Commission Regulation 1107/2009 Triazole Derived Metabolite: 1,2,4-Triazole Proposed revision to DT50 Summary, Scientific Evaluation and Assessment July 2011, revised September 2011 (after comments from MS and EFSA) and further revised January 2013 (minor clarifications added post-commenting) 2 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) CONTENTS Page Background 3 B.8 Environmental fate and behaviour B.8.1 Route and rate of degradation in soil B.8.1.1 Aerobic and anaerobic studies B.8.1.3 Field Dissipation studies B.8.10 References relied Appendix 1: List of endpoints June 2011 Appendix 2 : Copy of endpoints agreed at PRAPeR 12 meeting of 15 – 18 January 2007 3 3 3 21 68 70 72 3 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Background and summary A number of the triazole fungicides that were evaluated in the 2nd and 3rd stages of the review programme (as referred to in Article 8(2) of Council Directive 91/414/EEC) have metabolites in common. These metabolites (triazole alanine, triazole acetic acid and 1,2,4-triazole) are collectively known as the ‘Triazole Derivative Metabolites’. A group of Notifiers of triazole fungicides have formed a task force called the Triazole Derivative Metabolite Group (TDMG) to produce a common data package to cover the risk assessment for these metabolites. The UK is acting as RMS for a number of parent triazole active substances, and also for the evaluation of a number of metabolite assessments. This document presents a kinetic re-evaluation of previously submitted laboratory degradation data on the 1,2,4-triazole metabolite using the principles of the FOCUS Degradation Kinetics Guidance. In addition, the TDMG has generated field dissipation data to supplement the reassessment of the laboratory data. The endpoints derived are presented in Appendix I. Fate and behaviour endpoints for 1,2,4-triazole were previously discussed and agreed at the PRAPeR 12 meeting of 15 – 18 January 2007. The endpoints agreed at PRAPeR 12 are presented in Appendix II. CRD proposes that the revised endpoints for metabolite 1,2,4-triazole are agreed by Member States, and then MS should use these endpoints in the re-registration assessments of the triazole fungicide group of parent active substances. Note that the assessment of July 2011 (version 1) was amended in September 2011 (version 2) after comments from MS and EFSA, and again in January 2013 to provide further clarity of information (FINAL version 3). These latter amendments do not result in a change in the assessment or endpoints from those in version 2 and all changes are highlighted in turquoise. B.8 ENVIRONMENTAL FATE AND BEHAVIOUR B.8.1 Route and rate of degradation in soil (IIA 7.1.1, IIIA 9.1.1) B.8.1.1 Aerobic and anaerobic studies (II 7.1.1, IIIA 9.1.1) B.8.1.1.2 Soil rate of degradation studies - laboratory The study subject to re-evaluation is that of Slangen 2000 which was presented in the DARs of a number of triazole fungicide a.s. which have subsequently been listed on Annex I of Directive 91/414/EEC. For completeness, the previous UK evaluation of this study has been re-presented below (in italics) to aid the consideration of the reassessment. i) The degradation of metabolite 1,2,4-triazole under aerobic conditions was investigated according to SETAC (1995), CTB (1995), EPA (Subdivision N, part 1621, 1982) and BBA (part IV, 4-1, 1986) guidelines and GLP. The study is acceptable. [3,5-14C] 1,2,4-triazole (radiochemical purity >98%) was applied to three soils at a dose of 0.06 mg/kg of dry soil, based on an assumption of 750 g of fungicide active 4 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) substance/ha, incorporation in 5 cm of soil and a bulk density of soil of 1.5 g/ml, crop interception of 50 %, 50 mol% of the fungicide is converted to 1,2,4-triazole and that the ratio of the molar mass triazole to parent is 0.25. Portions of 100 g (dry weight) of treated soil were transferred into incubation vessels and adjusted to 40 % of the water holding capacity. The test vessels were connected to a humidified air supply (reduced in CO2 content) and to trapping vessels for volatiles (2-methoxyethanol) and carbon dioxide (2 x NaOH). Monitoring and adjustment of the water content in the soil was performed regularly. Temperature for the incubation was maintained at 20 ± 2°C under dark conditions (incubation vessels were made from brown glass). Details of the soils are given in Table B.8.1. Table B.8.1 Soils used in rate of degradation study on 1,2,4-triazole Identity and Soil type Texture analysis provenance (USDA) (%) Org. C (% ) sand silt clay Laacherhof pH holding capacity (CaCl2) 1.4 34.4 6.4 78.9 14.4 6.7 2.2 50.0 5.8 Laacherhof silt loam 36.9 51.1 12.0 0.98 36.4 6.7 BBA2.2 Hanhofen, Microbial biomass (mg C/kg soil) (g /100 g) 72.4 22.6 5.0 AXXa Germany sandy loam loamy sand Water init.:334 end: 198 init.:294 end: 138 Germany AIII Germany init.:252 end: 138 At each sample time, a single metabolism vessel with its traps was removed from the incubation system for analysis. Soil samples were extracted with methanol, methanol/water (x2) and finally with methanol under reflux conditions. After each extraction step residues were separated by centrifugation and volume and radioactivity of supernatants were determined. Analysis of extracts was performed by TLC combined with radiography scanning. The identity of metabolites and triazole was ascertained by co-chromatography with reference substances on selected samples. Residual radioactivity remaining in soil samples after extraction was determined by combustion in an oxidizer followed by LSC of the trapped carbon dioxide. The sodium hydroxide traps were weighed and their radioactivity determined in an aliquot by LSC. The identity of the radioactivity as 14C-CO2 was confirmed by precipitation as barium carbonate in the solutions from the first trap of the samples from day 120. Volatiles others than carbon dioxide were estimated by determining the weight of the 2-methoxyethanol and measuring its radioactivity in an aliquot. Results are presented in Tables B.8.2 – 4. Degradation rates calculated according to non-linear first order kinetics are presented in Table B.8.5. It should be noted that only data for the first 14 days of the incubation with the BBA 2.2 soil were used in the calculation. The absence of decline in residues of 1,2,4-triazole from day 30 onwards was considered by the study authors as being attributable to ‘reduced bioavailability’ and reduced microbial viability of the soil. 5 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.2 Results of aerobic soil incubation of 1,2,4-triazole in Laacherhof AXXa soil (% AR) 1,2,4triazole total extracts 92.79 89.18 61.84 37.39 30.82 25.76 1,2,4triazole shaking extracts 83.33 81.53 55.33 30.68 23.83 19.11 TAA Dihydro- CO2 shaking triazolone extracts shaking extracts Extr Unextr Mass balance *) day 0 93.72 7.50 101.22 day 0 89.18 9.60 98.78 day 1 2.95 0.04 65.56 31.27 96.87 day 3 4.75 0.39 42.13 58.40 100.92 day 7 6.30 0.89 37.12 56.14 94.15 day 6.93 0.61 7.38 33.31 51.59 92.27 14 day 29.58 22.03 5.58 5.70 35.16 60.82 101.68 14 day 19.83 16.35 1.29 0.52 5.34 22.15 71.86 99.34 30 day 12.49 9.77 0.55 1.03 12.53 14.60 74.64 101.78 61 day120 11.99 7.75 19.63 11.99 63.21 94.83 day120 11.94 8.18 11.10 11.94 66.16 89.20 *) Additionally one unknown with a maximum amount of 0.93 % found for day 0 Table B.8.3 Results of aerobic soil incubation of 1,2,4-triazole in BBA 2.2 Hanhofen soil (% AR) 1,2,4triazole total extracts 90.45 89.39 76.15 52.44 45.68 39.35 1,2,4triazole shaking extracts 80.39 80.25 58.80 43.09 34.47 29.26 TAA Dihydro- CO2 shaking triazolone extracts shaking extracts Extr Unextr Mass balance **) day 0 90.45 8.98 99.43 day 0 89.39 11.19 100.58 day 1 0.01 76.15 24.31 100.47 day 3 0.10 52.44 44.01 96.55 day 7 0.13 45.68 49.73 95.55 day 1.31 0.28 40.67 51.53 92.48 14 day 39.86 27.70 1.67 0.31 41.53 53.07 94.92 14 day 31.84 20.50 0.42 0.49 32.59 69.80 102.87 30 day 30.24 21.13 0.91 0.98 31.75 65.45 98.18 61 day120 32.12 21.00 1.42 32.12 54.13 87.67 day120 29.85 21.22 1.58 29.85 65.01 96.44 **) Additionally one unknown with a maximum amount of 0.63 % found for day 63 6 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.4 Results of aerobic soil incubation of 1,2,4-triazole in Laacherhof AIII soil (% AR) 1,2,4triazole total extracts 85.21 84.70 75.38 50.92 45.24 37.45 1,2,4TAA triazole shaking shaking extracts extracts 75.14 75.56 62.36 46.66 41.85 32.67 Dihydro- CO2 triazolone shaking extracts Extr Unextr Mass balance ***) day 0 85.21 16.87 102.08 day 0 84.70 13.22 97.92 day 1 0.1 75.38 24.72 100.20 day 3 0.57 50.92 44.19 95.68 day 7 1.84 45.24 49.38 96.45 day 2.61 0.28 40.06 47.74 88.07 14 day 38.74 32.40 1.85 1.25 40.59 49.31 91.15 14 day 18.29 16.41 0.27 2.22 19.42 20.99 61.64 102.0 30 day 6.58 5.52 1.50 19.65 8.08 49.90 77.62 61 day 3.74 3.56 24.65 3.74 59.33 87.53 83 day120 1.73 1.32 33.70 1.73 38.47 73.90 day120 2.39 1.61 32.22 2.39 41.79 76.40 ***) Additionally one unknown with a maximum amount of 0.19 % found for day 30 Table B.8.5 Degradation 1,2,4-triazole in three different soils under laboratory conditions (20˚C, 40% MWHC). Soil Soil type Laacherhof sandy loam AXXa BBA 2.2 loamy sand Laacherhof A III silt loam Organic carbon 1.4 % DT50 values 1st order, nonlinear one compartment 6.32 days 2.19 % 9.91 days*) 0.98 % 12.27 days Arithmetic mean 9.5 days *) Only 0-14 day data used for calculation. r2 0.75 0.81 0.95 The degradation rates calculated by the notifier have been confirmed as being acceptable by the Rapporteur. (Slangen 2000) Note that the final sentence relating to the kinetic calculations was in relation to the evaluation which was originally conducted prior to adoption of the FOCUS Degradation Kinetics Guidance Document. 7 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) The TDMG have now provided an updated kinetic assessment of the Slangen 2000 study. ii) The following assessment relates to re-evaluation of the Slangen 2000 study results according to FOCUS Degradation Kinetics guidance. As such, there is no requirement for this evaluation to comply with GLP as it comprises mathematical modelling, but the study appears to have been conducted appropriately. The applicant used the KinGUI 1.1 software with MatLab v 7.0.4.365 to perform the modelling analysis and statistical evaluation of kinetic fits. As the original data (see tables B.8.2 – 4) only had data from single replicate samples except for days 0 and 14, the two replicate data points from each of days 0 and 14 were averaged to give a single data point for each of these two sample times in the kinetic analysis. The fitting procedure started by free-fitting SFO to each data set. If the free-fitting approach was considered unacceptable, the fitting procedure was repeated but fixing the initial concentration. Following the SFO steps (free-fitting and fixed initial concentration), bi-phasic kinetics were considered. As the establishment of modelling parameters for 1,2,4-triazole was under consideration, HS and DFOP were tested for the Laacher Hof AXXa and BBA 2.2 datasets as the final concentrations were >10% and FOMC was used to model the Laacher Hof AIII dataset as final concentration was <10%. The data used in the fitting procedure is shown in Table B.8 6. It is noted that the FOCUS Degradation Kinetics guidance indicates that where concentrations do not reach 10% of the initial concentration at study end then HS or DFOP kinetics should be used. The study author has used both HS and DFOP for these soils. In addition, as DFOP appeared to represent the better kinetic for the Laacher Hof AXXa and BBA 2.2 soils, the Laacher Hof AIII dataset was also subject to analysis using DFOP