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BACK TO MAIN Laboratory Report-Revision 1 Analytical Report of Data for PFOS Dietary LC50 Study in Northern Bobwhite (Sera and Liver) Laboratory Report No. FACT-TOX-129 (U2723) Testing Laboratory 3M Environmental Technology & Safety Services 3M Environmental Laboratory Fluorine Analytical Chemistry Team (FACT) 2-3E-09 935 Bush Avenue, St. Paul, MN 55106 Laboratory Contact Kris Hansen, Ph.D. Bldg. 2-3E-09 P.O. Box 3331 St. Paul, MN 55133-3331 Phone: (651)778-6018 FAX: (651) 778-6176 Revision Date: March 19, 2001 Page 1 o f / J LAC O2jloi BACK TO MAIN 1 Introduction This study was designed to quantitatively assess perfluorooctane sulfonate (PFOS, C8F |7S 0 3\ CAS # 2795 39-3) levels in the sera and liver o f the northern bobwhite (Colinus virginianus) exposed to PFOS through diet. Details o f dosing conditions, clinical observations and other toxicity data were generated by the in-life testing facility (Wildlife International) and are not included in this report. Subsets of the eight groups o f test animals were sacrificed on day 8 and day 22, post-dose. Sera and liver collected from the sacrificed animals were sent to the 3M Environmental Laboratory and analyzed quantitatively for PFOS. Tissues from animals that died before the scheduled sacrifice date were collected and analyzed also; results are included in this report. Elucidation o f data trends was complicated by the sporadic nature o f the PFOS levels in sera. Within a particular dose group there was dramatic variation between individual animals. The relative standard deviation o f PFOS sera levels within a dose group ranged from greater than 20% to over 200%. By comparison, the results o f PFOS levels in the livers of the bobwhites were far more consistent within a particular dose level (group RSD between 10-55%) In general, both sera and liver levels o f PFOS increased with dose level, although due to animal to animal variability, this trend is not pronounced in the sera. With the exception o f group 4 animals, average levels o f PFOS in the sera o f the bobwhites generally increased or stayed the same between day 8 and day 22. In all dose groups, the group average for PFOS concentration in liver decreased dramatically between day 8 and day 22. Livers were collected from bobwhites that died between day 1 and day 8, prior to the scheduled sacrifice. In every case, PFOS levels in the livers o f these birds were over 110 pg/g wet weight. Although rigorous quality control measures and 3M Environmental Laboratory Standard Operating Procedures were followed, the acquisition o f this data was not necessarily collected according to applicable Good Laboratory Practices. Data presented here is the highest quality data available at this time. No data was collected for any analytes other than PFOS. It is recommended that bobwhite quail be avoided in future studies because the small amount o f tissue available causes analytical challenges that make verification difficult. 2 Sample Receipt ; - ' . ^ The Environmental Laboratory received 65 liver, 34 blood, and 34 serum specimens, frozen, on 05/19/99. Upon specimen receipt a 3M chain of custody was initiated. The Environmental Laboratory received 35 liver, 35 blood, 35 serum, and 6 bile specimens, frozen, on 06/29/99. Upon specimen receipt a 3M chain o f custody was initiated. The Environmental Laboratory received 30 pooled bobwhite quail serum tubes, frozen, on 06/30/99. Upon specimen receipt a 3M chain o f custody was initiated. A copy o f each form can be found in an Attachment to this report. 