Document NGdeXkz65Q7zYMQMb9brromMp

3M Environmental Technology and Services PO Box 33331 St. Paul, MN 55133-3331 612 778 6442 66( \ W & - 0 5 Protocol FACT-TOX-111 3M Study Title Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats PROTOCOL Author Lisa Clemen Date: June 8,1999 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification FACT-TOX-111 U2994 3M Environmental Laboratory 00952 Page 1 o f 10 Protocol #FACT-T0X-111 Study Identification Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats Test Material Sponsor Sponsor Representative Study Director Study Location(s) In vivo Testing Facility Analytical Testing Laboratory Perfluorooctane sulfonic acid potassium salt (T-6295) 3M Toxicology Services - Medical Department 3M Center, Building 220-2E-02 St. Paul, MN 55144-1000 Marvin T. Case, D.V.M., Ph.D. 3M Toxicology Services Telephone: 651-733-5180 Facsimile: 651-733-1773 Kristen J. Hansen, Ph.D. 3M Environmental Technology and Safety Services Building 2-3E-09 651-778-6018 Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 3M Environmental Laboratory 000953 Page 2 o f 10 Sub-Contract Laboratories Proposed Study Timetable Study Initiation Date Study Completion Date Protocol UFACT-TOX-111 Advanced Bioanalytical Services, Inc. 15 Catherwood Road Ithaca, NY 14850 Battelle Memorial Institute 505 King Avenue Columbus Ohio 43201-2693 June 8,1999 August 8, 2000 1. Study Oral (gavage) pharmacokinetic recovery study of potassium perfluorooctane sulfonic acid (PFOS) in rats. 2. Purpose This analytical study is designed to determine levels of potassium perfluorooctanesulfonate (PFOS) in specimens of liver and serum of rats. The in-life portion of this study was conducted at Argus Research Laboratories, study #418-015. All serum samples will be extracted and analyzed at Advanced Bioanalytical Services, Inc. and all liver samples extracted and analyzed at Battelle Memorial Institute. Additional analyses may be performed at the 3M Environmental Laboratory as methods are developed and validated.. If additional analyses are performed an amendment to this protocol will be written. 3. Regulatory Compliance This study will be conducted in accordance with the United States Food and Drug Administration, Good Laboratory Practices Standards, Final Rule 21 CFR 58, with the exception that analysis of the test material mixture for concentration, solubility, homogeneity, and stability will not be conducted, and is the responsibility of the Sponsor. 4. Quality A ssurance The 3M Environmental Laboratory Quality Assurance Unit will review the protocol and audit study conduct, data, and final report to determine compliance with Good Laboratory Practice Standards and with 3M Environmental Laboratory Standard Operating Procedures. The QA Unit at the sub-contract laboratory will audit their study conduct, data, and results report prior to submitting to the 3M Environmental Laboratory. 000954 3M Environmental Laboratory Page 3 of 10 Protocol UFACT-TOX-111 5. Test Material 5.1 Refer to Argus Research Laboratory protocol for study #418-015. 6. Control Matrices 6.1 Identification Rat liver and serum, and/or rabbit liver and serum traceability numbers will be recorded in the raw data and included in the final report 6.2 Source Argus Research and/or Sigma Chemical 6.3 Physical Description Rat liver and serum, and/or rabbit liver and serum 6.4 Purity and Stability Not applicable 6.5 Storage Conditions Frozen at -20 C 10 C or -50 C 10 C 6.6 Reserve Matrix A portion of the control matrix will be retained in the 3M archives for as long as the quality of the preparation affords evaluation, but not longer than ten years following the effective date of the final test rule (if applicable). 6.7 Disposition Matrices will be retained at the 3M Environmental Laboratory per GLP regulation. Certain matrices (feces, urine, and blood) may be disposed after QAU verification. 6.8 Safety Precautions Refer to MSDS for chemicals used. Wear appropriate laboratory attire, and follow adequate precautions for handling biological materials and preparing samples for analysis. 