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AR226-1898 T-6295.15 FR DT31 3M MEDICAL DEPARTMENT, CORPORATE TOXICOLOGY Low Level Oral Perfluorooctanesulfonate IPFOS) Dose Toxicokinetic Study in Rats: Serum and Liver PFOS Final Report Date: April 14, 2004 Study No. T-6295.15 Strategic Toxicology Study Number: DT-31 Research Client: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul MN 55133-3220 Sponsor: 3M Specialty Chemicals Division 3M Center, Building 236 Saint Paul MN 55133-3220 Study Location: 3M Strategic Alternative Toxicology Laboratory 3M Center, Building 270-SB-181 Saint Paul, MN 55133-3220 Study Director: Andrew M. Seacat Ph.D. Toxicology Specialist 3M Medical Dept. / Corporate Toxicology 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 Ph.: 651-575-3161, FAX: 651-733-1773 Study Toxicologist: Deanna Luebker M.S 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 Table of Contents Table of Contents............................................................................................................................................... 2 Summary.............................................................................................................................................................4 introduction.........................................................................................................................................................5 Methods............................................................................................................................................................... 5 R esults:................................................................................................................................................................ 7 Discussion......................................................................................................................................................... 10 Conclusion......................................................................................................................................................... 1 Signatures.......................................................................................................................................................... 12 Reviewed by...................................................................................................................................................... 12 References......................................................................................................................................................... 13 T a b le s .................................................................................................................................................................15 Table 1. Biological Parameters....................................................................................................................16 Table 2. Summary Serum PFOS T able.....................................................................................................17 Table 3. Summary Serum PFOS Table, LOQ/2, LOD/2 substituted....................................................... 18 Table 4. Summary of WinNonlin Noncompartmental Analysis of Mean Serum[PFOS].......................19 Table 5. Summary Liver [PFOS] (pg/g) and Percent (%) Dose in Liver................................................20 Table 6. Summary of WinNonlin Noncompartmental Analysis of Mean Liver [PFOS]...................... 21 Table 7. Summary Liver:Serum PFOS Concentration Ratio (pg/g:pg/g)................................................ 22 Figures...............................................................................................................................................................23 Figure 1. Average Serum PFOS Vs Time. Female rats. All dose groups................................................ 24 Figure 2. Average Serum PFOS Vs Time. Male rats. All dose groups....................................................24 Figure 3. Average Serum PFOS Vs Time. Female rats. 0 to 100 ug/Kg dose groups.............................25 Figure 4. Average Serum PFOS Vs Time. Male rats. 0 to 100 ug/Kg dose groups................................25 Figure 5. Liver PFOS Vs Time. Male and Female Rats. Control dose group..........................................26 Figure 6. Liver PFOS Vs Time. Male and Female Rats. 3 ug/Kg dose group........................................26 Figure 7. Liver PFOS Vs Time. Male and Female Rats. 10 ug/Kg dose group.......................................27 Figure 8. Liver PFOS Vs Time. Male and Female Rats. 30 ug/Kg dose group.......................................27 Figure 9. Liver PFOS Vs Time. Male and Female Rats. 100 ug/Kg dose group.....................................28 Figure 10. Liver PFOS Vs Time. Male and Female Rats. 5000 ug/Kg dose group............................... 28 Appendix 1: Biological Parameters. Individual and Summary Data Tables.............................................. 29 Appendix 2: Individual Sera PFOS and % of dose in sera............................................................................41 Appendix 3: Sera PFOS WinNonlin Model 200 Non Compartmental Analysis (NCA)............................ 49 Figure 11. NCA Avg Sera PFOS vs Time. Female rats. Control dose group..........................................49 Figure 12. NCA Avg Sera PFOS vs Time. Male rats. Control dose group............................................. 49 Figure 13. NCA Avg Sera PFOS vs Time. Female rats. 3 pg/Kg dose group.......................................... 50 Figure 14. NCA Avg Sera PFOS vs Time. Male rats. 3 pg/Kg dose group..............................................50 Figure 15. NCA Avg Sera PFOS vs Time. Female rats. 10 pg/Kg dose group........................................ 51 Figure 17. NCA Avg Sera PFOS vs Time. Female rats. 30 pg/Kg dose group........................................ 52 Figure 18. NCA Avg Sera PFOS vs Time. Male rats. 30 pg/Kg dose group.......................................... 52 Figure 19. NCA Avg Sera PFOS vs Time. Female rats. 100 pg/Kg dose group.....................................53 Figure 20. NCA Avg Sera PFOS vs Time. Male rats. 100 pg/Kg dose group........................................ 53 Figure 21. NCA Avg Sera PFOS vs Time. Female rats. 5000 pg/Kg dose group...................................54 Figure 22. NCA Avg Sera PFOS vs Time. Male rats. 5000 pg/Kg dose group...................................... 54 Appendix 4. Individual Liver PFOS, % dose in Liver and Liver/Sera PFOS amount (pg/pg) ratio..........79 Appendix 5 WinNonlin NCA Model 200 Liver PFOS................................................................................ 86 Figure 23. NCA of Average Liver PFOS Vs Time. Female Rats. Control dose group......................... 86 Figure 24. NCA of Average Liver PpOS Vs Time. Male Rats. Control dose group............................. 86 Figure 25. NCA of Average Liver PFOS Vs Time. Female Rats. 3 ug/Kg dose group.........................87 Figure 26. NCA o f Average Liver PFOS Vs Time. Male Rats. 3 ug/Kg dose group............................ 87 Figure 27. NCA of Average Liver PFOS Vs Time. Female Rats. 10 ug/Kg dose group...................... 88 Figure 28. NCA of Average Liver PFOS Vs Time. Male Rats. 10 ug/Kg dose group.......................... 88 Figure 29. NCA of Average Liver PFOS Vs Time. Female Rats. 30 ug/Kg dose group...................... 89 2 Figure 30. NCA of Average Liver PFOS VsTime. Male Rats. 30 ug/Kg dose group.......... 89 Figure 31. NCA of Average Liver PFOS VsTime. Female Rats. 100 ug/Kg dose group.....90 Figure 32. NCA of Average Liver PFOS VsTime. Male Rats. 100 ug/Kg dose group........ 90 Figure 33. NCA of Average Liver PFOS VsTime. Female Rats. 5000 ug/Kg dose group...91 Figure 34. NCA of Average Liver PFOS VsTime. Male Rats. 5000 ug/Kg dose group...... 91 Appendix 6: Statistical Analyses.................................................................................................................. 116 Male and Female sera PFOS LOQ/2 substituted data.......................................................................... 116 0, 3, 10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, days 1, 4 and 14 post-dose...120 0, 3, 10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, day 1 post-dose.......................122 0, 3, 10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, day 4 post-dose.......................123 0, 3, 10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, day 14 post-dose.................... 125 5000 ug/Kg dose group Male sera PFOS Statistics........ ........................................................................127 0. 3, 10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics days 1, 4 and 14 post-dose.... 129 0. 3, 10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 1 post-dose.............. 130 0. 3, 10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 4 post-dose.............. 133 0. 3, 10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 14 post-dose............ 135 5000 pg/Kg dose group Female Sera PFOS Statistics............................................................................ 137 10 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose.....................................139 30 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose....................................141 100 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose..................................143 5000 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose................................145 Male and female liver PFOS concentrations SAS analysis data:...........................................................148 0, 3, 10, 30 and 100 ug/Kg dose group Male Liver PFOS Statistics days 1,4, and 14 post-dose........ 152 5000 ug/Kg dose group Male Liver PFOS Statistics.............................................................................. 153 0, 3, 10, 30 and 100 ug/Kg dose group Female Liver PFOS Statistics days 1,4 and 14 post-dose.....155 5000 ug/Kg dose group Female Liver PFOS Statistics.......................................................................... 157 3 Summary This study was done to generate kinetic data on Perfluorooctanesulfonate (PFOS) in rats in the low dose range, which could be of value in the risk assessment procedure. Male and female Sprague-Dawley rats were given single oral doses of 3, 10, 30, 100, or 5000 pg/Kg PFOS (N=5/sex/dose group/time point). The 3, 10, 30 andlOO pg/Kg dose groups and their corresponding vehicle control group animals (N=5/sex/time point) were sacrificed on days 1, 4 and 14 post-dose. The 5000 pg/'Kg dose group animals and their corresponding vehicle control group animals (N=3/sex/time point) were sacrificed on days 1,4, 14 and 28 post-dose. Serum and liver were collected for analysis of PFOS levels at necropsy. The female rats generally had higher serum PFOS levels than the males in all dose groups and time points. The maximum serum PFOS concentration in the control animals was 60 ng/mL. The low doses (3, 10 and 30 pg/Kg) resulted in maximum serum PFOS concentrations of 50 ng/mL in female rats given a 30 ug/Kg dose, equivalent to control values. The 100 ug/Kg dose group had maximum serum PFOS concentrations of 200 ppb in female rats on day 4 post-dose, and 120 ppb in male rats on day 14 post-dose. The 5000 ug/Kg dose group had maximum serum concentrations of 18.88 8.00 ppm (12% of dose) in females and 13.36 2.56 (~ 9 % of dose) in males on day one post-dose. The serum PFOS half-life of elimination was estimated from the terminal elimination rate constant to be 16 days in female rats and 14 days in male rats. The male rats had higher liver concentrations of PFOS than females at all time points. Significant increases in liver PFOS concentrations compared to control values occurred in the 10, 30 and 100 ug/Kg dose group males and in the 100 pg/Kg dose group females. The average SD liver PFOS concentrations for the 5000 pg/Kg dose groups were in the 30 to 40 pg/g range in both males and females with no significant differences at any time. Liver PFOS concentration half-lives of elimination were determined to be 18 and 24 days for the male 100 ug/Kg and the female 5000 pg/Kg dose groups, respectively. The maximum liver PFOS concentrations and liver PFOS concentration areas under the curve (AUCs) were greater than the control values in the 10 ug/Kg dose group and above for the male rats and in the 30 pg/Kg dose group and above for the female rats. Liver to serum PFOS concentration ratios in the 30 pg/Kg and 5000 pg/Kg dose groups were ranged from 2:1 to 11:1 over time. In contrast, the liver to serum PFOS concentration ratios in the 100 pg/Kg dose group varied from 5:1 to 52:1 and increased in females and decrease in males over time. These results show that PFOS given at 5000 pg/Kg has a serum half-life of elimination of approximately 14-16 days in male and female rats, respectively, which are approximately 2-fold greater than the previously reported serum PFOS half-life of elimination for male rats dosed with an equivalent dose of 14C labeled PFOS. 4 Introduction This study was performed to determine the lowest doses that would elicit a measurable increase in PFOS levels in the liver and serum of rats. This data was necessary to differentiate between background levels of PFOS found in these tissues in control animals and dose-responses in other PFOS toxicity studies. In addition, this study was designed to provide a low-dose response PFOS tissue curve for other toxicokinetic studies on fluoropolymers that contained some residual non-polymerized fluorochemical monomers which are known to be metabolized to PFOS. Previous toxicokinetic studies on PFOS indicated that at least 95% of a single oral dose of [!4C]FC-95 (mean dose 4.2 mg/kg) administered to male rats was absorbed within 24 h, not metabolized, and the plasma elimination half-life of 14C was 7-5 days (Johnson and Ober 1979). Other studies had shown that PFOS could be dermally absorbed at high doses of 5000 mg/kg (O'Malley and Ebbens 1980), but not necessarily at a very low dose of 0.3 mg/kg which was possibly too low to make an assessment o f dermal absorption (Johnson 1995). Single i.v. doses (mean 4.2 mg/kg) of [14C]FC-95 in 0.9% NaCl were administered to male rats. By 89 days after dosing, 30.2% of the administered l4C had been excreted in the urine and 12.6% had been excreted in the feces (Johnson and Ober 1979). Fecal and total excretion of 14C were markedly increased in male rats administered cholestyramine (~ 2.7 g/kg/d) in their diet following single i.v. doses o f [I4C]FC-95. The results suggest that there was significant enterohepatic circulation of FC-95 (Johnson et al. 1980; Johnson et al. 1984). Methods Regulatory Compliance: This was a non-GLP study. Dose and Dosing Procedures; Two hundred and fourteen (107 male and 107 female) Sprague-Dawley rats from Harlan Laboratories, Inc. 6-8 weeks old and Weighing approximately 150-200g were used in this study. The original Protocol was amended twice to change and/or add dose groups. To strengthen the low-dose portion of the dose response curve, Dose Group 5 (originally 300 pg/Kg) was replaced with a 10 pg/'kg dose group. Dose Groups 6 and 7 were added to extend the time course and provide a 5 mg/Kg dose group as a reference dose for comparison to other studies. The final dose groups and scheduled sacrifice times were as follows: Dose Group Number of Animals GROUP I Control GROUP 2 3pg/kg GROUP 3 30pg/kg GROUP 4 100pg/kg GROUP 5 IQpg/kg_______ GROUP 6 Additional Control 15 male, 15 female 15 male, 15 female 15 male, 15 female 15 male, 15 female 15 male, 15 female 12 male, 12 female [PFOS] in dose (pg/mL) 0.6 6.0 20.0 2.0 Scheduled Sacrifice, N/sex (Day Post-Dose) 1,4 andl4, 5/sex 1,4 andl4, 5/sex 1,4 andl4, 5/sex 1,4 and 14, 5/sex 1,4 andl4, 5/sex 1,4,14 and 28, 3/sex 5 GROUP 7: 5000 ng/kg 20 male, 20 female 1000 I 1,4,14 and 28, S/sex A Img/mL stock solution of perfluorooctane sulfonic acid, potassium salt;(perfluorooctanesulfonate, PFOS, CAS # 2795-39-3, CgFi7OS02'K+) in 2% Tween 80 was prepared immediately prior to dosing. Serial dilutions of the stock solution was performed to reach the appropriate concentration for each dose level such that a single oral gavage dose volume of 5 ml dosing solution / kg body weight would deliver the desired dose. Doses were administered via oral gavage to rats on day zero o f the study. Dose Selection Justification: The doses selected for this low dose PFOS study were designed to yield serum PFOS levels that are at or near the limit of quantification o f approximately 15 parts per billion (ppb). Based on previous data, 1 mg o f ingested PFOS yielded approximately 5 ppm PFOS in the blood. Therefore, using linear extrapolation, 3 gg/kg would theoretically yield about 15 ppb PFOS in the blood. Three pg/kg was thus selected as the low dose. Method of Sample Collection and Analysis: At the designated times, animals were euthanized by CO2 and gross necropsy performed. During necropsy, systemic blood ( 5 ml) was collected via the abdominal aorta, transferred to blood collection tubes without anticoagulant and allowed to clot for a period of 15 to 30 minutes at room temperature then centrifuged at 1100 x g for 5 minutes. The serum was transferred to labeled 1.5 ml microfuge tubes and centrifuged again at 2000 x g to remove any remaining red blood cells, transferred to labeled polypropylene tubes and flash-frozen in liquid nitrogen. Livers were removed, weighed, placed individually into labeled tubes and flash frozen in liquid nitrogen. Serum and liver samples were stored in a freezer set to maintain -60 to -80C then packed in dry ice and shipped to: Kris Hansen, 3M Environmental Technology and Safety Services for analysis. Certain liver samples were further sent to and analyzed for PFOS concentration by Battelle Laboratories for PFOS concentration. (Battelle Laboratories, Columbus, Ohio). The analytical