3 Holding Times There is no holding time criteria associated with these samples for LC/ESMSMS analysis. Page 2 of4 3 LAC o BACK TO MAIN 4 Methods - Analytical and Preparatory Analytical methods ETS-8-4.1, Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollow ing exceptions: Due to the small volume of bobwhite sera available, standard curves were prepared in methanol instead o f from bobwhite sera. Four matrix spikes were prepared from the pooled bobwhite quail serum received on 06/30/99. PFOS lot 171 was used as reference material. FACT-M -1.1, Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollow ing exception: The extraction was performed following ETS-8-6.0; this method had been validated but not approved at the time o f extraction. PFOS lot 171 was used as reference material. ETS-8-6.0, Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollow ing exception: Due to the small amount o f bobwhite liver available, standard curves were prepared in methanol instead o f from bobwhite liver. In some cases, liver samples were very limited and two or more samples had to be combined for extraction. These combinations are indicated in the results. Twenty-two matrix spikes were prepared from the bobwhite liver. PFOS lot 171 was used as reference material. ETS-8-5.1, Analysis o f Potassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts Using HPLC-Electrospray/Mass Spectrometry. FACT-M-2.1, Analysis of Potassium Perfluorooctanesulfonate or Other Fluorochemicals on Liver Extracts Using HPLC-Electrospray/Mass Spectrometry with thefollow ing exception: The analysis was performed following ETS-8-7.0, which had been validated but not approved at the time of extraction. As described in the method, all results are reported in terms o f pg o f analyte per gram o f liver (wet weight). ETS-8-7.0, Analysis o f Potassium Perfluorooctanesulfonate or Other Fluorochemicals on Liver Extracts Using HPLC-Electrospray/Mass Spectrometry. Sample preparation - HPLC/ESMSMS: ion-pairing extraction Analyte is extracted from a sample matrix with ion pairing reagent [tetrabutyl ammonium hydrogen sulfate (TBA)] in a pH-controlled environment The cationic reagent selectively targets anionic fluorochemicals. Once the anion-TBA pair is formed, the analyte is transferred into a non-polar organic solvent (methyl-ierfbutyl ether), dried, and reconstituted in methanol for MS analysis. H PL C /E SM SM S In HPLC, an aliquot o f extract is injected and passed through a reverse phase liquid chromatographic column. Based on the affinity o f the analyte for the stationary phase in the column relative to the liquid mobile phase, die analyte is retained for a characteristic amount o f time. For example, in a standard solution PFOS may elute at 8.0 minutes. Retention times between a standard PFOS solution and die analyte extracted from tissue in this analysis were matched on the HPLC system to within 1%. Following HPLC separation, ESMSMS provides a rapid and accurate means for analyzing a wide range of organic compounds, including fluorochemicals. Electrospray, an ionization technique used primarily for detection o f molecular ions, is generally operated at relatively mild temperatures. Molecules are ionized, and a primary ion, characteristic o f the analyte, is selected. This ion is bombarded with high-energy gas; subsequent collisions create smaller secondary ionic fragments unique to the primary ion, which are detected. For example, for PFOS (CSF 17S 0 3`) analysis, ion 499 is selected as the characteristic primary ion. This ion is fragmented into other ions such as 80 amu (corresponding to