7. Reference Material 7.1 Identification Potassium perfluorooctanesulfonate (PFOS), lot #s 171,215, or 217 (equivalent lots) 7.2 Source 3M Specialty Chemicals 7.3 Physical Description White powder 7.4 Purity and Stability Purity of PFOS is 99% or greater. Stability has not been determined. 7.5 Storage Conditions Room temperature 7.6 Reserve Material A reserve sample from each batch o f PFOS used in this study will be retained as long as the quality of the preparation affords evaluation, but not longer than ten years following the effective date of the final test rule (if applicable). 7.7 Disposition Unused reference material will be retained for use by the 3M Environmental Laboratory and will be discarded when the quality of preparation no longer affords evaluation. CD =093 000055 3M Environmental Laboratory Page 4 of 10 Protocol UFACT-TOX-111 7.8 Safety Precautions Refer to MSDS for chemicals used. Wear appropriate laboratory attire, and follow adequate precautions for handling biological materials and preparing samples for analysis. 8. Test System Rats were used as the test system and were maintained and dosed as described in Argus Research protocol #418-015. The female rats will be given the test material or control once daily beginning 42 days prior to cohabitation until day 0 of presumed gestation. See table 1 for more dosage information. Dosage Group 1 2 3 Table 1 Dosage Levels, Concentration, and Volumes Number of female rats 8 8 8 (@ Dosage mg/kg/day 0 0.1 k)k Concentration mg/kg/day 0 0.02 0.32 . Dosage volume mL/kg 5 5 5 9. Specimen and Sample Receipt The 3M Environmental Laboratory will receive homogeneity samples and specimens of the following body tissues and fluids from the indicated points in the study. All specimens will be packed-on dry ice for shipping. See table 2 for more specimen information. Body tissue/fluid Table 2 Specimen Information Collected Serum - Dam and Pup animals Urine and Feces - Dam and Pup animals Liver - Dam and Pup animals Dam-Predose, Days 7, 14, 15, 21, and 22 Pup-Day 21 Predose, Days 7, 15,21, and 22 Dam-At the termination of the study Pup-Day21 Expected # of specimens 144 Dam 24 Pup (pooled) 120 Urine and 120 Feces 24 Dam 24 Pup (pooled) Total number of expected specimens: 456 Total number of test animals: 16 Total number of control animals: 8 cu OC0956 3M Environmental Laboratory Page 5 o f 10 Protocol UFACT-TOX-111 Specimens sent to 3M Environmental Laboratories will be received and tracked according to applicable Standard Operating Procedures. 10. Prepara tory Methods 10.1 FACT-M-1.1, Extraction of Potassium Perfluorooctanesulfonate or Other Anionic Fluorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry 10.2 ETS-8-4.1, Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum or Other Fluid for Analysis Using HPLCElectrospray/Mass Spectrometry 10.3 If preparatory methods other than those listed above are used, an amendment to this protocol will be written. Any deviations from these methods will be documented and included with the study data. 10.4 If analyses are sub-contracted to other laboratories, an amendment will be written to include their methods and copies of each method will be attached to this protocol. 11. Analytical Methods 11.1 FACT-M-2.1, Analysis of Fluorochemicals in Liver Extracts Using HPLCElectrospray/Mass Spectrometry 11.2 ETS-8-5.1, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum or Other Fluid Extracts Using HPLC-Electrospray/Mass Spectrometry 11.3 If analytical methods other than those listed above are used, an amendment to this protocol will be written. Any deviations from these methods will be documented and included with the study data. 11.4 If analyses are sub-contracted to other laboratories, an amendment will be written to include their methods and copies of each method will be attached to this protocol. 12. Data Quality Objectives The number of spikes/duplicates, use of surrogates, and information on other data quality indicators are included in the analytical methods. In addition, the following criteria will be met: 12.1 Linearity r2> 0.98 12.2 L im its o f d etection / quantitation 12.2.1 Method Detection Limit (MDL) for PFOS 'C o a) Serum: 1.75 ppb b) Liver: 15 ppb C 000957 3M Environmental Laboratory Page 6 o f 10 Protocol MFACT-TOX-111 12.2.2 Limit of Quantitation (LOQ) - Equal to the lowest acceptable standard in the calibration curve 12.3 Duplicate acceptable precision < 30% for the method 12.4 Spike acceptable recoveries 70% -130% 12.5 Use of confirmatory methods Indeterminate samples will be re-analyzed using a confirmatory method. If a confirmatory method is used, an amendment to this protocol will be written. 