methods used were equivalent to the methods reported elsewhere (Hansen et al. 2001). Results were reported to the 3M Medical department in an Excel file format from the 3M Environmental lab and as a letter report containing liver PFOS data from Battelle. A final analytical report was not issued and the primary data were archived by the 3M Environmental Lab as FACT Project No.: TOX 107 on October 8, 2003. Analysis of the Data The average serum PFOS values used for the statistical analyses and for the noncompartmental analyses (NCA on WinNonlin software, (NCA Model 200) were generated by substitution o f one-half the limit o f quantitation (i.e LOQ/2 = 0.00275 (ig/mL for serum PFOS) or one half the limit of detection (LOD/2 = 0.00089 pg/mL for serum PFOS) for all samples that were below those limits, in order to convert the non numeric endpoint into numeric values. Noncompartmental parameters were estimated using a linear trapezoidal rule with linear interpolation to integrate the data. WinNonlin estimated the rate constant, Xz, associated 6 with the terminal elimination phase. If could be estimated, the calculated parameters were extrapolated to infinity. WinNonlin repeats regressions using the last three, then the last four, five points etc, with non-zero concentrations. Points prior to Cmax were not used. The regression with the largest adjusted R2was selected, with these caveats: If the adjusted R2 does not improve, but is within 0.0001 of the largest adjusted R2 value, the regression with the larger number of points was used. Xz must be positive, and calculated from at least three data points. WinNonlin estimated /,z by performing a regression of the natural logarithm of the concentration values in this range on sampling time. is defined as: Xz = -1 (estimated slope). The total amount of PFOS in the liver was calculated based on measured liver weights. The total amount on PFOS in the serum was calculated based on a conversion factor of 32.3 mL of serum per Kg body weight in the rat (Alman and Dittmer 1971). Results: Biological Parameters There was no effect of treatment on body weight (BW) or liver weight (LW) (Table 1; Individual and Summary data are in Appendix 1). Serum PFOS The female rats generally had higher sera PFOS concentrations than the male rats at all dose levels. The average sera PFOS concentrations were below the LOQ for serum PFOS of 5.55 ng/mL for both the male and female rats at the 3 pg/Kg dose levels and essentially below the LOQ in male rats at the 10 and 30 pg/Kg dose levels (Table 2; Appendix 2). The 100 ug/Kg dose group had maximum serum PFOS concentrations of 200 ppb in female rats on day 4 post-dose, and 120 ppb in male rats on day 14 post-dose. The 5000 pg/Kg dose group had maximum serum concentrations on day one post-dose that were significantly higher in females (18.88 8.00 ppm, 12% of dose) than in males 13.36 2.56, ~ 9 % of dose) (Table 2; Appendix 6). Female serum PFOS concentrations were significantly higher than the male serum PFOS concentrations on day one post-dose in the 10, 30, 100 and 5000 ug/Kg dose groups by Dunnett's t-test method (p<0.05, Appendix 6). The other time points were not analyzed for significant differences between male and female serum PFOS concentrations. Statistically significant increases in serum PFOS concentrations compared by the Dunnett's t-test method to control values (LOQ/2 or LOD/2 substituted values) occurred in both male and female rats only in the 100 ug/Kg (Table 3) and 5000 ug/Kg (statistical analyses not shown) dose groups at all time points. Dunnett's t test was used to determine significantly different means from the control group mean (Dunnett 1964). Individual pairwise comparisons using Student's t test were also computed. Student's t test is sized for only an individual comparison. The serum PFOS concentrations for the female 30 pg/Kg dose group were significantly greater than the control values by Student's t-test on days one and four post-dose, demonstrating the difference in serum PFOS concentration response between genders at that dose level (Table 3, Appendix 6). 7 The average SD serum PFOS versus time graphs for female (Fig. 1) and male (Fig.2) rats illustrate the contrast between the 5000 pg/Kg dose group and the lower dose groups. The average SD serum PFOS versus time graphs for the 0, 3, 10 and 100 pg/Kg dose groups for female (Fig. 3) and male (Fig.4) rats contrasts male serum PFOS concentrations, with female serum PFOS levels in that dose range. The males given the 100 pg/Kg dose had serum PFOS concentrations that appeared to be increasing at the end of the 14-day observation period. In contrast, the female rats given the 100 pg/Kg dose had serum PFOS concentrations that decreased over thel4 day observation period. Individual sera PFOS data are shown in Appendix 2. WinNonlin Sera PFOS Noncomparmental analysis The average SD serum PFOS vs time were plotted in separate graphs for each dose group and gender using WinNonlin Noncompartmental Analysis (NCA), Model 200 (Appendix 3). The average serum PFOS values used for the NCA were generated by substitution of one half the limit of quantitation (LOQ/2 = 0.00275 pg/mL) or one-half the limit of detection (LOD/2 = 0.00089 pg/mL) for all samples that were below those limits, in order to convert the non-numeric endpoint into numeric values. The average maximum serum PFOS concentration (Cmax) occurred on day 1 post-dose for the female dose groups of 10 ug/Kg and higher, and on day 14 for the males in the 100 pg/Kg dose group and day 1 for the 5000 pg/Kg dose group for both genders (Table 4). The average serum PFOS half-life of elimination (HL_Lambda_z) was estimated to be 16 days for female rats, and 14 days for the male rats, given 5000 pg/Kg (Table 4). The 21-day serum half-life of elimination derived for the female 30 pg/Kg dose group was not considered a completely reliable half-life of elimination estimate given that there were detectable PFOS values in some of the control group serum samples that were in the same concentration range, however it may have some merit given that all the samples had serum PFOS values above the LOQ. The shorter half-lives of elimination of approximately 8 days derived for the 3ug/Kg group were excluded because the LOQ/2 substitutions had been used in those calculations, and the values were equivalent to levels of PFOS in the control serum. Liver PFOS Liver samples were analyzed at two different laboratories under different standard curves with different limits of quantification. The control (group 1), 3, 10, 30 and 100 pg/Kg dose-groups on day 14 post-dose were analyzed at the 3M Environmental Lab with an LOQ of 10 ng/g. The control (group 1), 3, 10, 30 and 100 pg/Kg dose-groups on days 1 and 4 post-dose were analyzed at the Battelle Lab with an LOQ o f 134 ng/g. On days 1 and 4 post-dose, the control (group 6) and 5000 pg/Kg dose groups were analyzed with an LOQ of 5.88 ng/g and on days 14 and 28 post-dose they were analyzed with an LOQ of 37 ng/g at the 3M Environmental Lab. The liver PFOS concentrations (pg/mL) were below the limit of quantitation (LOQ) for liver PFOS in the majority of both the male and female rats in the control and 3 pg/Kg dose groups and were below the LOQ in female rats at the 10 pg/Kg dose level on day one post-dose (Table 5; Individual data in Appendix 4). The average liver PFOS values were consistently higher than the LOQ in all males in the 10 pg/Kg and higher dose groups, and in all females in the 30 pg/Kg and higher dose groups. The percent of PFOS dosed that was present in the liver ranged from 18% to 244 % of the dose and tended to be higher in the lower dose groups. The background liver PFOS levels found in the control group animals averaged 40 ng/g in two out of 22 male control rat livers analyzed and 104 ng/g in two out of 27 female control rat livers analyzed. It should be noted that five of the male control livers and ten of the female control livers were analyzed under a standard curve that had a limit of quantification of 134 ng/g, and that the other livers were analyzed at much lower limits of quantification. Therefore, the average background liver PFOS concentrations derived from the substitution of the LOQ/2 values for these samples may be artificially high and/or some background PFOS may not have been detected in those liver samples (Appendix 6). The presence of the background PFOS may have contributed the average of 277 + 89% of dose found in the liver for two of the 3 ug/Kg dose group male rats. None o f the other dose groups had an average greater than 86% of the dose in the liver. The overall average SD percent of dose in the liver was 33 12 % in female rats, and 55 20 % in male rats, excluding the 3 pg/Kg dose group male rat results. Males had higher liver concentrations of PFOS than females at all time points. The average SD liver PFOS concentration versus time graphs for the 0, 3, 10, 30 and 100 pg/Kg dose groups on days 1, 4 and 14 post-dose (Figures 5 - 9 respectively) show a clear dose-response relationship above the 30 pg/Kg dose level. The liver PFOS concentrations for the 100 pg/Kg dose group were highest on day 1 post dose and were approximately 1.0 ppm in the females and 1.5 ppm in the males. Significant increases in liver PFOS concentrations compared to control values occurred in the 10, 30 and 100 ug/Kg dose group males and in the 100 pg/Kg dose group females when the 100 ug/Kg and lower dose groups were analyzed across all time points (Appendix 6). The average SD liver PFOS concentrations for the 5000 pg/Kg dose groups on days 1,4, 14 and 28 post-dose were in the 30 to 40 pg/g range and showed considerable overlap between the males and females at most time points (Figure 10). The 5000 pg/Kg dose group liver PFOS concentrations were considered to be very high compared to the control liver values and were therefore not analyzed by a Dunnett' t-test method for comparison to control liver PFOS values. The 5000 pg/Kg dose group liver PFOS concentrations were analyzed for changes over time, and no significant differences were found in the livers of male or female rats treated with 5000 ug/Kg PFOS at any time by a Students t-test. WinNonlin Liver PFOS Noncompartmental analysis The average SD liver PFOS concentration vs time graphs were plotted in separate graphs for each dose group and gender using WinNonlin NCA, Model 200 (Appendix 5). 9 The average liver PFOS values used for the NCA in WinNonlin were generated by substitution of one half the limit of quantitation for all samples that were below the LOQ. The maximum liver PFOS concentration (C max, ug/g) was on day 4 or 14 for most dose groups of 10 pg/Kg and higher (Table 6). Liver PFOS concentration half-lives of elimination were determined to be 18 and 24 days for the male 100 ug/Kg and the female 5000 jug/Kg dose groups, respectively. The liver PFOS Cmax and area under PFOS concentration versus time curves (AUCs, day*pg/mL) were greater than the control values in the 10 ug/Kg dose group and above for the male rats. The liver PFOS AUCs were consistently greater than the control values in the 30 pg/Kg dose group and above for the female rats. Liver PFOS Cmax and the area under the curve at 100 pg/'Kg dose group were approximately 10 and 15 fold greater than the control in male and female rats, respectively. Partition between liver and serum. The liver to serum PFOS concentration (pg/g:pg/mL) ratios exhibited a wide range of values overall, depending on dose (Table 6). Liver to serum PFOS concentration ratios ranged from 2 to 11 and remained relatively stable over the full time course of the study in the 30 pg/Kg and 5000 pg/Kg dose groups. In contrast, the liver to serum PFOS concentration ratios in the 100 pg/Kg dose group varied from 5 to 52 and appeared to increase in time in females and decrease in time in the males (Table 6). The total amount of PFOS in the serum and liver were calculated from the tissue PFOS concentrations based on a conversion factor of 32.3 mL serum per kilogram of body weight in the rat, and on the actual liver weights. The ratio o f total PFOS in liver to total PFOS in serum (pg:pg) was consistently and proportionally higher than the ratio o f PFOS concentrations in these tissues (Appendix 4). Discussion The analytical methods for serum and liver PFOS concentration determinations used in this study were basically the same as published methods that yield results that were considered quantitative to 30% (Hansen et al. 2001). The average matrix spike recoveries from quality control serum samples spiked with 250 ppb PFOS ranged from 7 6 l l % t o l l 2 l % (data not shown). The average matrix spike recoveries from quality control liver samples spiked with 250 ppb PFOS ranged from 89 1 % to 115 4% (data not shown). These extraction efficiencies are similar to the published values for the analytical methods used of 93 9% for serum PFOS and 100 13% for liver PFOS (Hansen et al. 2001). The low level of PFOS detected in less than 10% of the control group serum and liver samples (2 out of 27 each), may have been caused by contamination. The rats used in this study were from Harlan Laboratories and were raised on diets that did not contain fishmeal. That source of rats had previously been determined not to have any background PFOS in their serum or liver (Luebker et al. 2001). Furthermore, the data are inconsistent with serum and some o f the liver PFOS values being below the limit of quantification in the 3, 10 and 30 pg/Kg dose groups. 10 The serum and liver samples were analyzed in sets under different standard curves with various limits of quantification and limits of detection. The sera data was derived using appropriately low LOQs o f approximately 6 ng/mL for the majority o f the data or 37 ng/mL for some of the serum samples from the 5 mg/Kg dose group. The standard curve with the higher LOQ was employed to appropriately bracket the response from the sample set. Liver samples from day 14 and from all the 5 mg/Kg dose group rats were analyzed under a standard curve with an appropriately low limit o f quantification of 10 ng/mL. In contrast, the liver samples from days 1 and 4 post -dose were analyzed under a standard curve with a high LOQ of 134 ng/g. Valuable day 1 and day 4 post-dose data was probably lost from the lowest dose group liver datasets on these days because of the high LOQ, particularly from the female rat livers. Furthermore, detectable liver PFOS concentrations were found in all dose groups from day 14 post-dose analyzed under a standard curve with a lower limit of quantification. The low doses (3, 10 and 30 pg/Kg) resulted in maximum serum PFOS concentrations of 50 ng/mL in female rats given a 30 ug/Kg dose, were equivalent to the maximum serum PFOS concentration in the control animals of 60 ng/mL. The female rats generally had higher serum levels than the males in all dose groups and time points. The half-life of elimination in the liver of 18 days for the male and 24 days for the female rats were similar in duration to the half-life of elimination in the serum of 14 days for the male and 16 days for the female. The partition between liver and serum appear to be in dynamic equilibrium (defined as a constant distribution ratio between the two body compartments (Oberg et al. 2002)) for the samples in the 30 and 5000 pg/Kg dose groups, but not in the 100 pg/Kg dose group which a wide range of liver to serum PFOS ratios over the course of the study. Conclusion These results suggest that doses of 3 pg/Kg and 10 pg/Kg in adult female rats, and 3 pg/Kg, 10 pg/Kg, or 30 pg/Kg in male rats yielded serum PFOS levels that were not statistically different than control values. Only doses of 30 pg/Kg and higher in females and 100 pg/Kg in males (out to 14 days) increased serum PFOS levels above background levels in controls. Conversely, only doses of 100 pg/Kg and higher in females and 30 pg/Kg in males significantly increased liver PFOS levels over background levels in controls. These results showed that PFOS given at 5000 pg/Kg has a serum half-life of elimination of approximately 14-16 days in male and female rats, respectively, which are approximately 2-fold greater than the previously reported serum half-life of elimination for male rats dosed with an equivalent dose of ,4C labeled PFOS. The serum results and most of the liver results were of comparable quality to published data using equivalent methods. New methods could improve robustness of the data and lower the limits of detection, however the conclusions, at least for the serum data would not likely be changed even with more precise data. It is recommended that the liver samples be re analyzed under improved extraction and analytical methods now available. il Signatures Prepared by Deanna Luebker M.S (/ Advanced Research Toxicologist 0 ^ /o 9 /^ 0 0 < ~ / Date ( Q ^ M a ^j / Y V }, Andrew M. Seacat, Ph.D., DABT. Toxicology Specialist Study Director Reviewed by John L. Butenhoff, Ph.D., CIH, DABT Sponsor `s Representative 0+ i / j i .t i 1) Date f\-$ > ru d L ( Z , ' Z . & o Date References Alman, and Dittmer (1971). Blood and Other Body Fluids. .FASEB, 5. Dunnett, C. W. (1964). New tables for multiple comparisons with control. Biometrics 20,482-491. Hansen, K. J., Clemen, L. A., Ellefson, M. E., and Johnson, H. O. (2001). Compound-specific, quantitative characterization of organic fluorochemicals in biological matrices. Environmental Science and Technology 35, 766-770. Johnson, J. D. (1995). Final report analytical study Single -dose dermal absorption/toxicity study of T6049 in Rabbits. 3M Environmental Laboratory,, Saint Paul, MN. Johnson, J. D., Gibson, S. J., and Ober, R. E. (1980). Enhanced elimination of FC.