Chloropicrin (PC 081501) MRIDs 49854401/50167601 Analytical method for chloropicrin in ground, drinking, and surface water Reports: ECM: EPA MRID No.: 49854401. Todd, M. 2007. CHLOROPICRIN - VALIDATION OF METHODOLOGY FOR THE DETERMINATION OF RESIDUES OF CHLOROPICRIN IN GROUND, DRINKING AND SURFACE WATER. Study No.: SXC 0007/072387. Report prepared by Huntingdon Life Sciences Ltd., Cambridgeshire, England, and sponsored and submitted by European Chloropicrin Group, Washington, D.C.; 45 pages. Final report issued June 27, 2007. Document No. Guideline: Statements: Classification: PC Code: EFED Final Reviewer: CDM/CSS- Dynamac JV Reviewers: ILV: EPA MRID No. 50167601. Keenan, D. 2017. Independent Laboratory Validation of Chloropicrin in Ground, Surface, and Drinking Water. PTRL Project No.: 2854W. Report prepared by PTRL West (now doing business at EAG Laboratories), Hercules, California, sponsored and submitted by Chloropicrin Task Force, Niklor Chemical Co. Inc., Mojave, California; 78 pages. Final report issued January 18, 2017. MRIDs 49854401 & 50167601 850.6100 ECM: The study was conducted in accordance with OECD, UK and UK Department of Health Good Laboratory Practice (GLP) standards (p. 3; Appendix 7, p. 88 of MRID 49854401). Signed and dated No Data Confidentiality, GLP, and Quality Assurance statements were provided (pp. 2-4). A statement of authenticity was included with the QA statement. ILV: The study was conducted in accordance with German GLP standards, which are based on OECD GLP standards, which are accepted by European communities, the USA (FDA and EPA, FIFRA GLP standards, 40 CFR, Part 160) and Japan (p. 3 of MRID 50167601). Signed and dated No Data Confidentiality, GLP, Quality Assurance, and Authenticity statements were provided (pp. 2-5). This analytical method is classified as supplemental. An updated ECM should be submitted with the ILV modifications. The specificity of the method was not well-supported by the ECM representative chromatograms because of chromatogram readability. Two of the three ECM matrices were not characterized. 081501 James Lin Environmental Engineer LisaMuto, M.S. Environmental Scientist Joan Gaidos, Ph.D., Environmental Scientist Signature: Date: Signature: Date: Signature: Date: 09/14/2018 12/19/2018 08/15/2018 Page 1 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 This Data Evaluation Record may have been altered by the Environmental Fate and Effects Division subsequent to signing by CDM/CSS-Dynamac JVpersonnel. The CDM/CSS-Dynamac Joint Venture role does not include establishing Agency policies. Executive Summary The analytical method, Huntingdon Life Sciences Ltd. Study No. SXC 0007/072387, is designed for the quantitative determination of chloropicrin in water at the LOQ of 0.1 |ig/L using GC/ECD (primary) and GC/MS (confirmatory). The LOQ is less than the lowest toxicological level of concern in water. The ECM validated the method using characterized surface water and uncharacterized ground and drinking waters; the ILV validated the method using characterized surface, ground, and drinking water matrices. The ILV validated the method in the second trial with two modifications to the sample processing procedure to increase analyte recovery: shortened extraction time and use of a smaller vessel to reduce headspace. The first trial failed due to low recoveries; therefore, the ECM should be updated with the ILV modifications. All ILV and ECM data regarding repeatability, accuracy, precision, and linearity were satisfactory for chloropicrin, except for the ILV linearity for the GC/MS analysis of chloropicrin in drinking water. ILV representative chromatograms were satisfactory, but ECM representative chromatograms were very difficult to interpret. The LODs of the ECM and ILV differed. Table 1. Analytical Method Summary Analyte(s) by Pesticide MRID EPA Review Matrix Method Date (dd/mm/yyyy) Registrant Analysis Limit of Quantitation (LOQ) Environmental Chemistry Method Independent Laboratory Validation Chloropicrin 49854401 50167601 Water1-2 27/06/2007 European Chloropicrin Group Chloropicrin Task Force (Niklor Chemical Co. Inc.) GC/ECD & GC/MS 0.1 ng/L 1 In the ECM, surface (lake) water (pH 7.7; 180 mg equiv. CaCCh/L total hardness and alkalinity; 15.2 mg/L total organic carbon; 12.7 mg/L dissolved organic carbon) was collected from Diss Mere, Diss, Norfolk, and used in the study (p. 10 of MRID 49854401). The water sample was characterized in a separate study (not specified). The drinking water was obtained from a tap in the Residue Analysis Department, and the ground water was obtained from a source in the Huntingdon area by Anglian Water. The drinking and ground water were not characterized; the source of the ground water was not further specified. 