kinetics to determine whether the endpoints from the database of three soils could be expressed by a single consistent kinetic. Table B.8.6 1,2,4-triazole metabolite data used in fitting (taken from Slangen 2000, % AR) Laacher Hof AXXa DAA BBA 2.2 Laacher Hof AIII Residues % 0 90.99 89.92 84.96 1 3 7 14 30 61 83 120 61.84 37.39 30.82 27.67 19.83 12.49 76.15 52.44 45.68 39.60 31.84 30.24 Not determined Not determined 11.96 30.99 75.38 50.92 45.24 38.09 18.29 6.58 3.74 2.06 8 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) In terms of initial concentration used in the fitting procedure, the mass balance at day 0 was used as there were significant unextracted residues (10 – 15% AR) by the time that the day zero sample was taken after dosing. Radiochemical purity of the test compound was also >98% indicating that radiolabelled impurities would be insignificant at this initial sample time; this is confirmed by the information on the extracted fraction which virtually completely comprised 1,2,4-triazole. Correction of the day 0 residue to account for unextracted residues etc. is appropriate and in accordance with FOCUS Kinetics guidance (see section 6.1.6 of the Kinetics guidance). Fitting was unconstrained except for attempts to achieve an acceptable fit with SFO kinetics where the initial concentration was constrained. As these attempts did not result in acceptable fit, results of SFO fit with constrained initial concentration have not been shown. The results obtained for all three soils are collated below. DT50 / DT50-fast DT90 / DT50-slow [d] Resid. fit Model Visual ranking Table B.8.7 Statistical results of the Applicant kinetic fits for 1,2,4-triazole in three laboratory soils: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. χ2 test error T-test k1 T-test k2 Other Laacher Hof AXXa SFO 4.7 15.6 -- -- 29.7 0.028 HS 1.9 59.2 ++ ++ 8.5 <0.001 0.03 DFOP 0.9 59.2 ++ ++ 5.1 0.001 0.006 SFO 40.9 136.2 -- -- 26.4 0.061 HS 3.2 203.9 + ++ 6.2 <0.001 0.047 DFOP 1.5 247.6 ++ ++ 5.1 0.004 0.065 Tb: 3.2447 (Ttest tb: 0.030) “g”: 0.683 (P= <0.001) BBA 2.2 Tb: 4.0139 (Ttest tb: NA) “g”: 0.580 P= <0.001) Laacher Hof AIII SFO 9.9 33.0 - - 19.3 FOMC 22.1 (from DT90) + + 12.3 DFOP 0.8 ++ ++ 4.5 20.6 0.003 0.003 <0.001 “g”: 0.443 P= <0.001) Summary: Geomean all DFOP fits: DFOP 1.0 d 1 67.1 d “g”: 0.5691 = arithmetic mean The Applicants assessments for each individual soil are shown below in Table B.8.8 10 and Figures B.8.1 – 9. Note that for visual and residual assessment, the following key is used: ++ = Very good; + = Good; - = Marginal; - - = Poor 9 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Laacher Hof AXXa Table B.8.8 Statistical results of the Applicant kinetic fits for Laacher Hof AXXa soil: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of randomness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Kinetics DT50 DT90 Visual ranking Resids fit SFO Kinetics 4.7 DT50 -fast 1.9 DT50 -fast 0.9 15.6 DT50slow 59.2 DT50slow 59.2 -- -- Visual ranking ++ Visual ranking ++ Resids fit ++ Resids fit ++ HS Kinetics DFOP Tb 3.2447 g 0.683 χ2 test error 29.7 χ2 test error 8.5 2 χ test error 5.1 T-test k 0.028 T-test k1 <0.001 T-test k1 0.001 T-test k2 0.03 T-test k2 0.006 T-test tb 0.030 T-test g <0.001 10 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.1 Laacher Hof AXXa Applicant SFO visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 20 Parent 15 10 Residuals 5 0 -5 -10 -15 -20 0 20 40 60 Time 80 100 120 11 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.2 Laacher Hof AXXa Applicant HS visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 6 Parent 4 Residuals 2 0 -2 -4 -6 -8 0 20 40 60 Time 80 100 120 12 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.3 Laacher Hof AXXa Applicant DFOP visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 5 Parent 4 3 Residuals 2 1 0 -1 -2 -3 -4 0 20 40 60 Time 80 100 120 13 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) BBA 2.2 Table B.8.9 Statistical results of the Applicant kinetic fits for BBA 2.2 soil: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Kinetics DT50 SFO Kinetics 40.9 DT50 -fast 3.2 DT50 -fast 1.5 HS Kinetics DFOP DT90 136.0 DT50Tb slow 4.0139 203.9 DT50g slow 247.6 0.580 a = not available Visual ranking Resids fit -- -- Visual ranking + Visual ranking ++ Resids fit ++ Resids fit ++ χ2 test error 26.4 χ2 test error 6.2 2 χ test error 5.1 T-test k 0.061 T-test k1 <0.001 T-test k1 0.004 T-test k2 0.047 T-test k2 0.065 T-test tb NA a T-test g <0.001 14 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.4 BBA 2.2 Applicant SFO visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 40 Parent 30 Residuals 20 10 0 -10 -20 0 20 40 60 Time 80 100 120 15 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.5 BBA 2.2 Applicant HS visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 6 Parent 4 Residuals 2 0 -2 -4 -6 0 20 40 60 Time 80 100 120 16 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.6 BBA 2.2 Applicant DFOP visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 5 Parent 4 3 Residuals 2 1 0 -1 -2 -3 -4 -5 0 20 40 60 Time 80 100 120 17 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Laacher Hof AIII Table B.8.10 Statistical results of the Applicant kinetic fits for Laacher Hof AIII soil: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of randomness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Kinetics DT50 DT90 Visual ranking Resids fit SFO Kinetics 9.9 DT50 33.0 DT90 - - DT50 -fast 0.8 73.4 DT50slow 20.6 Visual ranking + Visual ranking ++ Resids fit + Resids fit ++ FOMC Kinetics DFOP g 0.443 χ2 test error 19.3 χ2 test error 12.3 χ2 test error 4.5 T-test k 0.003 T-test k1 0.003 T-test k2 <0.001 T-test g <0.001 18 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.7 Laacher Hof AIII Applicant SFO visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 20 Parent 15 10 Residuals 5 0 -5 -10 -15 -20 0 20 40 60 Time 80 100 120 19 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.8 Laacher Hof AIII Applicant FOMC visual and residual fits Measured & Predicted vs. Time 100 Parent 90 80 Concentration 70 60 50 40 30 20 10 0 0 20 40 60 Time 80 100 120 Residual Plot 12 Parent 10 8 Residuals 6 4 2 0 -2 -4 -6 -8 0 20 40 60 Time 80 100 120 20 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.9 Laacher Hof AIII Applicant DFOP visual and residual fits Measured & Predicted vs. Time 120 Parent 100 Concentration 80 60 40 20 0 0 20 40 60 Time 80 100 120 Residual Plot 4 Parent 3 2 Residuals 1 0 -1 -2 -3 -4 0 20 40 60 Time 80 100 120 The Applicant proposed the following DT50 values for use in modelling. It should be noted that the slow phase DFOP values are significantly longer than those agreed at PRAPeR 12. 21 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.11 Applicant proposed soil degradation DT50 values of 1,2,4-triazole. Location Laacher Hof AXXa BBA 2.2 Laacher Hof AIII Model fitted: DFOP DFOP DFOP “g” 0.683 0.580 0.443 DT50-fast [d] 0.9 1.5 0.8 DT50-slow [d] 59.2 247.6 20.6 1.0 67.1 Geometric mean: Arithmetic mean: 0.569 Note that no normalisation for moisture content has been performed. This is considered appropriately conservative for 1,2,4-triazole as a terminal metabolite as normalisation for moisture content will result in faster degradation. However, assuming that under conditions of constant moisture status (40% of water holding capacity) the ‘g’ value will be unchanged and fast and slow phases are equally affected, the RMS has calculated the following corrected DT50 values. Note that the values for BBA 2.2 are unchanged as the moisture content in the study was the same as the FOCUS recommended pF2 value for a loamy sand soil. Table B.8.11a RMS calculated DT50 values of 1,2,4-triazole corrected for moisture. Location Model fitted: “g” Laacher Hof AXXa BBA 2.2 Laacher Hof AIII DFOP DFOP DFOP 0.683 0.580 0.443 Geometric mean: Arithmetic mean: Moisture correction factor 0.624 1 0.531 DT50-fast [d] DT50-slow [d] 0.6 1.5 0.4 36.9 247.6 10.9 0.7 46.4 0.569 The Evaluator agrees with the use of the initial concentration of 100% for all three soils based on the mass balance data from the original study for all three soils. The Evaluator also considers that whilst DFOP would not be normally be used on data sets where the applied substance reached at least 10% of the initial dose within the study, e.g. the data set for the Laacher Hof AIII soil, DFOP is considered the preferred kinetic for all three soils, this being a pragmatic choice for consistent modelling parameters. Overall, the Evaluator agrees with this kinetic assessment of 1,2,4-triazole in laboratory soils. Chapple 2010a B.8.1.3 Field studies Field dissipation i) A field dissipation study was conducted on 1,2,4-triazole in accordance with BBA (Part IV, 4-1, 1986) and SETAC (1995) guidelines and to GLP. 22 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) The field dissipation study was conducted at four different locations, one each in Germany, UK, Italy and Spain. Details of the sites and soil properties are given in Table B.8.12 - 15. The study author stated that the sites had not been treated with chemicals which could influence the dissipation behaviour of 1,2,4-triazole or interfere with analysis. Site histories were presented, which indicated no apparent application of triazole fungicides in at least the previous three years at the German and Italian site. Site history was not actually available for the UK site although it was acknowledged that the site had been used for field testing of triazole fungicides previously. No history of previous pesticide use was available for the Spanish site. Untreated control plots were sampled at each site to determine background levels of 1,2,4-triazole. Table B.8.12 Soil characteristics of German field dissipation site 23 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.13 Soil characteristics of Italy field dissipation site Table B.8.14 Soil characteristics of UK field dissipation site 24 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.15 Soil characteristics of Spain field dissipation site Note: MWHC = maximum water holding capacity; organic matter content calculated from % organic carbon x 1.724 At all sites, a single application of 100 g 1,2,4-triazole/ha was applied to bare soil plots which had been pre-prepared to have a fine crumb structure. Following application, grass seed was sown on the bare soil plots at all sites except the Spanish site. The staff conducting the Spanish site considered that given the time of year of application, i.e. June, and the prevailing drought conditions at the time of application, there was no prospect of grass emergence, and so no grass seed was sown at this site. The study author noted that the substance was incorporated almost immediately after application to a depth of 8cm by the grass seed sowing operation. Accuracy of application was checked using filter papers laid on the soil surface. Control plots were also left unsprayed to check for any background levels of 1,2,4-triazole. Meteorological data were available for all four sites. For the German site, a weather station was available at the test location. For the Italian site, the weather station was located 200 m away from the site. For the UK, weather data was available for a weather station 32km away. For the Spanish site, weather data were available from a site 2.7 km away. Full daily weather data were not presented in the report, but meteorological data at selected dates and total precipitation and monthly mean temperature were available. Total precipitation and range of monthly mean temperature are presented with results of the residues for the individual sites in Tables B.8.20 – 23. It should be noted that daily resolution weather data were provided in another report (Chapple 2010b) detailing normalisation of field dissipation data. Soil sampling was conducted before and immediately after application at all sites. Day 0 samples were taken to a depth of 10cm (50 mm diameter, 40 cores per sample). On 11 subsequent occasions up to 454 days after application, samples were taken down to a maximum of 50cm depth. Dates and depths of samples are given in the following 25 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) results tables. Note that the Italian study was terminated at day 180 as the site became no longer available. Soil samples were deep frozen within 24 hours and stored frozen at -18ºC or below until preparation for analysis. Prior to analysis, cores were divided into 10cm segments and the sample from the appropriate segment homogenised. 