S 0 3'), and 230 amu (C^FSOf). Each o f these secondary fragments is detected and can be used to differentiate PFOS from other compounds that might have the same characteristic 499 amu primary ion, but different chemical compositions and secondary ion fragmentation patterns. Page 3 ofji? Uc 03 ilk. BACK TO MAIN 5 Analysis 5.1 Calibration A mid-level calibration check standard is analyzed every 5-10 samples to monitor instrumental drift Calibration check standards were compliant if instrumental response was within +/- 30% of the expected value. Quantitation o f the target analytes was based on linear regression analysis (weighted 1/x) o f two unextracted methanol curves bracketing each group of samples. Quantitation of each analyte was based on the response o f one or more specific daughter ions using the multiple response-monitoring mode o f the instrument 3M lot #217 (purity 86.9%) o f PFOS was used for calibration standards. As purity information was not available at the time o f data collection, reported results have not been corrected. 5.2 Extraction and Method Blanks A t least two extraction blanks, utilizing water as surrogate matrix, were extracted with each batch o f samples. Additionally, two method blanks, utilizing control sample matrix (if available), were extracted along with each batch o f samples and curves. Blanks were compliant if no target analyte was detected above the limit o f detection for a specific analyte. In this study, all blanks were compliant. 5.3 Surrogates Tetra-hydroperfluorooctanesulfonate was used as a surrogate in this analysis. Surrogate response was monitored to confirm gross instrumental failure, if necessary. Surrogate response was acceptable for all analyses. 5.4 Matrix Spikes Over the course o f the analysis, twenty-six matrix spikes were prepared in the bobwhite liver matrix. All of the matrix spikes were recovered at greater titan 50% o f the expected value with two exceptions. The two 50 ng/g matrix spikes that were prepared on September 24,1999 with the day 3-7 liver samples showed analyte recovery o f approximately 350% o f the expected value. These samples and matrix spikes were re extracted on October 08,1999. The re-extracted matrix spike recoveries were greater than 50% (104% 110%), therefore day 3-7 data are reported from extraction date October 08,1999 - refer to extraction sheet for animal numbers. Because the bobwhite sera were so limited, only four matrix spike samples were prepared. All matrix spikes showed analyte recovery of greater than 90% o f the expected value. 5.5 Laboratory Control Samples Laboratory Control Samples were not a component o f this study. 5.6 Sample Related Comments Samples were not analyzed for any residual or potential metabolite other than PFOS. Other matrices submitted with this study will be analyzed as needed upon written request from the requestor. In the event o f further analysis, an additional report will be generated. Page 4 of$ 1 Uc'O t/23loi BACK TO MAIN 6 Data Summary ;j Elucidation of data trends was complicated by the sporadic nature o f the PFOS levels in sera. Within a particular dose group there was dramatic variation between individual animals. The relative standard deviation o f PFOS sera levels within a dose group ranged from greater than 20% to over 200%. By comparison, the results o f PFOS levels in the livers of the bobwhites were far more consistent within a particular dose level (group RSD between 10-55%) In general, both sera and liver levels increased with dose level, although due to animal to animal variability, this trend is not pronounced