12.6 Demonstration of specificity Chromatographic retention time, mass spectral daughter ion characterization. 13. Sub-Contracted Analysis 13.1 All analyses as detailed in this protocol will be performed at 3M Environmental Laboratories, Building 2-3E-09, 935 Bush Avenue, St. Paul, MN 55106, at Advanced Bioanalytical Services, Inc., 15 Catherwood Road, Ithaca, NY 14850, or at Battelle Memorial Institute, 505 King Avenue, Columbus, Ohio 43201-2693. 13.2 An amendment to this protocol will be written if analyses are performed at laboratories other than the 3M Environmental Laboratories, Advanced Bioanalytical Services, Inc., or Battelle Memorial Institute. 14. Statistical Analysis Averages and standard deviations will be calculated. The statistical methods that will be used are described below: 14.1 Data transformations and analysis Data will be reported as the concentration (weight/weight or weight/vol) of PFOS or metabolite per tissue or fluid. 14.2 Statistical analysis Statistics used may include regression analysis of concentrations over time, and standard deviations calculated for the concentrations within each dose group. If necessary, simple statistical tests, such as Student's t test, may be applied to evaluate statistical difference. 15. Report A report containing all the results of the study will be prepared by the 3M Environmental Laboratory. If analyses are sub-contracted to other laboratories, each laboratory will prepare a report and submit it to the 3M Environmental Laboratory for inclusion in the 3M Environmental Laboratory report. Each report will include, but not be limited to, the following, when applicable: 15.1 Name and address of the facility performing the study 15.2 Dates upon which the study was initiated and completed C00D5S Exact/opy of Original y} M - 4-ftf initial Date 3M Environmental Laboratory Page 7 o f 10 Protocol #FACt-TOX-111 15.3 A statement of compliance by the Study Director addressing any exceptions to Good Laboratory Practice Standards 15.4 Objectives and procedures as stated in the approved protocol, including any changes in the original protocol 15.5 The test substance identification by name, chemical abstracts number or code number, strength, purity, and composition or other appropriate characteristics, if provided by the Sponsor 15.6 Stability and the solubility of the test substances under the conditions of administration, if provided by the Sponsor 15.7 A description of the methods used to conduct the test(s) 15.8 A description of the test system 15.9 A description of any circumstances that may have affected the quality or the integrity o f the data 15.10 The name of the Study Director and the names of other scientists, professionals, and supervisory personnel involved in the study 15.11 A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the analytical chemistry data, and a statement of the conclusions drawn from the analyses 15.12 Statistical methods used to evaluate the data, if applicable 15.13 The signed and dated reports of each of the individual scientists or other professionals involved in the study, if applicable 15.14 The location where raw data and the final report are to be stored 15.15 A statement prepared by the Quality Assurance Unit listing the dates that study inspections and audits were made, and the dates of any findings reported to the Study Director and Management If it is necessary to make corrections or additions to a report after it has been accepted, the changes will be made in the form of an amendment issued by the Study Director. The amendment will clearly identify the part of the report that is being amended, the reasons for the amendment, and will be signed by the Study Director. 