15 g subsamples of the homogenised soil were then extracted with acetonitrile/water (6:4 v/v) at 65-70ºC for one hour. Following centrifugation, an aliquot of the extract was derivitised using dansyl chloride in acetone and aqueous sodium sulphate to form dansyl triazole; derivitisation was stated to have been performed to enable analysis to be performed using a non-polar HPLC system and to allow production of more transitional ions in mass spectrometric analysis, aiding sensitivity and detection. Following addition of ammonium hydroxide solution, the derivatives were partitioned into ethyl acetate and dried with anhydrous sodium sulphate. The study author stated that possible matrix effects on the determination of dansyl triazole were eliminated by the use of an internal standard solution of a mixture of isotopically labelled reference items added to an aliquot of the extract. Extracts were concentrated to dryness and reconstituted in acetonitrile/water (1:1 v/v). Following filtering, the extracts were analysed by HPLC-MS/MS. The claimed LOQ was 3 µg/kg for each of the four test sites, and LOD was set at 1 µg/kg. Procedural recovery at the claimed LOQ was in the range 67 – 118% with a mean of 94%. Filter papers from day 0 application checks were also extracted and analysed in a similar method. Procedural recovery at a fortified level of 0.1 µg/cm2 was in the range 75 – 101% with a mean of 89%. In the context of the day 0 samples, the LOQ of 3 µg/kg represents: Germany 4.9 – 5.9% of day 0 sample concentrations Italy 6.0 – 8.6% of day 0 sample concentrations UK 6.9 – 7.3% of day 0 sample concentrations Spain 6.6 – 9.7% of day 0 sample concentrations Representative chromatograms were presented for this study, which demonstrated good separation of peaks and close agreement of peaks from actual residues and the internal standard. Further assessment of methods of analysis data are presented in B.8.1.3 iii); in summary, the methods of analysis are considered acceptable. Results of analysis were presented as both 1,2,4-triazole equivalents in both wet and dry weight of soil, but only dry weight concentrations are presented here, in line with normal practice. Residues below LOQ were treated in the following manner: Values between LOD and LOQ were set to measured values Values <LOD were set 0.5LOD for samples of the next deeper soil layer in case the detection had been >LOD for the previous soil layer. The curve was stopped after the first non detect of <LOD if no later value >LOD followed At day 0, values <LOD in deep horizons were set to 0 This procedure is in line with FOCUS kinetics guidance on treatment of values below LOQ and LOD. 26 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) The concentrations of 1,2,4-triazole in dry weight soil were also used to calculate g/ha doses in 10cm of soil at individual time points for the purpose of kinetic calculations. The bulk density of soil was assumed to be 1.5 g/cm3 for this calculation. The Evaluator confirms that this conversion is acceptable. The study author used MatLab with KinGUI to fit SFO, FOMC and DFOP kinetics to the experimental data. Results of residues in µg/kg (dry weight) are shown in Tables B.8.16 – 19, and results converted to g/ha in 10cm soil depth are presented in Tables B.8.20 – 23. Results of filter paper analysis are also shown in Table B.8.15a. Table B.8.15a Results of filter paper analysis as dose check (1,2,4-triazole) Germany Italy UK Spain ng/cm2 mg/m2 g/ha 873.9 901.0 869.9 804.9 8.739 0.010 8.699 8.049 87.4 90.1 87.0 80.5 Amount recovered % 87 90 87 81 27 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.16 Residues of 1,2,4-triazole in German field dissipation study (µg/kg). 28 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.17 Residues of 1,2,4-triazole in Italian field dissipation study (µg/kg). 29 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.18 Residues of 1,2,4-triazole in UK field dissipation study (µg/kg). 30 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.19 Residues of 1,2,4-triazole in Spanish field dissipation study (µg/kg). 31 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) It is noted that in the Italian, UK and Spanish sites, residues of 1,2,4-triazole were found at day 0 in untreated plots, this being particularly noticeable at the Italian site. Subsequent calculation of g/ha residues and kinetic assessments did not correct residues for the concentrations in the untreated plots. The RMS considers that the noncorrection of residues would not significantly influence DT50 values recorded. Table B.8.20 Residues of 1,2,4-triazole in German field dissipation study (g/ha) Total Rainfall 1381mm; Mean monthly air temperature range 0 – 18ºC Table B.8.21 Residues of 1,2,4-triazole in Italian field dissipation study (g/ha) Total rainfall 431mm; Mean monthly air temperature range 5 – 23ºC 32 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.22 Residues of 1,2,4-triazole in UK field dissipation study (g/ha) Total Rainfall 714mm; Mean monthly air temperature range 5 – 18ºC Table B.8.23 Residues of 1,2,4-triazole in Spanish field dissipation study (g/ha) Total Rainfall 600mm; Mean monthly air temperature range 3 – 23ºC Results of the kinetic assessment performed by the applicant are shown in Table B.8.24. Note this is on the basis of data not normalised to standard temperature and moisture conditions. The kinetics selected by the study author as the best fit are highlighted in bold text. Visual and residual fits are shown in Figures B.8.10 – 12 for the German trial, Figures B.8.13 – 15 for the Italian trial, Figures B.8.16 – 18 for the UK trial and Figures B.8.19 – 21 for the Spanish site. 33 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.24 Results of kinetic assessment on 1,2,4-triazole un-normalised field dissipation data Location Germany Kinetic Model SFO FOMC DFOP SFO FOMC Italy DFOP SFO FOMC UK DFOP SFO FOMC Spain DFOP DT50 [days] 22.9 7.8 DT90 [days] 75.9 366.7 α 0.4454 β 2.0966 11.3 241.6 k1 0.1149 k2 0.0051 g 0.6602 48.8 16.3 162.2 >1000 α 0.3883 β 3.2894 21.2 207.4 k1 0.3500 k2 0.0086 g 0.4000 21.8 8.1 72.3 188.4 α 0.5728 β 3.4434 6.8 109.3 k1 0.4863 k2 0.0154 g 0.4633 85.6 28.6 284.4 >1000 α 0.3618 β 4.9336 28.1 717.6 k1 0.0632 k2 0.0020 g 0.5732 Visual Chi2 Assess* 24.9 + 15.2 t-test O 18.5 O + 17.9 11.3 k1 0.0449 k2 0.1137 g 7.2x10-4 k 0.0026 - + 10.7 O + 25.4 20.2 + 17.8 O + 21.8 12.6 + 13.3 k 0.0037 - k1 0.0853 k2 0.0060 g 0.0018 k 0.0064 k1 0.0868 k2 0.0178 g 0.0024 k 0.0031 k1 0.0395 k2 0.0903 g 4.5x10-4 *Visual assessment: + = good O = medium -- = bad t-test: SFO, for rate constant; DFOP, for k1, k2 and g; FOMC, t-test not applicable, no confidence intervals for α or β parameters given Note α and β values for FOMC and k values for DFOP fast and slow phases and associated g values have been added for clarity if these are required for e.g. PECsoil calculation. The DT50 and DT90 values in each case refer to the whole decline curve. 34 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.10 SFO visual and residual fits for 1,2,4-triazole at German field dissipation site 35 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.11 FOMC visual and residual fits for 1,2,4-triazole at German field dissipation site Figure B.8.12 DFOP visual and residual fits for 1,2,4-triazole at German field dissipation site 36 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.13 SFO visual and residual fits for 1,2,4-triazole at Italian field dissipation site Figure B.8.14 FOMC visual and residual fits for 1,2,4-triazole at Italian field dissipation site 37 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.15 DFOP visual and residual fits for 1,2,4-triazole at Italian field dissipation site Figure B.8.16 SFO visual and residual fits for 1,2,4-triazole at UK field dissipation site 38 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.17 FOMC visual and residual fits for 1,2,4-triazole at UK field dissipation site Figure B.8.18 DFOP visual and residual fits for 1,2,4-triazole at UK field dissipation site 39 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.19 SFO visual and residual fits for 1,2,4-triazole at Spanish field dissipation site Figure B.8.20 FOMC visual and residual fits for 1,2,4-triazole at Spanish field dissipation site 40 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.21 DFOP visual and residual fits for 1,2,4-triazole at Spanish field dissipation site Based on the good visual fits, the RMS accepts the Applicant choice of best fit kinetics as reported in Table B.8.24 above. However, it should be noted that care should be exercised in the use of these endpoints as they only represent dissipation/disappearance of this substance which has been applied as the starting material. In the real agricultural field situation, formation will be occurring from the parent at the same time as degradation and other dissipation processes which would lead to a longer dissipation rate. Tarara 2010 ii) A modelling procedure was conducted on the results of the field dissipation studies described in Tarara 2010 to normalise the field dissipation DT50 and DT90 to standard temperature and moisture conditions of 20ºC and pF2. As this was a modelling exercise, GLP is not applicable, but the procedure appeared to have been conducted in an appropriate manner. The study author investigated the suitability of the field dissipation study data for normalisation to standard temperature and moisture conditions. It was noted that volatilisation may be an important route of loss for 1,2,4-triazole as the substance has a vapour pressure of 3.4 x 10-1 Pa at 25ºC (reported in the DAR for the substance difenoconazole; it is also noted from the DAR for difenoconazole that the Henry’s law constant is calculated to be 3 x 10-5 Pa.m3.mol-1 at 25ºC). In addition, it was noted that soil photolysis may be a potential route of degradation. 