Document 2gX9KBk53NdeGRQrw6jvQV9a

3M Strategic Toxicology Laboratory Study Title: Acute Inhalation T.oxicokinetic Study of Perfluoroctanesulfonyl Fluoride (POSF) T-7098.4 Test Number (sample): T-7098.4 (Perfluoroctanesulfonyl Laboratory Project Identifi'cation: ST64 In-Life Initiation Date: 5/1/2001 In-Life Completion Date: 5/14/2001 Research Client: 3M Specialty Materials Division 3M Center Building 236 Saint Paul, MN 55133-3220 Fluoride (POSF) Testing Laboratory: 3M Strategic Toxicology Laboratory 3M Center Building 270-SB- 181 Saint Paul, MN 55133-3220 Study Director: Andrew M. Seacat, Ph.D. Toxicologist Specialist 3M Medical Department/Toxicology 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 Services Study Toxicologist: Tommie Yvette Turner, Ph.D. Post Doctoral Research Fellow 3M Medical Department/Toxicology 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 Services J 2"-7098.4 Acute Inhalation Toxicokinetic Final Report Study of Perfluoroctanesulfonyl Fluoride (POSF) Summary: An acute inhalation toxicokinetic screen was conducted using 3 male rats for control (air-only) and 3 male rats exposed to perfluoroctanesulfonyl fluoride (POSF) at a nominal concentration of 1000 ppm for 4 hours. All animals gained weight during the recovery period and showed no clinical pathology. One animal (N=I) per time point from the POSF-treated animals was sacrificed on day 1, day 6 and day 14 post-exposure. All animals tolerated the 4-hour inhalation exposure and gained weight. Organ weights were normal and histological evaluation of the liver, lungs, bladder, kidney and brain revealed no significant findings. Perfluorooctanesulfonate (PFOS) was found in the serum from each animal (N=I) at 8.6, 9.3 and 5.0 ppm of days I, 6 and 14 post dose, respectively. The percentage of inhaled POSF represented by these serum PFOS concentrations were 0.18%, 0.23%, and 0.10%, respectively. Conclusion: This study demonstrates that there was a small percentage (<0.5%) of total POSF inhaled found as PFOS in serum collected over 14 days. Results show that there was no significant change in PFOS in serum concentration over time. POSF can be absorbed from the lung, and metabolized to PFOS, which is present in serum for greater than 14 days. I. Study Objective: This study was done to establish toxicokinetic parameters data on an acute inhalation of perfluoroctanesulfonyl fluoride (POSF). The goals of this study were to test the compound via inhalation in rats at 1000 ppm for 4 hours for distribution and metabolism, and to use the results as a guidance for a 13 week inhalation study. II. Test Articles: A. Identification: T# Name 7098.3 POSF Structure CsFtTSO2F MW (g/mole) Density (g/ml) 502 1.69 B. Purity and Stability: Responsibility of research client. This POSF sample was.lab fractionated and acid/water washed. Chemical characterization of CsFlTSO2F by IHNMR and 19F-NMR Spectroscopy was conducted by 3M Specialty Adhesives & Chemicals Analytical Laboratory/SMD to determine the purity of the nominal product and to characterize as many impurity components as possible. The sample was found to be a high purity form of the nominal POSF product (1). C. Handling and Storage: Upon receipt, the test article was stored tightly sealed at room temperature. / -;;, T . ]'-7098.4 AcuteInhalationToxicokJnctiSctudyofPcrfluoroctanesulfoFnlyuioride(POSF) _ Report o. D. Disposition of Test Material: Any remaining unused portion of the test article was _r--- returned to the research client after completion of the study. HI. Test System: Male rats were exposed to test compound in air for 4 hours for one day in a 13 liter glass exposure chamber (3 rats/chamber/compound). The flow-through atmosphere was generated by mixing test compound from a syringe pump into a stream of fresh air flowing at 3.0 L/rain into the chamber and exhausted through a vent. The syringe flow rate was calculated based on the Ideal Gas Law equation: ((target concentration)(MW)/(24.79L) ttl/mL)) x (1 g/10 i_tg) x (airflow rate) x (1/density) x (1000 Given: Target concentration (ppm) MW = g/mole 24.79L: The volume of one mole of an ideal gas at 25 C and 1"atmosphere conversion factor: (1 gram)/(1 x 106_tg) airflow rate: (L/rain) ..... density: (ml.Jg) conversion factor: 1000 l.tl/mL (760 mm Hg) The target concentrations of each test compound, the syringe flow rate and the airflow rate were calculated using the Ideal Gas Law and are provided in the following table: Dose Group C Syringe Flow Rate Airflow Rate Treated: POSF (ppm) I000 (laYmin) 30 (L/rain) 2.0 - 3.0 Toxicokinetic group Control: Air only 2.0 - 3.0 Standard target concentrations for each compound were prepared and analyzed by Fourier Transform Infrared Spectrometry (FTIR). Chamber air was monitored by FTIR for concentrations of test compound and oxygen (2). Airflow was increased when necessary to maintain a level of at least 1000 ppm. Control animals were exposed to room air under equivalent chamber conditions as the POSF dose group animals. The control rats used in this study were also used for the control group in the limit test of POSF (T-7098.3) conducted at the same time. Animals were handled in accordance with an animal usage protocol approved by LARC protocol # 2000-0068. The day of dosing was designated day zero of the study. Body weights were determined immediately prior to dosing and immediately prior to II f 2-7098.4Acute InhalationToxicokinctic Studyof PcrfluoroctancsulfonyFiluoride(POSF) F'malReport euthanasia. At the termination of the study, all animals were humanely euthanized by CO2 asphyxiation and necropsies were performed. Blood serum was collected from one animal/time point of the toxicokinetic group sacrificed at day I, day 6, and at day 14 post exposure for PFOS analysis. Analysis of PFOS levels by high-pressure liquid chromatography/tandem electrospray mass spectroscopy (HPLC-MS/MS) was performed according to published methods (3). Serum from the rat sacrificed day one post dose (Animal #RO1468) was initially extracted and quantified by LC/MS/MS based on a PFOS standard curve extracted from urine collected from the limit toxicity study in rats (T-7098.3). This was done to get an estimate of serum PFOS levels on day one post-dose assuming an equal extraction of urine PFOS std curve and serum PFOS. Serum from this animal was later extracted and quantified against a standard curve extracted from serum, and therefore this data was not used in any calculations, but it is included here for completeness (Table 4). Lungs, liver, and bladder were collected at sacrifice. A portion of each tissue was immediately, fixed in a 10% formalin solution for subsequent histological evaluation. The remaining sections were placed directly into propylene tubes filled with liquid nitrogen for possible future evaluation of POSF metabolites levels. IV. Results and Discussion: Test Atmosphere The POSF atmosphere generated in the exposure chamber, measured continuously for four hours by FTIR, averaged 932 + 93 ppm for the toxicokinetic group (4). The average measured concentration in the chamber was 93% of the target concentration for the toxicokinetic group. The airflow was increased from 2 to 3 liters/minutes to adjust chamber concentration. Control Group: The three control group male rats had no adverse clinical observations noted during the 4 hr air-only exposure. The average body weight was 378 + 12 g at the initiation of the study. The control group gained an average of 22 g (5.5%) of body weight during the 14 day study period. Liver weights averaged 14.1 __.1.0 g and the average liver to body weight ratio was 0.035 (Tablel). Necropsy findings and the histopathology report by Dr. Elden Lamprecht, 3M Surgical and Histopathology Services, is found in Appendix l. All rat lungs (both control and treated groups) had mild to moderate congestion of alveoli with occasional extravasation of blood around arterioles and bronchioles. This is possibly an agonal change associated with CO2 euthanasia. It was noted that some of the lungs were not 7 ; r _ * 3"-7098.4 Acute Inhalation F'ifial-Report .7=L:. Toxicokinetic Study of Perfluoroctanesulfonyl Fluoride (POSF) well insufflated. Other than these artifacts, histological analysis revealed no significant changes in lungs and urinary bladder in all three of the controls. POSF Toxicokinetic Group: The three POSF group male rats showed no adverse clinical observations during the in-life phase of the rats. All animals gained weight during the study. At necropsy, no abnormal gross observations were made. Liver weights were equivalent to the control liver weights. Histological analysis in one of the animals revealed that one macroscopic focus of atelectasis (collapsed alveoli) was present in the lung lobe. The change appeared chronic in duration and was not considered to be compound-related. There were no significant changes in the bladder. At necropsy for each