2 In the ILV, the ground water (PTRL ID 2706W-032; pH 7.3; 627 mg equiv. CaCCh/L hardness; 960 ppm total dissolved solids) obtained from Northwood, North Dakota, drinking (tap) water (PTRL ID 2706W-072/054; pH 7.5; 30 mg equiv. CaCCh/L hardness; 60 ppm total dissolved solids) obtained from Hercules, California, surface water (PTRL ID 2706W-069; pH 7.8; 1183 mg equiv. CaCCh/L hardness; total dissolved solids not determined) obtained from Hercules, California, were used in the study (pp. 15-16 of MRID 50167601). The water samples were provided by EAG-Hercules and previously characterized by Agvise Laboratories. Page 2 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 I. Principle of the Method Samples (20 mL) of water in 50-mL polypropylene tubes were fortified, as necessary (p. 11 of MRID 49854401). An aliquot (2 mL) of hexane was added, and the sample was shaken on a mechanical shaker for 30 minutes. After centrifugation (3500 rpm for 5 minutes), aliquots of the upper hexane phase were taken for analysis. Samples were analyzed using a Hewlett Packard 6890 series gas chromatograph coupled to an electron capture detector (ECD; p. 12 of MRID 49854401). The GC/ECD conditions consisted of a Phenomenex Zebron ZB-50 column (0.53 mm x 30 m, l-|-im), injector temperature 200°C, temperature program [40°C for 2 min. then 10°C/min. to 80°C then 30°C/min. to 200°C], carrier gas nitrogen, and 1 |aL injection volume. Expected retention time was ca. 4.3 minutes. For confirmation, samples were analyzed using a Varian 1200 series gas chromatograph coupled to a mass spectrometer (p. 13 of MRID 49854401). The GC/MS conditions consisted of a RTX- 5ms (0.25 mm x 30 m, 0.25-|am), injector temperature 200°C, temperature program [40°C for 3 min. then 10°C/min. to 60°C then 30°C/min. to 200°C], carrier gas helium, 1 |aL injection volume, and CI- ionization mode. One ion was monitored: m/z 119. Expected retention time was ca. 4 minutes. In the ILV, the ECM was performed as written, except for a shortened extraction time (1 minute with inversion and vortexing), use of a smaller vessel to reduce headspace, and use of a different analytical instrumentation (pp. 16-17, 19-21, 25; Figure 1, p. 31 of MRID 50167601). For GC/ECD analysis, an Agilent 6890 series GC [DB-17 column (0.53 mm x 30 m, l-|-im)] coupled to an ECD was used. All other GC/ECD parameters were the same as those of the ECM. For GC/MS analysis, an Agilent 7890A series GC [DB-5ms column (0.25 mm x 30 m, 0.25-|am)] coupled to a MS was used. All other GC/MS parameters were the same as those of the ECM. Expected retention times were ca. 4.7 minutes for GC/ECD and ca. 2.6 minutes for GC/MS. The Limit of Quantification (LOQ) for chloropicrin in water was 0.10 |ig/L in the ECM and ILV (pp. 8, 12-13 of MRID 49854401; pp. 12, 22 of MRID 50167601). The Limit of Detection (LOD) was 0.05 |ig/L in the ECM; in ILV, the LOD was 0.004 |ig/L for ground water and 0.01 |ig/L for drinking and surface water. Page 3 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 II. Recovery Findings ECM (MRID 49854401): Mean recoveries and relative standard deviations (RSDs) were within guideline requirements (mean 70-120%; RSD <20%) for analysis of chloropicrin in three water matrices at fortification levels of 0.1 |ig/L (LOQ) and 1.0 |ig/L (lOxLOQ; Tables 3-8, pp. 19- 24). Chloropicrin was identified using GC/ECD and GC/MS for primary and confirmatory analyses, respectively. For GC/MS, one ion was monitored. Performance data (recovery results) from primary and confirmatory analyses were fairly comparable, but more varied for drinking water. The surface (lake) water (pH 7.7; 180 mg equiv. CaCCb/L total hardness and alkalinity; 15.2 mg/L total organic carbon; 12.7 mg/L