41 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) The study author noted that in three of the four field dissipation sites (i.e. not the Spanish site), the substance was incorporated almost immediately after application to a depth of 8cm by the grass seed sowing operation. Reference to sections 3.3. and Appendix 5 of the Tarara 2010 study demonstrate that whilst grass was not sown at the Spanish site, an incorporation procedure was included at this site. The study author stated that in each study, four samples were taken at day 0, two before incorporation and two after. The study author analysed the day 0 data set for the four sites using ANOVA (analysis of variance) techniques, and whilst finding that the mean difference in residue before and after incorporation was small (mean residue before 46.4, mean residue after 44.9, units not stated but assumed to µg/kg dry weight), the difference was not statistically significant. Difference between sites was significant. From this, they concluded that any effect of volatilisation and photolysis would have been only slight or non-existent. The Evaluator notes that the residues measured at day zero were relatively scattered at all sites, and only at the Spanish site was there much of a difference apparent between the first two and the second two day 0 samples (see Table B.8.20); the magnitude of difference between the pre- and post-incorporation residues is similar to the magnitude of scatter at other sites, suggesting that differences between pre- and post-incorporation residues at the Spanish sites may not be a real reduction in residues over time but a chance arrangement of scattered data. The Evaluator also notes that there were no volatile substances other than CO2 captured in the Slangen 2000 laboratory study on 1,2,4-triazole which adds additional weight to the hypothesis that volatilisation would have been minimal. In addition, whilst the vapour pressure is relatively high, the Henry’s Law constant is relatively low suggesting that volatilisation from moist soil would be relatively low. Overall, the Evaluator concludes that significant losses via volatilisation and/or photolysis would have been minimal. The study author also noted that plant uptake may also influence the results from this procedure. However, whilst acknowledging that this normalisation procedure did not explicitly take plant uptake into consideration, they stated that any subsequent groundwater modelling should specifically set crop uptake for this metabolite to 0. The Evaluator notes that grass cropping was implemented in the subsequent PEARL procedure to simulate soil temperature and soil moisture, but this procedure did not actually simulate pesticide fate. Thus the study author is correct to make this specific recommendation to avoid crop uptake being double counted in any exposure assessment. Prior to kinetic fitting, the same pre-processing of raw field dissipation residues data was conducted as performed in the Tarara 2010 study. Consequently the residues data, expressed in g/ha and assumed to be in the top 10cm, and shown in Tables B.8.21 – 24, were used in the analysis. The Evaluator notes that soil residues from 0-30cm and on occasion 0-40cm were used to derive the residues in g/ha. It is also noted that FOCUS leaching models assume that degradation slows at increasing soil depth and thus if this was not taken into consideration in the normalisation procedure for residues reaching depth, there could be an over-estimation of speed of degradation. The general assumption in FOCUS models is that degradation slows by a factor of 0.5 at soil depths of 30-60cm; there is no depth-related reduction in degradation in the 0-30cm layer. In the 1,2,4-triazole residues data set, residues were only infrequently analysed for soil depths of 30-40cm, but these were present below LOD in all instances. On further questioning from the Evaluator, the Applicant stated that 88-98% of residues 42 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) were found in the 0-10cm layer and considered that any influence of change in temperature, moisture or depth dependent degradation rate would be relatively small over this soil depth. They also demonstrated that normalising the timesteps based on just the top 5cm depth compared to a 30cm depth had a very small impact on the normalised timesteps in the case of these four soils; this is shown in Table B.8.24a. The Evaluator considers that whilst the effect of moisture was not considered in this response, the fact that the majority of the residue was located in the top most layer means that effectively restricting consideration to the top 10cm in this case has little impact on the normalisation procedure. Table B.8.24a Effect of target soil depth (5 cm v. 30 cm) on timestep calculations for correction of field trial time scale to temperature (20 °C) and soil moisture (pF2) Days 0 1 3 8 14 28 59 91 121 185 364 451 Germany 5 cm 30 cm 0.0 0.3 0.9 3.3 5.8 12.8 31.5 58.4 76.0 96.3 136.9 190.5 0.0 0.3 0.9 3.1 5.6 12.4 29.7 56.2 73.9 94.5 133.9 186.8 Italy Days 5 cm 30 cm 0 1 3 7 14 25 60 90 118 180 0.0 0.8 2.9 7.3 11.9 21.9 56.4 76.5 92.9 112.7 0.0 0.9 2.8 7.0 12.0 22.1 57.4 78.4 94.4 114.7 UK Days 0 1 2 7 14 28 63 91 135 191 364 454 5 cm 30 cm Days Spain 5 cm 30 cm 0.0 0.3 0.7 3.4 8.1 14.0 35.1 57.0 78.6 98.3 145.6 189.4 0.0 0.4 0.7 3.2 7.8 14.0 34.3 55.8 77.4 97.1 143.9 186.2 0 1 3 8 16 28 64 93 140 210 330 415 0.0 0.7 1.9 4.8 9.6 17.3 44.7 64.1 81.6 98.9 133.8 192.1 0.0 0.8 2.4 5.8 10.4 18.5 45.0 63.0 80.9 98.1 133.0 191.0 43 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Free-fitting of the data was allowed. The four replicate samples taken at day 0 were averaged due to the variability of the residue values and because it was considered that the four replicates at this single time point compared to the single replicate at other times would inappropriately weight the fit to the day 0 data. The daily weather data from the local weather stations for the study period were used to derive the .met files for use in PEARL. Whilst the daily resolution weather data are not available in the Tarara 2010 field dissipation study report, the study author for the normalisation procedure has included the daily weather data as an appendix. Soil descriptions were input into PEARL using the available textural information from the Tarara 2010 report. Additional hydraulic parameters were then derived using the ROSETTA software. Soil profiles of 1m depth were described; the profile was discretised at 2.5cm intervals for the first 60cm and then at 5cm intervals for the remaining profile. The three sites where grass was sown had grass implemented as the crop using relevant development stages and leaf area indices taken from the FOCUSgw report. Using this information, PEARL was run for each soil to generate daily soil temperature and soil moisture information for each of the field dissipation sites. The daily values at 1.25, 3.75, 6.25 and 8.75cm were averaged to give a value for the top 10cm. The daily difference between predicted soil temperature and moisture and the standard conditions, i.e. 20ºC and pF2 were then calculated and used in a time step normalisation procedure to amend the sampling times used in kinetic analysis. Temperature normalisation used a Q10 value of 2.58 as recommended by the EFSA PPR Panel in their opinion of 2007. Moisture normalisation used a Walker exponent of 0.7 as recommended in the FOCUSgw report. The study author stated that volumetric water contents at pF2 were calculated for each soil. These values (shown below) are different to those quoted in the Tarara 2010 study. However, given that the soil moisture parameters used in PEARL were derived from ROSETTA, it was appropriate that the pF2 values for the simulated soils be calculated from ROSETTA parameters for consistency. This seems to be an acceptable approach in the opinion of the evaluator. In addition, the calculated values are, in most cases, quite close to the measured values (see Tables B.8.12 – 15 for measured values). Table B.8.25 Calculated pF2 values for field soils 44 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.26 Calculated timesteps for use in kinetic evaluation Following calculation of the timesteps, kinetic analysis was undertaken with these new values to derive kinetic parameters for use in modelling. The modelling was undertaken using KinGUI v 1.1 with MatLab v 7.0.4.365. The methodology used generally agreed with principles outlined in the FOCUS Degradation Kinetics guidance document. FOMC kinetics was not attempted for the Italian and Spanish sites because residues at the end of the study were >10% of the day 0 residue However, it is noted that they considered a t-test value of<0.1 to be acceptable for field studies. The Evaluator notes that this is not specifically stated in FOCUS Degradation Kinetics; FOCUS Degradation Kinetics uses a general assumption that t-test values <0.05 can be considered as indicating the parameter is significantly different from zero at a 5% significance level and that values between 0.05 and 0.1 can be considered as acceptable provided that there is further discussion and justification based on fit and weight of evidence. However, the Evaluator can understand the reasons for using this assumption given the general observation that field dissipation data can be more scattered than laboratory data. It is noted that the study author attempted to obtain DFOP kinetic endpoints for each trials site in order to maintain consistency for selection of mean input parameters but also to make use of the bi-exponential approaches for groundwater modelling as detailed in FOCUS Degradation Kinetics guidance. Results of statistics generated from the kinetic analyses are given in Tables B.8.27 – 30, and graphical fits in Figures B.8.22 – 35. 45 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.27 Statistical results of the Applicant kinetic fits for SFO model to normalised field dissipation data: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Table B.8.28 Statistical results of the Applicant kinetic fits for FOMC model to normalised field dissipation data: DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Note: t-test is not directly applicable to the α and β parameters; assessment of confidence intervals for both FOMC assessments indicates that the ranges include zero for the β parameter for both sites and therefore may be unreliable Table B.8.29 Statistical results of the Applicant kinetic fits for Hockey Stick model to normalised field dissipation data: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. 46 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.30 Statistical results of the Applicant kinetic fits for DFOP model to normalised field dissipation data: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. Errata: Note that the k1 t-test result for the Vilobi, Spain site above is incorrect, it is actually 0.082. In every case, the kinetic behaviour observed was bi-phasic rather than SFO. The Evaluator considers that for the UK site that DFOP is a slightly better fit than FOMC. In the case of the Italian and Spanish sites, there seems to be no advantage in choosing DFOP over HS except for the pragmatic choice of consistency and using DFOP in leaching models. In the study author’s view, the preferred kinetic at the German site was FOMC. In an attempt at obtaining consistency of modelling endpoints conforming to DFOP, a further assessment of the German site was conducted. In this, the initial data point was removed and the DFOP kinetics re-fitted. This resulted in an improved visual and residual fit and improved χ2 error test. However, the applicant considered that removal of the data point could not be justified and thus in a further analysis, the g parameter in the DFOP assessment was fixed at the value of 0.655, i.e. that achieved when the initial data point was removed, but the initial data point was re-instated in the dataset. This improved the visual and residual fits, but resulted in a poorer χ2 error test. The statistical results of this further assessment are shown below. Table B.8.31 Statistical results of the Applicant kinetic fits for DFOP model to normalised field dissipation data for German site: DT50 and DT90 [d], visual ranking and quality of fits, visual assessment of random-ness of residuals, scaled error ε to pass the 2 test, and T-test for relevant parameter fit. It is noted that relying solely on the outcome of the statistical evaluation in Table B.8.31 above indicates that the procedure fixing g to 0.655 results in a poorer χ2 value compared to the free fit to the German site dataset but improved t-test for the k1 value. However, whilst the procedure