Document vENKmMxexLMrQY8p9oDMr92m

M22G-OIH 3M M ED ICA L DEPARTM ENT, CO RPO RA TE TO X ICO LO G Y P rotocol fo r S tudy No. T-7098.1 PH A RM A CO K IN ETIC STUDY O F PO SF IN RATS 01/07/99 T-7098.1 POSFPK j 0 A "7 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 (PO SF) 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 (PFO S) based m aterials. 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, M N 55133-3220 Study Location: 3M Strategic Toxicology Laboratory 3M Center, Building 270-3S-06 room SB314 Saint Paul, M N 55133-3220 Study Director: Andrew M. Seacat, Ph.D. Sr. R esearch Toxicologist 3M M edical Dept. / C orporate 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 M edical D ept. / C orporate Toxicology 3M Center, Building 220-2E-02 Saint Paul, M N 55133-3220 Ph: 651-737-1374 FAX: 651-733-1773 Proposed Study Timeline (Assuming EHS& R approval on Jan. 5th.).* In-L ife S ta rt D ate: January 11th, 1999 In-L ife E n d D ate: February 9th, 1999 A nalytical C om pletion D ate: M arch 22nd, 1999 F in al R ep o rt C om pletion D ate: April 19th, 1999 004423 01/07/99 T-7098.1 POSFPK Regulatory Compliance: This study will be perform ed 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 H akes, Product Responsibility Liaison 3M Chemicals Division, will furnish highpurity POSF. Id e n tific a tio n : N am e: Perfluorooctane Sulfonyl Fluoride M olecular Form ula: To be provided. L o t N um ber: The lot numbers will be m aintained in the raw data. P urity: D ocum entation will be kept in on file. S ta b ility : D ocum entation will be kept on file. Storage C onditions: U pon receipt, test m aterial will be stored tightly sealed at room tem perature. C haracteristics: Inform ation on synthesis m ethods, com position o r other characteristics that define the test material will be kept on file. Anim als: Species: Rat S tra in : Sprague Dawley Source: H arlan Age a t in itiatio n o f treatm en t: 6-8 weeks W eight a t in itia tio n o f tre a tm e n t: approxim ately 150-250g N um ber an d 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 Xpost dose 5 5 mg/kg 5 day 4 post dose 6 5 mg/kg 5 day 29 post dose * Rats in groups 1-3 w ill be used concomitantly as control animals in studies T-7071.2, Pharmacokinetic Study o/M 556 in Rats, and T-7099.1, Pharmacokinetic Study o fFX- 845 in Rats. Id e n tific a tio n : AUA N um ber: ear tag w ith animal number or unique tail mark. 2154 004424 2 01/07/99 T-7098.1 POSFPK H usbandry: H ousing: Three specific rats from groups 3 and 6 will be housed individually in metabolism cages for portions o f the study (see Table 2). W hen 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. D iet/W ater: H arlan Teklad LM -485 M ouse/R at Sterilizable D iet, supplied by Harlan Teklad, M adison, W I, and tap w ater will be provided to all rats ad libitum throughout the study. Environm ent: Environm ental controls for the animal room will be set to maintain a tem perature 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: M ethod o f adm inistration/D ose p rep aratio n : A single 5mg/kg dose o f PO SF will be adm inistered via oral gavage to rats in groups 4-6 on day zero o f the study. The PO SF 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 w eight will be adm inistered to each rat. R e-suspension o f solids will be perform ed w ith 5 strokes o f the tissue grinder pestel before each sample is draw n-up in the syringe for dosing. A single 5 ml / kg body w eight dose o f 2% Tw een 80 w ill be administered via oral gavage to rats in groups 1-3 on day zero o f the study. Observation o fAnimals: C linical O bservations: Each animal will be observed daily for m ortality and m orbidity and notable findings will be recorded. Additional findings will be recorded as they are observed. Body W eights: Each animal will be weighed immediately prior to dosing, weekly thereafter and immediately prior to euthanasia. Specimen Collection: F requency (see Table 2): U rine and feces collections will be made on days 1 , 2 , 4 , 1 4 and 29 post dose. N ecropsies will be perform ed on days 1, 4 and 29 post dose. 004425 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 switch to met cages Jan 12 day 1 PD Collection Dy 1 PD sac Jan 19 day 8 PD Jan 26 day 15 PD Collection Switch to teg. cages. Feb 2 day 22 PD Feb 9 day 29 PD collection Dy 29 PD sac Jan 13 day 2 PD Collection Switch to teg cages. Jan 20 day 9 PD Jan 27 day 16 PD Feb 3 day 23 PD Jan 14 day 3 PD Switch to met cages. Jan 21 day 10 PD Jan 28 day 17 PD Feb 4 day 24 PD Jan 15 day 4 PD Collection Switch to teg cages. Dy 4 PD 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 POSFPK Method of Specimen Collection: U rine and feces will be collected from each metabolism cage at the designated times. The initial volum e o f urine will be recorded, the sides o f the urine collection apparatus will be washed w ith 10-20 ml deionized w ater and the final volume o f urine will be brought to 45 ml w ith additional deionized w ater. Daily feces w eight will be recorded for each animal. A t the designated times, animals will be euthanized by CO2 and gross necropsy perform ed. During necropsy, blood ( 6 ml) will be collected via the abdominal aorta and transferred to blood collection tubes w ithout anticoagulant. Blood samples will be allowed to clot for a period o f 15 to 30 m inutes at room tem perature, and the clot will be spun dow n in a centrifuge at 1100 x g for 5 minutes. The serum will be transferred to labeled 1.5 ml m icrofuge tubes and centrifuged again at 2000 x g to rem ove any remaining red blood cells. Each sera sample will then be transferred to a separate labeled polypropylene m icrofuge tube and flashfrozen in liquid nitrogen. Liver, kidneys and subcutaneous fat from each animal w ill be removed, weighed, flash frozen in liquid nitrogen and placed individually into labeled sterile sample bags. The rem ainder o f each carcass will be placed in a labeled ziplock bag and frozen (-70 C) until analysis. Specimen Handling: Specimens will tem porarily be stored in a freezer set to m aintain -60 to - 80C. F or m etabolite analysis, these specimens will be packed in dry ice and shipped to: 004426 4 Kris Hansen, Ph.D. 3M Environm ental 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 POSFPK The 3M Environmental Laboratory will m anage the extraction and analysis o f sera, liver, urine and feces for the parent compound, POSF, and its presum ed m etabolite, PFOS. All tissue sam ples will be retained for possible future analysis o f total organic fluorine (TO F) if deemed necessary by die Study D irector. 3M Environm ental Laboratory o r its designee would perform this analysis. All results w ill 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 S pecim ens day 1 post dose Serum 10 Liver 10 Kidneys 10 Subcutaneous fat 10 Carcass 10 U rine 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: D ata collected on tissue levels o f parent com pound and identifiable m etabolites will be analyzed for toxicokinetic param eters and for statistically significant differences betw een groups using Students T -test and/or ANOVA. Responsibilities: Deanna Nabbefeld and A ndrew Seacat will be responsible for dosing th e animals, collecting in-life specimens, perform ing the necropsy and collecting and sending tissue samples for analysis. K ris Hansen, 3M Environm ental, 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. 004427 5 Signatures: Study D irector Deanna NabbefeM^ M S Advanced Toxicologist Study Toxicologist Sponsor Representative 01/07/99 T-7098.1 POSFPK D ate D ate 004428 6