-95-14C and FC-14314C in rats with cholestyramine treatment. Riker Laboratories, Inc.,, St. Paul, MN. Johnson, J. D., Gibson, S. J., and Ober, R. E. (1984). Cholestyramine-enhanced fecal elimination of carbon-14 in rats after administration of ammonium [ 14C]perfluorooctanoate or potassium [14C]perfluorooctanesulfonate. Fundamental and Applied Toxicology 4, 972-6. Johnson, J. D., and Ober, R. F. (1979). Absorption of FC-95-14C in rats after a single oral dose. Riker Laboratories, Inc., St. Paul, MN. Luebker, D. J., Hansen, K. J., Clemen, L. A., Butenhoff, J. L., and Case, M. T. (2001). Background perfluorooctanesulfonate in experimental rats. Toxicologist 60, 232, Abstract ID: 1104. Oberg, M., Sjodin, A., Casabona, H., Nordgren, L, Klasson-Wehler, E., and Hakansson, H. (2002). Tissue distribution and half-lives of individual polychlorinated biphenyls and serum levels of 4-hydroxy2,3,3',4',5-pentachlorobiphenyl in the rat. Toxicol Sei 70, 171-82. O'Malley, K. D., and Ebbens, K. L. (1980). 28 Day percutaneous absorption study with FC-95 in albino rabbits. Riker Laboratories, Inc., St. Paul, MN. 13 14 Tables Table 1. Biological Parameters Summary Table 1: Biolo jical Parameters Day Post-Dose (day) Dose Sex (ug/Kg 1 BW BW L W 4 BW BW L W 14 BW ) (g) (* > (g) (g) j g ] _ (g) (g) 0 F Mean 1 9 0 191 7 .9 8 197 2 0 6 8 .1 6 184 SD 14 12 0 .4 3 11 9 0 .5 9 12 M Mean 2 4 9 2 5 2 11.4 2 4 9 271 11.4 2 2 9 SD 3 3 3 2 1.21 31 2 7 0 .9 7 6 3 F Mean 1 8 4 182 7 .2 8 191 2 0 2 8 .4 2 1 8 0 SD 7 5 0 .3 7 8 4 0 .3 8 8 M Mean 2 3 5 2 3 8 10 .9 2 2 6 2 4 9 10 .7 2 3 0 SD 8 8 0 .5 8 3 5 0 .8 3 8 10 F Mean 1 7 8 181 7 .3 4 182 191 7 .6 4 175 SD 7 9 0 .3 4 7 6 0 .2 5 7 M Mean 2 2 4 2 3 0 10.6 230 249 10.9 2 3 2 SD 12 10 0 .2 4 17 10 0 .9 0 10 30 F Mean 1 9 0 1 9 0 7 .8 6 183 191 7 .4 4 184 SD a 10 1 .1 4 8 9 0 .5 5 10 M Mean 2 3 0 2 3 3 10.3 232 254 10.6 233 100 SD 7 7 0 .3 5 5 4 0 .3 0 11 F Mean 1 8 6 185 7 .4 8 185 1 9 6 7 .8 2 1 8 9 SD 6 6 0.42 4 7 0.51 H M Mean 2 2 9 2 3 4 10 .8 2 2 8 2 5 6 1 1 .0 2 2 8 SD 5 6 0 .4 1 9 7 0 .7 8 3 5000 F Mean 2 0 2 199 8 .2 6 2 1 0 2 1 7 8 .7 2 2 0 5 SD 9 9 0 .7 3 6 10 0 .6 7 11 M Mean 2 8 8 2 9 0 12 .4 2 8 4 3 0 0 12 .8 2 8 7 SD 5 7 0.71 10 13 0.73 8 BWj;Initial Body Weight BWf:Final Body Weight LW: Liver Weight BW is l_ 212 13 294 20 215 9 295 22 207 8 307 7 213 11 301 16 224 16 299 7 230 13 352 15 L W (g) 8 .2 5 0 .5 9 13.1 1.39 8 .7 6 0 .4 2 12.7 2 .0 3 7 .8 4 0 .4 8 13.3 0 .4 6 7 .9 0 0 .6 7 13.2 0 .9 0 8 .9 6 1.13 1 3 .2 0 .5 5 8 .7 4 0 .5 0 14.7 1.31 28 BW BW L W (g) i s L _ (g) 205 239 9.70 16 25 1.47 272 351 14.5 9 17 0.71 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 206 241 8 .6 6 6 13 0 .5 6 287 398 15.2 9 22 1.07 16 Table 2. Summary Serum PFOS Table Dose fcS/Kg) 0 3 10 30 100 5000 Gender F Mean SD M Mean SD F Mean SD M Mean SD F Mean SD M "Mi "sL F Mean SD M Mean SD F Mean SD M Mean SD F Mean SD M Mean SD ______ 1 Sera [PFOS] (ug/mL) 0.01 0 00 % dose sera NA NA <LOQ NA <LOQ Sa-sriw4 <LOQ LOQ. <LOQ. 5.52 0.68 <LOQ NA 3 N*U 0.05 i 5.28 0.01 0.41 ! icT NA 11>fJ 0.17 0.08 5.52 2.64 0.03 0.91 0.00 0.16 18.88 8.00 11.96 5.04 13.36 8.65 2.56 1.62 Day _Post-Dose (day) 4 Sera %dose [PFOS] sera (ug/mL) <LOQ r - <LOQ NA NA 14 I Sera % [PFOS] dose (ug/mL) sera 0.06 NA llsBfi<LOQ 0.01 NA 28 Sera [PFOS] (ug/mL) 0.06 0.07 I % dose sera .A NA 0.01 " A <LOQ. ,N A LOQ w B S ii <LOQ m s m <LOQ im m m ,<LOQ- l i l l i i i <LOQ NA <LOQ NA* "NA" 'N A <LOQ rNA , 0.01 NA NA NA NA" A* NA NA 0.02 0.00 <LOQ~ 6.30 0.53 NA ' NA 0 01 |~' 3^37 NA S jg ll 'U 0.04 0.01 4 91 1 47 0 00 I 0 03 i 0 00 1 1 23 3 97 0 32 a" NA] <LOQ 31 NA <LOQ NA 0.20 0.08 t.79 2.76 0.01 0 55 B S H K B i 0 00 0 05 NA NAJ 0.03 0.96 0.12 4 93 A * NA | 0.01 0.20 0 03 1 08 NA NA 19.60 4.08 10.47 6.24 11.09 1.51 8.00 0.99 5.99 5.21 1.63 0.95 13.00 8.86 9.65 7.61 3.48 3.11 1.00 0.61 2.97 2.32 1.14 1.02 | Limit of Quantitation (LOQ) = 0.0055 ng/mL | NA: Not Applicable. 17 Table 3. Summary Serum PFOS Table, LOQ/2, LOD/2 substituted Gender M M M M M Day Dose N Mean Serum Post dose group PFOS substituted1 (ug/mL) 10 8 0.0020 13 5 0.0028 1 10 5 0.0028 1 30 5 0.0024 1 100 5 0.0276 Std Dev 0.00097 0.00000 0.00000 0.00084 0.00464 Comparison to control by T-test * M 40 8 0.0020 0.00097 M 43 5 0.0028 0.00000 M 4 10 5 0.0028 0.00000 M 4 30 5 0.0028 0.00000 M 4 100 5 0.0265 0.00515 * M M 14 0 8 0.0020 0.00261 M 14 3 5 0.0009 0.00000 M 14 10 5 0.0069 0.00331 M 14 30 5 0.0020 0.00103 M 14 100 5 0.1170 0.02737 * M F 10 8 0.0030 0.00245 F 13 5 0.0028 0.00000 F 1 10 5 0.0169 0.00219 F 1 30 5 0.0492 0.00504 F 1 100 5 0.1724 0.08294 * F 4 0 8 0.0020 0.00097 F 4 3 5 0.0028 0.00000 F 4 10 5 0.0186 0.00170 F 4 30 5 0.0439 0.01322 F 4 100 5 0.1984 0.07948 * F 14 0 8 0.0079 0.01997 F 14 3 5 0.0012 0.00084 F 14 10 5 0.0009 0.00000 F 14 30 5 0.0320 0.00238 F 14 100 5 0.0145 0.00145 * 1. One-half the limit of quantitation (LOQ/2 = 0.00275 mg/mL) or one half the limit of detection (LOD/2 = 0.00089 mg/mL) were substituted for all values that were below those limits. * Significantly different in comparison to control by Dunnetfs T-test ( p<0.05). Significantly different in comparison to control by Students T-test ( p<0.05). 18 Table 4. Summary of WinNonlin Noncompartmental Analysis of Mean Serum [PFOS] PFOS_ Dose (ug/kg) 0 3 Gender Tmax (day) F M F 28 28 1 M1 10 F 4 M 14 30 F 1 M' 4 100 F 4 M 14 5000 F 1 M1 NA: Not Applicable Cmax (ug/mL) 0.06 0.01 0.00 0.00 0.02 0.01 0.05 0.00 0.20 0.12 19.60 13.37 HL_Lamb da_z (day) NA NA 4r6-r-6(NA ) T^-(NA) NA NA 3i(NA) NA NA NA 16 14 Tlast (day) 28 28 14 14 14 14 14 14 14 14 28 28 Clast (ug/mL) 0.06 0.01 0.00 0.00 0.00 0.01 0.03 0.00 0.01 0.12 6.99 3.47 AUClast (day*ug/mL) 0.5 0.1 0.0 0.0 0.2 0.1 0.5 0.0 1.7 0.8 347.1 251.4 19 Table 5. Summary Liver [PFOS] (pg/g) and Percent (%) Dose in Liver. Day Post-Dose (day) 1 4 | 14 28 Dose Gende L[PFOS %dose LfPFOS %dos L[PFOS %dos L[PFOS %dos (ug/Kg r ] ] e] e] e ) 0 F N2 00 00 00 0 rA O o Mean 1.04 NA <LOQ NA <LOQ NA NA SD 1.119 NA <LOQ NA <LOQ NA <LOQ NA M N2 00 00 01 0 Mean 0.04 NA <LOQ NA <LOQ NA 0.03 NA SD 0.017 NA <LOQ NA <LOQ NA NA NA 3 F N0 00 05 5 NA NA Mean <LOQ NA <LOQ NA 0.02 37 NA NA SD <LOQ NA <LOQ NA 0.003 6.3 NA NA M N2 20 05 5 NA NA Mean 0.18 277 <LOQ NA 0.03 62 NA- NA SD 0.059 86.8 <LOQ NA 0.011 18.3 NA NA 10 F N 0 05 55 5 NA NA Mean <LOQ NA 0.15 62 0.05 22 NA NA SD <LOQ NA 0.003 3.0 0.007 2.2 NA NA M N5 55 55 5 NA NA Mean 0.16 78 0.18 86 0.08 46 NA NA SD 0.030 18.7 0.018 3.5 0.018 10.9 NA NA 30 F N 5 55 55 5 NA NA Mean 0.26 36 0.26 35 0.16 23 NA NA SD 0.033 3.7 0.054 6.7 0.006 1.2 NA NA M N5 55 55 5 NA NA Mean 0.41 61 0.45 68 0.33 63 NA NA SD 0.055 8.6 0.108 15.6 0.023 4.6 NA NA 100 F N5 55 55 5 NA NA Mean 0.86 35 0.90 38 0.53 25 NA NA SD 0.132 5.0 0.315 14.8 0.075 5.5 NA NA M N5 55 55 5 NA NA Mean 1.39 65 1.35 65 0.87 50 NA NA SD 0.216 8.0 0.107 2.4 0.128 5.7 NA NA 5000 F N5 55 55 55 5 Mean 37.66 31 40.25 34 40.14 34 20.96 18 SD 4.533 3.2 16.151 14.2 10.744 7.6 7.657 6.5 M N5 55 55 55 5 Mean 33.58 29 27.50 25 42.92 44 30.96 33 SD 5.104 5.5 5.767 4.6 17.467 20.3 2.700 1.7 LOQ = 10 ng/g for the control (group 1), 3, 10, 30 and 100 pg/Kg dose-groups on day 14 Post-dose ( 3M Environmental Lab). LOQ = 134 ng/g for the 0 (group 1), 3, 10, 30 and 100 pg/Kg dose-groups on days 1 and 4 post-dose (Battelle Lab). LOQ = 5.88 ng/g on days 1 and 4 post-dose and 37 ng/g on days 14 and 28 post-dose for the 0 (group 6) and 5000 pg/Kg dose groups (3M Environmental Lab). NA: Not Applicable_____________ _______________________________________ 20 Table 6. Summary of WinNonlin Noncompartmental Analysis of Mean Liver [PFOS] Dose Gender (ug/Kg) 0F M 3F M 10 F M 30 F M 100 F M 5000 F M NA: Not Applicable Mean Mean Mean Mean Mean Mean Mean Mean Mean Mean Mean Mean Tmax (day) 1 1 1 1 4 4 4 4 4 1 14 14 Cmax (ug/g) 0.29 0.05 0.07 0.11 0.15 0.18 0.26 0.45 0.90 1.39 40.14 42.92 HL_Lambda_z (day) 8 (NA) 4r8-(NA) -8-(NA) 8 (NA) NA NA NA NA NA 18 24 NA Tlast (day) 28 28 14 14 14 14 14 14 14 14 28 28 Clast (ug/g) 0.02 0.02 0.02 0.03 0.05 0.08 0.16 0.33 0.53 0.87 20.96 30.96 AUClast (day*ug/g) 1.10 0.65 0.68 0.83 1.34 1.91 3.01 5.37 10.24 15.90 913.68 977.67 21 Table 7. Summary Liver:Serum PFOS Concentration Ratio (ng/g:|ig/g). Variable = [liver]/ [sera] PFOS ratio (ppm/ppm) Dose (ug/Kg) 0 3 10 30 100 5000 Gender F M F M F M F M F M F M Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD Day_Post-Dose 1 NA NA NA NA NA NA NA NA NA NA NA NA 5 0.5 NA NA 9 11.4 52 15.0 2 1.3 3 0.8 (day) 4 NA NA NA NA NA NA NA NA 8 0.9 NA NA 6 1.1 NA NA 5 0.5 53 10.9 2 0.3 2 0.4 14 NA NA NA NA NA NA NA NA NA NA 10 3.5 5 0.4 NA NA 37 6.5 8 1.9 4 0.5 5 2.5 28 NA NA 4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 3 1.0 9 2.4 22 Figures 23 T-6295.15 FR DT31 Figure 1. Average Serum PFOS Vs Time. Female rats. All dose groups. Serum PFOS Vs time, low Dose PFOS. Gender=F -e- o ------ 0 Error -B - 3 ------ 3 Error -O- 10 ------ 10 Error - A - 30 ------ 30 Error - V 100 ------ 100 Error 5000 ------ 5000 Error User Chart (29-Mar-2004) Figure 2. Average Serum PFOS Vs Time. Male rats. All dose groups. Serum PFOS Vs time, low Dose PFOS. Gender=M User Chart <29-Mar-2004) -e- 0 -- 0 Error -a- 3 -- 3 Error 10 -- 10 Error --Ar~ 30 ------ 30 Error 100 ------ 100 Error 5000 ------ 5000 Error Serum [PFOS] (ug/mL) Figure 3. Average Serum PFOS Vs Time. Female rats. 0 to 100 ug/Kg dose groups. Average Serum [ PFOS] Vs Time, 0 to 100 ug/Kg Dose. Gender=F 0 -- 0 Error --F~?-- 10 -- 10 Error 30 -- 30 Error - A ~ 100 -- 100 Error Figure 4. Average Serum PFOS Vs Time. Male rats. 0 to 100 ug/Kg dose groups 0.3 r Average Serum [ PFOS] Vs Time, 0 to 100 ug/Kg Dose. Gender=M 0 -- 0 Error - B - 10 -- 10 Error 100 -- 100 Error Serum [PFOS] (ug/mL) 25 Figure 5. Liver PFOS Vs Time. Male and Female Rats. Control dose group. Liver PFOS Vs Time. Dose=0 - - Gender=F ----- Gender=F Error ~B~ Gender=M ------ Gender=M Error User Chart (27-Feb-2004) Figure 6. Liver PFOS Vs Time. Male and Female Rats. 3 ug/Kg dose group. Liver PFOS Vs Time. Dose=3 User Chart (27-Feb-2004) Q Gender=F ----- Gender=F Error -B ~ Gender=M ----- Gender=M Error 26 Figure 7. Liver PFOS Vs Time. Male and Female Rats. 10 ug/Kg dose group. Liver PFOS Vs Time. Dose=10 - 0 - Gender=F ------ Gender=F Error - 0 - Gender=M ------ Gender=M Error User Chart (27-Feb-2004) Figure 8. Liver PFOS Vs Time. Male and Female Rats. 30 ug/Kg dose group. Liver PFOS Vs Time. Dose=30 User Chart (27-Feb-2004) -0 - Gender=F ------ Gender=F Error *-0" Gender=M ------ Gender=M Error 27 Figure 9. Liver PFOS Vs Time. Male and Female Rats. 100 ug/Kg dose group. Liver PFOS Vs Time. Dose=100 o 2.0 - Q Gender=F ------ Gender=F Error - 0 - Gender=M ------ Gender=M Error User Chart (27-Feb-2004) Figure 10. Liver PFOS Vs Time. Male and Female Rats. 5000 ug/Kg dose group. Liver PFOS Vs Time. 70 - Dose=5000 - 0 " Gender=F ------ Gender=F Error - 0 - Gender=M ------ Gender=M Error User Chart (28-Mar-2004) 28 Appendix 1: Biological Parameters Individual and Summary Data Tables Dose (uglKg) 0 Gender F Day PostDose (day) 1 Animal# (8RO -xxxx) 1231 1233 232 3825 3826 3827 3828 3829 N NObs Mean SD Range Initial Body Weight (g) 204 209 207 176 181 176 184 187 8 8 190 13.96 33 Terminal BW (9) 203 201 210 180 184 179 186 187 8 8 191 11.58 31 Liver Weight (g) 8.00 8.10 8.70 7.70 7.40 7.50 8.10 8.30 8 8 7.975 0.430 1.30 4 1234 1235 1236 3830 3831 3832 3833 . 3834 N NObs Mean SD Range 198 213 211 185 188 200 184 194 8 8 197 11.13 29 202 220 210 197 200 220 196 207 8 8 206 9.47 24 8.00 8.10 8.80 7.50 7.70 9.10 7.60 8.50 8 8 8.163 0.585 1.60 14 **1492 **1493 1257 3835 3836 3837 3838 3839 N NObs Mean SD Range Missing Missing 205 175 176 175 185 191 6 8 184 12.04 30 194 201 230 218 215 196 216 223 8 8 212 13.08 36 7.70 8.20 8.70 8.60 8.20 7.20 8.30 9.10 8 8 8.250 0.593 1.90 28 1494 1495 1496 N NObs Mean SD 223 195 196 3 3 205 16.00 267 226 223 3 3 239 24.58 11.40 8.80 8.90 3 3 9.700 1.473 29 Range 28 44 2.60 Dose (ug/Kg) Gender M Day_PostDose {day) 1 Animal#_(8RO -xxxx) 1205 1206 1207 3778 3779 3780 3781 3782 N NObs Mean SD Range lnitial_Body_ Weight (g) 289 283 292 216 224 223 236 228 8 8 249 33.07 76 Terminal BW (9) 293 285 294 219 227 229 239 233 8 8 252 32.31 75 Liver Weight (9) 13.10 12.20 13.00 9.80 10.30 10.80 11.30 11.10 8 8 11.450 . 1.213 3.30 4 1208 1209 1210 3783 3784 3785 3786 3787 N NObs Mean SD Range 287 283 291 233 225 231 224 223 8 8 249 31.16 69 303 295 310 259 248 255 246 250 8 8 271 27.03 64 12.20 11.40 12.80 12.30 10.60 10.90 9.90 11.70 8 8 11.475 0.971 2.90 14 " I4 8 6 "1487 "1488 3788 3789 3790 3791 3792 N NObs Mean SD Range Missing Musing 224 229 225 239 227 5 8 229 5.81 14 270 268 283 301 328 300 306 301 8 8 294 19.99 60 11.70 11.80 12.60 12.80 16.20 13.20 13.20 13.10 8 8 13.075 1.395 4.50 28 1489 1490 1491 N NObs Mean SD Range 282 270 264 3 3 272 9.18 18 369 347 336 3 3 351 17.11 34 15.30 14.40 13.90 3 3 14.533 0.709 1.40 30 Dose (ug/Kg) 3 Gender F Day_PostDose (day) 1 Animal#_(8RO -xxxx) 3698 3699 3700 3701 3702 N NObs Mean SD Range lnitial_Body_ Weight ( g f 176 189 184 193 178 5 5 184 7.02 17 Terminal_BW (9) 179 184 184 188 174 5 5 182 5.19 13 Liver Weight (a) 7.30 6.80 7.80 7.40 7.10 5 5 7.280 0.370 1.00 4 3703 3704 3705 3706 3707 N NObs Mean SD Range 203 193 184 181 192 5 5 191 8.47 22 209 202 199 200 200 5 5 202 3.94 10 9.00 8.50 8.00 8.20 8.40 5 5 8.420 0.377 1.00 14 3708 3709 3710 3711 3712 N NObs Mean SD Range 190 173 177 173 186 5 5 180 7.89 17 225 200 213 220 218 5 5 215 9.33 25 8.80 8.10 8.70 9.20 9.00 5 5 8.760 0.416 1.10 31 Dose (ug/Kg) Gender M Day_PostDose (day) 1 A nim al# J8R O -xxxx) 3668 3669 3670 3671 3672 N NObs Mean SD Range lnitial_Body_ Weight (g) 240 240 223 231 241 5 5 235 7.96 18 Terminal BW -(g) 243 245 226 235 240 5 5 238 7.71 19 Liver Weight (g) 11.20 11.50 10.20 11.30 10.40 5 5 10.920 0.581 1.30 4 3673 3674 3675 3676 3677 N NObs Mean SD Range 228 224 229 222 228 5 5 226 3.22 7 254 243 250 245 252 5 5 249 4.60 11 11.60 10.30 11.30 9.50 10.70 5 5 10.680 0.832 2.10 14 3678 ' 3679 3680 3681 3682 N NObs Mean SD Range 234 240 220 233 223 5 5 230 8.09 19 294 333 278 286 285 5 5 295 21.82 55 12.10 16.30 11.30 11.70 12.20 5 5 12.720 2.033 5.00 32 Dose (ug/Kg) 10 Gender F Day_PostDose (day) 1 Animal#_(8RO -xxxx) 2855 3856 3857 3858 3859 N NObs Mean SD Range lnitial_Body_ Weight (g) 176 188 169 180 175 5 5 178 6.89 19 Terminal_BW Liver_Weight ~(g> 183 193 169 _______ (9 ,),........ 7.60 7.50 6.80 184 7.60 174 7.20 55 55 181 7.340 9.10 0.344 23 0.80 4 3860 3861 3862 3863 3864 N NObs Mean SD Range 185 175 192 175 181 '5 5 182 7.03 17 195 184 198 186 194 5 5 191 5.98 14 7.70 7.40 7.70 7.40 8.00 5 5 7.640 0.251 0.60 14 3865 3866 3867 3868 3869 N NObs Mean SD Range 171 184 170 182 171 5 5 175 6.74 14 210 219 202 207 198 5 5 207 8.04 21 8.40 8.00 7.50 7.20 8.10 5 5 7.840 0.483 1.20 33 Dose (ug/Kg) Gender M Day_PostDose (day) 1 Animal#_(8RO -xxxx) 3808 3809 3810 3811 3812 N NObs Mean SD Range lnitial_Body_ Weight (g) 229 203 230 231 229 5 5 224 12.05 28 Terminal_BW -(g) 238 211 235 234 232 5 5 230 10.46 26 Liver_Weight (9) 11.00 10.50 10.50 10.70 10.40 5 5 10.620 0.239 0.60 4 3813 3814 3815 3816 3817 N NObs Mean SD Range 227 258 232 217 216 5 5 230 16.85 42 252 249 263 237 243 5 5 249 9.62 26 11.00 10.30 12.20 9.90 11.30 5 5 10.940 0.896 2.30 14 3818 3819 3820 3821 3822 N NObs Mean SD Range 232 220 247 232 227 5 5 232 9.88 27 308 297 308 317 306 5 5 307 7.09 20 12.70 13.20 13.60 13.90 13.10 5 5 13.300 0.464 1.20 34 Dose (ug/Kg) 30 Gender F Day_PostDose (day) 1 Animal#_(8RO -xxxx) 3713 3714 3715 3716 3717 N NObs Mean SD Range lnitial_Body_ Weight (g) 198 190 174 201 187 5 5 190 10.63 27 Terminal BW (9) 204 187 177 193 187 5 5 190 9.85 27 Uver_Weght (9) 9.80 7.90 7.10 7.50 7.00 5 5 7.860 1.141 2.80 4 3718 3719 3720 3721 3722 N NObs Mean SD Range 177 186 177 180 195 5 5 183 7.64 18 183 202 183 189 198 5 5 191 8.87 19 6.90 8.00 6.90 7.40 8.00 5 5 7.440 0.550 1.10 14 3723 3724 3725 3726 3727 N NObs Mean SD Range 193 196 178 173 181 5 5 184 9.86 23 224 226 212 204 200 5 5 213 11.47 26 8.70 8.40 7.90 7.40 7.10 5 5 7.900 0.667 1.60 35 Dose (ug/Kg) Gender M Day_PostDose (day) 1 Animal#_(8RO -xxxx) *3685 3683 3684 3686 3687 N NObs Mean SD Range lnitial_Body_ Weight (g) 236 232 219 233 231 5 5 230 6.67 17 Terminal BW (g) 241 233 222 235 234 5 5 233 6.81 19 Liver_Weight (g) 10.40 9.70 10.30 10.50 10.60 5 5 10.300 0.354 0.90 4 3688 3689 3690 3691 3692 N NObs Mean SD Range 232 232 237 225 233 5 5 232 4.53 13 253 259 259 248 253 5 5 254 4.46 11 10.40 11.10 10.80 10.40 10.50 5 5 10.640 0.305 0.70 14 3693 3694 3695 3696 3697 N NObs Mean SD Range 232 250 228 221 232 5 5 233 10.68 29 297 327 294 285 301 5 5 301 15.90 43 12.90 14.60 12.60 12.40 13.60 5 5 13.220 0.896 2.20 36 Dose (ug/Kg) 100 Gender F Day_PostDose (day) 1 Animal#_(8RO -xxxx) 3840 3841 3842 3843 3844 N NObs Mean SD Range lnilial_Body_ Weight (g) 193 186 180 190 179 5 5 186 5.88 14 Terminal_BW _(g> 188 189 179 190 178 5 5 185 5.63 12 Liver Weight (g) 7.20 8.20 7.30 7.50 7.20 5 5 7.480 0.421 1.00 4 3845 3846 3847 3848 3849 N NObs Mean SD Range 192 182 184 185 180 5 5 185 4.28 11 204 190 201 197 187 5 5 196 7.20 17 8.00 7.10 8.30 8.20 7.50 5 5 7.820 0.507 1.20 14 3850 3851 3852 3853 3854 N NObs Mean SD Range 179 205 191 179 193 5 5 189 10.80 26 204 249 226 224 216 5 5 224 16.29 44 7.80 10.60 9.60 8.60 8.20 5 5 8.960 1.135 2.80 37 Dose (ug/Kg) Gender M Day_PostDose (day) 1 Animal#_(8RO -xxxx) 3793 3794 3795 3796 3797 N NObs Mean SD Range lnitial_Body_ Weight (g) 225 225 228 232 236 5 5 229 4.84 12 Termina!