fixing g results in a poorer χ2 value, the visual fit is improved for data points 5 onwards, i.e. from a residue of approx 30 g/ha or 35% of 47 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) the initial (contrast Figures B.8.29 for free fit and B.8.31 for fixed g). The Evaluator postulates that the reason why χ2 becomes higher in value with fixing of g is that the residuals for the first three data points have become much greater, indicating that the initial data points are fitted less well (indeed, estimated M0 for free fit is 86.96 g/ha vs. 73.60 g/ha for fit with g fixed; actual M0 was 87.5 g/ha). Thus the Evaluator considers that the study author conclusion that the overall fit of the curve was better as a result of fixing ‘g’ to 0.655 is subjective and that the choice of DFOP fits is marginal. Comparisons of k and g values for the two fits with the German site dataset and the difference in overall geometric mean parameters for all 4 soils based on DFOP kinetics are given in Tables B.8.32 and 33. Table B.8.32 Comparison of DFOP kinetic values for the German field dissipation site with free fitting of g and fixing g Fast phase DT50 0.21 days 2.5 days Free fit g Fixed g 1 Slow phase DT50 30.1 days 70.7 days g 0.4687 0.6550 Estimated M01 86.96 73.60 Actual M0 87.5 g/ha Table B.8.33 Comparison of geomean modelling parameters from all four field dissipation sites using alternative DFOP parameters from German site Free fit g Fixed g2 1 arithmetic mean Fast phase DT50 0.91 1.68 2 Slow phase DT50 48.8 60.5 g1 0.442 0.489 study author proposed values Overall, the Evaluator considers that there is little to choose between the mean parameters. Free fitting of ‘g’ at the German site leads to slightly faster degradation but less substance in the faster degrading compartment, whereas fixed ‘g’ at the German site leads to slightly slower degradation but a higher proportion of substance in the faster degrading compartment. Thus, taking all considerations into account, the Evaluator considers that the study authors choice of parameters based on the fixed g value for the German site is acceptable, particularly if 1,2,4-triazole is the terminal metabolite in the leaching assessment. The final presentation of the study author’s choice of modelling endpoints for 1,2,4-triazole is given below. 48 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.34 Proposed kinetic endpoints for 1,2,4-triazole for modelling (normalised to 20°C and pF2) a The proportion of the substance described by k1. bFixed to 0.655. cArith. mean 49 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Germany Figure B.8.22 SFO visual and residual fits for 1,2,4-triazole at German field dissipation site, timestep normalised 50 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.23 FOMC visual and residual fits for 1,2,4-triazole at German field dissipation site, timestep normalised 51 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.24 DFOP visual and residual fits for 1,2,4-triazole at German field dissipation site, timestep normalised 52 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.25 DFOP visual and residual fits for 1,2,4-triazole at German field dissipation site, timestep normalised, initial data point removed 53 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.26 DFOP visual and residual fits for 1,2,4-triazole at German field dissipation site, timestep normalised, initial data point included, ‘g’ fixed at 0.655 54 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Italy Figure B.8.27 SFO visual and residual fits for 1,2,4-triazole at Italian field dissipation site, timestep normalised 55 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.28 HS visual and residual fits for 1,2,4-triazole at Italian field dissipation site, timestep normalised 56 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.29 DFOP visual and residual fits for 1,2,4-triazole at Italian field dissipation site, timestep normalised 57 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) UK Figure B.8.30 SFO visual and residual fits for 1,2,4-triazole at UK field dissipation site, timestep normalised 58 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.31 FOMC visual and residual fits for 1,2,4-triazole at UK field dissipation site, timestep normalised 59 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.32 DFOP visual and residual fits for 1,2,4-triazole at UK field dissipation site, timestep normalised 60 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Spain Figure B.8.33 SFO visual and residual fits for 1,2,4-triazole at Spanish field dissipation site, timestep normalised 61 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.34 HS visual and residual fits for 1,2,4-triazole at Spanish field dissipation site, timestep normalised 62 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Figure B.8.35 DFOP visual and residual fits for 1,2,4-triazole at Spanish field dissipation site, timestep normalised Chapple 2010b Evaluator’s final note: The geomean slow phase DT50 from this evaluation is in reasonable agreement with that from the kinetic assessment of the laboratory studies. Overall, it is considered that the laboratory and the field data give strong support for 1,2,4-triazole demonstrating bi-phasic degradation. If 1,2,4-triazole is being considered as a terminal metabolite in a leaching assessment, it is appropriate to simulate degradation of this metabolite using two compartments, one for the rapid degradation phase and the other the slow degradation phase. The separate flows from 63 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) the precursor to the rapid and slow phases should be specified along a similar scheme as shown in chapter 8.3.3.2.2 and Box 8-2 of the FOCUS Degradation Kinetics guidance. Specifically, the proportion of flow to the rapid phase of degradation would be specified by ffM x g (i.e. formation fraction for precursor to 1,2,4-triazole multiplied by the DFOP ‘g’ parameter) and the proportion of flow from precursor to the slow phase would be specified by ffM x (1-g) (i.e. formation fraction for precursor to 1,2,4-triazole multiplied by 1-‘g’). Given that this method divides the formation fraction for the metabolite between fast and slowly degrading compartments using the ‘g’ parameter as part of the simulation, it is considered unnecessary to run the model twice as specified in FOCUS degradation kinetics (i.e. it is unnecessary to run the fast and slow compartments separately and add the results together). However, due to the 1/n value for 1,2,4-triazole being less than 1, the recommendation that the application rate in the simulation is doubled and the resulting leaching concentrations divided by 2 is supported. Note that no PEC calculations are considered in this evaluation document as the concentrations of 1,2,4-triazole will be dependent on the amount of formation from each separate triazole parent a.s. and the GAP details for the parent. iii) An assessment of freezer storage stability and analytical methods in support of these data is given below. Freezer Storage Stability of lH-l,2,4-Triazole[3,5-14C] in Soil; (report no. 108303) Samples of loamy sand soil from a site in Fresno, California, USA were fortified with [14C]-1, 2, 4-triazole at a concentration of 10.3 mg/kg and stored frozen at -25°C for up to 42 months. Duplicate samples were removed from the freezer and analysed for residues of 1, 2, 4-triazole at day 0 and after 1, 3, 6, 12, 21 and 42 months storage. A “control” sample, freshly fortified was also analysed at each time point Analysis for residues of 1, 2, 4-triazole was conducted as follows: Samples were extracted for 1 hour at room temperature (by magnetic stirrer) with methanol: water 4:1 (v/v) and centrifuged. The supernatant was decanted. The samples were then extracted for 30 minutes with two aliquots of methanol and centrifuged. Finally the samples were extracted for 2 hours with methanol: ammonium hydroxide (aq) 7:3 (v/v) and centrifuged. Radioactive residues were determined by LSC or LSC following combustion in the case of the post extraction solids. Aliquots of the extracts were filtered, concentrated under a stream of oxygen free nitrogen and analysed by TLC against certified reference standards. Further analyte confirmation was conducted by LC-MS (ESI mode). The results are shown below: The Evaluator considers that the results indicate that residues of 1, 2, 4-triazole are stable in samples of loamy sand soil stored frozen for up to 42 months. 64 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.35: Radioactive residues in samples of soil stored frozen for up to 42 months 65 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.36: Residues of 1,2,4 – triazole samples of soil stored frozen for up to 42 months of soil Shadrick et al, 1999 66 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Analytical method JA005-W04-01 was used in field dissipation studies. The method utilises derivation of 1, 2, 4-triaozle with dansyl chloride to give dansyl-1, 2, 4triazole. The following justification for using a derivatisation method was provided: “Derivatisation allowed triazole to be run in a non-polar HPLC system, similar to other fungicide metabolites. Also, derivatisation provided larger mass; therefore more transitional ions within the mass spectrometer environment, which aided sensitivity and detection.” This is considered acceptable by the Evaluator. Analytical Method for the Determination of Triazole in Soil and Sediment; Method JA005-W04-01 Samples of soil were extracted by ultrasonication for 1 hour with acetonitrile: water 6:4 (v/v) at 65-70°C. Samples of sediment were extracted in the same manner but using 1% sodium hydroxide: acetonitrile 6:4 (v/v). A 1ml aliquot of the extract was taken and derivatised with dansyl chloride to give dansyl 1, 2, 4-triazole. The derivatised extract was partitioned with ethyl acetate, filtered through anhydrous sodium sulphate, concentrated to dryness and reconstituted in acetonitrile: water 1:1 (v/v). Extracts were analysed by LC/MS/MS in positive ionisation mode; luna C13 column, 100 x 4.6mm, 5 µm i.d. Stable isotope [15N]-1, 2, 4-triazole was used as an internal standard by addition to the extract prior to derivatisation. Quantitative analysis was performed using reference standards of dansyl 1, 2, 4-triazole and stable isotope [15N]-dansyl 1, 2, 4-triazole. The ion transitions monitored were 303→170 for dansyl 1, 2, 4-triazole and 306→170 for the stable isotope internal standard. Results were expressed in terms of 1, 2, 4-triazole. The LOQ was stated to be 10µg/kg. Validation data were provided using soil from Saskatchewan, Canada as follows: Linearity: demonstrated in the range 2.5 – 100 ng/ml (equivalent to 5 – 200 µg/kg in the samples). R2 > 0.99. Precision: 7 determinations made at 10µg/kg. RSD = 1.84%. 