in the sera. With the exception of group 4 animals, average levels o f PFOS in the sera o f the bobwhites generally increased or stayed the same between day 8 and day 22. In all dose groups, the group average for PFOS levels in liver decreased dramatically between day 8 and day 22. Livers were collected from bobwhites that died between day 1 and day 7, prior to the scheduled sacrifice. In every case, PFOS levels in the livers of these birds were over 110 pg/g. Data Quality Objectives ah< ia lutei N o circumstances existed during the present study that would have adversely affected the quality or integrity o f the data. The following data quality objectives were followed during the study: % Linearity - The coefficient of determination (r^2) o f the standard curve was equal to or greater than 0.985 using 1/x weighting Limits o f quantitation - PFOS in sera is 0.010 pg/mL; PFOS in liver is approximately 0.070 pg/g Acceptable duplicate precision - < 30%, assessed by analysis o f the MS/MSDs. Acceptable spike recoveries - sera: 70% to 130%; liver: 50% to 150% Use o f confirmatory methods - Given the selectivity o f the analytical tool used (HPLC-ESMSMS) and lack o f a viable alternative for analysis, no confirmatory methods were used. Demonstration o f specificity - Specificity was demonstrated by chromatographic retention time (matched to standards to within 3%) and the response of at least one characteristic product ion arising from collisions o f an analyte-specific parent ion. % The characterization of the reference material (PFOS, lot #171), with respect to purity, stability, and homogeneity, was not complete on the date this report was issued. Once the characterization is complete, a copy o f the report will on file in die 3M Environmental Lab archives. Given the parameters listed above and assuming spike recoveries form a suitable indication o f endogenous analyte recovery, sera data are quantitative to +/-30 % and liver data are quantitative to +/-50 %. The validity o f these assumptions has not been verified by other techniques. If a greater degree o f accuracy is required, additional matrix should be provided so samples can be evaluated versus a standard curve extracted from the same matrix as the samples. It is recommended that bobwhite quail be avoided in future studies because the small amount o f tissue available causes analytical challenges. Page 5 of# 1 IAC. o j / j j /o, 8 Attachments Attachment A: Table of results of liver analysis Attachment B: Table o f results o f sera analysis Attachment C: Sample Receipt Attachment D: In-life protocol from Wildlife Int., LTD Attachment E: Extraction and analytical data BACK TO MAIN Page 6 off ? BACK TO MAIN 9 Signatures y 'C/r>.^';Xs <?; _ \4* ; *~iii . .vlzk * ?:.; <4 p j ^ Lisa Clemen, Advanced Chemist _____ -- ___________________________ Kris Hansen, PhD ., Principal Analytical Investigator 173/ 33)01 Date 3 I2 3 J0 I Date Quality Assurance Unit Representative Date Page 7 of^ BACK TO MAIN 9 Signatures A/ *i *__ C1 ii.rtv.''_________________________________________________________________QM bt Ullur UlcX_______________ Lisa Clemen, Advanced Chemist Date - b k i .-------Kris Hansen, Ph.D., Principal Analytical Investigator V --...... -- Quality Assurance Unit Representative 0C / o t /< ro Date Of - *if~C>o Date f * e ,w v 0, \ ^ ' BACK TO MAIN Laboratory Report Analytical Report of Data for PFOS Dietary LC50 Study in Northern Bobwhite (Sera and Liver) Laboratory Report No. FACT-TOX-129 (U2723) Attachment A Table of Results - Liver Analysis Day 3-7 Day 8 Day 22 FACT-TOX-129 BACK TO MAIN Study: Product Numbcr<Test Substance): Matrix: Method/Re vision: Analytical Equipment System N umber Instrument Software/Version: Date o f Extraction/Analyst: Date o f Analysis/Analyst Date o f Data Reduction/Analyst Sample Data . . 