animal, there was a small percentage of POSF measured in the serum for each individual animal (Table 3). Perfluorooctanesulfonate (PFOS) was found in the serum from each animal (N-l) at 8.6, 9.3 and 5.0 ppm of days 1, 6 and 14 post dose, respectively. The percentage of inhaled POSF represented by these serum PFOS concentrations were 0.18%, 0.23%, and 0.10%, respectively. These data suggest that the serum PFOS concentrations were at or near their apparent peak concentrations for the first week following exposure and then PFOS began to clear from the serum over the following week. Although there is no statistical power in the present study, the data are consistent with a plasma elimination half-life of 14Cfollowing single oral administration of [14C]FC-95 (mean dose 4.2 mg/kg) to male rats of 7,5 days (5). Rat # RO1468 was sacrificed on day 1 post dose and sera was collected and analyzed for PFOS. The calculation of serum PFOS was based on urine PFOS standard curve, assuming equal extraction (Figure 1). The amount of PFOS in serum averaged over time, collected on days 1, 6, and 14 post-exposur c was 25.90 +1.10 ppm (Table 4). Serum from this animal was later extracted and quantified against a standard curve extracted from serum. The data dervived from the PFOS standard curve (shown in table 4) extracted from urine was not used in any calculations, but it is included here for completeness. This toxicokinetic study confirmed early findings of a POSF 4-hour rat LCs0 greater than 1000 ppm in an inhalation limit toxicity study of POSF (6). In the limit study, rats were exposed to 1000 ppm POSF for four hours. All animals survived the duration of the study. The concentration of PFOS that was found in the serum on day 13 post --dose was 4.57 +0.70 ppm (N=3) which represented approximately 0.10 + 0.01% of the inhaled dose of POSF. The average amount of PFOS in the day-one overnight urine was 3.08 + 1.60 ppm (N=3) which represented approximately 0.05 + 0.03% on the inhaled dose of POSF (6). Thus, the data from the POSF limit test and the data from the toxicokinetic test presented herein are in agreement with each other and support the same conclusions. . ,.o _'-7098.4 Acute Inhalation Fin/d'.R_/pon Toxicokinetic Study of Perfluoroctanesulfonyl Fluoride (POSF) V. Conclusion: This study demonstrates that there was a small percentage (<0.5%) of total POSF inhaled found as PFOS ifi serum collected over 14 days. Results show that there was no significant change in POSF in serum concentration over time. 0 T-7098.4 Acute InhalationToxicokineticStudyof PerfluoroctanesulfonyFlluoride(POSF) ' Hiial_Report ".;.7"-. VI. References: 1. Kestner, T. ChEmical Characterization of POSF (FM-3270) by IH-NMR & 19F-NMR Spectroscopy. 3M Specialty Materials & Manufacturing Division Analytical Laboratory. Request No. 6370. May 15,2001. 2. Modified EPA Method 320. Pace Analytical. 3. Hansen, K.J., Clemen, L.A., Ellefson, M.E. and Johnson, H.O, (2001) Compound Specific, Quantitative Characterization of Organic FluorochEmicals in Biological MatricEs. Environ Sci and Tech., 35, 766-770. 4. Gutzkow, T. POSF Animal Exposure Test Report. Pace Analytical Services, Inc. 3M Laboratory Request No. E01-0648.3M ET&SS Field Analytical Services and TechnologiEs. 5. Johnson, J. D. and Ober, R._._.F1F9.79. Absorption of FC-95-14C in rats after a single oral dose. Project No. 8900310200, Riker Laboratories, Inc., St. Paul, MN. 6. SE:;;cat, A. and Turner, T. (2001) Acute Inhalation Limit Toxicity Study of Perfluooctanesulfonyl Fluoride (POSF) T-7098.3. .. " -' :i_i'_" .: 8"c.7098.4AcuteInhalationToxiokineticS. tudyof PerfluoroctanesulfonyFlluoride(POSF) FitialReport VL. < Signature Page // p "<.-Tommie Yvette Turner, Ph.D./[ q._.._- Date Post Doctoral Research Fello_/ Study Toxicologist v Andrew Seacat, Ph.D. Date Toxicologist Specialist Study Director Reviewed by: .. ___.._..._......_..-._,./,._./o_ John L, Butenhoffr.. 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" .... ....- T-7098.4AcuteInJudadoTno. xicokincficStudyof PerfluorocmnesulfoFnlyu]oride(POSF) FinalReport Apuendix I Histopatholoey Results Necropsy 513101 All rats lungs have generalized mild to moderate congestion of alveoli with occasional extravasation of blood around arterioles and bronchioles. This is possiblyanagonalchangeassociatewdithC02 euthanasiaS.ome lungswerenot wellinsufflated. Animal # Control RO1462: No significant changes in lung and bladder. Toxicokinetic RO1468: No significant changes in lung and bladder. Toxicokinetic RO 1471: One macroscopic focus of atelcctasis (collapsed alveoli) lobe. The change was chronic in duration. No significant bladder. is present in one lung change is present is Toxicokinetic RO1472: No significant changes in lung and bladder. o+ .. _= + =, ? o" T-7098.4 Acute Inhalation Toxicokinetic Study of Perfluooctanesulfonyl Fluoride (POSF) Final Report .. 