5 determinations made at 100 ppb. RSD = 2.29% Accuracy: Mean recovery values at fortification levels of 10µg/kg and 100 µg/kg for several different soil types were in the range 79-105%. Further details are given in the table below: 67 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.37 Average recoveries of 1,2,4-traiazole in soils The Evaluator considers that the method is suitably validated/fit for purpose. I. M. Murphy, 2004 Determination of the Residues of 1,2,4-Triazole in/on soil after spraying of 1,2,4Triazole (1000 TS) in the field in Germany, Italy, Great Britain and Spain Analytical Phase; report PF/04/004. Samples were analysed according to the method JA005-W04-01 above. The filter paper samples were extracted in the same way as the soil samples but extracts were diluted by 1:20 with acetonitrile: water 6:4 (v/v) before an aliquot was taken for derivatisation. The LOQ was stated to be 3µg/kg for soil and 0.1µg/cm2 for filter papers. A separate validation study for the method was provided (S Jones 2005. Report CX/05/035). The results are summarised in the table below 68 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Table B.8.38 Validation of Analytical Method for the Determination of 1,2,4-Triazole in Soil and Filter Papers. Matrix Method Analyte Soil HPLC– MS/MS Dansyl 1,2,4triazole Filter paper HPLC– MS/MS Dansyl 1,2,4triaozle LOQ (µg/kg) Recovery fortificati on level (µg/kg) 3.0 Recoveries % range (mean*) 65-100 (82.6) Repeatabi lity % RSD (n) 11.7 (12) 64-79 (72.0) 4.7 (12) Linearity 3.0 30 0.1 µg/cm2 0.1 µg/cm2 1.0 µg/cm2 75-101 (89) 0.75-100 ng/ml r2 = 0.9996 10.9 (6) 80-89 (83) Some of the control samples used for fortification had apparent residues at levels >30% of the LOQ – recoveries were corrected to take these residues into account. The potential residues in the blank samples may explain the variable recoveries. Whilst some individual recoveries are outside of the range 70-110%, the mean recoveries are within the acceptable range and the precision (repeatability) is acceptable. It has been demonstrated that the derivative is formed reproducibly by the use of an external standard of the derivate. The method is considered suitably validated. Further procedural recoveries in the soil at fortification levels of 3µg/kg were in the range 67-118% with a mean of 94% and for a fortification level of 100µg/kg were in the range 70-80% with a mean of 75%. These are considered acceptable by the Evaluator. S Jones 2005; M Fitzmaurice 2010 69 1,2 4-Triazole– Revision of DT50 B.8.10 July 2011 (revised September 2011) References relied on Annex point Author Year Title Source (where different from company) Company, Report No. GLP or GEP status (where relevant) Published or Unpublished Data protection claimed Y/N Owner B.8.1.1.2 Slangen, P.J. 2000 Yes TDMG B.8.1.1.2 Chapple, A. 2010a Yes TDMG B.8.1.3 Tarara, G. 2010 Yes TDMG B.8.1.3 Chapple, A. 2010b Yes TDMG B.8.1.3 Shadrick, B.A; Bloomberg, A.M; Helfrich, K.K. 1999 Yes TDMG B.8.1.3 Murphy, I.M. 2004 Degradation of 1,2,4-triazole in three soils under aerobic conditions NOTOX Safety & Environmental Research B.V., 's-Hertogenbosch, Netherlands Bayer CropScience AG, Report No.: 278336, Date: 2000-05-08 GLP, unpublished Kinetic evaluation of the degradation in soil of 1,2,4triazole under laboratory conditions Bayer CropScience AG Report No. MEF-10/556 Date: 2011-02-09 Non-GLP, unpublished Determination of the residues of 1,2,4-triazole in/on soil after spraying of 1,2,4-tiazole (1000 xx) in the field in Germany, Italy, Great Britain and Spain Bayer CropScience AG Report No.: RA-2145/04 Date: 2010-03-04 GLP, unpublished Kinetic evaluation of the dissipation in soil of 1,2,4-triazole under field conditions Bayer CropScience AG Report No.: MEF-10/069 Date: 2010-03-17 Non-GLP, unpublished Freezer storage stability of 1H1,2,4-triazole [3,5-14C] in soil Bayer CropScience AG Report No.: 108303 Date: 1999-12-22 GLP, unpublished Analytical method for the determination of triazole in soil and sediment Bayer CropScience AG Report No. 200516 Method No. JA005-W04-01 Date: 2004-02-20 GLP, unpublished Yes TDMG 70 1,2 4-Triazole– Revision of DT50 July 2011 (revised September 2011) Annex point Author Year Title Source (where different from company) Company, Report No. GLP or GEP status (where relevant) Published or Unpublished Data protection claimed Y/N Owner B.8.1.3 Jones, S. 2005 Yes TDMG B.8.1.3 Fitzmaurice, M. 2010 Validation of analytical method for the determination of triazole in soil and filter papers Battelle UK Ltd, Ongar, Essex, UK Bayer CropScience AG Date: 2005-06-20 GLP, unpublished Determination of the residues of 1,2,4-triazole in/on soil after spraying of 1,2,4-triazole (1000 TS) in the field in Germany, Italy, Great Britain and Spain, analytical phase Battelle UK Ltd, Ongar, Essex, UK Bayer CropScience AG Date: 2010-02-23 GLP, unpublished Yes TDMG 1,2 4-Triazole– Revision of DT50 – APPENDIX I 71 July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State United Kingdom July 2011 1,2 4-Triazole Fate and behaviour in the environment Appendix I – List of end points July 2011 Rate of degradation in soil (Annex IIA, point 7.1.1.2, Annex IIIA, point 9.1.1) Laboratory studies – modelling kinetic parameters 1,2,4-triazole (applied as parent) Aerobic conditions Soil type X1 pH t. oC / % MWHC DT50 fast DT50 (d) phase/DT5 20C 0 slow pF2/10kPa phase(d)/g St. (r2) Method of calculation Sandy loam 6.4 20 oC / 40 % 0.9/59.2/ 0.683 DFOP Loamy sand 5.8 20 oC / 40 % 1.5/247.6/ 0.580 DFOP Silt loam 6.7 20 oC / 40 % 0.8/20.6/ 0.443 DFOP 1.0/67.1/ 0.569 DFOP Geometric mean/median Field studies ‡ 1,2,4-triazole (applied as parent) Aerobic conditions, kinetics calculated for ambient conditions. Bare soil with grass sown immediately after application (with exception of Spain site where no grass sown). Soil type Location (indicate if bare (country or or cropped soil USA state). was used). 1 X1 pH Dept h (cm) DT50 (d) actual DT90( St. DT50 Method d) of (χ2) (d) actual Norm. calculati on Silt loam Germany 6.4 0-30 7.8 366.7 15.2 FOMC Silty clay loam Italy 7.6 0-40 21.2 207.4 10.7 DFOP Sandy loam UK 7.4 0-40 6.8 109.3 17.8 DFOP Loam Spain 5.8 0-30 28.1 717.6 13.3 DFOP X This column is reserved for any other property that is considered to have a particular impact on the degradation rate. 1,2 4-Triazole– Revision of DT50 – APPENDIX I 72 July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State United Kingdom July 2011 1,2 4-Triazole Fate and behaviour in the environment Geometric mean/median 1,2,4-triazole (applied as parent) Aerobic conditions, kinetics calculated timestep normalised to 20ºC and pF2 moisture. Bare soil with grass sown immediately after application (with exception of Spain site where no grass sown). Soil type Location pH Dept h (cm) DT50 (d) DT50 (d) Fast phase Slow phase ‘g’ St. (χ2) Method of calculati on Silt loam Germany 6.4 0-30 2.5 70.7 0.65 18.8 5 DFOP Silty clay loam Italy 7.6 0-40 1.4 59.8 0.36 10.6 4 DFOP Sandy loam UK 7.4 0-40 0.5 25.1 0.45 18.1 8 DFOP Loam Spain 5.8 0-30 4.6 126.0 0.48 12.7 9 DFOP 1.68 60.5 DFOP Geometric mean (‘g’ value is arithmetic mean) Repeat for as many metabolites as necessary pH dependence ‡ (yes / no) (if yes type of dependence) No 0.48 9 73 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Appendix 2 – copy of endpoints agreed at PRAPeR 12 meeting of 15 – 18 January 2007 Route of degradation (aerobic) in soil (Annex IIA, point 7.1.1.1.1) Mineralization after 100 days ‡ Bitertanol 1,2,4-triazole 1.6-52% after 90-120 days triazole label (n=6) Epoxiconazole - Metabolite [3,5-14C] 1,2,4-triazole: Study duration 360 days, (CO2 % TAR) soil 1, pre-adapted 24 % after 90 days 39.1 % after 180 days soil 2, pre-adapted 51.6 % CO2 after 90 days soil 3, non-adapted: 15/14 % CO2 after 90 days - Metabolite [3,5-14C] 1,2,4-triazole, rate study: Sandy loam 20/11 % CO2 after 120 days (study end) Loamy sand: 1.4/1.6 % CO2 after 120 days (study end) Silt loam: 34/32 % CO2 after 120 days (study end) Triadimenol 1,2,4-triazole 1.6-52% after 90-120 days triazole label (n=6) Agreed End Point (PRAPeR 12) Non-extractable residues after 100 days ‡ Bitertanol 1,2,4-triazole 38-67% after 90-120 days triazole label (n=6) Epoxiconazole - Metabolite [3,5-14C] 1,2,4-triazole (% TAR): soil 1, pre-adapted - max. 75 % after 28 days, 43 % after 360 d (study end) soil 2, pre-adapted - max. 53.5 % after 28 days, 74 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment 33.7 % after 360 d (study end) Triadimenol 1,2,4-triazole 38-67% after 90-120 days triazole label (n=6) Agreed End Point (PRAPeR 12) Metabolites requiring further consideration ‡ - name and/or code, % of applied (range and maximum) Bitertanol 1,2,4-triazole 44% at 62 d (n=1) declining to 36% at study end (120 days) Cyproconazole Triazole ( triazole label), 17.36 % at 140 d (n= 1), max 17.36%, day 140 Triazole acetic acid (triazole label), 0-6.7 % at 140 d (n= 1). Bromuconazole 1,2,4-triazole is no major metabolite in soil, No end points derived. Difenoconazole 1, 2, 4-triazole (CGA 71019) max. 20.6-23.4% after 190/271 d [14C-triazole]-label (n=2) Epoxiconazole Investigated: triazole 14C-labelled epoxiconazole (% TAR): Study duration 343 d, only study end data: Loamy sand: 5.0 % (4.6/5.4 %) 1,2,4-triazole 2.4 % others after 343 d Sandy loam: 1.9 % 1,2,4-triazole after 343 d 3.3 % others after 343 d Sandy loam: 6,6 % (5.3/7.9 %) 1,2,4-triazole after 175 d. (study end, only study end data). Fenbuconazole RH-0118 (1,2,4-triazole) peaked at 12.4% AR after 363 d [14C-TR] label (25°C) Flunquiconazole Triazole – 9.0-18.9% after 119-182 d, [14C]Fluquinconazole – triazolyl label (n = 3 soils at 20 C) Paclobutrazol 75 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment 1,2,4-triazole is no major metabolite in soil, however some end points were derived. Tetraconazole Laboratory study (dark): no metabolite formation Laboratory study under sunlight: TAA (Triazolyl acetic acid): max: 4.55 and 4.89% after 60 and 112 days (n=1) Triadimenol Triazole ring not labelled; in absence of other information, 1,2,4-triazole assumed to be formed at 100% for environmental exposure assessment Route of degradation in soil - Supplemental studies (Annex IIA, point 7.1.1.1.2) Anaerobic degradation ‡ Bitertanol Anaerobic degradation ‡ Triadimenol Anaerobic degradation ‡ Agreed end point 1,2,4-triazole Mineralisation 1.3% after 126 d triazole label (n=1) Non extractable residues max 21% 64 days declining to 16% at study end 126d triazole label (n=1) Major metabolite Triazole acetic acid 50% at study end 126 days triazole label (n=1) 1,2,4-triazole Mineralisation 1.3% after 126 d triazole label (n=1) Non extractable residues max 21% 64 days declining to 16% at study end 126d triazole label (n=1) Major metabolite Triazole acetic acid 50% at study end 126 days triazole label (n=1) 76 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Rate of degradation in soil (Annex IIA, point 7.1.1.2, Annex IIIA, point 9.1.1) Laboratory studies ‡ Bitertanol Laboratory studies (range or median, with n value, with r2 value) ‡ DT50lab (20C, aerobic): ‡ 1,2,4-triazole 20C: 6.3-12.3 d (n=3, r2= 0.75-0.95) triazole acetic acid 20C: 6-11 d (n=3, r2= 0.76-0.9) For FOCUS modelling geometric mean first order 20ºC DT50lab normalised to -10kPa soil moisture: 1,2,4-triazole: 7.4 d DT90lab (20C, aerobic): ‡ 1,2,4-triazole 20C: 21-41 d (n=3, r2= 0.75-0.95) triazole acetic acid 20C: 20-37 d (n=3, r2= 0.76-0.9) Cyproconazole Triazole Soil type pH © Sandy loam f. f. DT50 (d) kdp/k 20C f pF2/10kPa St. (r2) Method of calculation 20 oC / 34.4 % 6.32 5 0.75 SFO 5.8 20 oC / 50 % 9.91 13 0.81 SFO 6.7 20 oC / 36.4 % 12.27 8 0.95 SFO 9.15/9.91 8.06/8.2 (Hanhofen) Silt loam DT50/ DT90 (d) 6.4 (Laacherhof AXXa) Loamy sand t. oC / % MWHC (Laacherhof A III) Geometric mean/median 77 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Difenoconazole CGA 71019 Aerobic conditions 1,2,4-triazole Soil type pH t. oC / % MWHC sandy loam loamy sand silt loam Geometric mean Median 6.4 5.8 6.7 20 / 40 20 / 40 20 / 40 DT50/ f. f. DT90 kdp/ (d) kf 6.3 / 21 9.9 / 33 12 / 41 9.1 / 30.5 9.9 / 33 Epoxiconazole Laboratory studies (range or median, with n value, with r2 value) ‡ DT50 (d) 20 C pF2/10kPa 4.3 7.6 7.5 6.3 7.5 St. (r2) Method of calculation 0.75 0.81 0.95 SFO SFO SFO DT50lab (20C, aerobic): standardised to field capacity pF2 if test soil moisture < field capacity, -10kPa, with Q10 2.2, Walker-equation exponent 0.7. r² > 0.7 Metabolite 2 = 1,2,4-triazole 20 °C, moisture not standardised Loamy sand 10 d Sandy loam 6d Silt loam 12 d geometric mean DT50lab: 9d Fenbuconazole