)) EFSA Journal 2015;13(8):

17 7. List of studies to be generated, still ongoing or available but not peer reviewed This is a list of data gaps identified during the peer review process, including those areas where a study may have been made available during the peer review process but not considered for procedural reasons (without prejudice to the provisions of Article 56 of Regulation (EC) No 1107/2009 concerning information on potentially harmful effects). Residue monitoring method for ground water able to quantify all the components of the residue definition: isoproturon and its metabolites 1-OH-isoproturon, 2-OH-isoproturon, didesmethylisoproturon and propanoic acid-isoproturon (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Sections 1, 2 and 4). Further toxicological data on plant metabolites 2-OH-isoproturon and OH-monodesmethylisoproturon observed in significant proportions and levels in the plant metabolism studies are required (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 2 and 3). Investigation of potential endocrine-mediated effects of isoproturon, following the OECD Conceptual Framework (OECD, 2012), using a stepwise approach from Level 2 for mammalian toxicology (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Sections 2). Residue trials using pre-emergence application where samples are analyse for isoproturon, 2-OHisoproturon and OH-monodesmethyl-isoproturon are required (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 3). Information on the stability of the metabolite RPA is required (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 3). At least one further reliable aerobic laboratory soil DegT 50 endpoint for metabolite didesmethylisoproturon would be needed, to satisfy the EU data requirements (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 4). A standard batch soil adsorption study for the metabolite didesmethyl-isoproturon ensuring an appropriate range of environmentally relevant soil ph are investigated, (considering the different tautomers and possible zwitterions of didesmethyl-isoproturon) is not available (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 4). A supplemental report to the aqueous photolysis study by Gorman (1995) that includes a description of what was done in relation to attempting to identify the photodegradates > 10 % is not available (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 4). The potential for exposure to and if relevant risk to aquatic organisms from exposure to aqueous photolysis metabolites has not been adequately addressed (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 4). A satisfactory literature search on published peer reviewed scientific literature for the metabolites of isoproturon that reach levels that trigger assessment in environmental compartments was not available (relevant for all representative uses evaluated; submission date proposed by the applicant: unknown; see Section 4). EFSA Journal 2015;13(8):