dissolved organic carbon) was collected from Diss Mere, Diss, Norfolk, and used in the study (p. 10). The water sample was characterized in a separate study (not specified). The drinking water was obtained from a tap in the Residue Analysis Department, and the ground water was obtained from a source in the Huntingdon area by Anglian Water. The drinking and ground water were not characterized; the source of the ground water was not further specified. ILV (MRID 50167601): Mean recoveries and RSDs were within guideline requirements for analysis of chloropicrin in three water matrices at fortification levels of 0.1 |ig/L (LOQ) and 1.0 |ig/L (lOxLOQ; Table 1, p. 27). Chloropicrin was identified using GC/ECD and GC/MS for primary and confirmatory analyses, respectively. For GC/MS, one ion was monitored. Performance data (recovery results) from primary and confirmatory analyses were fairly varied, but more comparable for surface water. The ground water (PTRL ID 2706W-032; pH 7.3; 627 mg equiv. CaCCb/L hardness; 960 ppm total dissolved solids) obtained from Northwood, North Dakota, drinking (tap) water (PTRL ID 2706W-072/054; pH 7.5; 30 mg equiv. CaCCb/L hardness; 60 ppm total dissolved solids) obtained from Hercules, California, surface water (PTRL ID 2706W-069; pH 7.8; 1183 mg equiv. CaCCb/L hardness; total dissolved solids not determined) obtained from Hercules, California, were used in the study (pp. 15-16). The water samples were provided by EAG-Hercules and previously characterized by Agvise Laboratories. The method was validated in the second trial with two modifications to the sample processing procedure to increase analyte recovery: shortened extraction time (1 minute with inversion and vortexing) and use of a smaller vessel to reduce headspace (pp. 11, 16-17, 19-21, 25). The first trial failed due to low recoveries; therefore, the ECM should be updated with the ILV modifications. Page 4 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Table 2. Initial Validation Method Recoveries for Chloropicrin in Water Analyte Fortification Level (jig/L) Number of Tests Recovery Range (%) Mean Recovery (%) Standard Deviation (%)4 Relative Standard Deviation (%) GC/ECD Ground Water Chloropicrin 0.1 5 82-90 86 3.0 3.5 1.0 5 78-84 82 2.4 3.0 Drinking Water Chloropicrin 0.1 5 80-96 92 7.0 7.6 1.0 5 84-100 94 7.1 7.6 Surface Water Chloropicrin 0.1 5 78-93 87 6.2 7.1 1.0 5 77-93 85 6.2 7.3 GC/MS (m/z 119) Ground Water Chloropicrin 0.1 5 92-95 94 1.1 1.2 1.0 5 81-88 85 2.6 3.1 Drinking Water Chloropicrin 0.1 5 76-89 80 5.2 6.4 1.0 5 75-83 80 3.4 4.3 Surface Water Chloropicrin 0.1 5 75-89 83 6.2 7.5 1.0 5 76-87 80 4.3 5.4 Data (uncorrected recovery results; p. 14) were obtained from Tables 3-8, pp. 19-24 of MRID 49854401. * The surface (lake) water (pH 7.7; 180 mg equiv. CaCCh/L total hardness and alkalinity; 15.2 mg/L total organic carbon; 12.7 mg/L dissolved organic carbon) was collected from Diss Mere, Diss, Norfolk, and used in the study (p. 10). The water sample was characterized in a separate study (not specified). The drinking water was obtained from a tap in the Residue Analysis Department, and the ground water was obtained from a source in the Huntingdon area by Anglian Water. The drinking and ground water were not characterized; the source of the ground water was not further specified. Page 5 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Table 3. Independent Validation Method Recoveries for Chloropicrin in Water Analyte Fortification Level (jig/L) Number of Tests Recovery Range (%) Mean Recovery (%) Standard Deviation (%)4 Relative Standard Deviation (%) GC/ECD Ground Water Chloropicrin 0.1 5 88-91 90 1 1 1.0 5 85-89 88 2 2 Drinking Water Chloropicrin 0.1 5 88-97 94 4 4 1.0 5 81-85 83 2 2 Surface Water Chloropicrin 0.1 5 73-77 75 1 2 1.0 5 83-86 85 1 2 GC/MS (m/z 119) Ground Water Chloropicrin 0.1 5 71-76 74 2 3 1.0 5 66-82 72 8 11 Drinking Water Chloropicrin 0.1 5 108-117 110 4 3 1.0 5 61-89 79 11 14 Surface Water Chloropicrin 0.1 5 70-77 72 3 4 1.0 5 73-84 79 5 6 Data (uncorrected recovery results, pp. 20-21) were obtained from Table 1, p. 27 of MRID 50167601. * The ground water (PTRL ID 2706W-032; pH 7.3; 627 mg equiv. CaCCh/L hardness; 960 ppm total dissolved solids) obtained from