18 Appendix 2 Serum PFOS To: Andrew Seacat/US-Corporate/3M/US @3M-Corporate Tommie Tumer/US-Corporate/3M/U S cc: John L. Butenhoff/US-Corporatel3M/US@3M-Corporate Paul Lieder/U S-Corporate/3M/US @3M-Corporate Sue Tanaka/US-Corporate/3M/US @3M-Co rporate Kathy Thompson/US-Corporate/3M/U S Deanna J. Luebker/US-Corporate/3M/US@3M-Corporate Subject: PFOS Levels from rats exposed to POSF via inhalationT-7098.3 Andrew and Tommie: The I to 100 estimate for the serum levels of PFOS was right on target. The following results are based on the rat serum being diluted 1:100 prior to extraction. The extraction was carried out at pH 3.0 using a single shake out in Ethyl Acetate. The EA was transferred to a clean polypropylenetube and blown to dryness using nitrogen gas. The resultant residue was re-dissolved in 100 uL of 25% acetonitrile and 75% 10 mm ammonium acetate. Of this solution20 uL were injected onto a Betasil C18 reversed phase HPLC column (50 x 2 mm, 5 micron particle size). The flow rate was maintained at 0.5 ml/min. A gradientflow of acetonitrile was used to elute the compounds of interest startingwith 15% acetonitrile and ending with 650 acetonitrile over 5.5 minutes. Return to initialoperational parameters and column equilibration requires and additional2.5 minutesfor a run time cycle of 8.0 minutes per run. The MS was operated in the Electro Spray negative ionization mode with a constant 3.0 kV potential applied to the source. The method used was an MS/MS method involvingthe transitionof the following ions: Compound Q-2 Offset Voltage Base Peak MS/MS Ion Intemal Standard (PFHS) 60 volts 299 amu 80 ainu 60 volts PFOS 499 ainu 80 amu The capillary heater was held at a constant 300 degrees C. All quantitative calculationswere based on the MS/MS ion rations between the compound of interest (PFOS) and the internal standard (PFHS). The standard curve ranged from 5 ng/mL (ppb) to 250 ng/mL (ppb). A quadraticcurve fit was used with the standard value weighted 1/X. This curve fit produced an R squared value of 0.9935. 18 _'_:::.. T-.:/098.4. AcuteLt,halation ToxicokJnctJcS, tudyof Pcrt'iaooctanesuifonyRluorid (POSF) 19 - .:._e first tube of two available for control animal number 1 R01464 was diluted and ,::runwith the initial extractions. This tube (1:100 dilution)had a PFOS level of 2.3 -i_Pm. Repeat of this tube dilutionproduced comparable results (2.1 ppm). The ._=cond tube labeled 1R01464 was diluted 1:100 and extracted. The second tube on "_iS control animal had a result of less than the lowest standard (5.0 ppb on the '_lilL_tedserum). Only one serum tube was available for animal 1R01465, unfortunately this tube was labeled 1R0465 with the correct date and T # recorded. I diluted and extracted this tube and recorded results under the animal number 1R01465. The sample for 1R01468 was noted as hemolyzed (2+ hemolysis). The initial result of 1R01472 appeared to be significantly lower than 1R01468 and 1R01471. This '_ _ sample was also re-extracted and the analysis repeated. No significant difference was noted on re-extraction. Results: Rat # Control PFOS Level 1R01462 yes 1R01463 yes 1R01464 yes < 5.0 <5.0 <5.0 ppb (on diluted serum) " " 1R01465 no 1R01466 no 1R01467 no 1R01468 no 1R01471 no 1R01472 no 5.1 ppm .... 4.8 ppm 3,8 ppm 8.6 ppm 9.3 ppm 5.0 ppm (5.1 ppm on duplicate) dje ,, _. ...... .................... David J. Ehresman - 3M Corporate Toxicology . _ Bldg: 236,-IB-22 " Phone 651-733-5070 FAX 651-737-4754 djehresman @mmm.com :, _ '-' " " "" ....... ]9 SUBJECT PAGE APPENDIX E - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY E-1 to E-3 APPENDIX F - CERTIFICATE OF ANALYSIS F-1 to F-3 APPENDIX G - ANALYTICAL AND BIOANALYTICAL REPORT G-1 to G-153 APPENDIX H - TEMPERATURE ANFD RELATIVE HUMIDITY REPORT H- 1 APPENDIX I- POSITIVE CONTROL DATA I-1 to I-4 APPENDIX J - HISTOPATHOLOGY REPORT J-I to J-105 APPENIDX K - HEMATOLOGY REPORTS AND CLINICAL CHEMISTRY K-I to K-150 APPENDIX L - STATEMENT OF THE STUDY DIRECTOR L-1 APPENIDIX M - QUALITY ASSURANCE STATEMENT M-1 to M-2 ix