Document 5LwBppy82aJnN9d6JX9z89w1R

IZ jjL b - Y S Laboratory R eport-R evision 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 1o f / IAG O220\ G 006 1 Introduction This study was designed to quantitatively assess perfluorooctane sulfonate (PFOS, C8F |7S 0 3\ CAS # 279539-3) levels in the sera and liver o f die northern bobwhite (Colinus virginianus) exposed to PFOS ihrough diet. Details of dosing conditions, clinical observations and other toxicity data were generated by die 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 of data trends was complicated by the sporadic nature of the PFOS levels in sera. Within a particular dose group there was dramatic variation between individual animals. The relative standard deviation of PFOS sera levels within a dose group ranged from greater than 20% to over 200%. By comparison, the results of 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 of 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 die 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 of 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 o f 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 of custody was initiated. The Environmental Laboratory received 30 pooled bobwhite quail serum tubes, frozen, on 06/30/99. Upon specimen receipt a 3M chain of custody was initiated. A copy of 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 o f ^ 1 IAC oi/ailo- 4 Methods - Analytical and Preparatory Analytical methods ETS-8-4.1, Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollowing exceptions: Due to the small volume of bobwhite sera available, standard curves were prepared in methanol instead of from bobwhite sera. Four matrix spikes were prepared from die pooled bobwhite quail serum received on 06/30/99. PFOS lot 171 was used as reference material. FACT-M-1.1, Extraction of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollowing 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 of Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry with thefollowing exception: Due to the small amount of bobwhite liver available, standard curves were prepared in methanol instead of 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 of 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 thefollowing 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 of pg of analyte per gram o f liver (wet weight). ETS-8-7.0, Analysis of 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-fertbutyl ether), dried, and reconstituted in methanol for MS analysis. HPLC/ESMSMS In HPLC, an aliquot o f extract is injected and passed through a reverse phase liquid chromatographic column. Based on the affinity of the analyte for the stationary phase in die column relative to the liquid mobile phase, the analyte is retained for a characteristic amount of 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 of molecular ions, is generally operated at relatively mild temperatures. Molecules are ionized, and a primary ion, characteristic of 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 (C8F|7S 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 (CjFsSOj-). Each of 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. Page3of^ 7 UC- i\ l j / lO f 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% o f the expected value. Quantitation o f the target analytes was based on linear regression analysis (weighted 1/x) of two unextracted methanol curves bracketing each group of samples. Quantitation of each analyte was based on the response of one or more specific daughter ions using the multiple response-monitoring mode o f the instrument 3M lot #217 (purity 86.9%) ofPFOS was used for calibration standards. As purity information was not available at the time of data collection, reported results have not been corrected. 5.2 Extraction and Method Blanks At least two extraction blanks, utilizing water as surrogate matrix, were extracted with each batch of samples. Additionally, two method blanks, utilizing control sample matrix (if available), were extracted along with each batch of samples and curves. Blanks were compliant if no target analyte was detected above die limit of 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 of die analysis, twenty-six matrix spikes were prepared in the bobwhite liver matrix. All of the matrix spikes were recovered at greater than 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 of approximately 350% of 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% of the expected value. 