454-103 Bobwhite Quail LC50 PFOS Bobwhite Quail Liver ETS-8-6.0 & ETS-8-7.0 Amelia 062498 Masslynx 3.3 10/08/99 SEE/MCH 10/21/99,01/18/00 IAS/MMH 10/22/99,01/19/00 IAS/MMH Filename: R-Squared Value: Slope: Y-Intercept See Attachments See Attachments See Attachments See Attachments BOBWHITE LIVER DAY 3-7 Group Dose Sample # Method Blk BWL10089-H20 Blk-11 QC * 50 ppb BWL10089-H20 Blk-12 R342--0-Day8-50ppb-MS-11 R342-0-Day8-50ppb-MSD-l 1 QC - lOOOppb B439-0-Day8-lppm-MS-12 B439-0-Day8-1ppm-MSD-12 Groupl R342-0-Day8 0.0 mg/kg food GroupS 146 mg/kg food B439-0-Day8 G355-Day7 Group6 R352, B450-Day5 293 mg/kg food Y417, R353-Day6 G357, G358-Day7 B449, Y416-Day7 Group7 586 mg/kg food AL354-Day3 R354-Day4 R355, Y401, B452-Day5 Y418, B451,G360-Day6 G359, AL355-Day7 Group8 AL356, R357, Y419-Day3 1171 mg/kg food AL357, B453, G361-Day4 B454, G362-Day4 R356, Y420-Day4 PFOS = Perfluorooctanesulfonate Limit of Quantitation (LOQ): PFOS is approximately 0.070 ug/g NA = not applicable G1 Day 8 data is included to support QC NE = not extracted Date Entered/By: 01/18/00, 01/26/00 LAC Date Verified/By: 04/12/00 KJH PFOS Calc. Cone, ng/g 0.00 0.00 68.7 64.6 980 1034 229 383 110527 248693 201523 153461 123521 NE . NE 125957 235179 111299 133852 131788 122934 124333 Concentration of PFOS ug/g or % Ree. <LOQ <LOQ 110% 104*/. 82% 86% 0.229 0.383 111 249 202 153 124 NE NE 126 235 111 134 132 123 124 Mean PFOS ug/g or %Rec <LOQ 107% 84% 0.306 111 182 157 128 RSD (%) Std. Dev.(ug/g) MS/MSD RPD NA 6% 5% 35.7 0.109 NA 30.2 55.0 43.0 67.7 4.21 5.40 FACT-M-2.1 Excel Version 5/95 TOX129 liver-3.xls 3/8/01 9:40 AM FACT-TOX-129 BACK TO MAIN Study: Product Number{Test Substance): Matrix: Method/Revision: Analytical Equipment System N umber Instrument Software/Version: Date of Extraction/Analyst: Date of Analysis/Analyst; Date of Data Reduction/Analyst: Sample Data * , 454-103 Bobwhite Quail LC50 PFOS Bobwhite Quail Liver FACT-M-1.1 and FACT-M-2.1 Amelia 062498 Masslynx 3.2 06/30/99 SAH 07/02/99.07/22/99,09/01/99 MEE/PTF 07/20/99, 07/23/99, 09/02/99 DRB/MEE/PTF Filename: R-Squartd Value: Slope: Y-Interccpt: See Attachments See Attachments See Attachments See Attachments BOBWHITE LIVER DAY 8 Group Dose Sample # Method Blk BWL-H20 Blk-6/30-1 BWL-H20 Blk-6/30-2 , BWL-H20 Blk-6/30-3 BWL-H20 Blk-6/30-4 QC - 1000 ppb G347-MS-1 G347-MSD-I Y405-MS-2 Y405-MSD-2 QC - 50 ppb G346-0-MS11-1 G348-0-MS11-2 G347-0-MS11-3 Y405-0-MS11-4 QC - 1000 ppb G346-0-MS11-1 G348-0-MS11-2 G347-0-MS11-3 Y405-0-MS11-4 Groupl G347 0.0 mg/kg food B438/AL341 Y405 AL343 B440 Y406 Y407 R341 R342 B439 G346/R346 Day6 G348/AL342 Group2 B442/AL345 18.3 mg/kg food R340 G350/Y409 Group3 Y411 36.6 mg/kg food B444/R347 AL347/G352 Group4 R348/B446 73.2 mg/kg food G354/AL349 Y413 Groups AL351/Y415 146 mg/kg food G356/B448 Group 6 293 mg/kg food AL353-Day8 PFOS = Perfluorooctanesulfonate Limit o f Quantitation (LOQ): PFOS is approximately 0.070 ug/g NA = not applicable NH = not extracted Date Entered/By: 07/21/99, 07/23/99, 09/03/99 LAC Date Verified/By: 08/09/99 GML, 04/12/00 KJH PFOS Calc. Cone. ng/g 0.00 0.00 0.00 0.00 977 972 920 872 47.2 72.1 42.8 51.7 841 955 680 804 0.00 34.7 0.00 86.2 39.1 0.00 60.6 9.72 16.5 0.00 147 0.00 22780 15691 17080 28909 20463 28049 43414 48741 39880 65404 75293 NE Concentration of PFOS ug/g or % Ree. <LOQ <LOQ <LOQ <LOQ 82*/. 