16. Location of Ra w Data, Records, and Final Report Original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including those items listed below, will be retained in the archives of 3M Environmental Laboratory for at least a period of time as specified by regulation, and as established by 3M Environmental Laboratory Standard Operating Procedures. 000959 Initial Date 3M Environmental Laboratory Page 8 o f 10 Protocol UFACT-TOX-111 16.1 The following raw data and records will be retained in the study folder in the study/project archives according to 3M Environmental Laboratory Standard Operating Procedures: 16.1.1 Approved protocol and amendments 16.1.2 Study correspondence 16.1.3 Shipping records 16.1.4 Raw data 16.1.5 Approved final report (original signed copy) 16.1.6 Electronic copies of data 16.2 The following supporting records will be retained separately from the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures: 16.2.1 Training records 16.2.2 Calibration records 16.2.3 Instrument maintenance logs 16.2.4 Standard Operating Procedures, Equipment Procedures, and Methods 17. Specimen Retention Specimens will be maintained in the 3M Environmental Laboratory specimen archives for a period of time as specified by regulation or as long as the quality of the preparation affords evaluation, but not longer than ten years following the effective date of the final test rule (if applicable), and as established by 3M Environmental Laboratory Standard Operating Procedures. 18. Protocol Amendments and deviations Planned changes to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. All changes to the protocol will be indicated in the final report. Any other changes will be in the form o f written deviations, signed by the Study Director and filed with the raw data. 19. A ttachments 19.1 Attachment A Preparatory and analytical methods 3M Environmental Laboratory CC09G0 Page 9 o f 10 20. Signatures arvin T. Case, D.V.M., Ph.D., Sponsor Representative Protocol ttFACT-TOX-111 Y?<? Date l L tk Kristen J. Hansen, Ph.D., 3M Environmental Laboratory Study Director Date iginal 3M Environmental Laboratory CO0961 Page 10 o f 10 Study Title Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats PROTOCOL AMENDMENT NO. 1 Amendment Date: August 12,1999 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification ET&SS FACT-TOX111 LIRNU2994 Exacts!qpy of Original 3M Environmental Laboratory CC03G2 Protocol FACT-TOX111 Amendment No. 1 This amendment modifies the following portion(s) of the protocol: 1. PROTOCOL reads: Section 10.0 and 11.0 list the following methods to use for extraction and analysis: FACT-M-1.1 "Extraction of Potassium Perfluorooctanesulfonate or Other Anionic Fluorochemical Surfactant from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" FACT-M-2.1 "Analysis of Fluorochemicals in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" AMEND to read: The extraction and analytical methods FACT-M-1.1 and FACT-M-2.1, respectively, were updated on 07/22/99 to: ETS-8-6.0 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-7.0 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" REASON: The extraction and analytical methods FACT-M-1.1 and FACT-M-2.1, respectively, were updated on 07/22/99 to ETS-8-6.0 and ETS-8-7.0. These methods were updated to replace the extraction solvent ethyl acetate with a different extraction solvent MTBE (methyl tert butyl ether), POAA and Monoester were removed from the standard mix, and M556 was added to the standard mix. The analytical method was updated to include linear regression with 1/x weighting and a few minor changes in the HPLC 1100 instrument parameters. 2. PROTOCOL reads: Section 10.4 and 11.4 state that if the analyses are sub-contracted to other laboratories an amendment will be written to include these methods. A mend to read: The extraction and analytical methods to follow at Advanced Bioanalytical Services will be attached to the protocol. The extraction and analytical method to follow at Battelle Memorial Institute is: "Method for Analysis of Perfluorooctane Sulfonate (PFOS) in Rat Sera-by LC/MS/MS, Version& CPLw REASON: The analytical methods at the sub-contract laboratories were not included in the original protocol. IA C 0009G3 3M Environmental Laboratory Protocol FACT-TOX111 Amendment No. 1 3. PROTOCOL reads: Section 12.2.1 b) lists liver method detection