Document 1ywqr0EmbJB61dV7v5Q0NrkVm

3M MEDICAL DEPARTMENT, CORPORATE TOXICOLOGY Protocol for Study No. T-7098.1 PHARM ACOKINETIC STUDY OF POSF IN RATS 01/07/99 T-7098.1 POSF PK Study Objective: The objective o f this study is to assess the potential for oral absorption, urinary and fecal clearance and biological persistence o f perfluorooctane sulfonyl fluoride (POSF) in male Sprague Dawley rats after a single oral dose. The POSF compound is the starting material for the synthesis o f a wide variety o f perfluorooctane sulfonate (PFOS) based materials. The purpose o f this study is to understand the rate o f metabolism o f POSF to PFOS by the liver. This study will provide data for proper risk characterization o f POSF. Research Client: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul, M N 55133-3220 Sponsor: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul, MN 55133-3220 Study Location: 3M Strategic Toxicology Laboratory 3M Center, Budding 270-3 S-06 room SB314 Saint Paul, M N 55133-3220 Study Director: Andrew M. Seacat, Ph D. Sr. Research Toxicologist 3M Medical Dept. / Corporate Toxicology 3M Center, Building 220-2E-02 Saint Paul, M N 55133-3220 Ph.: 651-575-3161 FAX: 651-733-1773 Study Toxicologist: Deanna Nabbefeld, MS Advanced Research Toxicologist 3M Medical Dept. / Corporate Toxicology 3M Center, Building 220-2E-02 Saint Paul, MN 55133-3220 Ph: 651-737-1374 FAX: 651-733-1773 Proposed Study Timeline (Assuming EHS&R approval on Jan. 5th); In-Life S ta rt D ate: January 11th, 1999 In-Life E nd Date: February 9th, 1999 A nalytical C om pletion D ate: March 22nd, 1999 F inal R eport C om pletion D ate: April 19th, 1999 000103 01/07/99 T-7098.1 Regulatory Compliance: POSFPK This study will be performed in the 3M Strategic Toxicology Laboratory under a defined protocol and classified as a "Class B Study" as explained in TOX SOP 0950, Strategic Toxicology Lab GLP Program Procedure. Test Material: Dan Hakes, Product Responsibility Liaison 3M Chemicals Division, will furnish highpurity POSF. Identification: Name: Perfluorooctane Sulfonyl Fluoride Molecular Formula: To be provided. Lot Number: The lot numbers will be maintained in the raw data. Purity: Documentation will be kept in on file. Stability: Documentation will be kept on file. Storage Conditions: Upon receipt, test material will be stored tightly sealed at room temperature. Characteristics: Information on synthesis methods, composition or other characteristics that define the test material will be kept on file. Animals: Species: Rat Strain: Sprague Dawley Source: Harlan Age at initiation of treatment: 6-8 weeks Weight at initiation of treatment: approximately 150-250g Number and sex: 30 males Table 1 - Dose Groups Group Dose N Euthanasia *1 0 mg/kg 5 day 1 post dose *2 0 mg/kg 5 day 4 post dose *3 0 mg/kg 5 day 29 post dose 4 5 mg/kg 5 day 1 post dose 5 5 mg/kg 5 day 4 post dose 6 5 mg/kg 5 day 29 post dose * Rats in groups 1-3 will be used concomitantly as control animals in studies T-7071.2, Pharmacokinetic Study o fMS56 in Rats, and T-7099.1, Pharmacokinetic Study o fFX- 845 in Rats. Identification: AUA Number: ear tag with animal number or unique tail mark. 2154 000104 2 01/07/99 T-7098.1 POSFPK Husbandry: Housing: Three specific rats from groups 3 and 6 will be housed individually in metabolism cages for portions o f the study (see Table 2). When not in metabolism cages, these rats will be group housed in standard cages. All other rats will be group housed in standard cages throughout the study. Diet/Water: Harlan Teklad LM-485 Mouse/Rat Sterilizable Diet, supplied by Harlan Teklad, Madison, WI, and tap w ater will be provided to all rats ad libitum throughout the study. Environment: Environmental controls for the animal room will be set to maintain a temperature o f 72 3F, humidity o f 30-70%, a minimum o f 10 exchanges o f room air per hour and a 12 hour light/dark cycle. Dose and Dosing Procedures: Method of administration/Dose preparation: A single 5mg/kg dose o f POSF will be administered via oral gavage to rats in groups 4-6 on day zero o f the study. The POSF will be prepared as a 1% (1 mg/ml) uniform suspension in 2% Tween 80 using a 15 ml tissue grinder. A volume o f 5 ml suspension / kg body weight will be administered to each rat. Re-suspension o f solids will be performed with 5 strokes o f the tissue grinder pestel before each sample is drawn-up in the syringe for dosing. A single 5 ml / kg body weight dose o f 2% Tween 80 will be administered via oral gavage to rats in groups 1-3 on day zero o f the study. Observation o fAnimals: Clinical Observations: Each animal will be observed daily for mortality and morbidity and notable findings will be recorded. Additional findings will be recorded as they are observed. Body Weights: Each animal will be weighed immediately prior to dosing, weekly thereafter and immediately prior to euthanasia. Specimen Collection: Frequency (see Table 2): Urine and feces collections will be made on days 1, 2, 4, 14 and 29 post dose. Necropsies will be performed on days 1, 4 and 29 post dose. 000105 3 Table 2 - Schedule Jan 10 Jan 11 day 0 DOSING Jan 17 day 6 PD Jan 24 day 13 PD Jan 18 day 7 PD Jan 25 day 14 PD Switch to met cages. Jan 31 day 20 PD Feb 7 day 27 PD Feb 1 day 21 PD Feb 8 day 28 PD sw itch to met cages Jan 12 day 1 PD C o llectio n Dy 1 PD sac Jan 19 day 8 PD Jan 26 day 15 PD C o llectio n Switch to reg. cages. Feb 2 day 22 PD Feb 9 day 29 PD collection Dy 29 PD sac Jan 13 day 2 PD C ollection Switch to reg cages. Jan 20 day 9 PD Jan 27 day 16 PD Feb 3 day 23 PD Jan 14 day 3 PD Sw itch to met cages. Jan 21 day 10 PD Jan 28 day 17 PD Feb 4 day 24 PD Jan 15 day 4 PD C o llectio n Switch to reg cages. Dy4PD sac. Jan 22 day 11 PD Jan 29 day 18 PD Feb 5 day 25 PD Jan 16 day 5 PD Jan 23 day 12 PD Jan 30 day 19 PD Feb 6 day 26 PD 01/07/99 T-7098.1 POSF PK Method of Specimen Collection: Urine and feces will be collected from each metabolism cage at the designated times. The initial volume o f urine will be recorded, the sides o f the urine collection apparatus will be washed with 10-20 ml deionized water and the final volume o f urine will be brought to 45 ml with additional deionized water. Daily feces weight will be recorded for each animal. At the designated times, animals will be euthanized by C 0 2 and gross necropsy performed. During necropsy, blood ( 6 ml) will be collected via the abdominal aorta and transferred to blood collection tubes without anticoagulant. Blood samples will be allowed to clot for a period o f 15 to 30 minutes at room temperature, and the clot will be spun down in a centrifuge at 1100 x g for 5 minutes. The serum will be transferred to labeled 1.5 ml microfuge tubes and centrifuged again at 2000 x g to remove any remaining red blood cells. Each sera sample will then be transferred to a separate labeled polypropylene microfuge tube and flashfrozen in liquid nitrogen. Liver, kidneys and subcutaneous fat from each animal will be removed, weighed, flash frozen in liquid nitrogen and placed individually into labeled sterile sample bags. The remainder o f each carcass will be placed in a labeled ziplock bag and frozen (-70 C) until analysis. Specimen Handling: Specimens will temporarily be stored in a freezer set to maintain -60 to 80C. For metabolite analysis, these specimens will be packed in dry ice and shipped to: 000106 4 Kris Hansen, Ph.D. 3M Environmental Technology and Safety Services 935 Bush Avenue St. Paul, M N 55133-3331 Ph: 612-778-6081, FAX: 612-778-6176. 01/07/99 T-7098.1 POSF PK The 3M Environmental Laboratoiy will manage the extraction and analysis o f sera, liver, urine and feces for the parent compound, POSF, and its presumed metabolite, PFOS. All tissue samples will be retained for possible future analysis o f total organic fluorine (TOF) if deemed necessary by the Study Director. 3M Environmental Laboratory or its designee would perform this analysis. All results will be provided for inclusion in the final report. The number, type and date o f collection o f specimens to be generated for analysis are as follows: Table 3 - Specimens Specimens day 1 post dose Serum 10 Liver 10 Kidneys 10 Subcutaneous fat 10 Carcass 10 Urine 6 Feces 6 day 2 post dose 6 6 day 4 post dose 10 10 10 10 10 6 6 day 15 post dose 6 6 day 29 post dose 10 10 10 10 10 6 6 Total 30 30 30 30 30 30 30 Data Analysis: Data collected on tissue levels o f parent compound and identifiable metabolites will be analyzed for toxicokinetic parameters and for statistically significant differences between groups using Students T-test and/or ANOVA. Responsibilities: Deanna Nabbefeld and Andrew Seacat will be responsible for dosing the animals, collecting in-life specimens, performing the necropsy and collecting and sending tissue samples for analysis. Kris Hansen, 3M Environmental, will be responsible for analytical analysis. Andrew Seacat will draft a final report and ensure the report receives appropriate 3M review before a final report is issued. 00107 5 Signatures: Study Director Deanna Nabbefera, MS Advanced Toxicologist Study Toxicologist Sponsor Representative 01/07/99 T-7098.1 POSF PK Date Date 00108 6