81% 76% 72% 79% 120% 71% 86% 70% 79% 57% 67% <LOQ <LOQ <LOQ 0.0862 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.147 <LOQ 22.8 15.7 17.1 28.9 20.5 28.0 43.4 48.7 39.9 65.4 75.3 NE Mean PFOS ug/g or %Rec <LOQ 81% 74% 89% 68% <LOQ 18.5 25.8 44.0 70.3 NA RSD (%) Std. Dev.(ue/e) MS/MSD RPD NA 0% 5% 21% 9% NA NA 20.3 3.76 18.0 4.65 10.1 4.46 9.94 6.99 NA FACT-M-2.1 Excel Version 5/95 TOX129 liver-3.xls 3/8/01 9:01 AM FACT-TOX-129 BACK TO MAIN Study: Product NumbeKTest Substance): Matrix: M eth od/R evis io n: Analytical Equipment System Number: Instrument Software/Version: Date o f Extraction/Analyst: Date o f Analysis/Analyst: Date o f Data Reduction/Analyst: Sample Data . BOBWHITE LIVER DAY 22 Group Dose Sample # Method Blk BWL07209-H20 Blk-9-1 BWL07209-H2O Blk-9-2 BWL07209-H20 Blk-9-3 QC - 1.00 ppm BWL07209-H20 Blk-9- B435-MS-9-1 B435-MSD-9-I G343-MS-9-2 G343-MSD-9-2 100ppb R344-MS-9-3 R344-MSD-9-3 Groupl AL338 0.0 mg/kg food B435 G343 R344 Sample spilled lost -3 mL Y402 AL339 B436 G344 R338 Y403 AL340 B437 G345 R339 Y404 Group2 AL344 18.3 mg/kg food B441 G349 R345 Y408 Group3 AL346 36.6 mg/kg food B443 G351 R343 Y410 Group4 73.2 mg/kg food AL348 B445 G353 R348 Y412 Groups AL350 146 mg/kg food B447 R349 Y414 Group6 293 mg/ieg food AL352 PFOS = Perfluorooctanesulfonate Limit o f Quantitation (LOQ): PFOS is approximately 0.070 ug/g NA = not applicable Date Entered/By: Date Verified/By: 07/21/99,07/23/99 LAC 08/09/99 GML, 04/12/00 KJH 454-103 Bobwhite Quail LC50 PFOS Bobwhite Quail Liver FACT-M-1.1 and FACT-M-2.1 Soup 020199 Masslynx 3.2 07/20/99 MCH 07/22/99, 07/23/99 DRB 07/23/99,07/26/99 DRB Filename: R-Squared Value: Slope: Y-Intercept: See Attachments See Attachments See Attachments See Attachments PFOS Calc. Cone. ng/g 0.00 0.00 0.000 0.00 1026 943 1134 1082 136 143 190 164 266 136 153 141 265 285 235 215 494 177 130 141 230 2290 3015 3132 4518 3271 2518 4817 4126 921 2215 4324 11905 10253 10755 12618 33391 17719 23167 29026 39326 Concentration of PFOS ug/g or / Ree. <LOQ <LOQ <LOQ <LOQ 85% 78% 95% 91% 113% 119% 0.190 . 0.164 0.266 0.136 0.153 0.141 0.265 0.285 0.235 0.215 0.494 0.177 0.130 0.141 0.230 2.29 3.01 3.13 4.52 3.27 2.52 4.82 4.13 0.921 2.22 4.32 11.9 10.3 10.8 12.6 33.4 17.7 23.2 29.0 39.3 Mean PFOS ug/g or */.Rec <LOQ 82% 93% 116% 0.215 3.25 2.92 10.0 25.8 NA RSD (%) Std. Dev.(ug/g) MS/MSD RPD NA 8% 5% 5% 43.4 0.093 24.8 0.806 53.4 1.56 33.0 3.29 26.5 6.84 NA FACT-M-2.1 Excel Version 5/95 TOX129 liver-3.xls 3/8/01 9:01 AM BACK TO MAIN Laboratory Report Analytical Report of Data for PFOS Dietary LC50 Study in Northern Bobwhite (Sera and Liver) Laboratory Report No. FACT-TOX-129 (U2723) Attachment B Table o f Results - Sera Analysis Day 8 Day 22 Results of MS/MSD Analysis FACT-T0X-129 Study: Product Numberf Test Substance) Matrix: Method/Revision: Analytical Equipment Systeiri Number Instrument Software/Vcrsion: Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst: Dates of Analysis/Analyst: Date of Data Reduction/Analyst: Sample Data BOBWH1TE SERUM Day 8 Group Dose Sam ple # Method Blk H20 Blk-62899-11-1 H20 Blk-62899-11-2 H20 Blk-62899-11-3 Matrix Blk H20 Blk-62899-11-4 RBS Blk-062899-IM RBS Blk-062899-11-2 RBS BIk-062899-11-3 RBS Blk-062899-11-4 G roupl G347 0.0 mg/kg food B438 Y405 AL343 B440 Y406 Y407 R341 R342 AL342 B439 G346 G348 AL341 Group2 18.3 mg/kg food B442 R340 G350 Y409 AL345 Group3 Y411 36.6 mg/kg food B444 R347 AL347 G352 Group4 R348 73.2 mg/kg food B446 G354 AL349 Y4I3 Group5 146 mg/kg food AL351 Y415 G356 B448 G rou p 293 mg/kg food AL353 Limit of Quantitation (LOQ): PFOS 0.010 ug/mL PFOS Dietary LC50 Study with Northern Bobwhites PFOS Bobwhile Serum ETS-8-4.1 