limit as 15 ppb. AMEND to read: The liver method detection limit is 8.50 ppb (ng/g). REASON: The validation supporting methods ETS-8-6.0 and ETS-8-7.0 included a lower method detection limit for PFOS. Amendment Approval arvin Case, D.V.M., Ph.D., Sponsor Representative n * Date Kris J. Hansen, Ph.D., Study Director Exactb)y of Original 3M Environmental Laboratory C00964 Study Title Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats PROTOCOL AMENDMENT NO. 2 Amendment Date: September 30,1999 Performing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project identification ET&SS FACT-TOX111 LIRNU2994 3M Environmental Laboratory 000965 Protocol FACT-TOX111 Amendment No. 2 This amendment modifies the following portion(s) of the protocol: 1. Protocol reads: Section 2 states the study is designed to determine levels of potassium perfluorooctanesulfonate (PFOS) in specimens of liver and sera in rats. Amend to read: The study is designed to determine levels of potassium perfluorooctanesulfonate (PFOS) in specimens of liver, sera, and urine in rats. Reason: The urine analytical methods were validated and approved after approval of the original protocol. 2. Protocol reads: Section 10.0 and 11.0 list the following methods to use for extraction and analysis: ETS-8-4.1 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-6.0 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-5.1 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical ' Compounds in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" ETS-8-7.0 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" Amend to read:: ETS-8-4.1 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-6.0 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" ETS-8-96.0 "Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Urine for Analysis Using HPLC-Electrospray/Mass Spectrometry/Mass Spectrometry" ETS-8-5.1 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry" ETS-8-7.0 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds in Liver Extracts Using HPLC-Electrospray/Mass Spectrometry" ETS-8-97.0 "Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds in Urine Extracts Using HPLC-Electrospray/Mass Spectrometry/Mass Spectrometry" REASON: The extraction and analytical methods ETS-8-96.0 and ETS-8-97.0, were approved after approval of the original protocol. C00966 3M Environmental Laboratory Exact/iOiiy of Original Protocol FACT-TOX111 Amendment No. 2 3. Protocol reads: Section 6.1 lists control matrices as rat liver and serum or rabbit liver and serum. Source Argus and/or Sigma Chemical. Amend to read: Control matrices identification rat liver and serum, rabbit liver and serum, human urine and/or rat urine. Source Argus, Sigma Chemical, Lampire Biologicals, Biological Specialty Corp. and/or Golden West Biologicals. Reason: Addition of control urine specifications. 4. Protocol reads: Section 12.2.1 a) lists sera method detection limit as 1.75 ppb and b) lists liver method detection limit as 15 ppb. Amend to read: The method detection limits for all compounds and matrices will be taken from the methods used for extraction and analysis. Reason: The method detection limits listed are specific to the 3M Environmental Laboratory. Statement was added to allow for sub-contracted analyses and/or revised methods. Original *O< 3M Environmental Laboratory C009G7 Protocol FACT-TOX111 Amendment No. 2 5. Protocol reads: Section 16 states that the original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including: approved protocol and amendments, study correspondence, shipping records, raw data, approved final report, electronic copies of data, training records, calibration records, instrument maintenance logs and standard operating procedures, equipment procedures, and methods will be retained in the archives of the 3M Environmental Laboratory. Amend to read: Section 16 states that the original data, or copies thereof, will be available at the 3M Environmental Laboratory to facilitate audits of the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including: approved protocol and amendments, study correspondence, shipping records, raw data, approved final report, and electronic copies of data will be retained in the archives of the 3M Environmental Laboratory. All corresponding training records, calibration records, instrument maintenance logs, standard operating procedures, equipment procedures, and methods will be retained in the archives of the facility performing each analysis. Reason: Clarification of the disposition of archived records if analyses are performed at a sub contract laboratory. 