Aerobic conditions 1,2,4triazole Soil type pH t. oC / % MWHC (USDA) (CaCl2 ) Sandy loam 6.4 Loamy sand 5.8 Silt loam 6.7 Arithmetic mean 20oC / 40 % MWHC 20oC / 40 % MWHC 20oC / 40 % MWHC DT50 /DT90 (d) 1st order 20 °C, pF2 10 d 5d 9d 8d Method of calculation 6.32 / 21.0 DT50 (d) St. (r2) 20C pF2/10kP a 5.0 0.75 9.91 / 33.0 9.9 0.81 SFO 12.27 / 40.8 8.2 0.95 SFO 7.7 SFO 78 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Fluquinconazole Laboratory studies (range or median, with n value, with r2 value) Aerobic studies: Triazole DT50lab (20 C, aerobic, sfo non-linear regression): 6.32, 9.91, 12.27 d, mean = 9.5 d (n = 3 soils, r2 = 0.75-0.95) Triazole DT50lab (20 C, aerobic, multicompartment model): 2.34, 9.34, 12.27 d, mean = 7.98 d (n = 3 soils, r2 = 0.95-0.99) Triazole DT90lab (20 C, aerobic, sfo non-linear regression): 21.0, 32.9, 40.8 d, mean =31.6 d (n = 3 soils, r2 = 0.75-0.95) For FOCUS gw modelling: Triazole DT50lab (aerobic, 1st order kinetics): mean = 7.0 d (normalized to 10kPa, 20 C with Q10 of 2.2 and B-value of 0.7) Tetraconazole Triazolyl acetic Aerobic conditions acid (TAA) Soil type X1 pH t. oC / % MWHC DT50/ DT90 F.F. DT50 (d) kdp/k 20C pF2/10kPa 2 Method of calculation f (d) Sand 5.2 (0.01M CaCl2) 20°C / 60% (pF=2.5) 9.6/ 31.8 10.9 12.0 SFO Loamy sand 5.6 (0.01M CaCl2) 20°C / 60% (pF=2.5) 8.4/ 27.8 8.6 14.1 SFO Sandy loam 6.3 (0.01M CaCl2) 20°C / 60% (pF=2.5) 18.7/ 62.1 16.7 9.5 SFO DT50: 11.6 / 10.9 Geometric mean/median 11.5/ 9.6 79 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Triadimenol Laboratory studies (range or median, with n value, with r2 value) ‡ Aerobic DT50: 1,2,4-triazole 20C: 6.3-12.3 d (n=3, r2= 0.75-0.95) triazole acetic acid 20C: 6-11 d (n=3, r2= 0.76-0.9) Aerobic DT90: 1,2,4-triazole 20C: 21-41 d (n=3, r2= 0.75-0.95) triazole acetic acid 20C: 20-37 d (n=3, r2= 0.76-0.9) Agreed end point 1,2,4-triazole Aerobic conditions Soil type (USDA) pH Sandy loam 6.4 Loamy sand 5.8 Silt loam 6.7 Geometric mean (CaCl2 ) t. oC / % MWHC DT50/ DT90 (d) 20oC / 40 % MWHC 20oC / 40 % MWHC 20oC / 40 % MWHC 6.32 / 21.0 f. f. DT50 (d) kdp/ 20C kf pF2/10kPa 5.0 St. (r2) Method of calculation 0.75 SFO 9.91 / 33.0 9.9 0.81 SFO 12.27 / 40.8 8.2 0.95 SFO 7.4 Agreed End-point for calculating PEC soil for EU assessments 12 days (Not normalised). 80 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Laboratory studies ‡ anaerobic Bitertanol DT50lab (20C, anaerobic): ‡ 1,2,4-triazole 58 d (n=1, r2= 0.77.) Cyproconazole Triazole Soil type Silt loam pH t. oC / % MWHC DT50/ DT90 (d) 81/291 f. f. kdp/ kf ---- 7.31 20/40 (KC l) Geometric mean/median 81/291 Difenoconazole Laboratory studies 1,2,4- Triazole‡ Anaerobic conditions g/ha pH t. oC / % DT50 / DT50 (d) 1 MWHC DT90 (d) 20 C pF2/10kPa 128 7.2 20 / flooded stable - DT50 (d) 20C pF2/10kPa ------ St. (r2) Method of calculation 0.972 SFO St. (r2) Method of calculation - - 1 Test concentration re-calculated into corresponding g a.s./ha dose for comparison with the representative uses. Triadimenol Anaerobic: 1,2,4-triazole 20˚C: DT50 = 58 d (n=1, r2= 0.77.) Agreed end point 81 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Soil adsorption/desorption (Annex IIA, point 7.1.2) Bitertanol Kf /Koc ‡ Kd ‡ pH dependence (yes / no) (if yes type of dependence) ‡ 1,2,4-triazole Kfoc:43-120 ml/g, 1/n 0.83-1.016 (n=4) No evidence of pH dependence Arithmetic mean values for FOCUS modelling 89ml/g, 1/n 0.91 Cyproconazole 1,2,4-Triazole‡ Soil Type Alpaugh Silty clay Hollister Clay loam Lawrenceville Silty clay Pachappa Sandy loam OC % 1.2 3.0 1.2 1.4 Soil pH 8.8 6.9 7.0 6.9 Kd Arithmetic mean pH dependence (yes or no) Difenoconazole CGA 71019 (1,2,4-Triazole)‡ Soil Type silty clay clay loam silty clay loam sandy loam Arithmetic mean Median pH dependence (yes or no) Koc Kf Kfoc 0.833 120 0.748 43 0.722 104 0.719 89 0.7555 89 1/n 0.897 0.827 0.922 1.016 0.91 No. OC % 0.70 1.74 0.70 0.81 Soil pH 8.8 6.9 7.0 6.9 Kd Koc Kf Kfoc (mL/g) (mL/g) (mL/g) (mL/g) 0.83 120 0.75 43 0.72 104 0.72 89 0.75 89 0.74 82 No 1/n 0.90 0.83 0.92 1.02 0.91 0.91 82 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Epoxiconazole metabolite 1,2,4-triazole Koc: 43-120, arithmetic mean 89 (n=4) soil pH Koc Kf 1/n Sandy loam 6.9 89 0.720 1.016 (62 % sand, 21 % silt, 17 % clay, 1.4 % org. matter, Corg 0.812 %.) Clay loam 6.9 43 0.748 0.827 (26 % sand, 46 % silt, 28 % clay, 3.0 % org. matter, Corg 1.74 %.) Silty clay 8.8 120 0.833 0.897 (11 % sand, 44 % silt, 45 % clay, 1.2 % org.C matter, Corg 0.696) Silty clay loam 7.0 104 0.722 0.922 (9 % sand, 62 % silt, 29 % clay, 1.2 % org. matter, Corg 0.696) Kf /Koc ‡ pH dependence (yes / no) (if yes type of dependence) ‡ yes (metabolite), increasing sorption with increasing pH Fenbuconazole 1,2,4-triazole Soil Type (USDA) OC % Soil pH (CaCl2) Silty clay 0.70 8.8 Clay loam 1.74 6.9 Sand 0.12 4.8 Silty clay loam 0.70 7.0 Sandy loam 0.81 6.9 Mean1 1 Results from the sand soil excluded as an outlier Fluquinconazole Kf/Koc Kd pH dependence (yes/no) (if yes type of dependence) Kf Kfoc 1/n r2 0.833 0.748 0.234 0.722 0.720 0.756 120 43 202 104 89 89 0.897 0.827 0.885 0.922 1.016 0.916 0.996 0.997 0.997 0.998 0.997 - Triazole: Kf: 0.234-0.833 mL/g (mean = 0.651 mL/g, 5 soils) Kfoc: 43-202 mL/g (mean = 111.6 mL/g, 5 soils) 1/n: 0.827-1.016 (mean = 0.909, 5 soils) Kd: Not determined No pH dependence for Fluquinconazole or its metabolite, dione. 83 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Paclobutrazol Metabolite 1,2-4 triazole ‡ Soil Type OC % Soil pH Kd Koc (mL/g) (CaCl2) (mL/g) 8.8 0.833 120 6.9 0.748 43 4.8 0.234 202 Kf Kfoc 1/n (mL/g) (mL/g) 0.833 120 0.748 43 0.2341 2021 0.897 0.827 0.8851 Lakeland 0.70 1.74 0.12 Lawrenceville 0.70 7.0 0.722 104 0.722 104 0.922 Pachappa 0.81 6.9 0.719 89 0.720 59 1.016 Alpaugh Hollister 0.7561 Arithmetic mean pH dependence (yes or no) 891 0.91551 Yes. Positive correlation with organic carbon, pH and clay content 1 Results from the Lakeland soil were excluded on the basis of being an outlier Tetraconazole TAA(Triazole acetic acid) Soil Type OC % Soil pH Kd Koc Loamy sand 14.42 3.38 0.150 1.04 (0.01M CaCl2) Kf Kfoc 1/n adsorb: 0.903; desorb: 0.786 Clay 0.89 7.55 0.178 20 (0.01M CaCl2) adsorb: 0.911; desorb: 0.865 Silt loam 2.13 5.16 0.448 21 (0.01M CaCl2) adsorb: 0.926; desorb: 0.820 Geometric mean/median 0.229/ 0.178 7.59/ 20.0 adsorb: 0.913/ 0.911; desorb: 0.823/ 0.820 pH dependence, Yes or No no 84 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Triadimenol Kf /Koc ‡ 1,2,4-triazole Kfoc:43-120 ml/g, 1/n 0.83-1.016 (n=4) No evidence of pH dependence Kd ‡ pH dependence (yes / no) (if yes type of dependence) ‡ Agreed end point Metabolite 1,2-4 triazole ‡ Soil Type(USDA) Silty clay Clay loam OC % Soil pH Kd Koc (mL/g) (CaCl2) (mL/g) 8.8 6.9 4.8 Kf Kfoc 1/n (mL/g) (mL/g) 0.833 120 0.748 43 0.234 202 0.897 0.827 0.8851 Sand 0.70 1.74 0.12 Silty clay loam 0.70 7.0 0.722 104 0.922 Sandy loam 0.81 6.9 0.720 89 1.016 Arithmetic mean (of 4 values excluding the very low OC sand that 0.756 was considered not representative of agricultural soils) pH dependence (yes or no) 89 0.9155 No Mobility in soil (Annex IIA, point 7.1.3, Annex IIIA, point 9.1.2) Tetraconazole Column leaching ‡ Tetraconazole metabolites column leaching in slightly humous sand: elution: 200 mm time period : 2 days Tetraconazole metabolites column leaching in slightly humous sand: Triazole: 59.04% (of applied Triazole) TAA: 95.53% (of applied TAA) 85 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Tetraconazole Aged residues leaching ‡ Laboratory study (dark): Aged for (d): 30 d Time period (d): 2 d Elution (mm): 200 mm Laboratory study under sunlight: Aged for (d): 53 d (samples D2 and D3), 48 d (sample D1) Time period (d): 2 d Elution (mm): 200 mm Soil residues post ageing Laboratory study (dark): % of 14C-Tetraconazole: 96.21%AR % of bound residues: 2.32%AR Laboratory study under sunlight: % of bound residues: 8.50%AR (53 d); 8.38%AR (48d) Characterization of soil residue before leaching was performed just for the 53 d aged sample; Tetraconazole in D2 sample (53 d): 41.80 %AR M14360-alcohol in D2 sample (53 d):8.07 %AR Triazole in D2 sample (53 d):1.97 %AR DFA in D2 sample (53 d):9.97 %AR M14360-acid in D2 sample (53 d): 2.74 %AR TAA in D2 sample (53 d): 4.11 %AR Analysis of leachate Laboratory study: tetraconazole equivalents in the leachate: 0.15%AR bound residues: 2.13%AR Study under sunlight: Total residues/radioactivity: 33.15%AR Active substance: 0.00%AR Metabolites: 0.45%AR (M14360-alcohol), 1.88%AR (Triazole), 10.01%AR (DFA), 4.12%AR (M14360-acid), 5.69%AR (TAA) Radioactivity retained in soil segment: 12.93%AR in top 5 cm; 15.29%AR in top 30 cm 86 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment PEC (soil) (Annex IIIA, point 9.1.3) Cyproconazole Metabolite I Method of calculation Triazole Molecular weight relative to the parent: 0.236 (69.1 g/mol) DT50 (d): 12.3 days. Kinetics: SFO, Lab worst case Difenoconazole CGA 71019 (1,2,4-Triazole) Method of calculation Epoxiconazole Method of calculation Initial PECs= Max parent PECs x Max. metabolite in soil x Mol. Wt fraction. where: Max. parent PECs: 0.016 mg/kg (seed treatment); 0.136 mg/kg (apples); 0.096 mg/kg (carrots) Max. CGA 71019 in soil: 23% Molecular weight fraction: 0.170. PEC: 1st order kinetic, DT50Lab worst case standardised to 15 °C of 3 studies: 18 d, 5 cm soil layer, 1.5 kg/L bulk density, with interception (FOCUS) for cereal scenario f = 0.5 (BBCH 25) and 0.7 (BBCH 61). Interval between applications 21 d. ModelMaker, k1_deg = 1st order rate constant BAS 480 F to metabolite 1 1,2,4-triazole 0.0038/d, , k2_deg = 1st order rate constant BAS 480 to metabolite 0.039/d, molar mass correction BAS 480 F to metabolite 1,2,4-triazole 0.211 (69.1 g/mol/329.8g/mol). Fluquinconazole Triazole Method of calculation Kinetics: first order DT50: 12.27 d (worst case first order laboratory value) Soil depth: 5 cm 87 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Paclobutrazol Metabolite II: 1,2,4-triazole (CGA 71019) Method of calculation Tetraconazole TAA Method of calculation Peak formation of 1,2,4-triazole from paclobutrazol of 3% AR from laboratory studies (correction factor from parent paclobutrazol = 0.007 taking into account differences in molecular weights). Molecular weight relative to the parent: MWM = 127.1; MWP = 372.16 DT50 (d): 15 days Kinetics: 1st order, best fit according to Slide Write Plus software Field or Lab: representative worst case from laboratory studies. Triadimenol Method of calculation Application rate For M04, a conservative approach has been taken for PECsoil calculation, assuming 100% formation. The longest laboratory DT50 at 20˚C and pF2 with an r2 value ≥0.85 is 8.2 days. Molecular weight correction factor is 0.234. With multiple application scenarios and metabolites, there is uncertainty with respect to peak metabolite formation, and as a worst case, PECsoil for this metabolite is calculated on the basis of the maximum total dose of triadimenol. The metabolite 1,2-dihydro-triazolone was formed at maximum of 30.8% AR in a study on degradation of M04. Therefore the initial PECsoil has been calculated on the basis of the maximum total dose of triadimenol, molecular weight correction factor of 0.288 and 30.8% AR formation. The initial PECsoil for this metabolite is 0.009 mg/kg. 88 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Agreed end point Metabolite I Method of calculation Molecular weight relative to the parent: DT50 (d): x days Kinetics: SFO Field or Lab: representative worst case from field studies. Route and rate of degradation in water (Annex IIA, point 7.2.1) Hydrolytic degradation of the active substance and metabolites > 10 % ‡ pH 5: 1,2,4-triazole: stable at 25 ° pH 