Northwood, North Dakota, drinking (tap) water (PTRL ID 2706W-072/054; pH 7.5; 30 mg equiv. CaCCh/L hardness; 60 ppm total dissolved solids) obtained from Hercules, California, surface water (PTRL ID 2706W-069; pH 7.8; 1183 mg equiv. CaCCh/L hardness; total dissolved solids not determined) obtained from Hercules, California, were used in the study (pp. 15-16). The water samples were provided by EAG-Hercules and previously characterized by Agvise Laboratories. III. Method Characteristics The LOQ for chloropicrin in water was 0.10 |ig/L in the ECM and ILV (pp. 8, 12-13, 16 of MRID 49854401; pp. 12, 22; Tables 2-4, pp. 28-30 of MRID 50167601). In the ECM and ILV, the LOQ was defined as the lowest fortification level where an acceptable mean recovery is obtained. The LOD was reported as 0.05 |ig/L (equivalent to 0.05 ng/mL) in the ECM; in ILV, the LOD was 0.004 |ig/L for ground water and 0.01 |ig/L for drinking and surface water. In the ILV, the LOD was calculated for each matrix using the following equation: LOD = (to.99 x SD) Where, to.99 is the one-tailed t statistic for n = 5 (3.747) and SD is the standard deviation of the analyte recovery measurements at the target LOQ. No calculations or comparisons to background levels were reported to justify the LOQ for the method in the ECM or ILV; no calculations or comparisons to background levels were reported to justify the LOD for the method in the ECM. Page 6 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Table 4. Method Characteristics Analyte1 Chloropicrin GC/ECD (Primary) GC/MS (Confirmation) Limit of Quantitation (LOQ) ECM 0.10 ng/L ILV Limit of Detection (LOD) ECM 0.05 ng/L ILV 0.004 |ig/L (ground) 0.01 |ig/L (drinking and surface) Linearity (calibration curve r2 and concentration range) ECM r2 = 0.9990 (0.5-100 ng/mL) r2 = 0.9990 (0.5-80 ng/mL) ILV r2 = 0.9993 (ground, drinking & surface) r2 = 0.99503422 (ground) r2 = 0.99202763 (drinking) r2 = 0.99689992 (surface) (0.5-100 ng/mL Repeatable ECM1 Yes at LOQ and lOxLOQ [characterized surface (lake) water matrix and uncharacterized drinking and ground water matrices] ILV2-3 Yes at LOQ and lOxLOQ [characterized surface, drinking (tap), and ground water matrices] Reproducible Yes at LOQ and lOxLOQ Specific ECM No matrix interferences were observed or quantified; however, chromatograms were very faint. Some non-interfering matrix contaminants were observed in the drinking water. Matrix interferences appeared to be <10% of the LOQ peak (based on peak height); however, chromatograms were very difficult to interpret. ILV Matrix interferences were ca. 7% of the LOQ (based on peak area) in drinking water. No matrix interferences were observed or quantified in ground and surface water. No matrix interferences were observed or quantified. Data were obtained from pp. 8, 12-13, 15-16 (specificity andLOQ/LOD); Tables 1-2, p. 18 (calibration data); Tables 3-8, pp. 19-24 (recovery data); Figures 3-14, pp. 27-32 (chromatograms) of MRID 49854401; pp. 12, 22; Tables 2-4, pp. 28-30 (LOQ/LOD); Table 1, p. 27 (recovery data); Figures 15-19, pp. 45-49 and Figure 32-34, pp. 62-64 (calibration data); Figures 2-14, pp. 32-44; Figures 20-31, pp. 50-61 (chromatograms) of MRID 50167601. 1 In the ECM, surface (lake) water (pH 7.7; 180 mg equiv. CaCCh/L total hardness and alkalinity; 15.2 mg/L total organic carbon; 12.7 mg/L dissolved organic carbon) was collected from Diss Mere, Diss, Norfolk, and used in the study (p. 10 of MRID 49854401). The water sample was characterized in a separate study (not specified). The drinking water was obtained from a tap in the Residue Analysis Department, and the ground water was obtained from a source in the Huntingdon area by Anglian Water. The drinking and ground water were not characterized; the source of the ground water was not further specified. 2 In the ILV, the ground water (PTRL ID 2706W-032; pH 7.3; 627 mg equiv. CaCCh/L hardness; 960 ppm total dissolved solids) obtained from Northwood, North Dakota, drinking (tap) water (PTRL ID 2706W-072/054; pH 7.5; 30 mg equiv. CaCCh/L hardness; 60 ppm total dissolved solids) obtained from Hercules, California, surface water (PTRL ID 2706W-069; pH 7.8; 1183 mg equiv. CaCCh/L hardness; total dissolved solids not determined) obtained from Hercules, California, were used in the study (pp. 15-16 of MRID 50167601). The water samples were provided by EAG-Hercules and previously characterized by Agvise Laboratories. 