6. Protocol reads: Section 17 states that specimens will be maintained in the 3M Environmental Laboratory specimen archives. Amend to read: Specimens will be maintained in the 3M Environmental Laboratory specimen archives. AJ1 specimens sent to sub-contract laboratories will be returned to the 3M Environmental Laboratory upon completion of analysis and submission of the sub-contract laboratory(s) final report. The specimens will be returned with the following documentation: the signed original chain of custody and records of storage conditions while at the sub-contract facility. Reason: Clarification of the disposition of specimens and documentation for analyses performed by a sub-contract laboratory. 3M Environmental Laboratory CC03G8 Amendment Approvai Protocol FACT-TOX111 Amendment No. 2 Marvin Case, D.V.M., Ph.D., Sponsor Representative jgk Kristen J. Hansen, Ph.D., Study Director S Ocr m f Date O O tGzouJ*' 3M Environmental Laboratory 00969 Study Title Analytical Laboratory Report on the Determination of the Presence and Concentration of Potassium Perfluorooctanesulfonate (CAS Number 2759-39-3) in the Serum, Liver, and Urine of Crl:CDBR VAF/Plus Rats Exposed to PFOS Via Gavage PROTOCOL AMENDMENT NO. 3 Amendment Date: 20 January 2000 Performing Laboratories Urine Analyses 3M Environmental Technology and Safety Services Fluorine Analytical Chemistry Team Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Performing Laboratories Liver Analyses Battelle Memorial Institute 505 King Avenue Columbus, OH 43201-2693 Serum Analyses Advanced Bioanalytical Services, Inc. 15 Cathenvood Road Ithaca, NY 14850 Laboratory Project Identification ET&SS LRN-U2994 FACT TOX-111 Argus Study: 418-015 3M Medical Department Study: T-6295.14 3M Environmental Laboratory 000370 Protocol LRN-U2994 Amendment Number 3 This amendment modifies the following portion(s) of the protocol: 1. Protocol reads: The study director for the present study was identified in the protocol as Kristen J. Hansen, Ph.D. Amend to read: The role of study director for the present study was reassigned to Marvin T. Case, D.V.M., Ph.D., as of 20 January 2000. The previous study director, Kristen J. Hansen, has been reassigned to the role of Principle Analytical Investigator. Reason: The role of study director was reassigned in an effort to ensure compliance with Good Laboratory Practice Standards that outline study personnel requirements (refer to 21 CFR Part 58). 2. Protocol reads: The sponsor for the present study was identified as Marvin T. Case, D.V.M., Ph.D. Amend to read: The role of sponsor for the present study was reassigned to John L. Butenhoff, Ph.D., as of 20 January 2000. Reason: To ensure that the study director does not also carry the duties of study sponsor, the sponsor role was reassigned. In this manner, personnel responsibilities and workload are more evenly balanced. Initial OZ3J 3M Environmental Laboratory 000971 Amendment Approval Protocol LRN-U2994 Amendment Number 3 JohnL. Butenhoff, PhD., Sponsor Representative Date L,j~As<- ----- -------------------------------- Kristen J Hansen, PhD., Outgoing Study Director / / - & >- z< m o Date /l IzJyM uy Marvin T. Case, D.V.M., PhD., Incoming Study Director Date 3M Environmental Laboratory 000372 Study Title Oral (Gavage) Pharmacokinetic Recovery Study of PFOS in Rats PROTOCOL AMENDMENT NO. 4 Amendment Date: 20 April 2000 Performing Laboratories Urine Analyses 3M Environmental Technology and Safety Services Fluorine Analytical Chemistry Team Building 2-3E-09, 935 Bush Avenue St. Paul, MN 55106 Feces Analyses Centre Analytical Laboratories, Inc. 3048 Research Drive State College, PA 16801 Liver Analyses Battette Memorial Institute 505 King Avenue Columbus, OH 43201-2693 Serum Analyses Advanced Bioanalytical Services, Inc. 15 Catherwood Road Ithaca, NY 14850 Laboratory Project Identification ET&SS LRN-U2994 FACT TOX-111 Argus Study: 418-015 3M Medical Department Study: T-6295.14 5? 