7: stable at 25 ° Photolytic degradation of active substance and metabolites above 10 % ‡ pH 9: 1,2,4-triazole: stable at 25 ° Bitertanol metabolites formed (triazole label, natural water) 1,2,4-triazole max 86% AR at 6 test system days Epoxiconazole Metabolite: 1,2,4-triazole, 14C-labelled 80 mg/L triazole in distilled water containing humic acid (Fluka). No photochemical loss after 30 days (natural sun light). absorption coefficient < 10 L/mol x cm Fluquinconazole [Triazole is not expected to photodegrade in water under environmental conditions, with mean molar extinction coefficients for triazole determined at <0.1 L mol-1 cm-1] Agreed end point 89 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Degradation in water / sediment Fluquinconazole Degradation in water/sediment - DT50 water - DT90 water Water phase (two different multi-compartment modelling methods used): - DT50 whole system - DT90 whole system Metabolites: Triazole (aerobic) DT50 = 41.9-190 d, (1st order compartmental model TopFit v1, n = 2, r2 = 0.8714-0.9620), DT50 = 11.6-92.5 d, (1st order 5-compartmental model TopFit v2, n = 2, r2 = 0.752-0.936) Distribution in water / sediment systems (metabolites) Whole system: Metabolites: not determined Water phase: Triazole (aerobic) = 0.0-2.3% (day 0), 3.9-5.1% (day 1), 28.8% and 31.6% (day 14 and 63, respectively), 4.5-21.4% (day 100) (n = 2 systems) SN 616368 (aerobic) = max. 2.1% (day 2), 0.6% (day 26), not detected (day 100) Sediment phase: Triazole (aerobic) = 0.0-0.9% (day 0), 0.0-0.5% (day 1), 21.5% and 37.4% (day 26 and 63, respectively), 26.3-33.7% (day 100) (n = 2 systems) Triazole: Maximum of 37.4% applied radioactivity in sediment after 100 d. Triazole (aerobic) DT50 in sediment = 105.36931.5 d, (1st order 5-compartmental model TopFit v2, n = 2, r2 = 0.937-0.977) For FOCUS surface water modelling a DT50 value in sediment of was set to 730 days. 90 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Paclobutrazol 1,2-4 triazole Max in water 9.4% A.R. after 84 d. Water / sediment system pH pH water sed phase Basing (14C 7.6 triazole label) t. oC DT50DT90 whole sys. N/R 22 N/R St. (r2) DT50DT90 r2 water N/R N/R DT50- St. DT90 (r2) sed N/R N/R N/R Method of calculatio n - Geometric mean/median Triadimenol Distribution in water / sediment systems (metabolites) ‡ No metabolites in water or sediment at >1.7% at any time. For environmental exposure assessment, M-04 assumed to form at 100% AR. Agreed end point PEC (surface water) and PEC sediment (Annex IIIA, point 9.2.3) Bitertanol Method of calculation FOCUS step 2 ‘STEPS 1-2 in FOCUS’, ‘FOCUS surface water tool version 1.1’ 1,2,4-triazole Soil DT50 7.4 d Soil formation fraction 44 % (x69/337) Whole aquatic system DT50 *999 d Aquatic system formation *100% (x69/337) fraction 89 ml/g Kfoc Parameters used in FOCUSsw step 3 Cyproconazole Metabolite Triazole Molecular weight:69.1 Parameters used in FOCUSsw step 1 and Water solubility (mg/L):1,250,000 2 Soil or water metabolite:Soil Koc/Kom (L/kg): 89 DT50 soil (d): 8.6 days [Average Lab value, FOCUS normalised] DT50 water/sediment system (d): 300 DT50 water (d):300 DT50 sediment (d):300 Crop interception (%): For Step 3, the model performs this calculation 91 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Parameters used in FOCUSsw step 3 (if performed) Difenoconazole CGA 71019 (1,2,4-triazole) Parameters used in FOCUSsw step 1 and 2 Epoxiconazole Method of calculation Vapour pressure:2.2 x10-4 Pa Kom/Koc: 89 1/n: 0.92 (Freundlich exponent general) Metabolite kinetically generated in simulationyes: Formation fraction in soil (kdp/kf): (If formation degradation of metabolite is kinetically simulated by PRZM) For FOCUS calculations the maximum occurrence of triazole with respect to parent was set at 90 % Molecular weight (g/mol): 69 Water solubility (mg/L): 730 Soil or water metabolite: Both KOC (mL/g): 89 (mean value) DT50 soil (d): 6.45 (arithmetic mean of normalised lab values) DT50 water/sediment system (d): 1000 (worst case assumption) DT50 water (d): 1000 (worst case assumption) DT50 sediment (d): 1000 (worst case assumption) Simulated together with parent compound: Crop interception (%): 0 (seed treatment); 70 (apples and carrots) "No drift" option used for seed treatment scenario. Max. occurrence observed (%): Water/Sediment: 9.6 (worst case assumption calc. by RMS) Soil: 23.4 not required, maximum 1.7 % in water phase of water/sediment study Fenbuconazole Parameter Mol wt. (g/mol) Water solubility (mg/l) Max. observed in soil studies (%) Max. observed in water/sediment studies Kfoc (ml/g) 1,2,4-triazole 69.1 1000* 12.4 0 89 92 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment DT50 soil (d) DT50 water/sediment (d) DT50 water (d) DT50 sediment (d) Application 7.7 999* 999* 999* Single application of 7.12g/ha based on total dose of parent of 4 x 70g a.s./ha used on pome fruit *worst case assumptions Flunquinconazole Triazole Parameters used in FOCUSsw step 1 and 2 Parameters used in FOCUSsw step 3 (if performed) Tetraconazole TAA (triazol acetic acid) Parameters used in FOCUSsw step 1: calculation of initial PECsw Molecular weight (g/mol): 69.07 Water solubility (mg/l): 50.0 mg/l Koc/Kom (ml/g): Koc = 89.0 (mean of 4 values) 1/n: 0.91 (mean of 4 values) DT50 soil (d): 7 d (geometric mean) DT50 water/sediment system (d): Not applicable DT50 water (d): 52.1 d (mean value) DT50 sediment (d): 730 d (worst-case value) Maximum occurrence observed (%): 65% (soil), 59.0% (sediment) Not applicable Molecular weight: 372.16 Water solubility (mg/l): not provided Soil or water metabolite: soil metabolite Koc (L/kg): 11.7 DT50 soil (d): not used DT50 water/sediment system (d): not used DT50 water (d): not used DT50 sediment (d): not used Crop interception (%):not used Maximum occurrence observed (% molar basis with respect to the parent): 14.11 % in soil Ratio of field to water body: 10 Effective sediments depth of the surface water: 1cm Sediment bulk density (kg/L): 0.8 Sediment organic carbon content (%): 5 93 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Sediment depth of the surface water: 5 cm Equations from FOCUS Step 1 were used Triadimenol Step 2 – M04 (1,2,4-Triazol) Summary of chemical property parameters input to FOCUS Step 2 (version 1.1) for triadimenol and M04 surface water modelling Input parameter Unit M04 (1,2,4 – triazol) Physico-chemical parameters Molecular mass g.mol-1 69.1 Water solubility mg.l-1 700000 Soil degradation parameters $ Geometric mean Half-life days 9.9*** Max. observed formation % 100%** Sediment/water degradation parameters @ Mean Half-life (whole days 999 system) Max. observed formation % 100%** Sorption parameters £ Median KfOC cm3.g-1 89+ @ arbitrary high value as measured data not available, also set for degradation DT50 in water and sediment compartments $ half lives were from field dissipation studies normalised to 20˚C and field capacity moisture content (-10kPa), as recommended by FOCUS guidance. £ see section B.8.2.4. * also set for degradation DT50 in water and sediment compartments; value from 22˚C study, NOT normalised to 20˚C. **extreme worst case assumption *** notifier stated that DT50 soil used was 7 days, geometric mean of temperature and moisture normalised lab data. In practice, value used was 9.9 days, the median uncorrected DT50. + mean value 94 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Agreed end point 1,2,4-triazole Parameters used in FOCUSsw step 1 and 2 Parameters used in FOCUSsw step 3 (if performed) Molecular weight: Water solubility (mg/L): Soil or water metabolite: Koc/Kom (L/kg): (if necessary, soil metabolites) DT50 soil (d): x days (If necessary, Lab or field. In accordance with FOCUS SFO) DT50 water/sediment system (d): (representative worst case from sediment water studies) DT50 water (d): DT50 sediment (d): Crop interception (%): Maximum occurrence observed (% molar basis with respect to the parent) Water: Sediment: Vapour pressure: Kom/Koc: 1/n: (Freundlich exponent general or for soil ,susp. solids or sediment respectively) Metabolite kinetically generated in simulation (yes/no): Formation fraction in soil (kdp/kf): (If formation degradation of metabolite is kinetically simulated by PRZM) PEC (groundwater) (Annex IIIA, point 9.2.1) Bitertanol Method of calculation and type of study (e.g. modelling, monitoring, lysimeter ) FOCUSPELMO 3.2.2 modelling for scenarios pertinent to top fruit and cereals. 1,2,4-triazole Geometric mean Soil DT50 (10kPa 20ºC Soil formation fraction Kfoc 1/n 7.4 d 100 % 89 ml/g 0.91 95 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment Cyproconazole Method of calculation and type of study (e.g. modelling, field leaching, lysimeter) PECgw were calculated using the FOCUS PELMO model Metabolites Triazole Data as for parent (cyproconazole) with the following exceptions: Median parent DT50 field 8.6 d (normalisation to 10 kPa or pF2, 20°C with Q10 of 2.2). Koc: triazole, mean 89, 1/n= 0.92. Assuming a 90 % transformation rate from parent to metabolite. Difenoconazole Method of calculation and type of study (e.g. modelling, field leaching, lysimeter ) Model used: FOCUS PEARL 2.2.2 Scenarios: Difenoconazole and the metabolites CGA71019 and CGA 205375 were simulated in separate model runs. CGA 71019 (1,2,4-triazole): DT50 soil (d): 6.45 (arithmetic mean of normalised lab values) Koc (mL/g): 89 (mean value) 1/n: 0.9 (mean value) Epoxiconazole Method of calculation and type of study (e.g. modelling, monitoring, lysimeter ) Fenbuconazole Method of calculation and type of study (e.g. modelling, field leaching, lysimeter ) Fluquinconazole Method of calculation and type of study (e.g. modelling, monitoring, lysimeter) FOCUS-PELMO 2.2.2 and FOCUS-Macro 3.3.1 Metabolite 1,2,4-triazole: DT50 lab 8 (20°C, pF2 standardised). Koc 43, 1/n 0.827, water sol. 700 mg/L,pH independent, TSCF (crop uptake default 0.5) Mean 1,2,4-triazole Kfoc: 89 ml/g Mean 1,2,4-triazole 1/n: 0.9 (default) Modelling using FOCUS groundwater scenarios Model used: FOCUS PELMO 3.3.2 including the FOCUS shell and simulation model. Triazole: calculations were based on a mean DT50 value of 7.0 d (determined from a 96 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment laboratory study, corrected to pF2 moisture and 20 C where appropriate). Averaged Freundlich sorption data used were Kfoc 89.0 ml/g and 1/n = 0.91. A temperature correction exponent (Q10) of 2.2 and a moisture correction exponent (B) of 0.7 were used. A transformation rate of 65% (Fluquinconazole to triazole) was used, based on a calculated formation fraction. Tetraconazole TAA Method of calculation and type of study (e.g. modelling, field leaching, lysimeter ) For FOCUS gw modelling, values used – Model(s) used: FOCUS-PELMO 3.3.2 TAA DT50 lab :9.7 (mean value, not normalized) Koc: 11.7 (worst case mean value); 1/nads = 0.9 (default value) Triadimenol M04 Parameters Molecular weight Solubility in water (mg/l at 20˚C) Vapour pressure (Pa at 20˚C) First order DT50 (days) Reference temperature (˚C) Reference soil moisture (pF) Activation energy (kJ/mole) Moisture exponent Kom-value (ml/g) Exponent of the Freundlich isotherm Formation fraction Agreed end point Method of calculation and type of study (e.g. modelling, field leaching, lysimeter ) 69.1 7 20 2 54 0.7 51.7 0.92 1 Geometric mean or median DT50lab/field x d (normalisation to 10kPa or pF2, 20 C with Q10 of 2.2). KOC: parent, arithmetic mean or median x, 1/n= y. 97 1,2 4-Triazole– Revision of DT50 - APPENDIX II July 2011 List of end points (based on EPCO Manual E4 - rev. 4 (September 2005)) Rapporteur Member Month and year Active Substance (Name) State EFSA January 2007 Triazole fungicides and their metabolites Fate and behaviour in the environment