3 The method was validated in the second trial with two modifications to the sample processing procedure to increase analyte recovery: shortened extraction time (1 minute with inversion and vortexing) and use of a smaller vessel to reduce headspace (pp. 11, 16-17, 19-21, 25 of MRID 50167601). The first trial failed due to low recoveries; therefore, the ECM should be updated with the ILV modifications. Page 7 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Linearity is satisfactory when r2 >0.995. IV. Method Deficiencies and Reviewer's Comments 1. An updated ECM should be submitted with the ILV modifications. The ILV validated the method in the second trial with two modifications to the sample processing procedure to increase analyte recovery: shortened extraction time (1 minute with inversion and vortexing) and use of a smaller vessel to reduce headspace (pp. 11, 16-17, 19-21, 25 of MRID 50167601). The first trial failed due to low recoveries. 2. In ILV, the linearity was not satisfactory for the GC/MS analysis of chloropicrin in drinking water (r2 = 0.99202763; Figures 15-19, pp. 45-49 of MRID 50167601). 3. The specificity of the method was not well-supported by the ECM representative chromatograms. GC/ECD chromatograms were very faint, and GC/MS chromatograms were very difficult to interpret due to poor resolution and the over-laying of the peak area on the analyte peak. 4. Two of the three ECM water matrices were not characterized: drinking and ground water matrices (p. 10 of MRID 49854401). 5. The estimations of LOQ and LOD in ECM and ILV were not based on scientifically acceptable procedures as defined in 40 CFRPart 136 (pp. 8, 12-13, 16 of MRID 49854401; pp. 12, 22; Tables 2-4, pp. 28-30 of MRID 50167601). In the ECM and ILV, the LOQ was defined as the lowest fortification level where an acceptable mean recovery is obtained. The LOD was reported as 0.05 |ig/L (equivalent to 0.05 ng/mL) in the ECM; in ILV, the LOD was 0.004 |ig/L for ground water and 0.01 |ig/L for drinking and surface water. In the ILV, the LOD was calculated for each matrix using the following equation: LOD = (to.99 x SD), where, to.99 is the one-tailed t statistic for n = 5 (3.747) and SD is the standard deviation of the analyte recovery measurements at the target LOQ. No calculations or comparisons to background levels were reported to justify the LOQ for the method in the ECM or ILV; no calculations or comparisons to background levels were reported to justify the LOD for the method in the ECM. Detection limits should not be based on arbitrary values. 6. In the ECM, the storage stability of the final sample extracts of chloropicrin was determined to be 3 days when stored at -18°C (n = 2; p. 16; Tables 3-8, pp. 19-24 of MRID 49854401). 7. The matrix effects were determined to be insignificant in the ECM (p. 16; Tables 3-8, pp. 19-24 of MRID 49854401). 8. The ILV study author reported that no communications between the ILV and ECM occurred (p. 25 of MRID 50167601). 9. It was reported for the ILV that one sample set (10 fortified samples, 2 matrix controls, 1 solvent control, and 1 reagent blank) required ca. 12 hours (p. 22 of MRID 50167601). Page 8 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Preparation of standard solutions and samples each required ca. 4 hours. GC/ECD analysis/data processing and GC/MS analysis/data processing each required ca. 2 hours. V. References U.S. Environmental Protection Agency. 2012. Ecological Effects Test Guidelines, OCSPP 850.6100, Environmental Chemistry Methods and Associated Independent Laboratory Validation. Office of Chemical Safety and Pollution Prevention, Washington, DC. EPA 712-C-001. 40 CFR Part 136. Appendix B. Definition and Procedure for the Determination of the Method Detection Limit-Revision 1.11, pp. 317-319. Page 9 of 10 ------- Chloropicrin (PC 081501) MRIDs 49854401/50167601 Attachment 1: Chemical Names and Structures Chloropicrin IUPAC Name: CAS Name: CAS Number: SMILES String: CI CI — i — I Page 10 of 10 T ri chl oronitrom ethane Not reported 76-06-2 Not found ------- |