_BW (9) 233 227 233 236 242 5 5 234 5.66 16 Liver_Weight (9) 11.30 10.50 10.30 10.80 11.10 5 5 10.800 0.412 1.00 4 3798 3799 3800 3801 3802 N NObs Mean SD Range 231 234 224 214 238 5 5 228 9.33 24 261 258 255 245 262 5 5 256 6.54 16 11.80 10.10 11.60 10.20 11.10 5 5 10.960 0.783 1.70 14 3803 3804 3805 3806 3807 N NObs Mean SD Range 227 229 231 226 225 5 5 228 2.51 6 291 291 305 302 305 5 5 299 7.23 14 13.10 12.40 13.00 13.60 13.80 5 5 13.180 0.550 1.40 38 Dose (ug/Kg) 5000 Gender F Day_PostDose (day) 1 Animal#_(8RO -xxxx) 1237 1238 1239 1240 1241 N NObs Mean SD Range lnitial_Body_ Weight (g) 191 214 205 205 195 5 5 202 9.11 23 Terminal_BW Liver Weight - (g )_______ 186 212 0 ) ______ 7.30 8.50 201 9.20 199 7.80 196 8.50 55 55 199 8.260 9.32 0.730 26 1.90 4 1242 1243 1244 1245* 1246* N NObs Mean SD Range 211 220 205 207 207 5 5 210 6.00 15 222 232 204 213 215 5 5 217 10.43 28 9.00 9.60 8.00 8.90 8.10 5 5 8.720 0.669 1.60 14 1247 1248 1249 1250 1251 N NObs Mean SD Range 197 222 195 204 209 5 5 205 10.83 27 226 253 223 224 226 5 5 230 12.63 30 8.50 9.50 8.40 9.00 8.30 5 5 8.740 0.503 1.20 28 1252 1253 1254 1255 1256 N NObs Mean SD Range 197 206 213 207 206 5 5 206 5.72 16 259 234 248 225 239 5 5 241 13.02 34 9.20 8.20 9.30 8.10 8.50 5 5 8.660 0.559 1.20 39 Dose (ug/Kg) Gender M Day_PostDose (day) 1 Animal#_(8RO -xxxx) 1211 1212 1213 1214 1215 N NObs Mean SD Range Initial Body Weight (g) 290 292 292 285 279 5 5 288 5.37 13 Terminal BW _(g) 291 293 296 289 278 5 5 290 6.78 18 Liver Weight (9) 12.30 13.50 12.00 12.40 11.60 5 5 12.360 0.709 1.90 4 1216 1217 1218 1219 1220 N NObs Mean SD Range 291 275 271 292 289 5 5 284 9.84 21 309 290 284 313 305 5 5 300 12.76 29 12.70 12.50 11.90 13.90 12.80 5 5 12.760 0.727 2.00 14 1221 1222 1223 1224 1225 N NObs Mean SD Range 288 288 287 298 275 5 5 287 8.17 23 348 363 347 370 332 5 5 352 15.01 38 13.30 16.40 14.20 15.70 13.80 5 5 14.680 1.314 3.10 28 1226 1227 1228 1229 1230 N NObs Mean SD Range 281 290 275 300 287 5 5 287 9.45 25 379 424 373 417 399 5 5 398 22.35 51 14.30 16.70 14.30 16.00 14.90 5 5 15.240 1.071 2.40 40 T-6295.15 FR DT31 Appendix 2: Individual Sera PFOS and % of dose in sera. Dose (ug/Kg) 0 Day_ PostDose (day) 1 4 14 Gende r Animal#_(8R0- Sera [PFO Sera_PFOS %dose in xxxx) S] (ug) sera (ug/mL) F 1231 1233 1232 3825 3826 3827 3828 3829 N NObs Mean SD 0.01 Missing Missing Missing Missing Missing Missing Missing 1 8 0.009 Missing 0.06 Missing Missing Missing Missing Missing Missing Missing 1 8 0.057 Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing M 1205 1206 1207 3778 3779 3780 3781 3782 N NObs Mean SD Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing F 1234 1235 1236 3830 3831 3832 3833 3834 N NObs Mean SD Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing M 1208 1209 1210 3783 3784 3785 3786 3787 N NObs Mean SD F **1492 **1493 Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Page 3 of 3 Dose (ug/Kg) 3 1257 3835 3836 3837 3838 3839 N NObs Mean SD Missing Missing Missing 0.06 Missing Missing 1 8 0.057 Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing M 1486 1487 1488 3788 3789 3790 3791 3792 N NObs Mean SD 0.01 Missing Missing Missing Missing Missing Missing Missing 1 8 0.008 Missing 0.07 Missing Missing Missing Missing Missing Missing Missing 1 8 0.072 Missing Missing Missing Missing Missing Missing Missing Missing Missing 0 8 Missing Missing 28 F 1494 1495 1496 N NObs Mean SD 0.14 0.02 0.01 3 3 0.055 0.070 1.17 0.11 0.10 3 3 0.462 0.613 Missing Missing Missing 0 3 Missing Missing M Day Po stDose (day) 1 Gende r F 1489 1490 1491 N NObs Mean SD Animal# xxxx) (8RO- 0.01 0.01 0.02 3 3 0.010 0.006 Sera [PFO S] (ug/mL) 0.08 0.08 0.19 3 3 0.115 0.063 Sera PFOS (ug> Missing Missing Missing 0 3 Missing Missing %dose in sera 3698 3699 3700 3701 3702 N NObs Mean SD Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing M 3668 3669 3670 3671 3672 N NObs Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing 0 5 42 Dose (ug/Kg) 10 Mean SD Missing Missing Missing Missing Missing Missing 4 F 3703 3704 3705 3706 3707 N NObs Mean SD Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing M 3673 3674 3675 3676 3677 N NObs Mean SD Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing 14 F M Day Po stDose (day) 1 Gende r F 3708 3709 3710 3711 3712 N NObs Mean SD 3678 3679 3680 3681 3682 N NObs Mean SD Animal# xxxx) (8RO- Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Sera [PFO S] (ug/mL) Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Sera PFOS (ug) Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing %dose in sera 3855 3856 3857 3858 3859 N NObs Mean SD 0.01 0.02 0.02 0.02 0.02 5 5 0.017 0.002 0.09 0.10 0.08 0.11 0.11 5 5 0.098 0.013 4.89 5.45 4.84 6.16 6.26 5 5 5.521 0.676 M 3808 3809 3810 3811 3812 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 43 Dose (ug/Kg) 30 N NObs Mean SD 0 5 Missing Missing 0 5 Missing Missing 0 5 Missing Missing 4 F 3860 3861 3862 3863 3864 N NObs Mean SD 0.02 0.02 0.02 0.02 0.02 5 5 0.019 0.002 0.10 0.12 0.13 0.11 0.11 5 5 0.114 0.010 5.49 6.90 6.64 6.29 6.18 5 5 6.299 0.535 M 3813 3814 3815 3816 3817 N NObs Mean SD Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing 14 F M Day Po St" Dose (day) 1 Gende r F 3865 3866 3867 3868 3869 N NObs Mean SD 3818 3819 3820 3821 3822 N NObs Mean SD Animal# xxxx) (8R0- Missing Missing Missing Missing Missing 0 5 Missing Missing 0.01 0.01 0.01 0.01 Missing 4 5 0.008 0.003 Sera [PFO S] (ug/mL) Missing Missing Missing Missing Missing 0 5 Missing Missing 0.08 0.11 0.06 0.06 Missing 4 5 0.078 0.025 Sera PFOS (ug) Missing Missing Missing Missing Missing 0 5 Missing Missing 3.33 5.12 2.40 2.64 Missing 4 5 3.372 1.229 %dose in sera 3713 3714 3715 3716 3717 N NObs Mean SD 0.04 0.05 0.05 0.06 0.05 5 5 0.049 0.005 0.28 0.30 0.27 0.35 0.30 5 5 0.300 0.030 4.65 5.29 5.23 5.78 5.43 5 5 5.277 0.409 M 3683 3684 3685 Missing Missing Missing Missing Missing Missing Missing Missing Missing 44 Dose (ug/Kg) 100 3686 3687 N NObs Mean SD Missing Missing 0 5 Missing Missing Missing Missing 0 5 Missing Missing Missing Missing 0 5 Missing Missing 4 F 3718 3719 3720 3721 3722 N NObs Mean SD 0.06 0.05 0.04 0.02 0.05 5 5 0.044 0.013 0.34 0.31 0.26 0.13 0.31 5 5 0.270 0.082 6.34 5.61 4 .87 2.46 5.29 5 5 4.913 1.471 M 3688 3689 3690 3691 3692 N NObs Mean SD Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing Missing Missing Missing Missing Missing 0 5 Missing Missing 14 F M Day Po stDose (day) 1 Gende r F 3723 3724 3725 3726 3727 N NObs Mean SD 3693 3694 3695 3696 3697 N NObs Mean SD Animal# xxxx) (8RO- 0.03 0.03 0.03 0.03 0.03 5 5 0.032 0.002 Missing Missing Missing Missing Missing 0 5 Missing Missing Sera [PFO S] (ug/mL) 0.23 0.24 0.24 0.19 0.20 5 5 0.220 0.024 Missing Missing Missing Missing Missing 0 5 Missing Missing Sera PFOS (ug) 3.97 4.16 4.40 3.59 3.75 5 5 3.973 0.321 Missing Missing Missing Missing Missing 0 5 Missing Missing %dose in sera 3840 3841 3842 3843 3844 N NObs Mean SD 0.23 0.21 0.23 0.03 0.17 5 5 0.172 0.083 1.37 1.28 1.30 0.18 0.97 5 5 1.021 0.493 7.08 6.88 7.24 0.97 5.41 5 5 5.515 2.643 M 3793 0.04 0.27 1.18 45 Dose (ug/Kg) 5000 3794 3795 3796 3797 N NObs Mean SD 0.03 0.02 0.03 0.03 5 5 0.028 0.005 0.20 0.17 0.20 0.20 5 5 0.208 0.034 0.90 0.77 0.85 0.85 5 5 0.908 0.158 4 F 3845 3846 3847 3848 3849 N NObs Mean SD 0.21 0.15 0.20 0.32 0.11 5 5 0.198 0.079 1.35 0.94 1.31 2.03 0.65 5 5 1.258 0.520 7.06 5.17 7.16 10.98 3.60 5 5 6.793 2.762 M 3798 3799 3800 3801 3802 N NObs Mean SD 0.02 0.02 0.03 0.03 0.02 5 5 0.026 0.005 0.19 0.18 0.25 0.26 0.20 5 5 0.217 0.037 0.83 0.7.7 1.13 1.22 0.84 55 0.959 0.203 14 F M Day Po stDose (day) 1 Gende r F 3850 3851 3852 3853 3854 N NObs Mean SD 3803 3804 3805 3806 3807 N NObs Mean SD Animal# xxxx) (8RO- 0.01 0.01 0.02 0.01 0.02 5 5 0.015 0.001 0.09 0.16 0.11 0.13 0.10 5 5 0.117 0.027 Sera [PFO S] (ug/mL) 0.08 0.12 0.11 0.10 0.11 5 5 0.105 0.014 0.85 1.49 1.10 1.22 0.95 5 5 1.123 0.250 Sera PFOS (ug) 0.47 0.57 0.60 0.53 0.58 5 5 0.551 0.051 3.74 6.51 4.75 5.42 4.23 5 5 4.929 1.079 %dose in sera 1237 1238 1239 1240 1241 N NObs Mean SD 17.40 19.00 6.89 28.80 22.30 5 5 18.878 8.001 104.32 129.70 44.57 184.54 140.45 5 5 120.719 51.505 10.92 12.12 4.35 18.00 14.41 5 5 11.961 5.039 46 M 4F M 14 F M 28 F 1211 1212 1213 1214 1215 N NObs Mean SD 1242 1243 1244 1245* 1246 N NObs Mean SD 1216 1217 1218 1219 1220 N NObs Mean SD 1247 1248 1249 1250 1251 N NObs Mean SD 1221 1222 1223 1224 1225 N NObs Mean SD 1252 1253 1254 1255 1256 N NObs Mean SD 10.30 12.90 12.00 14.60 17.00 5 5 13.360 2.560 24.90 18.10 20.10 0.28 15.20 5 5 19.601 4.084 12.80 12.90 14.50 11.70 13.10 5 5 13.000 1.000 11.90 9.23 10.50 10.60 13.20 5 5 11.086 1.513 14.20 10.80 7.19 7.02 9.06 5 5 9.654 2.972 4.43 7.13 6.66 8.63 8.12 5 5 6.994 1.632 96.61 121.83 114.26 136.01 152.29 5 5 124.199 21.176 177.59 135.27 132.23 1.89 105.08 5 5 110.413 65.979 127.40 120.34 132.51 117.96 128.78 5 5 125.396 6.062 86.41 75.13 75.50 76.32 96.23 5 5 81.917 9.268 159.03 126.27 80.43 83.68 96.80 5 5 109.241 33.193 36.95 53.70 53.08 62.52 62.39 5 5 53.726 10.421 6.67 8.34 7.83 9.53 10.90 5 5 8 .655 1.622 16.83 12.30 12.90 0.18 10.15 5 5 10.473 6.238 8.76 8.75 9.78 8.08 8.91 5 5 8.856 0.608 8.77 6.77 7.74 7.48 9.21 5 5 7.995 0.988 11.04 8.77 5.60 5.62 7.04 5 5 7.615 2.316 3.75 5.21 4.98 6.04 6.06 5 5 5.209 0.947 47 M 1226 1227 2.51 2.60 30.60 35.49 2.18 2.45 1228 3.06 36.79 2.68 1229 1230 3.97 5.24 53.26 67.24 3.55 4.69 N 555 NObs Mean SD 5 3.476 1.143 5 44.675 15.226 5 3.107 1.021 * Sera PFOS value for animal # 8RO-1245 was considered an outlier. Worksheet: Sheet! 48 Appendix 3: Sera PFOS WinNonlin Model 200 Non Compartmental Analysis (NCA). Figure 11. NCA Avg Sera PFOS vs Time. Female rats. Control dose group. Mean (ug/mL) 0.100 ; Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=0, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting O 0.010 y fO 0.001 -i---- 0 5 ^ - 0 " Observed ------ Predicted 10 15 20 25 30 Day_Post_Dose (day) Figure 12. NCA Avg Sera PFOS vs Time. Male rats. Control dose group. Mean (ug/mL) 0.100 t Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=0, Gender=M Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting 0.010 t !o o 0.001 -i-------------- 05 o 10 15 20 Day_Post_Dose (day) o - 0 - Observed ------ Predicted 25 30 49 Mean (ug/mL) Figure 13. NCA Avg Sera PFOS vs Time. Female rats. 3 ng/Kg dose group. 0.010 - Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=3, Gender=F Rsq=0.9514 Rsq_adjusted=0.9029 HL_Lambda_z= 10.5828 (day) (3 points used in calculation) Uniform Weighting - - Observed ------ Predicted 0.001 ----------- ----------- ----------- '---------- ----------- `---------- :---------- 0 2 4 6 8 10 12 14 Day_Post_Dose (day) Figure 14. NCA Avg Sera PFOS vs Time. Male rats. 3 (ig/Kg dose group. Average Sera [PFOS] Vs time. Low Dose PFOS 0.0100 t PFOS_Dose=3, Gender=M Rsq=0.9514 Rsq_adjusted=0.9029 HL_Lambda_z=7.2798 (day) (3 points used in calculation) Uniform Weighting 0.0010 r - 0 - Observed ------ Predicted 0.0001 0 2 4 6 8 10 12 14 Day_Post_Dose (day) Mean (ug/mL) 50 Mean (ug/mL) Figure 15. NCA Avg Sera PFOS vs Time. Female rats. 10 pg/Kg dose group. Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=10, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting 0.1000 t 0.0100 y 0.0010 I 0.0001 ----------- - 02 O - 0 - Observed ------ Predicted 4 6 8 10 12 14 Day_Post_Dose (day) 16. NCA Avg Sera PFOS vs Time. Male rats. 10 pg/Kg dose group. 0.010 Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=10, Gender=M Rsq=Undefined Rsq_adjusted=UndeFmed HL_Lambda_z=Undefmed (day) (0 points used in calculation) Uniform Weighting o oo - 0 - Observed ------ Predicted 0.001 -- .................... ............... ---------- `---------- r---------- ----------- , 0 2 4 6 8 10 12 14 Day_Post_Dose (day) = Mean (ug/mL) 51 Mean (ug/mL) Figure 17. NCA Avg Sera PFOS vs Time. Female rats. 30 (jg/Kg dose group. Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=30, Gender=F Rsq=0.9985 Rsq_adjusted=0.9971 HL_Lambda_z=21.1307 (day) (3 points used in calculation) Uniform Weighting 0.10 r G0 - 0 - Observed ------ Predicted 0.01 ---------------------- '---------- >-----------'-----------1-----------1-----------1 0 2 4 6 8 10 12 14 Day_Post_Dose (day) Figure 18. NCA Avg Sera PFOS vs Time. Male rats. 30 fig/Kg dose group. Average Sera [PFOS] Vs time. Low Dose PFOS 0.010 t PFOS_Dose=30, Gender=M Rsq=Undefined Rsq_adjusted=Undeflned HL_Lambda_z=Undefined (day) (0 points used In calculation) Uniform Weighting OO - - Observed ------ Predicted 0.001 :----------------------------- --------------- -------------- i-------------- 1------------- 7-------------- : 0 2 4 6 8 10 12 14 Day_Post_Dose (day) Mean (ug/mL) 52 Figure 19. NCA Avg Sera PFOS vs Time. Female rats. 100 ng/Kg dose group. Average Sera [PFOSJ Vs time. Low Dose PFOS PFOS_Dose=100, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_lambda_z=Undefined (day) (0 points used in caiculation) Uniform Weighting 1.00 7 Eo -2- 0.10 7 o Mean 0.01 1----------'---------- '---------- - 024 6 _______ ,_______ 8 10 12 Day_Post_Dose (day) - - Observed ------ Predicted o 14 Figure 20. NCA Avg Sera PFOS vs Time. Male rats. 100 jzg/Kg dose group. 1.00 7 Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=100, Gender=M Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting Mean (ug/mL) 0.10 I 0.01 ------------- 02 O 4 6 8 10 Day_Post_Dose (day) 12 Observed ------ Predicted 14 53 Mean (ug/mL) Figure 21. NCA Avg Sera PFOS vs Time. Female rats. 5000 pg/Kg dose group. Average Sera [PFOS] Vs time. Low Dose PFOS 100 r PFOS_Dose=5000, Gender=F Rsq=0.9755 Rsq_adjusted=0.951 HLJ.ambda_z=16.3858 (day) (3 points used in calculation) Uniform Weighting " - - Observed ------ Predicted 1 j-------------.-------------1------- :---- '-------------i-------------'-------------1 0 5 10 15 20 25 30 Day_Post_Dose (day) Figure 22. NCA Avg Sera PFOS vs Time. Male rats. 5000 pg/Kg dose group. 100 T Average Sera [PFOS] Vs time. Low Dose PFOS PFOS_Dose=5000, Gender=M Rsq=0.9414 Rsq_adjusted=0.912 HL_Lambda_z= 13.7635 (day) (4 points used in calculation) Uniform Weighting - 0 - Observed ------ Predicted 1 ------------- ----------------------------------------'-------------.------------ 1 0 5 10 15 20 25 30 Day_Post_Dose (day) Mean (ug/mL) 54 Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=0 Gender=F Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301055 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 0.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best, fit for lambda z Summary Table Time day Cone. ug/mL 0.0000 @ 1.000 4.000 14.00 28.00 0.0000 0.003026 0.002045 0.007930 0.05513 Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.0000 0.001513 0.009119 0.05899 0.5004 0.0000 0.001513 0.01832 0.6143 12.20 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda z Lambda_z Lambda_z_lower Lambda_z upper HL Lambda z Hag Tmax Cmax Tlast Clast AUClast AUCall 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 28.0000 0.0551 28.0000 0.0551 0.5004 0.5004 55 AUCINF_obs AUCINF_D_obs AUC_%Extrap_obs Vz_F_obs C1_F obs AUCINF_pred AUCINF_D_pred AUC_%Extrap_pred Vz_F_pred Cl_F_pred AUMClast AUMCINF_obs AUMC_%Extrap_obs AUMCINF_pred AUMC_%Extrap_pred MRTlast MRTINF_obs MRTINFjored day*ug/mL day *kg *ug M L /ug % mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/raL % day*day*ug/mL % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 12.1963 Missing Missing Missing Missing 24.3735 Missing Missing Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=0 Gender=M Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 0.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day Cone. ug/mL 0.0000 @ 1.000 4.000 14.00 28.00 0.0000 0.002045 0.002045 0.002030 0.01030 Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.0000 0.001023 0.007158 0.02753 0.1138 0.0000 0.001023 0.01636 0.1994 2.417 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast 1/day day day day day day ug/mL day ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 28.0000 0.0103 28.0000 0.0103 0.1138 57 AUCall AUCINF_obs AUCINF_D_obs AUC_%Extrap_obs Vz_F_obs Cl_F_obs AUCINF_pred AUCINF_D_pred AUC_%Extrap_pred Vz_F_pred Cl_F_pred AUMClast AUMCINF_obs AUMC_%Extrap_obs AUHCINF_pred AUMC_%Extrap_pred MRTlast MRTINF_obs MRTINF_pred day*ug/mL day*ug/mL day*kg*ug/mL/ug mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL day day day 0.1138 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 2.4171 Missing Missing Missing Missing 21.2320 Missing Missing Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=3 Gender=F Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 3.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 3 1.000 * 4.000 * 14.00 * Cone. ug/mL 0.0000 0.002750 0.002750 0.001246 Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.002995 - .0002453 0.002461 .0002891 0.001278 - .235e-005 0.0000 0.001375 0.009625 0.02961 0.0000 0.001375 0.02200 0.1642 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC_%Extrap obs l/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL day*ug/mL day*kg*ug/mL/ug % 0.9514 0.9029 -0.9754 3 0.0655 1.0000 14.0000 10.5828 0.0000 1.0000 0.0028 14.0000 0.0012 0.0296 0.0296 0.0486 0.0162 39.1203 59 Vz F obs CI F obs AUCINF pred AUCINF_D_pred ' AUC %Extrap_pred Vz F pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap_obs AUMCINF_pred AUMC %Extrap_pred MRTlast METINF obs MRTINF_pred mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day 941.9015 61.6920 0.0491 0.0164 39.7324 932.4319 61.0717 0.1642 0.7210 77.2234 0.7355 77.6710 5.5470 14.8267 14.9719 Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwoj PFOS_Dose=3 Gender=M Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 3.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day Cone. ug/mL Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.0000 0 1.000 * 4.000 * 14.00 * . 0.0000 0.002750 0.002750 0.0008700 0.003114 -0.0003636 0.002340 0.0004100 0.0009030 -3.303e-005 0.0000 0.001375 0.009625 0.02773 0.0000 0.001375 0.02200 0.1379 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall ACTCINF obs AUCINF D obs AUC_%Extrap__obs l/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL day*ug/mL day*kg*ug/mL/ug % 0.9514 0.9029 -0.9754 3 0.0952 1.0000 14.0000 7.2798 0.0000 1.0000 0.0028 14.0000 0.0009 0.0277 0.0277 0.0369 0.0123 24.7874 61 Vz F obs Cl_F_obs AUCINF pred AUCINF_D_pred AUC %Extrap_pred Vz F pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap_pred MRTIast MRTINF_obs MRTINFjored mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day 854.7389 81.3842 0.0372 0.0124 25.4885 846.7712 80.6256 0.1379 0.3618 61.8833 0.3703 62.7582 4.9739 9.8145 9.9514 Input File: Descriptive Statistics Workbook - (Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=10 Genaer=F Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 10.