03 of Original Date 3M Environmental Laboratory C00973 Protocol LRN-U2994 Amendment Number 4 This amendment modifies the following portion(s) of the protocol: 1. Protocol reads: The amended section 2.0 text states that this study is designed to determine PFOS in specimens of rat liver, serum, and urine. Amend to read: This study is designed to determine PFOS in specimens of rat liver, serum, feces, and urine. Reason: The analysis of fecal tissue for the target chemical and/or its analytes was added to th scope of the study following the issuance of the protocol. Feces extraction and analytical methods were not validated and approved prior to protocol approval. 2. Protocol reads: The amended section 6.0 lists rat or rabbit liver, serum, and urine. Amend to read: Add: rat or rabbit feces with a physical description of rat or rabbit feces. Reason: Analysis of fecal tissue for the target chemical and/or its analytes was added to the scope of the study following the issuance of the original protocol. 3. Protocol Reads: Section 13.1 lists all of the laboratories that will be conducting analyses for this study. Amend to read: Add: Centre Analytical Laboratories, Inc., 3048 Research Drive, State College, PA 16801 Reason: Feces analyses were added to the scope of this study. The sub-contract laboratory performing analyses was not in the original protocol. 4 . Protocol Reads: Sections 10.4 and 11.4 state that if the analyses are sub-contracted to other laboratories an amendment will be written to include these methods. Amend to read: The feces extraction and analytical method used by Centre Analytical Laboratories will be; OOM-023--003 (Revision 2), "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS." Reason: The sub-contract laboratory performing feces analyses was added to the scope of this study; this method was not validated and approved prior to protocol approval. 3M Environmental Laboratory 000974 Protocol LRN-U2994 Amendment Number 4 This amendment modifies the following portion(s) of the protocol: 1. Protocol reads: The amended section 2.0 text states that this study is designed to determine PFOS in specimens of rat liver, serum, and urine. Amend to read: This study is designed to determine PFOS in specimens of rat liver, serum, feces, and urine. Reason: The analysis of fecal tissue for the target chemical and/or its analytes was added to the scope of the study following the issuance of the protocol. Feces extraction and analytical methods were not validated and approved prior to protocol approval. 2. Protocol reads: The amended section 6.0 lists rat or rabbit liver, serum, and urine. Amend to read: Add: rat or rabbit feces with a physical description of rat or rabbit feces. Reason: Analysis of fecal tissue for the target chemical and/or its analytes was added to the scope of the study following the issuance of the original protocol. 3. Protocol Reads: Section 13.1 lists all of the laboratories that will be conducting analyses for this study. Amend to read: Add: Centre Analytical Laboratories, Inc., 3048 Research Drive, State College, PA 16801 Reason: Feces analyses were added to the scope of this study. The sub-contract laboratory performing analyses was not in the original protocol. 4. P rotocol Reads: Sections 10.4 and 11.4 state that if the analyses are sub-contracted to other laboratories an amendment will be written to include these methods. Amend to read: The feces extraction and analytical method used by Centre Analytical Laboratories will be; 00M-O23-O03 (Revision 2), "Determination of Fluorochemical Residues in Monkey/Rat Feces by LC/MS/MS." Reason: The sub-contract laboratory performing feces analyses was added to the scope of this study; this method was not validated and approved prior to protocol approval. ITB jUBj7SlTQ10 3M Environmental Laboratory 000975 Amendment Approval Protocol LRN-U2994 Amendment Number 4 John L. Butenhoff, Ph.D,, Sponsor Representative A fif' / /C^ r -Z.0 )<> Date U*VT GlsuMarvin T. Case, D.V.M., Ph.D., Study Director A/Cf / u J ^ W ) Date OJ 3M Environmental Laboratory 000376