and ETS-8-5 1 Amelia 062498, Soup 020199 Masslynx 3.2 See Attachments See Attachments See Attachments See Attachments 06/28/99, 08/24/99 SEE/SRP 07/23/99,07/26/99, 08/24/99,09/01/99 DRB/MEE/PTF 07/26/99, 07/27/99, 09/02/99, DRB/MEE/PTF C on cen tration of PFOS ug/m L <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.136 <LOQ <LOQ <LOQ 0.200 3.51 8.41 0.0542 <LOQ NA 57.7 9.38 76.3 21.4 25.2 114 21.6 5.18 43 6 Mean PFOS u g /m L RSD % Std. Dev. (ug/m L) <LOQ NA <LOQ NA <LOQ NA NA <LOQ 1 outlier NA 129 3.04 3.92 75.6 41.2 31.2 119 41.6 49.4 NA NA NA " Not Applicable PFOS * Periluorooctanesulfonate Date Entered/By: Dale Verified/ By: 07/26/99,07/27/99,09/03/99 LAC 08/10/99 GML, 04/12/00 KJH BACK TO MAIN ETS-8-5 1 Excel Version 5.0/95 TOX129 sera-3.xls 3/8/01 FACT-TOX-129 Study Product NumberfTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Sofrware/Venion Filename: R-Squarcd Value: Slope: Y-Intercept: Dates of Extraction/Analyst: . Dates of Analysis/Analyst: Date of Data Reduction/Analyst Sample Data PFOS Dietary LC50 Study with Northern Bobwhites PFOS Bobwhite Serum ETS-8-4.1 and ETS-8-5 1 Soup 020199, Amelia 062498 Masslynx 3.2 See Attachments See Attachments See Attachments See Attachments 07/14/99, 08/24/99 SEE 07/16/99, 07/20/99, 08/25/99 GML/MEE/SAH 07/19/99, 07/21/99, 09/02/99 DRB/MEE BOBWHITE SERUM Day 22 G rou p Dose Method Blk Matrix Blk G rou pl 0.0 mg/kg food Group2 18.3 mg/kg food Group3 36.6 mg/kg food G roup4 73.2 mg/kg food G roupS 146 mg/kg food G roop6 293 mg/kg food Sam ple # H20 Blk-71499-11-1 H20 Blk-71499-11-2 H20 Blk-71499-11-3 H20 Blk-71499-11-4 RBS Blk-071499-11-1 RBS Blk-071499-11-2 RBS Blk-071499-11-3 RBS Blk-071499-1M AL338 B435 G343 R344 Y402 AL339 B436 G344 R338 Y403 AL340 B437 G345 R339 Y404 AL344 B441 G349 R345 Y408 AL346 B443 G351 R343 Y410 AL348 B445 G353 R349 Y412 AL350 B447 R350 Y414 AL352 C o n ce n tr a tio n o f PFOS ug/m L or / Ree <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.0326 0.0284 0.0874 0.125 0.0303 0.0338 0.0293 0.0478 0.0393 0.945 <LOQ 0.0232 0.0502 0.0282 4.56 5.12 5.25 7.28 7.25 2.77 9.59 6.71 13.1 4.19 9.31 16.7 16.6 69.6 18.8 60.0 36.5 27.0 38.4 112 Mean PFOS ug/m L RSD % Std. D ev. (ug/m L ) <LOQ NA <LOQ NA 0.115 - Two Outliers 217 0.251 21.7 5.89 1.28 57.3 7.27 4.17 93.6 26.2 24.5 34.5 40.5 14.0 NA NA Limit of Quantitation (LOQ): PFOS " 0 010 ug/mL NA - Not Applicable PFOS - Periluorooctanesulfonate Date Entered/By: 07/26/99, 07/27/99, 09/03/99 LAC Date Verified/ By: 08/10/99 GML, 04/12/00 KJH BACK TO MAIN ETS-8-5.1 Excel Version 5.0/95 TOX129 sera-3.xls 3/8/01 FACT-TOX-129 Study: Product Number(Test Substance): PFOS Dietary LC50 Study with Northern Bobwhites PFOS Matrix: Method/Revision: Analytical Equipment System t>iumber Bobwhite Serum ETS-8-4.1 and ETS-8-5.1 Amelia 062498 Instrument Software/Veriian: Masslynx 3.3 Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst Dates of Analysis/Analyst Date of Data Reduction/Analyst: See Attachments See Attachments See Attachmenu See Attachments 11/24/99 SEE 11/30/99 HOJ 12/3/99 GML Sample Data BOBWHITE SERUM MS/MSD'i Group Dose Sample# Concentration of PFOS ug/mL or % Ree Mean PFOS ug/mL or %Rec RSD % Std. Dev. % Method Blk Matrix Blk H20 Blk-11249-11-1 BWS Blk-11249-11-1 BWS Blk-11249-11-1 <LOQ <LOQ <LOQ <LOQ <LOQ NA NA QC - 50 ppb BWS11249-MS-11-1 96% BWS11249-MSD-l 1-1 104% 250 ppb BWS11249-MS-11-1 91% BWS11249-MSD-11-1 103% 97% 12% Limit of Quantitation (LOQ): PFOS - 0.010 ug/mL NA - Not Applicalbe Date Entered/By: 12/3/99 GML Date Verified/ By: 04/12/00 KJH BACK TO MAIN ETS-8-5.1 Excel Version 5.0/95 TOXI29 sera-3.xls 3/8/01