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day Cone, ug/mL 0.0000 @ 1.000 4.000 14.00 0.0000 0.01685 0.01857 0.0008700 Pred. ug/mL Residual AUC AUMC ug/mL day*ug/mL day*day*' 0.0000 0 .008424 0.06156 0.1588 0.0000 0.008424 0.1451 0.5775 Weight @) Note - the concentration at dose time was added for extrapolation *) Starred values were included in the estimation o f Lambda z . *** Warning 14530: Lambda z could not be estimated No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda_z_lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast 1/day day day day day day ug/mL day ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.0186 14.0000 0.0009 0.1588 63 AUCall AUCINF obs AUCINF_D_obs AUC_%Extrap_obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap_pred Vz F_pred Cl F pred AUMClast AUMCINF_obs A(JMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred day*ug/mL day*ug/mL day*kg*ug/mL/ug % mL/kg niL/day/kg day*ug/mL day*kg*ug/mL/ug % raL/kg raL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day 0.1588 Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0.5775 Missing Missing Missing Missing 3.6373 Missing Missing Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=10 Gender=M Date: Time: 3/29/2004 11:04:05 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2Q03 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 10.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 0 1.000 4.000 14.00 Cone. ug/mL 0.0000 0.002750 0.002750 0.006872 Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.0000 0.001375 0.009625 0.05774 0.0000 0.001375 0.02200 0.5580 0) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 14.0000 0.0069 14.0000 0.0069 0.0577 0.0577 65 AUCINF cbs AUCINF_D_obs AUC %Extrap_obs Vz F obs Cl F obs AUCINF_pred AUCINF_D_prea AUC %Extrap_pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap_obs AUMCINF_pred AUMC %Extrap_pred MRTlast MRTINF obs MRTINF pred day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*ug/'mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing - Missing 0.5580 Missing Missing Missing Missing 9.6655 Missing Missing Input File: Descriptive Statistics Workbook - (Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=30 Gender=F Date: Time: 3/29/2004 11:04:06 Average Sera (PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 30.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 * 4.000 * 14.00 * Cone. ug/mL 0.0000 0.04922 0.04387 0.03197 Pred. ug/mL Residual ug/mL 0.04886 0.0003563 0.04428 -0.0004163 0.03190 6.961e-005 AUC AUMC Weight day*ug/mL day*day*ug/mL 0.0000 0.02461 0.1642 0.5434 0.0000 0.02461 0.3616 3.477 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Hast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC_%Extrap_obs 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL day*ug/mL day*kg*ug/mL/ug % 0.9985 0.9971 -0.9993 3 0.0328 1.0000 14.0000 21.1307 0.0000 1.0000 0.0492 14.0000 0.0320 0.5434 0.5434 1.5180 0.0506 64.2014 67 Vz F obs C1 F obs AUCINF pred AUCINF_D_pred AUC_%Extrap_pxed Vz F pred C1 F pred AUMClast AUMCINF_obs ACJMC Extrap obs AUMCINF_pred AUMC_%Ext rap_p red MRTlast MRTINF_obs MRTINF pred mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day'*'day*ug/niL % day day day 602.4910 19.7635 1.5158 0.0505 64.1513 603.3344 19.7911 3.4767 46.8295 92.5758 46.7351 92.5608 6.3980 30.8505 30.8314 Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=30 Gender=M Date: Time: 3/29/2004 11:04:06 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 30.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 14.00 Cone. ug/mL 0.0000 0.002374 0.002750 0.001998 Pred. ug/mL Residual AUC AUMC Weight ug/mL day*ug/mL day*day*ug/mL 0.0000 0.001187 0.008873 0.03261 0.0000 0.001187 0.02125 0.2161 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda_z Lambda z Lambda_z_Lower Lambda z_upper HL Lambda z T-lag Tmax Cmax Tlast Clast AUClast AUCall 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.0028 14.0000 0.0020 0.0326 0.0326 69 AUCINF obs AUCINF_D_obs AUC %Extrap obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC %Extrap_pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap_pred MRTlast MRTINF obs MRTINF pred day*ug/mL day*kg*ug/mL/ug % ml/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL, % day*day*ug/mL % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 0.2161 Missing Missing Missing Missing 6.6264 Missing Missing Input File: Descriptive Statistics Workbook - (Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=100 Genaer=F Date: Time: 3/29/2004 11:04:06 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 100.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 14.00 Cone. ug/mL 0.0000 0.1724 0.1984 0.01452 Pred. ug/mL Residual ug/mL AUC ACJMC day*ug/mL day*day*i 0.0000 0.08621 0.6425 1.707 0.0000 0.08621 1.535 6.521 @) Note - the concentration at dose time was added. for extrapolation *) Starred values were included in the estimation of Lambda_z. *** Warning 14530 : Lambda z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tinax Cmax Tlast Clast AUClast AUCall 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.1984 14.0000 0.0145 1.7073 1.7073 Weight 71 AUCINF obs AUCINF_D_obs AUC %Extrap obs Vz F_obs Cl F obs AUCINF pred AUCINF_D_pred AUC %Extrap pred Vz F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTINF_obs MRTINF-pred day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 6.5206 Missing Missing Missing Missing 3.8192 Missing Missing Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PF0S_Dose=100 Gender=M Date: Time: 3/29/2004 11:04:06 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 100.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 14.00 Cone. ug/mL 0.0000 0.02756 0.02646 0.1170 Pred. ug/mL Residual ug/mL AUC AUMC day*ug/mL day*day*' 0.0000 0.01378 0.09480 0.8121 0.0000 0.01378 0.2139 8.934 @) Note - the concentration at dose time was added for extrapolation *) Starred values were included in the estimation of Lambda z. *** Warning 14530 : Lambda z could not be estimated No-parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tiast Clast AUClast AUCall 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 14.0000 0.1170 14.0000 0.1170 0.8121 0.8121 Weight 73 AUCINF obs AUCINF_D_obs AUC %Extrap obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs ACJMC_%Extrap obs AUMCINF_pred - AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 8.9339 Missing Missing Missing Missing 11.0003 Missing Missing Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo] PFOS_Dose=5000 Genaer=F Date: Time: 3/29/2004 11:04:06 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 5000.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 * 14.00 * 28.00 * Cone. ug/mL 0.0000 18.87 19.60 11.08 6.993 Pred. ug/mL 18.57 12.16 6.727 Residual ug/mL 1.033 -1.078 0.2654 MJC AUMC Weight day*ug/mL day*day* ug/mL 0.0000 9.434 67.14 220.6 347.1 0.0000 9.434 155.3 1323. 3780. 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. Final Parameters Rsq Rsq adjusted Corr_XY No_points lambda z Lambda z Lambda_z lower Lambda_z_uppe r HL_Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL day*ug/mL day*kg*ug/mL/ug 0.9755 0.9510 -0.9877 3 0.0423 4.0000 28.0000 16.3858 0.0000 4.0000 19.6006 28.0000 6.9925 347.0999 347.0999 512.4007 0.1025 75 AUC %Extrap obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap pred Vz F pred Cl F pred AMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred % mL/kg mL/day/kg day*ug/mL day*kg*ug/mL/ug % mL/kg mL/day/kg day*day*ug/mL day*day*ug/mL % day*day*ug/mL % day day day 32.2601 230.6756 9.7580 506.1277 0.1012 31.4205 233.5346 9.8789 3780.0447 12316.1237 69.3082 11992.1913 68.4791 10.8904 24.0361 23.6940 Input File: Descriptive Statistics Workbook - [Sera low dose PFOS descriptive statistics table wo 1245 outlier 3-29-04.pwo) PFOS_Dose=5000 Gender=M Date: Time: 3/29/2004 11:04:06 Average Sera [PFOS] Vs time. Low Dose PFOS WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 5000.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear .Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 e 1.000 * 4.000 * 14.00 * 28.00 Cone. ug/mL 0.0000 13.37 12.98 9.666 3.474 Pred. ug/mL 15.01 12.91 7.802 3.855 Residual ug/mL -1.649 0.07581 1.864 -0.3801 AUC AUMC Weight day*ug/mL day*day*u:g/mL 0.0000 6.683 46.21 159.5 251.4 0.0000 6.683 104.6 1041. 2669. 1.000 1.000 1.000 1.000 (?) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs 1/day day day day day day ug/mL day ug/mL day*ug/mL day*ug/mL day*ug/mL day*kg*ug/mL/ug 0.9414 0.9120 -0.9702 4 0.0504 1.0000 28.0000 13.7635 0.0000 1.0000 13.3659 28.0000 3.4745 251.4467 251.4467 320.4377 0.0641 77 AUC %Extrap_obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC %Extrap_pred Vz F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %xtrap_pred MRTlast MRTINF obs MRTINF pred % mL/kg mb/day/kg day*ug/'mb day*kg*ug/mb/ug % mL/kg mb/day/kg day*day*ug/mb day*day*ug/mb % day*day*ug/mb % day day day 21.5302 309.8347 15.6037 327.9857 0.0656 23.3361 302.7044 15.2446 2669.2167 5970.8873 55.2961 6332.1102 57.8463 10.6154 18.6335 19.3061 Appendix 4. Individual Liver PFOS, % dose in Liver and Liver/Sera PFOS amount (M-g^M-g) ratio. Dose (ug/Kg) 0 Day_ PostDose (day) X Gender Animal# (8R0xxxx) F 1231 1233 232 3825 3826 3827 3828 3829 N Mean SD M 1205 1206 1207 3778 3779 3780 3781 3782 N Mean SD 4F 1234 1235 1236 3830 3831 3832 3833 3834 N Mean SD M 1208 1209 1210 3783 3784 Liver [PFOS] (ug/g) <LOQ <LOQ <LOQ 1.83 0.25 <LOQ <LOQ <LOQ 2 1.04 1.12 0.03 0.05 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 2 0.04 0.02 <L0Q <LOQ <LOQ <L0Q <LOQ <L0Q <LOQ <LOQ 0 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ Liver PFOS_ (ug) NA NA NA 14.09 1.83 NA NA NA 2 7.96 8.67 %dose _in_ liver NA NA NA NA NA NA NA NA 0 NA NA Liver. PFOS_ ratio (ug/ug) NA NA NA NA NA NA NA NA 0 NA NA 0.33 0.59 NA NA NA NA NA NA 2 0.46 0.19 NA NA NA NA NA NA NA NA 0 NA NA NA NA NA NA NA NA NA NA 0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 000 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 79 14 F 3785 3786 3787 N Mean SD **1492 **1493 1257 3835 3836 3837 3838 3839 N Mean SD M 1486 1487 1488 3788 3789 3790 3791 3792 N Mean SD 28 F 1494 1495 1496 N Mean SD M 1489 1490 1491 N Mean SD Dose (ug/Kg) 3 Day PostDose (day) 1 Gender F Animai# (8R0xxxx) 3698 3699 3700 3701 3702 N Mean SD M 3668 3669 3670 <L0Q <L0Q <L0Q 0 <L0Q <L0Q <L0Q <LOQ <L0Q <LOQ <L0Q <LOQ <L0Q <LOQ 0 <L0Q <L0Q <L0Q <L0Q <LOQ <L0Q <LOQ <L0Q <L0Q <L0Q 0 <L0Q <L0Q <LOQ <L0Q <L0Q 0 <LOQ <L0Q <L0Q 0.03 <LOQ 1 0.03 <L0Q Liver [PFOS] (ug/g) <L0Q <LOQ <L0Q <LOQ <L0Q 0 <L0Q <LOQ 0.14 <L0Q 0.22 NA NA NA NA NA NA NA NA NA 000 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 000 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 000 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 000 NA NA NA NA NA NA NA 0.36 NA 1 0.36 NA NA NA NA 0 NA NA NA 4.69 NA 1 4.7 NA Liver PFOS (ug> NA NA NA NA NA 0 NA NA %dose in liver NA NA NA NA NA 0 NA NA Liver/Sera PFOS ratio (ug/ug) NA NA NA NA NA 0 NA NA 1.56 NA 2.26 216.04 NA NA NA 338.78 NA 80 3671 3672 N Mean SD 4F 3703 3704 3705 3706 3707 N Mean SD M 3673 3674 3675 3676 3677 N Mean SD 14 F 3708 3709 3710 3711 3712 N Mean SD M 3678 3679 3680 3681 3682 N Mean SD Dose (ug/Kg) 10 Day PostDose (day) 1 Gender F Animai# (8ROxxxx) 3855 3856 3857 3858 3859 N Mean SD M 3808 3809 3810 3811 3812 <L0Q <LOQ 2 Q.18 0.06 <L0Q <L0Q <LOQ <L0Q <LOQ 0 <L0Q <L0Q <L0Q <LOQ <L0Q <LOQ <L0Q 0 <L0Q <L0Q 0.02 0.02 0.02 0.03 0.02 5 0.02 0.00 0.03 0.03 0.05 0.04 0.02 5 0.03 0.01 Liver [PFOS] <ug/g) <LOQ <LOQ <LOQ <LOQ <LOQ 0 <LOQ <L0Q 0.14 0.21 0.17 0.16 0.14 NA NA 2 1.91 0.50 NA NA NA NA NA 0 NA NA NA NA NA NA NA 0 NA NA 0.19 0.19 0.20 0.25 0.18 5 0.20 0.03 0.42 0.49 0.55 0.46 0.22 5 0.43 0.12 Liver PFOS (ug) NA NA NA NA NA 0 NA NA 1.53 2.23 1.79 1.73 1.42 NA NA 2 277.41 86.79 NA NA 0 NA NA NA NA NA NA NA NA NA NA NA NA 00 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 00 NA NA NA NA 33.52 36.23 36.93 48.09 32.22 5 37.40 6.28 NA NA NA NA NA 0 NA NA 60.16 67.75 83.74 65.36 33.39 5 62.08 18.30 NA NA NA NA NA 0 NA NA %dose in liver NA NA NA NA NA 0 NA NA Liver/Sera PFOS_ ratio (ug/ug) NA NA NA NA NA 0 NA NA 66.65 109.76 77.54 75.20 62.30 NA NA NA NA NA 81 4F M 14 F M Dose (ug/Kg) 30 Day PostDose (day) 1 Gender F M N Mean SD 3860 3861 3862 3863 3864 N Mean SD 3813 3814 3815 3816 3817 N Mean SD 3865 3866 3867 3868 3869 N Mean SD 3818 3819 3820 3821 3822 N Mean SD Animal# (8ROxxxx) 3713 3714 3715 3716 3717 N Mean SD 3683 3684 3685 3686 3687 N Mean 5 0.16 0.03 0.15 0.14 0.14 0.15 0.15 5 0.15 0.00 0.18 0.21 0.16 0.19 0.18 5 0.18 0.02 0.04 0.05 0.05 0.06 0.05 5 0.05 0.01 0.09 0.08 0.05 0.09 0.09 5 0.08 0.02 Liver [PFOS] (ug/g) 0.24 0.27 0.22 0.29 0.29 5 0.26 0.03 0.38 0.41 0.50 0.35 0.42 5 0.41 5 1.74 0.31 1.16 1.05 1.11 1.08 1.19 5 1.12 0.06 1.94 2.14 1.95 1.88 1.99 5 1.98 0.10 0.33 0.41 0.40 0.40 0.43 5 0.39 0.04 1.17 1.08 0.66 1.23 1.15 5 1.06 0.23 Liver PFOS (ug) 2.32 2.09 1.53 2.15 2.06 5 2.03 0.30 3.64 4.19 5.16 3.69 4.42 5 4.22 5 78.29 18.65 0 NA NA 62.37 60.05 57.84 61.60 65.97 5 61.56 3.01 11.35 8.70 8.71 9.80 10.68 5 9.8 1.2 85.36 83.14 84.21 86.60 91.99 5 86.26 3.45 NA NA NA NA NA 0 NA NA 19.18 22.09 23.29 22.26 25.21 5 22.41 2.19 NA NA NA NA NA 0 NA NA 50.58 49.10 26.66 52.90 50.90 5 46.03 10.91 15.20 9.59 11.12 20.01 NA 4 14.0 4.7 %dose in liver 39.14 36.82 29.43 35.75 36.76 5 35.58 3.66 Liver/Sera PFOS ratio (ug/ug) 8.41 6.96 5.63 6.19 6.77 5 6.8 1.0 52.35 63.87 72.80 52.68 63.76 5 61.09 NA NA NA NA NA 0 NA 82 4F M 14 F M Dose (ug/Kg) 100 Day PostDose (day) 1 Gender F M SD 3718 3719 3720 3721 3722 N Mean SD 3688 3689 3690 3691 3692 N Mean SD 3723 3724 3725 3726 3727 N Mean SD 3693 3694 3695 3696 3697 N Mean SD Animai# (8ROxxxx) 3840 3841 3842 3843 3844 N Mean SD 3793 3794 3795 3796 3797 N Mean SD 0.06 0.31 0.25 0.30 0.17 0.28 5 0.26 0.05 0.64 0.42 0.36 0.40 0.42 5 0.45 0.11 0.16 0.15 0.16 0.16 0.17 5 0.16 0.01 0.36 0.32 0.32 0.30 0.35 5 0.33 0.02 Liver [PFOS] (ug/g) 1.04 0.90 0.77 0.89 0.70 5 0.86 0.13 1.06 1.36 1.66 1.42 1.45 5 1.39 0.22 0.62 2.12 2.02 2.04 1.27 2.24 5 - 1.94 0.38 6.60 4.65 3.91 4.14 4.38 5 4.74 1.08 1.37 1.27 1.29 1.15 1.18 5 1.25 0.09 4.66 4.66 4.08 3.76 4.71 5 4.37 0.43 Liver PFOS (ug) 7.49 7.39 5.60 6.67 5.04 5 6.44 1.09 11.98 14.28 17.10 15.34 16.10 5 14.96 1.96 8.64 NA 39.94 6.30 36.17 ' 6.45 38.46 7.90 23.53 9.55 38.29 7.24 55 35.28 7.5 6.70 1.3 95.09 66.85 54.94 61.43 62.67 5 68.20 15.63 -NA NA NA NA NA 0 NA NA 23.75 21.58 24.07 22.27 21.73 5 22.68 1.16 5.99 5.19 5.48 6.21 5.79 5 5.7 0.4 66.88 62.15 59.68 56.72 67.52 5 62.59 4.63 NA NA NA NA NA 0 NA NA %dose in liver 38.82 39.64 31.05 35.15 28.11 5 34.55 4.95 Liver/Sera PFOS ratio (ug/ug) 5.48 5.76 4.29 36.27 5.19 5 11.4 13.9 53.14 63.58 75.12 66.10 68.17 5 65.22 8.00 45.16 70.68 98.19 77.95 80.34 5 74.5 19.3 83 4F 3845 3846 3847 3848 3849 N Mean SD M 3798 3799 3800 3801 3802 N Mean SD 14 F 3850 3851 3852 3853 3854 N Mean SD M 3803 3804 3805 3806 3807 N Mean SD Dose (ug/Kg) 5000 Day PostDose (day) 1 Gender F Animai# (8ROxxxx ) 1237 1238 1239 1240 1241 N Mean SD M 1211 1212 1213 1214 1215 N Mean SD 4F 1242 1243 0.85 0.85 0.90 1.39 0.51 5 0.90 0.32 1.22 1.45 1.25 1.43 1.40 5 1.35 0.11 0.48 0.66 0.50 0.56 0.47 5 0.53 0.07 0.98 1.03 0.81 0.77 0.76 5 0.87 0.13 Liver [PFOS] (ug/'g) 33.50 41.50 32.90 42.90 37.50 5 37.66 4.53 36.50 38.90 32.10 34.80 25.60 5 33.58 ' 5.10 30.30 30.30 6.81 6.03 7.50 11.40 3.82 5 7.11 2.77 35.55 33.07 40.84 61.54 21.18 5 38.44 14.79 14.40 14.65 14.50 14.59 15.54 5 14.73 0.46 62.24 62.69 64.76 68.13 65.35 5 64.63 2.36 3.78 6.94 4.82 4.77 3.87 5 4.84 1.27 21.08 33.87 25.27 26.65 20.11 5 25.39 5.48 12.89 12.77 10.48 10.42 10.50 5 11.41 1.30 56.89 55.75 45.30 46.12 46.61 5 50.13 5.68 Liver PFOS (ug) %dose in liver 244.55 352.75 302.68 334.62 318.75 5 310.67 41.37 25.61 32.97 29.53 32.65 32.69 5 30.69 3.17 448.95 525.15 385.20 431.52 296.96 5 417.56 84.18 31.02 35.96 26.38 30.24 21.26 5 28.97 5.50 272.70 25.85 290.88 26.44 5.04 6.39 5.71 5.61 5.88 5 5.7 0.5 75.22 80.93 57.15 55.77 77.81 5 69.4 12.0 44.81 59.38 42.20 49.87 34.56 5 46.2 9.2 15.21 8.57 9.53 8.51 11.02 5 10.6 2.8 Liver/Sera PFOS ratio (ug/ug) 2.34 2.72 6.79 1.81 2.27 5 3.2 2.0 4.65 4.31 3.37 3.17 1.95 5 3.5 1.1 1.54 2.15 84 1244 1245 1246* N Mean SD 36.30 64.10 4 40.25 16.15 290.40 570.49 NA 4 356.12 143.17 28.33 55.12 NA 4 33.94 14.16 2.20 302.08 NA 4 77.0 150.1 M 1216 37.00 1217 25.80 1218 28.50 1219 23.40 1220 22.80 N5 Mean 27.50 SD 5.77 469.90 322.50 339.15 325.26 291.84 5 349.73 69.36 32.30 23.45 25.03 22.28 20.20 5 24.65 4.62 3.69 2.68 2.56 2.76 2.27 5 2.8 0.5 14 F 1247 1248 1249 1250 1251 N Mean SD 43.50 34.00 33.10 32.50 57.60 5 40.14 10.74 369.75 323.00 278.04 292.50 478.08 5 348.27 80.61 37.54 29.10 28.52 28.68 45.75 5 33.92 7.63 4.28 4.30 3.68 3.83 4.97 5 4.2 0.5 M 1221 44.60 1222 57.90 1223 27.70 1224 61.70 1225 22.70 N5 Mean 42.92 SD 17.47 593.18 949.56 393.34 968.69 313.26 5 643.61 305.61 41.19 65.94 27.41 65.01 22.78 5 44.47 20.34 3.73 7.52 4.89 11. 5a 3.24 5 6.2 3.4 28 F 1252 1253 1254 1255 1256 N Mean SD 12.30 23.20 30.10 13.80 25.40 5 20.96 7.66 113.16 190.24 279.93 111.78 215.90 5 182.20 71.55 11.49 18.47 26.28 10.80 20.96 5 17.60 6.54 3.06 3.54 5.27 1.79 3.46 5 3.4 1.2 M 1226 32.10 459.03 32.67 15.00 1227 26.90 449.23 30.98 12.66 1228 31.00 443.30 32.24 12.05 1229 30.50 488.00 32.53 9.16 1230 34.30 511.07 35.61 7.60 N5 555 Mean 30.96 470.13 32.81 11.3 SD 2.70 28.62 1 . 7 1 2.9 * Liver PFOS value for animal # 8RO-1246 was considered an outlier. 85 Appendix 5 WinNonlin NCA Model 200 Liver PFOS. Figure 23. NCA of Average Liver PFOS Vs Time. Female Rats. Control dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=Q, Gender=F Rsq=0.4923 Rsq_adjusted=0.2385 HL_Lambda_z=8.2547 (day) (4 points used in calculation) Uniform Weighting 1.000 T n 0.100 | - c 03 0.010 T 2CD 0.001 i -- 0 tT 5 O 10 15 20 Day_Post-Dose (day) ) - - Observed ------ Predicted 25 30 X vs. Observed Y and Predicted Y (27-Feb-2004) Figure 24. NCA of Average Liver PFOS Vs Time. Male Rats. Control dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=0, Gender=M Rsq=0.3936 Rsq_adjusted=0,0904 HL_Lambda_z=17.9946 (day) (4 points used in calculation) Uniform Weighting -OS)' 0.10 :T /-v 3 ; Q --- Q _ _ _ ____________ TO T "" " ~ ' ' --- -------- O I 0 .0 1 --------------- !---------------!---------- O-r------------------------------i--------------- 0 5 10 15 20 25 30 Day_Post-Dose (day) - 0 ~ Observed ------ Predicted X vs. Observed V and Predicted Y (27-Feb-20Q4) 86 Figure 25. NCA of Average Liver PFOS Vs Time. Female Rats. 3 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=3, Gender=F Rsq=0.9514 Rsq_adjusted=0.9029 HL_l_ambda_z=7.?932 (day) (3 points used in calculation) Uniform Weighting -O3S)' 0.10 t:; rc<aa 0.01 r: o- -JCL 6 8 10 Day_Post-Dose (day) - - - Observed ------ Predicted 12 14 X vs. Observed Y and Predicted Y (27-Feb-2004) Figure 26. NCA of Average Liver PFOS Vs Time. Male Rats. 3 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=3, Gender=M Rsq=0.9501 Rsq_adjusted=0.900t HL_Lambda_z=8.1134 (day) (3 points used in calculation) Uniform Weighting -S' 1.00 or f 0.10 cs 5 0.01 o- -o- 6 8 10 Day_Post-Dose (day) -O - 0 - Observed ------ Predicted 12 14 X vs. Observed Y and Predicted Y (27-Feb-2004) 87 Figure 27. NCA of Average Liver PFOS Vs Time. Female Rats. 10 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=10, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting -S' 1.00 t CO ; f" 0.10 | CO ; o o I 0.01 - 0 24 6 O - - Observed ------ Predicted 8 10 12 14 Day_Post-Dose (day) X vs. Observed V and Predicted Y (27-Feb-2004) Figure 28. NCA of Average Liver PFOS Vs Time. Male Rats. 10 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=10, Gender=M Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting 5 > 1.00 C3) ifTO" 0.10 0.01 0 O 2 oo 4 6 8 10 12 14 Day_Post-Dose (day) X vs. Observed Y and Predicted Y {27-Feb'2004) Observed Predicted 88 Figure 29. NCA of Average Liver PFOS Vs Time. Female Rats. 30 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=30, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting 1.0 T 0.1 o o Q - - Observed ------ Predicted 4 6 8 10 12 14 Day_Post-Dose (day) X vs. Observed Y and Predicted Y (27-Feb-2004) Figure 30. NCA of Average Liver PFOS Vs Time. Male Rats. 30 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=30, Gender=M Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting OS)> 1.0 o 02 OO - 0 - Observed ------ Predicted 4 6 8 10 12 14 Day_Post-Dose (day) X vs. Observed Y and Predicted Y (27-Feb-20G4) 89 Figure 31. NCA of Average Liver PFOS Vs Time. Female Rats. 100 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Oose=100, Gender=F Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_z=Undefined (day) (0 points used in calculation) Uniform Weighting o-OS)' 1.0 := 0.1 o O - - Observed Predicted 4 6 8 10 12 14 Day_Post-Dose (day) X vs. Observed Y and Predicted Y (27-Feb-2004) Figure 32. NCA of Average Liver PFOS Vs Time. Male Rats. 100 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=100, Gender=M Rsq=0.9725 Rsq_adjusted=0.945 HL_Lambda_z=18.1985 (day) (3 points used in calculation) Uniform Weighting O) 10.0 T 3 C l ,u i as o U. i 0 w ---------------- 0 . ....................................................................-- ------------------------------------ ----------------- Q 2 4 6 8 10 12 14 Day_Post-Dose (day) Observed ------ Predicted X vs. Observed Y and Predicted Y (27-Feb-2004) 90 Figure 33. NCA of Average Liver PFOS Vs Time. Female Rats. 5000 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 100 r Dose=5000, Gender=F Rsq=0.8307 Rsq_adjusted=0.6614 HL_Lambda_z=24.3878 (day) (3 points used in calculation) Uniform Weighting 5TcO - 0 - Observed ------ Predicted 10 -------------------- '--------- !--------- 1------------------- 1 0 5 10 15 20 25 30 Day_Post-Dose (day) X vs. Observed Y and Predicted Y (28-Mar-2004) Figure 34. NCA of Average Liver PFOS Vs Time. Male Rats. 5000 ug/Kg dose group. Liver PFOS Vs Time. Low Dose PFOS in Rats. Dose=5000, Gender=M Rsq=Undefined Rsq_adjusted=Undefined HL_Lambda_2=Undefined (day) (6 points used in calculation) Uniform Weighting 100 CD C :O CO I 10 O o 10 15 20 Day_Post-Dose (day) O Observed ------ Predicted 25 30 X vs. Observed Y and Predicted Y (27-Feb-2004) 91 Input File: Descriptive Statistics Workbook - [Untitled27] Dose=0 Gender=F Date: Time: 3/28/2004 23:03:07 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 0.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 * 4.000 * 14.00 * 28.00 * Cone. ug/g 0.0000 0.2859 0.04304 0.01006 0.01850 Pred. ug/g 0.09594 0.07458 0.03221 0.009940 Residual ug/g 0.1899 -0.03154 -0.02214 0.008560 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.1429 0.6363 0.9019 1.102 0.0000 0.1429 0.8300 2.395 7.007 1.000 1.000 1.000 1.000 0) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z . Final Parameters Rsq Rsq adjusted Corr_XY No points lambda z Lambda_z Lambda_z_lower Lambda_z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g 0.4923 0.2385 -0.7017 4 0.0840 1.0000 28.0000 8.2547 0.0000 1.0000 0.2859 28.0000 0.0185 1.1018 1.1018 1.3221 92 AUCINF_D_obs AUC_%Extrap_obs Vz_F_obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AOMC %Extrap obs AUMCINF_pred AUMC_%Extrap pred MRTlast MRTINF obs MRTINF pred day*kg*ug/g/ug % g/kg g/day/kg day*ug/g day* kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g day day day Missing 16.6641 0.0000 0.0000 1.2202 Missing 9.7016 0.0000 0.0000 7.0073 15.7999 55.6497 11.7316 40.2696 6.3599 11.9506 9.6147 Input File: Descriptive Statistics Workbook - [Untitled27] Dose=0 Gen.der=M Date: Time: 3/28/2004 23:03:08 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 0.00 Weighting for larabda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 * 4.000 * 14.00 * 28.00 * Cone. ug/g 0.0000 0.05156 0.04304 0.01006 0.02067 Pred. Residual ug/g ug/g 0.03943 0.03513 0.02390 0.01394 0.01212 0.007910 -0.01384 0.006729 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.02578 0.1677 0.4332 0.6483 0.0000 0.02578 0.3614 1.927 6.963 1.000 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC_%Extrap_obs 1/day day day day day day ug/g day ug/g day*ug/g' day*ug/g day*ug/g day*kg*ug/g/ug % 0.3936 0.0904 -0.6274 4 0.0385 1.0000 28.0000 17.9946 0.0000 1.0000 0.0516 28.0000 0.0207 0.6483 0.6483 1.1848 Missing 45.2834 94 Vz F obs Cl F obs AQCINF pred AUCINF_D_pred AUC_%Extrap_pred Vz_F pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred g/kg g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g day*day*ug/g % day day day 0.0000 0.0000 1.0101 Missing 35.8204 0.0000 0.0000 6.9633 35.9144 80.6113 26.4878 73.7112 10.7411 30.3124 26.2226 Input File: Descriptive Statistics Workbook - [Untitled27J Dose=3 Gender=F Date: Time: 3/28/2004 23:03:08 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 3.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 * 4.000 * 14.00 * Cone. ug/g 0.0000 0.06710 0.06710 0.02290 Pred. ug/g Residual ug/g 0.07535 -0.008254 0.05771 0.009394 0.02371 -0.0008110 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.03355 0.2349 0.6849 0.0000 0.03355 0.5368 3.482 1.000 1.000 1.000 0) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC %xtrap obs Vz F obs 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g day*kg*ug/g/ug % g/kg 0.9514 0.9029 -0.9754 3 0.0889 1.0000 14.0000 7.7932 0.0000 1.0000 0.0671 14.0000 0.0229 0.6849 0.6849 0.9423 0.3141 27.3229 35.7942 96 Cl F obs AUCINF pred AUCINF__D_pred AUC %Extrap_pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AMC %xtrap obs AUMCINF_pred AUMC %Extrap_pred MRTlast MRT1NF obs MRTINF pred g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g /day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day 3.1836 0.9514 0.3171 28.0195 35.4512 3.1531 3.4818 9.9811 65.1162 10.2113 65.9026 5.0840 10.5921 10.7325 Input File: Descriptive Statistics Workbook - [Untitled27] Dose=3 Gender=M Date: Time: 3/28/2004 23:03:08 Liver PFQS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2QQ3 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 3.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation- Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 0 1.000 * 4.000 * 14.00 * Cone. ug/g 0.0000 0.1125 0.06710 0.03422 Pred. ug/g 0.1004 0.07773 0.03308 Residual ug/g 0.01203 -0.01063 0.001142 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.05623 0.3256 0.8322 0.0000 0.05623 0.6275 4.365 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper SL Lambda z Tlag Ttnax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC %Extrap obs Vz F obs l/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g day*kg*ug/g/ug % g/kg 0.9501 0.9001 -0.9747 3 0.0854 1.0000 14.0000 8.1134 0.0000 r.ooo o 0.1125 14.0000 0.0342 0.8322 0.8322 1.2327 0.4109 32.4932 28.4862 98 Cl F obs AUCINF 'pred ADCINF_D_pred AUC_%Extrap pred Vz F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF. pred AUMC %Extrap pred MRTlast MRXINF obs MRTINF pred g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day 2.4336 1.2194 0.4065 31.7535 28.7983 2.4603 4.3649 14.6611 70.2280 14.3177 69.5137 5.2452 11.8933 11.7420 Input File: Descriptive Statistics Workbook - (Untitled27J Dose=10 Gender=F Date: Time: 3/28/2004 23:03:08 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 10.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day o.oooo e 1.000' 4.000 14.00 Cone. ug/g 0.0000 0.06710 0.1462 0.05036 Pred. ug/g Residual ug/g AUC AUMC Weight day*ug/g' day*day*ug/g 0.0000 0.03355 0.3535 1.336 0.0000 0.03355 1.011 7.461 0) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in'the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda_z Lambda_z Lambda z lower Lambda_z_upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.1462 14.0000 0.0504 1.3363 1.3363 Missing 100 AUCINF_D_obs AUC_%Extrap_obs Vz F obs Cl. F obs AUCINF pred AUCINF_D_pred AUC %Extrap pred Vz F pred Cl F pred AUMClast AUMCINF_obs AUMC %Extitop obs AUMCIN F_pred AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred day*kg*ug/g/ug % g/kg g/day/kg dav*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing 7.4606 Missing Missing Missing Missing 5.5830 Missing Missing Input File: Descriptive Statistics Workbook - [Untitled27] Dose=I0 Gender=M Date: Time: 3/28/2004 23:03:08 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 10.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 0 1.000 4.000 14.00 Cone. ug/g 0.0000 0.1640 0.1820 0.07980 . Pred. ug/g Residual ug/g AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.08200 0.6010 1.910 0.0000 0.08200 1.420 10.65 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda_z Lambda z Lambda z_lower Lambda z_upper HL_Lambda z Tlag Tmax Cmax Hast Clast AUClast AUCall AUCINF_obs 2/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.1820 14.0000 0.0798 1.9100 1.9100 Missing 102 AUCINF_D_obs AUC_%Extrap_obs Vz_F_obs Cl_F_obs AUCINF_pred AUCINF_D_pred AUC_%Extrap_pred Vz_F_pred Cl_F_pred AUMClast AUMCINF_obs AUMC_%Extrap_obs AUMCINFjpred AUMC_%Extrap_pred MRTlast MRTINF_obs MRTINF_pred day*kg*ug/g/ug g/kg g/day/kg day*ug/g day*kg*ug/g/ug g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing 10.6460 Missing Missing Missing Missing 5.5738 Missing Missing' Input File: Descriptive Statistics Workbook - [Untitled27] Dose=30 Gender=F Date: Time: 3/28/2004 23:03:08 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 30.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 14.00 Cone. ug/g 0.0000 0.2598 0.2614 0.1588 Pred. ug/g Residual ug/g AUC AUMC day*ug/g day*day*ug/g 0.0000 0.1299 0.9117 3.013 0.0000 0.1299 2.088 18.43 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda_z Lambda z Lambda_z lower Lambda_z_upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs 1/day day day day day day ug/g day ' ug/g day*ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.2614 14.0000 0.1588 3.0127 3.0127 Missing 104 AUCINF_D_obs AOC %Extrap obs Vz_F_obs Cl F obs AUCINF pred AOCINF_D_pred AUC_%Extrap_pred Vz F pred Cl F pred ACJMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTXNF obs MRTINF pred day*kg*ug/g/ug % g/kg gtday/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing 18.4320 Missing Missing Missing Missing 6.1181 Missing Missing Input File: Descriptive Statistics Workbook - [Untitled27] Dose=30 Gender=M Date: Time: 3/28/2004 23:03:09 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 30.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda_z Summary Table Time day 0.0000 9 1.000 4.000 14.00 Cone. ug/g 0.0000 0.4092 0.4462 0.3306 Pred. ug/g Residual ug/g AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.2046 1.488 5.372 0.0000 0.2046 3.496 35.56 @) Note - the concentration at dose time was added for extrapolation purposes. ') Starred values were included in the estimation of Lambda z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda z Lambda z Lambda_z_lower Lambda_z_upper HL Lambda z Tlag Tmax Cmax Hast Clast AUClast AUCall AUCINF obs 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.4462 14.0000 0.3306 5.3717 5.3717 Missing 106 AUCINF_D_obs AUC_%Extrap_obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap_pred Vz_F_pred Cl F pred AUMClast AUMCIMF_obs AUMC %Extrap obs AUMCINF_pred AQMC %Extrap pred MRTlast MRTINF obs MRTINF pred day*kg*ug/g/ug % g/kg g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % aay*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing 35.5616 Missing Missing Missing Missing 6.6202 Missing Missing Input File: Descriptive Statistics Workbook - fU'ntitled27] Dose=100 Gender=F Date: Time: 3/23/2004 23:03:09 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build'200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 100.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 0 1.000 4.000 14.00 Cone. ug/g 0.0000 0.8594 0.9006 0.5336 Pred. ug/g Residual ug/g AUC AUMC day*ug/g day*day* 0.0000 0.4297 3.070 10.24 0.0000 0.4297 7.122 62.49 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq adjusted Corr_XY No_points_lambda_z Lambda_z Lambda_z_lower Lambda_z_upper HL_Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 4.0000 0.9006 14.0000 0.5336 10.2407 10.2407 Missing 108 AUCINF_D_obs AUC_%Extrap_obs Vz F_obs Cl F obs AUCINF pred AUCINF_D_pred AUC %Extrap pred Vz F pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC %Extrap pred MRTlast MRTINF obs MRTINF pred day*kg*ug/g/ug % g/kg g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g '% day*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing 62.4864 Missing Missing Missing Missing 6.1018 Missing Missing Input File: Descriptive Statistics Workbook - [Untitled27] Dose=100 Gender=M Date: Time: 3/28/2004 23:03:09 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 3 Dose time: 0.00 Dose amount: 100.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 * 4.000 * 14.00 * Cone. ug/g 0.0000 1.390 1.350 0.8694 Pred. ug/g Residual ug/g 1.442 -0.05237 1.287 0.06337 0.8791 -0.009700 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.6950 4.805 15.90 0.0000 0.6950 10.88 98.74 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in-the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda' z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC %Extrap obs Vz_F obs l/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g day*kg*ug/g/ug % g /kg 0.9725 0.9450 -0.9861 3 0.0381 1.0000 14.0000 18.1985 0.0000 1.0000 1.3900 14.0000 0.8694 15.9020 15.9020 38.7280 0.3873 58.9393 67.7931 110 Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap_pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred AMC %Extrap pred MRTlast MRTINF obs MRTINF pred g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day 2.5821 38.9827 0.3898 59.2076 67.3502 2.5652 98.7380 1017.5985 90.2970 1027.8505 90.3937 6.2092 26.2755 26.3668 Input File: Descriptive Statistics Workbook - [Untitled27] Dose=5000 Gender=F Date: Time: 3/28/2004 23:03:09 Liver PFOS Vs Time. Low Dose PFOS in rats, MCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 5000.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda z method: Find best fit for lambda z Summary Table Time day 0.0000 @ 1.000 4.000 * 14.00 * 28.00 * Cone. ug/g 0.0000 37.66 40.25 40.14 20.96 Pred. ug/g 44.65 33.60 22.57 Residual ug/g -4.398 6.538 -1.612 AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.0000 18.83 18.83 135.7 316.8 537.6 3932. 965.3 1..197e+0Q4 1.000 1.000 1.000 @) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda z. Final Parameters Rsq Rsq adjusted Corr XY No points lambda z Lambda z Lambda z lower Lambda z upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall AUCINF obs AUCINF D obs AUC_%Extrap obs l/day day day day day day ug/g day ug/g day*ug/g day*ug/g day*ug/g day*kg*ug/g/ug % 0.8307 0.6614 -0.9114 3 0.0284 4.0000 28.0000 24.3878 0.0000 4.0000 40.2500 28.0000 20.9600 965.3450 965.3450 1702.8065 0.3406 43.3086 112 Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AOC_%Extrap pred Vz_F_pred Cl F pred AUMClast ACJMCINF_obs AUMC %Extrap obs AUMCINF_pred AUMC gExtrap pred MRTlast MRTINF obs MRTINF pred g/kg g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day 103.3125 2.9363 1759.5074 0.3519 45.1355 99.9832 2.8417 11973.500 58569.4342 79.5567 62152.0371 80.7351 12.4033 34.3958 35.3235 Input File: Descriptive Statistics Workbook. - [Untitled27] Dose=5000 Gender=M Date: Time: 3/28/2004 23:03:09 Liver PFOS Vs Time. Low Dose PFOS in rats, NCA 200 WINNONLIN NONCOMPARTMENTAL ANALYSIS PROGRAM Version 4.1 Build 200307301056 Core Version 16May2003 Settings Model: Plasma Data, Extravascular Administration Number of nonmissing observations: 4 Dose time: 0.00 Dose amount: 5000.00 Weighting for lambda_z calculation: Uniform weighting Calculation method: Linear Trapezoidal with Linear Interpolation Lambda_z method: Find best fit for lambda_z Summary Table Time day 0.0000 @ 1.000 4.000 14.00 28.00 Cone. ug/g 0.0000 33.58 27.50 42.92 30.96 Pred. ug/g Residual ug/g AUC AUMC Weight day*ug/g day*day*ug/g 0.0000 0.0000 16.79 16.79 108.4 232.2 460.5 3787 . 977.7 1..406e+004 0) Note - the concentration at dose time was added for extrapolation purposes. *) Starred values were included in the estimation of Lambda_z. *** Warning 14530: Lambda_z could not be estimated. No parameters could be extrapolated to infinity. Final Parameters Rsq Rsq_adjusted Corr_XY No points lambda z Lambda_z Lambda_z_lower Lambda_z_upper HL Lambda z Tlag Tmax Cmax Tlast Clast AUClast AUCall 1/day day day day day day ug/g day ug/g day*ug/g day*ug/g Missing Missing Missing 0 Missing Missing Missing Missing 0.0000 14.0000 42.9200 28.0000 30.9600 977.6700 977.6700 114 AUCINF obs AUCINF_D_obs AUC_%Extrap_obs Vz F obs Cl F obs AUCINF pred AUCINF_D_pred AUC_%Extrap pred Vz_F_pred Cl F pred AUMClast AUMCINF_obs AUMC %Extrap obs AUMCINF_pred ?,UMC_%Extrap_pred MRTlast MRTINF_obs MRTINF_pred day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*ug/g day*kg*ug/g/ug % g/kg g/day/kg day*day*ug/g day*day*ug/g % day*day*ug/g % day day day Missing Missing Missing Missing Missing Missing Missing Missing Missing Missing 14060.8800 Missing Missing Missing Missing 14.3820 Missing Missing Appendix 6: Statistical Analyses Male and Female sera PFOS LOQ/2 substituted data DosePFOS(ug/Kg) Group Gender Animal# Day Sera PFOS (ug/mL) 0 1F 8R03825F 1 0.00275 0 1F 8R03826F 1 0.00275 0 1F 8R03827F 1 0.00275 0 1F 8R03828F 1 0.00275 0 1F 8R03829F 1 0.00275 0 1F 8R03830F 4 0.00275 0 1F 8R03831F 4 0.00275 0 1F 8R03832F 4 0.00275 0 1F 8R03833F 4 0.00275 0 1F 8R03834F 4 0.00275 0 1F 8R03835F 14 0.00087 0 1F 8R03836F 14 0.00087 0 1F 8R03837F 14 0.05735 0 1F 8R03838F 14 0.00087 0 1F 8R03839F 14 0.00087 0 6F 9R01231F 1 0.00872 0 6F 9R01232F 1 0.00087 0 6F 9R01233F 1 0.00087 0 6F 9R01234F 4 0.00087 0 6F 9R01235F 4 0.00087 0 6F 9R01236F 4 0.00087 0 6F 9R01257F 14 0.00087 0 6F 9R01492F 14 0.00087 0 6F 9R01493F 14 0.00087 0 6F 9R01494F 28 0.13551 0 6F 9R01495F 28 0.01556 0 6F 9R01496F 28 0.01431 3 2F 8R03698F 1 0.00275 3 2F 8R03699F 1 0.00275 3 2F 8R03700F 1 0.00275 3 2F 8R03701F 1 0.00275 3 2F 8R03702F 1 0.00275 3 2F 8R03703F 4 0.00275 3 2F 8R03704F 4 0.00275 3 2F 8R03705F 4 0.00275 3 2F 8R03706F 4 0.00275 3 2F 8R03707F 4 0.00275 3 2F 8R03708F 14 0.00087 3 2F 8R03709F 14 0.00087 3 2F 8R03710F 14 0.00275 3 2F 8R03711F 14 0.00087 3 2F 8R03712F 14 0.00087 10 5F 8R03855F 1 0.01459 10 5F 8R03856F 1 0.01646 10 5F 8R03857F 1 0.01498 10 5F 8R03858F 1 0.01871 10 5F 8R03859F 1 0.0195 10 5F 8R03860F 4 0.01618 10 5F 8R03861F 4 0.02043 10 5F 8R03862F 4 0.01996 10 5F 8R03863F 4 0.01841 10 5F 8R03864F 4 0.01789 10 5F 8R03865F 14 0.00087 10 5F 8R03866F 14 0.00087 10 5F 8R03867F 14 0.00087 10 5F 8R03868F .14 0.00087 10 30 30 30 30 30 30 30 30 30 30 30 30 30 30 30 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 0 0 0 0 0 0 0 0 0 0 0 5F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 3F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 4F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 7F 1M 1M 1M 1M 1 .M 1M 1M 1M 1M 1M 1M 8R03869F 14 0.00087 8R03713F 1 0.04204 8R03714F 1 0.04993 8R03715F 1 0.04766 8R03716F 1 0.05596 8R03717F 1 0.0505 8R03718F 4 0.05714 8R03719F 4 0.04799 8R03720F 4 0.04388 8R03721F 4 0.02185 8R03722F 4 0.04847 8R03723F 14 0.03186 8R03724F 14 0.03358 8R03725F 14 0.03448 8R03726F 14 0.02829 8R03727F 14 0.03163 8R03840F 1 0.2257 8R03841F 1 0.2113 8R03842F 1 0.2263 8R03843F 1 0.03013 8R03844F 1 0.1687 8R03845F 4 0.2062 8R03846F 4 0.1542 8R03847F 4 0.2025 8R03848F 4 0.3212 8R03849F 4 0.1081 8R03850F 14 0.01279 8R03851F 14 0.01464 8R03852F 14 0.01572 8R03853F 14 0.01333 8R03854F 14 0.01611 9R01237F 1 17.3632941 9R01238F 1 19.0302 9R01239F 1 6.88593 9R01240F 1 28.804 9R01241F 1 22.2596154 9R01242F 4 24.9496 9R01243F 4 18.11 9R01244F 4 20.0964 9R01245F 4 0.27645 9R01246F 4 15.2462388 9R01247F 14 11.918 9R01248F 14 9.233 9R01249F 14 10.48 9R01250F 14 10.58 9R01251F 14 13.2104167 9R01252F 28 4.42975 9R01253F 28 7.125 9R01254F 28 6.656 9R01255F 28 8.63 9R01256F 28 8.12183544 8R03778M 1 0.00275 8R03779M 1 0.00275 8R03780M 1 0.00275 8R03781M 1 0.00275 8R03782M 1 0.00275 8R03783M 4 0.00275 8R03784M 4 0.00275 8R03785M 4 0.00275 8R03786M 4 0.00275 8R03787M 4 0.00275 8R03788M 14 0.00087 0 -1 M 8R03789M 14 0.00087 0 1M 8R3790M 14 0.00087 0 1M 8R03791M 14 0.00087 0 1M 8R03792M 14 0.00087 0 6M 9R01205M 1 0.00087 0 6M 9R01206M 1 0.00087 0 6M 9R01207M 1 0.00087 0 6M 9R01208M 4 0.00087 0 6M 9R01209M 4 0.00087 0 6M 9R01210M 4 0.00087 0 6M 9R01486M 14 0.00827 0 6M 9R01487M 14 0.00275 0 6M 9R01488M 14 0.00087 0 6M 9R01489M 28 0.00676 0 6M 9R01490M 28 0.00686 0 6M 9R01491M 28 0.01728 3 2M 8R03668M 1 0.00275 3 2M 8R03669M 1 0.00275 3 2M 8R03670M 1 0.00275 3 2M 8R03671M 1 0.00275 3 2M 8R03672M 1 0.00275 3 2M 8R03673M 4 0.00275 3 2M 8R03674M 4 0.00275 3 2M 8R03675M 4 0.00275 3 2M 8R03676M 4 0.00275 3 2M 8R03677M 4 0.00275 3 2M 8R03678M 14 0.00087 3 2M 8R03679M 14 0.00087 3 2M 8R03680M 14 0.00087 3 2M 8R03681M 14 0.00087 3 2M 8R03682M 14 0.QQ087 10 5M 8R03808M 1 0.00275 10 5M 8R03809M 1 0.00275 10 5M 8R03810M 1 0.00275 10 5M 8R03811M 1 0.00275 10 5M 8R03812M 1 0.00275 10 5M 8R03813M 4 0.00275 10 5M 8R03814M 4 0.00275 10 5M 8R03815M 4 0.00275 10 5M 8R03816M 4 0.00275 10 5M 8R03817M 4 0.00275 10 5M 8R03818M 14 0.00779 10 5M 8R03819M 14 0.01182 10 5M 8R03820M 14 0.00598 10 5M 8R03821M 14 0.00602 10 5M 8R03822M 14 0.00275 30 3M 8R03683M 1 0.00275 30 3M 8R03684M 1 0.00275 30 3M 8R03685M 1 0.00087 30 3M 8R03686M 1 0.00275 30 3M 8R03687M 1 0.00275 30 3M 8R03688M 4 0.00275 30 3M 8R03689M 4 0.00275 30 3M 8R03690M 4 0.00275 30 3M 8R03692M 4 0.00275 30 3M 8R03693M 4 0.00275 30 3M 8R03693M 14 0.00275 30 3M 8R03694M 14 0.00275 30 3M 8R03695M 14 0.00087 30 3M 8R03696M 14 0.00275 30 3M 8R03697M 14 0.00087 100 4M 8R03793M 1 0.03537 100 100 100 100 100 100 100 100 100 100 100 100 100 100 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 4M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 7M 8R03794M 8R03795M 8R03796M 8R03797M 8R03798M 8R03799M 8R03800M 8R03801M 8R03802M 8R03803M 8R03804M 8R03805M 8R03806M 8R03807M 9R01211M 9R01212M 9R01213M 9R01214M 9R01215M 9R01216M 9R01217M 9R01218M 9R01219M 9R01220M 9R01221M 9R01222M 9R01223M 9R01224M 9R01225M 9R01226M 9R01227M 9R01228M 9R01229M 9R01230M 1 1 1 1 4 4 4 4 4 14 14 14 14 14 1 1 1 1 1 4 4 4 4 4 14 14 14 14 14 28 28 28 28 28 0.02765 0.02322 0.02589 0.02565 0.02279 0.02179 0.0309 0.03307 0.02374 0.09057 0.15946 0.11196 0.12584 0.09723 10.3082 12.9238 12.0304 14.6014 16.9656 12.7824 12.8664 14.5372 11.6738 13.065 14.24275 10.82325 7.189 7.01575 9.059 2.5125 2.59785714 3.055 3.97 5.237 0, 3,10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, days 1, 4 and 14 post dose. Sera PFOS (ug/mL) By DosePFOS(ug/Kg) 0.1 !> ; o.o - -Si-^r DosePFOS(ug/Kg) o i ..(.)-.. Durmett's 0 .0 5 120 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.542851 0.520551 0.019327 0.012233 87 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.03637312 82 0.03063072 86 0.06700384 Mean Square 0.009093 0.000374 0.000779 F Ratio 24.3432 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 27 0.002958 3 15 0.002123 10 15 0.004124 30 15 0.002374 100 15 0.057009 Std Error 0.00372 0.00499 0.00499 0.00499 0.00499 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 27 0.002958 0.003485 0.00067 15 0.002123 0.000917 0.00024 15 0.004124 0.002679 0.00069 15 0.002374 0.000778 0.00020 15 0.057009 0.046441 0.01199 Dif=Mean{i]-Mean] 100 10 0 30 3 Means Comparisons 100 10 0.000000 0.052885 -0.05288 0.000000 -0.05405 -0.00117 -0.05463 -0.00175 -0.05489 -0.002 0 0.054051 0.001166 0.000000 -0.00058 -0.00083 30 0.054635 0.001750 0.000584 0.000000 -0.00025 3 0.054885 0.002001 0.000834 0.000251 0.000000 Alpha= 0.05 Comparisons with a control using Dunnetfs Method 2.51855 Abs(Dif)-LSD 100 10 0 30 3 0 0.038375 -0.01451 -0.01325 -0.01509 -0.01484 Positive values show pairs of means that are significantly different. 121 0 ,3 ,1 0 , 30 and 100 ug/Kg dose group Male sera PFOS Statistics, day 1 post-dose. Sera PFOS (ug/mL) By DosePFOS(ug/Kg) - 0.030 0.025 f 0.020 13 to O 0.015 a " 0.010 0.005 0.000 Oo . 0 il 3 l 10 ! 30 Each Pair 100 With Control Student's t Dunnett's DosePFOS{ug/Kg) 0.05 0.05 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.964443 0.958259 0.00204 0.006911 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.00259656 23 0.00009573 27 0.00269230 Mean Square 0.000649 0.000004 0.000100 F Ratio 155.9600 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 8 0.002045 3 5 0.002750 10 5 0.002750 30 5 0.002374 100 5 0.027556 Std Error 0.00072 0.00091 0.00091 0.00091 0.00091 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.002045 0.000973 0.00034 5 0.002750 0.000000 0.00000 5 0.002750 0.000000 0.00000 5 0.002374 0.000841 0.00038 5 0.027556 0.004644 0.00208 Dif=Mean[i]-Mean[j] 100 3 10 30 Means Comparisons 100 3 0.000000 0.024806 -0.02481 0.000000 -0.02481 0.000000 -0.02518 -0.00038 10 0.024806 0.000000 0.000000 -0.00038 30 0.025182 0.000376 0.000376 0.000000 0 0.025511 0.000705 0.000705 0.000329 122 0 -0.02551 -0.00071 -0.00071 -0.00033 0.000000 Alpha= Abs(Dif)-LSD 100 3 10 30 0 0.05 Comparisons for each pair using Student's t t 2.06864 100 3 10 -0.00267 0.022137 0.022137 0.022137 -0.00267 -0.00267 0.022137 -0.00267 -0.00267 0.022513 -0.00229 -0.00229 0.023105 -0.0017 -0.0017 30 0.022513 -0.00229 -0.00229 -0.00267 -0.00208 0 0.023105 -0.0017 -0.0017 -0.00208 -0.00211 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method Id! 2.65107 Abs(Dif)-LSD 0 100 0.022428 3 -0.00238 10 -0.00238 30 -0.00275 0 -0.0027 Positive values show pairs of means that are significantly different. 0, 3,10, 30 and 100 ug/Kg dose group Male sera PFOS Statistics, day 4 post-dose. ___________________ Sera PPOS (ug/mL) By DosePFOS(ug/Kg)__________________ 0.035 0.030 0.025 : o i0 3O) g 0.015 LL CL I 0.010 </} 0. 0.000 iiir a With Control 0 3 10 30 100 Dunnett's DosePFOSfug/Kg) 0.05 123 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations {or Sum Wgts) 0.954402 0.946472 0.002214 0.006782 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.00235909 23 0.00011271 27 0.00247180 Mean Square 0.000590 0.000005 0.000092 F Ratio 120.3524 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 8 0.002045 3 5 0.002750 10 5 0.002750 30 5 0.002750 100 5 0.026458 Std Error 0.00078 0.00099 0.00099 0.00099 0.00099 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.002045 0.000973 0.00034 5 0.002750 0.000000 0:00000 5 0.002750 0.000000 0.00000 5 0.002750 0.000000 0.00000 5 0.026458 0.005150 0.00230 Dif=Mean[i]-Mean] 100 3 10 30 0 Means Comparisons 100 3 0.000000 0.023708 -0.02371 0.000000 -0.02371 0.000000 -0.02371 0.000000 -0.02441 -0.00071 10 0.023708 0.000000 0.000000 0.000000 -0.00071 30 0.023708 0.000000 0.000000 0.000000 -0.00071 0 0.024413 0.000705 0.000705 0.000705 0.000000 Alpha= 0.05 Comparisons with a control using Dunnett's Method |d| 2.65107 Abs(Dif)-LSD 100 3 10 30 0 0 0.021067 -0.00264 -0.00264 -0.00264 -0.00293 Positive values show pairs of means that are significantly different. 124 0, 3 ,1 0 ,3 0 and 100 ug/Kg dose group Male sera PFOS Statistics, day 14 post-dose. Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations {or Sum Wgts) 0.94553 0.936057 0.011593 0.023214 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.05366259 23 0.00309140 27 0.05675399 Mean Square 0.013416 0.000134 0.002102 F Ratio 99.8122 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 8 0.002030 3 5 0.000870 10 5 0.006872 30 5 0.001998 100 5 0.117012 Std Error 0.00410 0.00518 0.00518 0.00518 0.00518 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.002030 0.002606 0.00092 5 0.000870 0.000000 0.00000 5 0.006872 0.003311 0.00148 5 0.001998 0.001030 0.00046 5 0.117012 0.027367 0.01224 Dif=Mean[i]-Mean[j] 100 10 0 30 3 Means Comparisons 100 10 0.000000 0.110140 -0.11014 0.000000 -0.11498 -0.00484 -0.11501 -0.00487 -0.11614 -0.006 0 0.114982 0.004842 0.000000 -0.00003 -0.00116 30 0.115014 0.004874 0.000032 0.000000 -0.00113 3 0.116142 0.006002 0.001160 0.001128 0.000000 Alpha= 0.05 Comparisons with a control using Dunnett's Method |d| 2.65107 Abs(Dif)-LSD 0 100 0.097460 10 -0.01268 0 -0.01537 30 -0.01749 3 -0.01636 Abs(Dif)-LSD 100 10 0 30 3 Positive values show pairs of means that are significantly different. Comparisons for each pair using Student's t t 2.06864 100 10 0 30 3 -0.01517 0.094972 0.101310 0.099846 0.100974 0.094972 -0.01517 -0.00883 -0.01029 -0.00917 0.101310 -0.00883 -0.01199 -0.01364 -0.01251 0.099846 -0.01029 -0.01364 -0.01517 -0.01404 0.100974 -0.00917 -0.01251 -0.01404 -0.01517 126 Positive values show pairs of means that are significantly different. Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.815667 0.781105 2.107108 9.872815 20 Source Model Error C Total Analysis of Variance DF Sum of Squares 3 314.34272 16 71.03845 19 385.38117 Mean Square 104.781 4.440 20.283 F Ratio 23.5998 Prob>F <.0001 Means for Oneway Anova Level Number Mean 1 5 13.3659 4 5 12.9850 14 5 9.6660 28 5 3.4745 Std Error 0.94233 0.94233 0.94233 0.94233 Level 1 4 14 28 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 5 13.3659 2.54032 1.1361 5 12.9850 1.02348 0.4577 5 9.6660 2.99223 1.3382 5 3.4745 1.14257 0.5110 Dlf=Mean[i]-Mean[j] 1 4 14 28 Means Comparisons 14 0.00000 0.38092 -0.38092 0.00000 -3.69993 -3.31901 -9.89141 -9.51049 14 3.69993 3.31901 0.00000 -6.19148 28 9.89141 9.51049 6.19148 0.00000 Alpha= 0.05 Comparisons for each pair using Student's t t 2.11989 Abs(Dif)-LSD 1 4 14 1 -2.82508 -2.44416 0.87485 4 -2.44416 -2.82508 0.49393 14 0.87485 0.49393 -2.82508 28 7.06633 6.68541 3.36640 28 7.06633 6.68541 3.36640 -2.82508 Positive values show pairs of means that are significantly different. 0. 3,10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics days 1 ,4 and 14 post dose. Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.497889 0.473396 0.046067 0.034905 87 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.17255803 82 0.17402099 86 0.34657902 Mean Square 0.043140 0.002122 0.004030 F Ratio 20.3277 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 27 0.009977 3 15 0.002249 10 15 0.012097 30 15 0.041684 100 15 0.128461 Std Error 0.00887 0.01189 0.01189 0.01189 0.01189 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 27 0.009977 0.027437 0.00528 15 0.002249 0.000861 0.00022 15 0.012097 0.008382 0.00216 15 0.041684 0.010700 0.00276 15 0.128461 0.104147 0.02689 Dif=Mean[i]-Mean[j] 100 30 Means Comparisons 100 30 0.000000 0.086777 -0.08678 0.000000 10 0.116364 0.029587 0 0.118484 0.031707 3 0.126213 0.039435 129 10 -0.11636 -0.02959 0.000000 0.002120 0.009849 0 -0.11848 -0.03171 -0.00212 0.000000 0.007729 3 -0.12621 -0.03944 -0.00985 -0.00773 0.000000 Alpha= 0.05 Comparisons with a control using Dunnett's Method 2.51855 Abs(Dif)-LSD 100 30 10 0 3 0 0.081121 -0.00566 -0.03524 -0.03158 -0.02963 Abs(Dif)-LSD 100 30 10 0 3 Positive values show pairs of means that are significantly different. Comparisons for each pair using Student's t t 1.98933 100 30 10 0 3 -0.03346 0.053314 0.082901 .0.088972 0.092749 0.053314 -0.03346 -0.00388 0.002195 0.005972 0.082901 -0.00388 -0.03346 -0.02739 -0.02361 0.088972 0.002195 -0.02739 -0.02494 -0.02178 0.092749 0.005972 -0.02361 -0.02178 -0.03346 Positive values show pairs of means that are significantly different. 0. 3,10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 1 post-dose. Sera PFOS (ug/mL) By DosePFOS(ug/Kg) DosePFOS(ug/Kg) 0-05 0.05 130 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.796351 0.760934 0.034689 0.043943 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.10822718 23 0.02767662 27 0.13590380 Mean Square 0.027057 0.001203 0.005033 F Ratio 22.4849 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 8 0.003026 3 5 0.002750 10 5 0.016848 30 5 0.049218 100 5 0.172426 Std Error 0.01226 0.01551 0.01551 0.01551 0.01551 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.003026 0.002452 0.00087 5 0.002750 0.000000 0.00000 5 0.016848 0.002193 0.00098 5 0.049218 0.005039 0.00225 5 0.172426 0.082936 0.03709 Dif=Mean[i]-MeanQ] 100 30 10 0 3 Means Comparisons 100 30 0.000000 0.123208 -0.12321 0.000000 -0.15558 -0.03237 -0.1694 -0.04619 -0.16968 -0.04647 10 0.155578 0.032370 0.000000 -0.01382 -0.0141 0 0.169400 0.046192 0.013822 0.000000 -0.00028 3 0.169676 0.046468 0.014098 0.000276 0.000000 Alpha= Abs(Dif)-LSD 100 30 10 0 3 0.05 Comparisons for each pair using Student's t + 100 -0.04538 0.077823 0.110193 0.128491 0.124291 2.06864 30 0.077823 -0.04538 -0.01301 0.005283 0.001083 10 0.110193 -0.01301 -0.04538 -0.02709 -0.03129 0 0.128491 0.005283 -0.02709 -0.03588 -0.04063 3 0.124291 0.001083 -0.03129 -0.04063 -0.04538 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method HI 2.65107 Abs(Dif)-LSD 0 100 0.116973 30 -0.00623 10 -0.0386 0 -0.04598 3 -0.05215 131 Positive values different. show pairs of means that are significantly 132 0. 3,10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 4 post-dose. _____________________ Sera PFOS (ug/mL) By DosePFOS(ug/Kg)_________________________ Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.847777 0.821304 0.033611 0.047661 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.14470536 23 0.02598254 27 0.17068790 Mean Square 0.036176 0.001130 0.006322 F Ratio 32.0237 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 8 0.002045 3 5 0.002750 10 5 0.018574 30 5 0.043866 100 5 0.198440 Std Error 0.01188 0.01503 0.01503 0.01503 0.01503 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.002045 0.000973 0.00034 5 0.002750 0.000000 0.00000 5 0.018574 0.001702 0.00076 5 0.043866 0.013220 0.00591 5 0.198440 0.079475 0.03554 Dif=Mean[i]-Mean|j] 100 Means Comparisons 100 30 0.000000 0.154574 10 0.179866 3 0.195690 0 0.196395 133 30 -0.15457 0.000000 0.025292 0.041116 0.041821 10 -0.17987 -0.02529 0.000000 0.015824 0.016529 3 -0.19569 -0.04112 -0.01582 0.000000 0.000705 0 -0.19639 -0.04182 -0.01653 -0.00071 0.000000 Alpha= Abs(Dif)-LSD 100 30 10 3 0 0.05 Comparisons for each pair using Student's t t 2.06864 100 30 10 -0.04397 0.110600 0.135892 0.110600 -0.04397 -0.01868 0.135892 -0.01868 -0.04397 0.151716 -0.00286 -0.02815 0.156758 0.002184 -0.02311 3 0.151716 -0.00286 -0.02815 -0.04397 -0.03893 0 0.156758 0.002184 -0.02311 -0.03893 -0.03476 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnetfs Method |d| 2.65107 Abs(Dif)-LSD 100 30 10 3 0 0 0.145598 -0.00898 -0.03427 -0.05009 -0.04455 Positive values show pairs of means that are significantly different. 134 0. 3,10, 30 and 100 ug/Kg dose group Female Sera PFOS Statistics - Day 14 post-dose. Sera PFOS (ug/mL) By DosePFOS(ug/Kg) Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or SumWgts) 0.540594 0.460697 0.011083 0.010945 28 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 0.00332427 23 0.00282502 27 0.00614929 Mean Square 0.000831 0.000123 0.000228 F Ratio 6.7662 Prob>F 0.0009 Means for Oneway Anova Level Number Mean 0 8 0.007930 3 5 0.001246 10 5 0.000870 30 5 0.031968 100 5 0.014518 Std Error 0.00392 0.00496 0.00496 0.00496 0.00496 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 8 0.007930 0.019969 0.00706 5 0.001246 0.000841 0.00038 5 0.000870 0.000000 0.00000 5 0.031968 0.002375 0.00106 5 0.014518 0.001448 0.00065 Means Comparisons Dif=Meanp]-Mean[j] 30 100 0 3 10 135 30 0.000000 0.017450 0.024038 0.030722 0.031098 100 -0.01745 0.000000 0.006588 0.013272 0.013648 0 -0.02404 -0.00659 0.000000 0.006684 0.007060 3 -0.03072 -0.01327 -0.00668 0.000000 0.000376 10 -0.0311 -0.01365 -0.00706 -0.00038 0.000000 Alpha= Abs(Dif)-LSD 30 100 0 3 10 0.05 Comparisons for each pair using Student's t t 2.06864 30 100 0 -0.0145 0.002950 0.010968 0.002950 -0.0145 -0.00648 0.010968 -0.00648 -0.01146 0.016222 -0.00123 -0.00639 0.016598 -0.00085 -0.00601 3 0.016222 -0.00123 -0.00639 -0.0145 -0.01412 10 0.016598 -0.00085 -0.00601 -0.01412 -0.0145 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method id! 2.61332 Abs(Dif)-LSD 10 30 0.012780 100 -0.00467 0 -0.00945 3 -0.01794 10 -0.01832 Positive values show pairs of mes different. 136 5000 pg/Kg dose group Female Sera PFOS Statistics. Sera PFOS (ug/mL) By Day Student's t Day 5000 ug/Kg dose group Female Rats Oneway Anova Summary of Fit RSquare RSquareAdj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.394958 0.281512 6.24891 13.17029 20 Source Model Error C Total Analysis of Variance DF Sum of Squares 3 407.8433 16 624.7821 19 1032.6253 Mean Square 135.948 39.049 54.349 F Ratio 3.4815 Prob>F 0.0407 Means for Oneway Anova Level Number Mean 1 5 18.8686 4 5 15.7357 14 5 11.0843 28 5 6.9925 Std Error 2.7946 2.7946 2.7946 2.7946 Level 1 4 14 28 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 5 18.8686 8.00137 3.5783 5 15.7357 9.33779 4.1760 5 11.0843 1.52170 0.6805 5 6.9925 1.63212 0.7299 Dif=Mean[i]-Mean[j] 1 4 14 28 Means Comparisons 14 0.0000 3.1329 -3.1329 0.0000 -7.7843 -4.6515 -11.8761 -8.7432 14 7.7843 4.6515 0.0000 -4.0918 28 11.8761 8.7432 4.0918 0.0000 Alpha= 0.05 Comparisons for each pair using Student's t t Abs(Dif)-LSD 1 4 14 28 2.11989 1 -8.37816 -5.24529 -0.59383 3.49793 4 -5.24529 -8.37816 -3.72670 0.36506 14 -0.59383 -3.72670 -8.37816 -4.28639 28 3.49793 0.36506 -4.28639 -8.37816 Positive values show pairs of means that are significantly different. Male Vs Female Serum PFOS Statistics 10 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose. Sera PFOS (ug/mL) By Gender 0.018 0.016 - 0.014 - e 0.012 - 3* c/3 0.010 o ^ 0.008 - W 0.006 - 0.004 - OO , ___ ,, F Gender M ..o Each P h r * * ' Student's t 0.05 o With C o M **S Dunnett's 0.05 139 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.962725 0.958065 0.001551 0.009799 10 Estimate Std Error Lower 95% Upper 95% t-Test Difference 0.014098 0.000981 0.011836 0.016360 t-Test OF Prob>|t| 14.374 8 <.0001 Assuming equal variances Analysis of Variance Source DF Sum of Squares Model 1 0.00049688 Error 8 0.00001924 C Total 9 0.00051612 Mean Square 0.000497 0.000002 0.000057 Means for Oneway Anova Level Number Mean F 5 0.016848 M 5 0.002750 Std Error 0.00069 0.00069 F Ratio 206.6188 Prob>F <.0001 Level F M Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 5 0.016848 0.002193 0.00098 5 0.002750 0.000000 0.00000 Means Comparisons Dif=Mean[i]-Mean[j] F F 0.000000 M -0.0141 M 0.014098 0.000000 Alpba= 0.05 Comparisons for each pair using Student's t t 2.30603 Abs(Dif)-LSD FM F -0.00226 0.011836 M 0.011836 -0.00226 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method |d| 2.30595 Abs(Dif)-LSD F M M 0.011836 -0.00226 Positive values show pairs of means that are significantly different. 140 30 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose. 0.06 ' Sera PFOS (ug/mL) By Gender 0.05 - - E ?> 0.03 - 0.02 ' 0.01 ' 0.00 --------- 1------- - FM Gender Each Fair Student's t 0 .0 5 With Contro} Dunnett's 0 .0 5 141 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or SumWgts) 0.981328 0.981265 0.003242 0.025796 300 Estimate Std Error Lower 95% Upper 95% t-Test Difference 0.046844 0.000374 0.046107 0.047581 t-Test 125.146 DF 298 Prob>|t| <.0001 Assuming equal variances Analysis of Variance Source DF Sum of Squares Model 1 0.16457703 Error 298 0.00313151 C Total 299 0.16770853 Mean Square 0.164577 0.000011 0.000561 F Ratio 15661.45 Prob>F <.0001 Means for Oneway Anova Level Number Mean F 150 0.049218 M 150 0.002374 Std Error 0.00026 0.00026 Level F M Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 150 0.049218 0.004522 0.00037 150 0.002374 0.000755 0.00006 Means Comparisons Dif=Mean[i]-MeanQ] F F 0.000000 M -0.04684 M 0.046844 0.000000 Alpha= 0.05 Comparisons for each pair using Student's t t 1.96798 Abs(Dif)-LSD FM F -0.00074 0.046107 M 0.046107 -0.00074 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method !d| 1.96796 Abs(Dif)-LSD F M M 0.046107 -0.00074 Positive values show pairs of means that are significantly different. 42 100 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose. Sera PFOS (ug/mL) By Gender 143 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.655294 0.654949 0.052588 0.099991 1000 Estimate Std Error Lower 95% Upper 95% t-Test Difference 0.144870 0.003326 0.138343 0.151397 t-Test 43.557 DF 998 Prob>|t| <.0001 Assuming equal variances Analysis of Variance Source DF Sum of Squares Model 1 5.2468292 Error 998 2.7600026 C Total 999 8.0068318 Mean Square 5.24683 0.00277 0.00801 F Ratio 1897.221 Prob>F <.0001 Means for Oneway Anova Level Number Mean F 500 0.172426 M 500 0.027556 Std Error 0.00235 0.00235 Level F M Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 500 0.172426 0.074255 0.00332 500 0.027556 0.004158 0.00019 Means Comparisons Dif=Mean[i]-Mean[j] F F 0.000000 M -0.14487 M 0.144870 0.000000 Alpha= 0.05 Comparisons for each pair using Student's t t 1.96238 Abs(Dif)-LSD FM F -0.00653 0.138343 M 0.138343 -0.00653 Positive values show pairs of means that are significantly different Comparisons with a control using Dunnett's Method |d| 1.96234 Abs(Dif)-LSD F M M 0.138343 -0.00653 Positive values show pairs of means that are significantly different. 5000 ug/Kg dose group male vs female Serum PFOS statistics, day 1 post-dose. Sera PFOS (ug/roL) By Gender 30 * 25 20 * O 15- 10 - Gender Each Pair Student's t 0.05 With Control Dunnett's 0.05 145 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.211689 0.211673 5.309536 16.11724 50000 Estimate Std Error Lower 95% Upper 95% t-Test Difference 5.50273 0.04749 5.40965 5.59581 t-Test DF 115.871 49998 Prob>|t| 0.0000 Assuming equal variances Analysis of Variance Source DF Sum of Squares Model 1 378500.2 Error 49998 1409502.2 C Total 49999 1788002.4 Mean Square 378500 28 36 F Ratio 13426.2 Prob>F 0.0000 Means for Oneway Anova Level Number Mean F 25000 18.8686 M 25000 13.3659 Std Error 0.03358 0.03358 Level F M Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 25000 18.8686 7.15678 0.04526 25000 13.3659 2.27217 0.01437 Means Comparisons Dif=Mean[iJ-Mean[j] F F 0.00000 M -5.50273 M 5.50273 0.00000 Alpha= 0.05 Comparisons for each pair using Student's t t 1.96005 Abs(Dif)-LSD FM F -0.09308 5.40965 M 5.40965 -0.09308 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method |d| 1.95996 Abs(Dif)-LSD M F 5.40965 M -0.09308 Positive values show pairs of means that are significantly different. 146 147 Male and female liver PFOS concentrations SAS analysis data: results KJH10.15.99 liverPFOS for Wirmonlin LOD substitued Dose(ug/Kg) Group Day Gender Animal# LPFOS (ug/g) 0 1 1M 3778 BLOQ 0 1 1M 3779 BLOQ 0 1 1M 3780 BLOQ 0 1 1M 3781 BLOQ 0 1 1M 3782 BLOQ 0 1 4M 3783 NA 0 1 4M 3784 NA 0 1 4M 3785 NA 0 1 4M 3786 NA 0 1 4M 3787 NA 0 1 14 M 3788 <LOD 0 1 14 M 3789 <LOD 0 1 14 M 3790 <LOD 0 1 14 M 3791 <LOD 0 1 14 M 3792 <LOD 3 2 1M 3668 0.139 3 2 1M 3669 BLOQ 3 2 1M 3670 0.222 3 2 1M 3671 BLOQ 3 2 1M 3672 BLOQ 3 2 4M 3673 BLOQ 3 2 4M 3674 BLOQ 3 2 4M 3675 BLOQ 3 2 4M 3676 BLOQ 3 2 4M 3677 BLOQ 3 2 14 M 3678 0.0349 3 2 14 M 3679 0.0299 3 2 14 M 3680 0.049 3 2 14 M 3681 0.039 3 2 14 M 3682 0.0183 30 3 1M 3683 0.375 30 3 1M 3684 0.407 30 3 1M 3685 0.496 30 3 1M 3686 0.351 30 3 1M 3687 0.417 30 3 4M 3688 0.635 30 3 4M 3689 0.419 30 3 4M 3690 0.362 30 3 4M 3691 0.398 30 3 4. M 3692 0.417 30 3 14 M 3693 0.361 30 3 14 M 3694 0.319 30 3 14 M 3695 0.324 30 3 14 M 3696 0.303 30 3 14 M 3697 0.346 100 4 1M 3793 1.060 100 4 1M 3794 1.360 100 4 1M 3795 1.660 100 4 1M 3796 1.420 100 4 1M 3797 1.450 ' 100 4 4M 3798 1.220 100 4 4M 3799 1.450 100 4 4M 3800 1.250 100 4 4M 3801 1.430 100 4 4M 3802 1.400 100 4 14 M 3803 0.984 100 4 14 M 3804 1.030 LPFOScloq 0.07 0.07 0.07 0.07 0.07 ? ? ? ? ? 0.005 0.005 0.005 0.005 0.005 0.139 0.07 0.222 0.07 0.07 0.07 0.07 0.07 0.07 0.07 0.0349 0.0299 0.049 0.039 0.0183 0.375 0.407 0.496 0.351 0.417 0.635 0.419 0.362 0.398 0.417 0.361 0.319 0.324 0.303 0.346 1.06 1.36 1.66 1.42 1.45 1.22 1.45 1.25 1.43 1.4 0.984 1.03 148 100 100 100 10 10 10 10 10 10 10 10 10 10 1010 10 10 10 o o o o o o o o o o o o 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 0 0 0 0 0 0 0 0 o o o o 14 M 14 M 14 M 1M 1M 1M 1M 1M 4M 4M 4M 4M 4M 14 M 14 M 14 M 14 M 14 M 1M 1M 1M 4M 4M 4M 14 M 14 M 14 M 28 M 28 M 28 M 1M 1M 1M 1M 1M 4M 4M 4M 4M 4M 14 M 14 M 14 M 14 M 14 M 28 M 28 M 28 M 28 M 28 M 1F 1F 1F 1F 1F 4F 4F 4F 4F 4F 14 F 14 F 3805 3806 3807 3808 3809 3810 3811 3812 3813 3814 3815 3816 3817 3818 3819 3820 3821 3822 1205 1206 1207 1208 1209 1210 1486 1487 1488 1489 1490 1491 1211 1212 1213 1214 1215 1216 1217 1218 1219 1220 1221 1222 1223 1224 1225 1226 1227 1228 1229 1230 3825 3826 3827 3828 3829 3830 3831 3832 3833 3834 3835 3836 0.806 0.766 0.761 0.139 0.212 0.170 0.162 0.137 0.176 0.208 0.160 0.190 0.176 0.092 0.082 0.048 0.088 0.088 0.025 0.049 <10Q <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ <LOD 36.5 38.9 32.1 34.8 25.6 37 25.8 28.5 23.4 22.8 44.6 57.9 27.7 61.7 22.7 32.1 26.9 31 30.5 34.3 1.83 0.247 BLOQ BLOQ BLOQ BLOQ BLOQ BLOQ BLOQ BLOQ <LOQ <LOD 0.806 0.766 0.761 0.139 0.212 0.17 0.162 0.137 0.176 0.208 0.16 0.19 0.176 0.092 0.082 0.048 0.088 0.088 0.025 0.049 0.003 0.003 0.003 0.003 0.019 0.019 0.019 0.019 0.019 0.019 36.5 38.9 32.1 34.8 25.6 37 25.8 28.5 23.4 22.8 44.6 57.9 27.7 61.7 22.7 32.1 26.9 31 30.5 34.3 1.83 0.247 0.07 0.07 0.07 0.07 0.07 0.07 0.07 0.07 0.005 0.005 149 o^ o o o ^o o -o^ o o- o" ok o o^ o` A o^o oo, a oo- ` oo- * -oo, ioo, u oo-J ,oo- 4 oo" 4 oo' x oo- * oo- i oo*J `oo' Aoo-a oW oW oC OoO oJ WoWoWoW o( Ao) WoWoOo) CoA oJ W W ( J O ) 0 ) W W W O H * J W O J Q 0 3 W W W O O O U I\Ul\l lU 1lOit^liyilA^^-t-^4.A4i`A ^ ^ ^ Ji.ii^ W W W C )W 0 3 W W W W W W W W C O rO h JfO fO N )N )M N JN JN fO N M fO N i' 4*.4*>4* 4^ .4* -t*4* -I** 4* 4* 4* -4- 4* 4* 4*.4* 4*.A 4>. 4x 4^ 4^ 4 4. 4> 4^ 4* 4 * . ............. 4*. 4^ 4> T i - n T i - n ,T i - T i * n - T i - n ' n T i "n - n T i - n - T 'n * T i - n * n ' n "n ' n 7 m i T T T n - n T i - n ' n - n `n " T i * m m T i - n T i - n ' m i i i T ) - r ] - n - n * n ` m i - n ' n i i * n T i T n m - n ' n OOOOOOOO' bcnocjbiconco-^-^A^i CJ) N) O (D <0 O) NJ NO 05 -*1 t_/> o OOOoooo b 4x -*4 "4 NO en en en en b roo en en 4* CO 4- en o CD -* O CO b CD o 4 oPopOO CO o '**4 CD CO b 4 CD. e-an CO CD C*O0 oA 4 O !a O) O) O y en en pp 1a -A Oco en p 1a en 03 ooo N00O *- -4 co NO O) ooo k> co io en o e NO -"4 4* pK00o Kpo -* o Ko o> oOo rCoONbObro o bro o b NO *vl CD en *"4 *"4 NO CO Pb NO -a en NO -O l1 'J N `bJ N bS N Nb N bS QbiObenOben 10 0 0 0 0 0 0 0 o0 0 0 0 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5000 5 14 F 6 1F 6 1F 6 1F 6 4F 6 4F 6 4F 6 14 F 6 14 F 6 14 F 6 28 F 6 28 F 6 28 F 7 1F 7 1F 7 1F 7 1F 7 1F 7 4F 7 4F 7 4F 7 4F 7 4F 7 14 F 7 14 F 7 14 F 7 14 F 7 14 F 7 28 F 7 28 F 7 28 F 7 28 F 7 28 F 3869 1231 232 1233 1234 1235 1236 1257 **1492 **1493 1494 1495 1496 1237 1238 1239 1240 1241 1242 1243 1244 1245* 1246* 1247 1248 1249 1250 1251 1252 1253 1254 1255 1256 0.0531 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 33.50 41.50 32.90 42.90 37.50 30.30 30.30 36.30 64.10 0.51 43.50 34.00 33.10 32.50 57.60 12.30 23.20 30.10 13.80 25.40 0.0531 0.00294 0.00294 0.00294 0.00294 0.00294 0.00294 0.0185 0.0185 0.0185 0.0185 0.0185 0.0185 33.5 41.5 32.9 42.9 37.5 30.3 30.3 36.3 64.1 0.51 43.5 34 33.1 32.5 57.6 12.3 23.2 30.1 13.8 25.4 151 0, 3,10, 30 and 100 ug/Kg dose group Male Liver PFOS Statistics days 1,4, and 14 post' dose. L PFOS (ug/g) LOQ/2 By Dose(ug/Kg) Oneway Anova Summary of Fit RSquare RSquareAdj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.919401 0.915214 0.130889 0.338684 82 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 15.047737 77 1.319149 81 16.366886 Mean Square 3.76193 0.01713 0.20206 F Ratio 219.5877 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 22 0.02614 3 15 0.07281 10 15 0.14187 30 15 0.39533 100 15 1.20313 Std Error 0.02791 0.03380 0.03380 0.03380 0.03380 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 22 0.02614 0.026569 0.00566 15 0.07281 0.049758 0.01285 15 0.14187 0.050837 0.01313 15 0.39533 0.082704 0.02135 15 1.20313 0.285072 0.07361 Dif=Mean[i]-Mean[j] 100 30 10 3 Means Comparisons 100 30 0.00000 0.80780 -0.80780 0.00000 -1.06127 -0.25347 -1.13033 -0.32253 10 1.06127 0.25347 0.00000 -0.06906 3 1.13033 0.32253 0.06906 0.00000 0 1.17700 0.36920 0.11573 0.04667 152 0 -1.17700 -0.36920 -0.11573 -0.04667 0.00000 Alpha= Abs(Dif)-LSD 100 30 10 3 0 0.05 Comparisons for each pair using Student's t t 1.99126 100 30 10 -0.09517 0.71263 0.96610 0.71263 -0.09517 0.15830 0.96610 0.15830 -0.09517 1.03516 0.22736 -0.02611 1.08973 0.28193 0.02846 3 1.03516 0.22736 -0.02611 -0.09517 -0.04060 0 1.08973 0.28193 0.02846 -0.04060 -0.07858 Positive values show pairs of means that are significantly different. Comparisons with a control using Dunnett's Method |d| 2.51374 Abs(Dif)-LSD 100 30 10 3 0 0 1.06683 0.25903 0.00556 -0.06350 -0.09920 Positive values show pairs of means that are significantly different. 5000 ug/Kg dose group Male Liver PFOS Statistics _____________ L PFOS (ug/g) LOQ/2 By Day_____________ Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response 0.305761 0.175591 9.639723 33.74 153 Observations (or Sum Wgts) 20 Source Model Error C Total Analysis of Variance DF Sum of Squares 3 654.8200 16 1486.7880 19 2141.6080 Mean Square 218.273 92.924 112.716 F Ratio 2.3489 Prob>F 0.1111 Means for Oneway Anova Level Number Mean 1 5 33.5800 4 5 27.5000 14 5 42.9200 28 5 30.9600 Std Error 4.3110 4.3110 4.3110 4.3110 Level 1 4 14 28 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 5 33.5800 5.1036 2.2824 5 27.5000 5.7671 2.5791 5 42.9200 17.4672 7.8116 5 30.9600 2.6996 1.2073 Dif=Mean[i]-Mean[j] 14 1 28 4 Means Comparisons 14 1 0.0000 9.3400 -9.3400 0.0000 -11.9600 -2.6200 -15.4200 -6.0800 28 11.9600 2.6200 0.0000 -3.4600 4 15.4200 6.0800 3.4600 0.0000 Alpha= 0.05 Comparisons for each pair using Student's t + Abs(Dif)-LSD 14 1 28 4 2.11989 14 -12.9244 -3.5844 -0.9644 2.4956 1 -3.5844 -12.9244 -10.3044 -6.8444 28 -0.9644 -10.3044 -12.9244 -9.4644 4 2.4956 -6.8444 -9.4644 -12.9244 Positive values show pairs of means that are significantly different. 154 0, 3,10, 30 and 100 ug/Kg dose group Female Liver PFOS Statistics days 1 ,4 and 14 post dose. L PFQS (ug/g) LOQ/2 By Dose(ug/Kg) _Cor 1' :O COO _ <$> O i~ O J--------- ------ ------ j----- i----- 0 3 10 30 100 Dose(ug/Kg) Dunnett's 0.05 155 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.569623 0.548629 0.224716 0.227654 87 Source Model Error C Total Analysis of Variance DF Sum of Squares 4 5.4805073 82 4.1407832 86 9.6212905 Mean Square 1.37013 0.05050 0.11188 F Ratio 27.1326 Prob>F <.0001 Means for Oneway Anova Level Number Mean 0 27 0.103357 3 15 0.054300 10 15 0.088853 30 15 0.226667 100 15 0.764533 Std Error 0.04325 0.05802 0.05802 0.05802 0.05802 Level 0 3 10 30 100 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 27 0.103357 0.348784 0.06712 15 0.054300 0.023026 0.00595 15 0.088853 0.042971 0.01109 15 0.226667 0.060218 0.01555 15 0.764533 0.252676 0.06524 Dif=Mean[i]-Mean[j] 100 30 0 10 3 Means Comparisons 100 30 0.000000 0.537867 -0.53787 0.000000 -0.66118 -0.12331 -0.67568 -0.13781 -0.71023 -0.17237 0 0.661176 0.123310 0.000000 -0.0145 -0.04906 10 0.675680 0.137813 0.014504 0.000000 -0.03455 3 0.710233 0.172367 0.049057 0.034553 0.000000 Alpha= 0.05 Comparisons with a control using Dunnett's Method |d| 2.51855 Abs(Dlf)-LSD 100 30 0 10 0 0.478920 -0.05895 -0.15403 -0.16775 3 -0.1332 Positive values show pairs of means that are significantly different. 156 5000 ug/Kg dose group Female Liver PFOS Statistics ______________L PFOS (ug/g) LOQ/2 By Day_____________ Student's t Day 0.05 Oneway Anova Summary of Fit RSquare RSquare Adj Root Mean Square Error Mean of Response Observations (or Sum Wgts) 0.278393 0.143092 13.28613 32.7655 20 Source Model Error C Total Analysis of Variance DF Sum of Squares 3 1089.6202 16 2824.3401 19 3913.9603 Mean Square 363.207 176.521 205.998 F Ratio 2.0576 Prob>F 0.1463 Means for Oneway Anova Level Number Mean 1 5 37.6600 4 5 32.3020 14 5 40.1400 28 5 20.9600 Std Error 5.9417 5.9417 5.9417 5.9417 Level 1 4 14 28 Std Error uses a pooled estimate of error variance Means and Std Deviations Number Mean Std Dev Std Err Mean 5 37.6600 4.5330 2.027 5 32.3020 22.6155 10.114 5 40.1400 10.7444 4.805 5 20.9600 7.6572 3.424 Dif=Mean[l]-Meanfj] 14 1 4 28 Means Comparisons 14 1 0.0000 2.4800 -2.4800 0.0000 -7.8380 -5.3580 -19.1800 16.7000 4 7.8380 5.3580 0.0000 -11.3420 28 19.1800 16.7000 11.3420 0.0000 Alpha= 0.05 Comparisons for each pair using Student's t t 157 Abs(Dif)-LSD 14 1 4 28 2.11989 14 -17.8132 -15.3332 -9.9752 1.3668 1 -15.3332 -17.8132 -12.4552 -1.1132 4 -9.9752 -12.4552 -17.8132 -6.4712 28 1.3668 -1.1132 -6.4712 -17.8132 Positive values show pairs of means that are significantly different. 158