5.5 Laboratory Control Samples Laboratory Control Samples were not a component of 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 off f U c or/inloi G O O ^- 6 Data Summary - Elucidation of data trends was complicated by the sporadic nature of the PFOS levels in sera. Within a particular dose group there was dramatic variation between individual animals. The relative standard deviation of PFOS sera levels within a dose group ranged from greater than 20% to over 200%. By comparison, the results of 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 die exception of group 4 animals, average levels o f PFOS in the sera o f die 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 o f these birds were over 110 pg/g. No circumstances existed during the present study that would have adversely affected die quality or integrity o f the data. The following data quality objectives were followed during the study: % Linearity - The coefficient of determination (r*2) of die standard curve was equal to or greater than 0.985 using 1/x weighting Limits of 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 of the MS/MSDs. Acceptable spike recoveries - sera: 70% to 130%; liver: 50% to 150% Use o f confirmatoiy methods - Given the selectivity o f the analytical tool used (HPLC-ESMSMS) and lack of a viable alternative for analysis, no confirmatory methods were used. Demonstration of 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 of 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 of 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 of tissue available causes analytical challenges. Page 5 of# 7 , U C O J / j ) / 0i 00^9 8 Attachments'r ! Attachment A: Table o f 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 Page6of^i G 0 0 e. V ? 0 9 Signatures ...C A ___Ch*t& ________________________________ o i h i i pi Lisa Clemen, Advanced Chemist Date z iiz io i Kris Analytical Investigator Date Quality Assurance Unit Representative Date Page 7 of^ lAC' o i / i j 01 0 -0 0 ^ 7 ' 9 Signatures (^K A ` ______________________________________ O ulnL -lcto______________ Lisa Clemen, Advanced Chemist Date .. -- _______________________ Kris Hansen, Ph.D., Principal Analytical Investigator 0C / o t /0 -0 Date Quality Assurance Unit Representative Q> * * i f - c > o Date 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 o f Results - Liver Analysis Day 3-7 Day 8 Day 22 GCO FACT-TOX-129 Study; Product Numbet(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst Date of Analysis/Analyst Date of Data Reduction/Analyst: Sample Data 454-10] Bobwhite Quail LC50 PFOS Bobwhite Quail Uvei ETS-8-6.0 & ETS-8-7.0 Amelia 062491 Masslynx J. 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-inlercept See Attachments See Attachments See Attachments See Attachments BOBWHITE LIVER DAY 3-7 Group Dote Sample a Method Bile BWL10089-H20 Blk-11 BWLI0089-H2O Blk-12 QC - 50 ppb QC-lOOOppb R342-O-Day8-50ppb-MS-l 1 R342-0-Day8-50ppb-MSD-l 1 B439-0-Day8-lppm-MS-12 B439-0-Day8-lppm-MSD-12 Groupl 0.0 mg/kg food Group! 146 mg/kg food R342-0-Day8 B439-0-Day8 G355-Day7 Group6 R3S2, B450-Day5 293 mg/kg food Y417, R353-Day6 G357, G358-Day7 B449, Y416-Day7 Group7 586 mg/kg food AU54-Day3 R354-Day4 R355, Y401, B452-Day5 Y418, B451, G360-Day6 G359, AL355-Day7 Groups 1171 mg/kg food AL356, R357. Y419-Day3 AU$7. 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 LAC Date Verified/By: 04/12/00 JOH PFOS Calc. Cone. "ST 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. Dcv.(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 TOX129livcr-3.xls 3/8/01 9:40 AM FACT-TOX-129 Study: Product NumbctfTcst Substance): Mitri*: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Date or 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 ! 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-Squared Value: Slope: Y-Intercept: See Attachments See Attachments See Attachments See Attachments BOBWHITE LIVER DAY 8 Croup Dose Sample * Method Blk BWL-H20 Blk-6/30-l BWL-H20 Blk-600-2 BWL-H20 Blk-6/30-3 BWL-H20 Blk-600-4 QC-lOOOppb G347-MS-1 G347-MSD-1 Y405-MS-2 Y405-MSD-2 QC - SOppb G346-0-MS11-1 G348-0-MS11-2 G347-0-MSI1-3 Y405-0-MSI1-4 QC - lOOOppb G346-0-MS11-I G348-0-MS11-2 G347-0-MS11-3 Y405-0-MSH-4 Groupl 0.0 mg/kg food G347 B438/AL341 Y405 AL343 B440 Y406 Y407 R341 R342 B439 G346/R346 Day6 G348/AL342 Group2 18.3 mg/kg food B442/AL345 R340 G350/Y409 Group3 36.6 mg/k| food Y411 B444/R347 AL347/G352 Group4 73.2 mg/kg food R348/B446 G354/AL349 Y413 Group5 146 mg/kg food Croup 6 293 mg/kg food AL351/Y415 G356/B448 AL353-Day8 PFOS - Perfluorooctanesulfonate Limit of Quantitation (LOQ): PFOS is approximately 0.070 ug/g NA = not applicable ME " not extracted Date Entered/By: Date Verified/By: 07/21/99,07/23/99,09/03/99 LAC 08/09/99 QML, 04/12/00 KJH PFOS Calc. Cone. "V 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 CoDcentratioa of PFOS ug/g or % Ree. <LOQ <LOQ <LOQ <LOQ 82% 81% 76% 72% 79% 120% 71% 86% 70% 79% 57% 67% <l o q <LOQ <LOQ 0.0862 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.147 <LOQ 22.8 15.7 17.1 28.9 20.3 28.0 43.4 48.7 39.9 65.4 75.3 NE Meaa PFOS ug/g or #/Rcc <LOQ 81% 74% 89% 68% <LOQ 18.5 25.8 44.0 70.3 NA RSD (%) Std. Dev.fug/f) MS/MSD RPD NA 0% 5% 21% 9% NA NA 20.3 3.76 18.0 4.6$ 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 -0 0 0 7 :. FACT-TOX-129 Study: Product NumbeKTcst Substance): Matrix: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reducdon/Analyst: Sample Data 454-103 Bobwhile Quail LC50 PFOS Bobwhile Quail Liver FACT-M-1.1 and FACT-M-2 1 Soup 020199 Masslyn* 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 BOBWHITE LIVER DAY 22 Group Dose Sample# Method Blk BWL07209-H20 Blk-9-1 BWL07209-H2O BIk-9-2 BWL07209-H2O Blk-9-3 QC -1.00 ppm BWL07209-H20 Blk-9-4 B43S-MS-9-1 B43-MSD-9-1 G343-MS-9-2 G343-MSD-9-2 lOOppb R344-MS-9-3 R344-MSD-9-3 Group! AL338 0.0 mg/kg food B43S 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 R34S Y408 Group3 AL346 36.6 mg/kg food B443 G351 R343 Y4I0 Group4 73.2 mg/kg food AL348 B445 G353 R348 Y4I2 Group5 AL350 146 mg/kg food B447 R349 Y414 Group6 293 mg/kg food AL352 PFOS Perfluoroocttnesulfonatc Limit of 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 PFOS Calc. Cone. ng/| 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 <l o q <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 %Rcc <LOQ 82% 93% 116% 0.215 3.25 2.92 10.0 25.8 NA RSD (%) Std. Dev.(ug/g) MS/MSD RFD 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 5/9$ TOX129 livcT~3.xls 3/8/01 9:01 AM 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 o f MS/MSD Analysis 000* FACT-TOX-129 Study: Produo NumbcrCTcn Substance): Matrix: M ethodicvision: Analytical Equipment Syateni Number Instrument Software/Vcrskxv Filename ((Squared Value: Slope: Y-lniertept Dates of Extraction/Analyst: Dates of Analysis/Analyst Date of Data Reduction/Analyst Sample Data BOBWHITE SERUM Day Group Dose Sample# Method Blk H20 m-6M 99-\ ].| H20 BOt'62899-l 1-2 H20 BIk-2l99-U-3 H20 BRt-62899-IM Matrix Blk RBS Blk-062199-It1 rbs m - a tim - u - i RBS BQc~062S99*U-3 RBS Blk-062199-IM Groupl 0347 0.0 mg/kg food B438 Y403 AL343 B440 Y406 Y407 R34I R342 AL342 B439 G346 G348 AL341 Groupl B442 18.3 mg/kg food R340 G350 Y409 AL345 GronpS 36.6 mg/kg food Y411 B444 RJ47 AL347 G352 Grenp4 73.2 mg/kg food R348 B446 G354 AL349 Y4I3 GroapS AL351 146 mg/kg food Y4J3 G336 B448 Grop6 293 mg/kg food AL353 Limit of Quantitation (LOQ): PFOS 0.010 ug'mL PFOS Dietary LC50 Study with Northern BobwhitcS PFOS Bobwhite Serum ETS-8-4. I and ETS-8-3.1 Amelia 062491, Soup 020199 Maulyna 3.2 See Attachments See Attachments See Attachments See Attachments 06/28/99,01/24/99 SEE/SRP 07/23/99,07/26/99, 08/24/99,09/01/99 DRfi/MEE/PTF 07/26/99,07/27/99,09/02/99, DRB/MEE/PTF CeaceutratitB of PFOS ... 'L <LOQ <LOQ <LOQ <LOO <LOQ <LOQ <LOQ <LOO <LOQ <LOQ <L0Q <LOQ <LOQ <L0Q <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.136 <LOQ <LOQ <LOQ 0.200 3.51 8.41 0.0542 <LOQ NA J7.7 9.38 76.3 21.4 25.2 114 21.6 5.18 43.6 M aaa PFOS ug/mL H tS D % Std. Dev. (ugriaL) <LOQ NA <LOQ NA <LOQ NA NA <LOQ 1outlier NA 129 304 3.92 73.6 41.2 31.2 119 41.6 49.4 NA NA NA Not Applicable PFOS PerfluorooetaMSulfonate Date Entered/By: 07/26/99,07/27/99. 09/03/99 LAC Date Verified/By: 08/10/99 GML, 04/12/00 KJH ETS-8.5 1 Excel Version 5.0/95 TOX129 sera-J xls FACT-TOX-129 Study: Product NumberiTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Softwarc/Version: Filename: R-Squared Value: Slope: Y-Imercept. Dates of ExtracnkWAnalyst: Dates of Analysis/Analyst: Date of Data Reductioo/Analyst Sample Data FFOS Dietary LC50 Study with Northern Bobwhites PFOS Bobwhite Serum ETS-l-4 landETS-I-5I Soup 020199, Amelia 06249 Masalynx 3 2 See Attachmeats See Attachments See Attachments See Attachments 07/14/99,01/24/99 SEE 07/16/99,07/20/99,08/23/99 GMVMEE/SAH 07/19/99.07/21/99,09/02/99 DRB/MEE BOBWHITE SERUM Day 22 Group Doae Method Blk Sample # H 70B 1k-7U -IM H20 Blk-71499-11-2 Ceoceutratfoa fP F O S ug/mL o r % Rec <LOQ <LOQ Mean PFOS ag/nL. RSD% Std. Dev. (ug/mL) Matrix Blk H20 Blk-71499*11*3 H 20 Blk-7149911-4 RBS Blk-071499-11 1 <LOQ <LOQ ^COQ <LOO NA Grvupl 0.0 mg/kg food RBS Blk-071499-11-2 RBS Blk-071499-11-3 RBS BIk-071499-11-4 aESS B435 G343 R344 Y402 AL339 B436 G344 <LOQ <LOQ <LOQ <LOQ 0.0326 0.0284 0.0874 0.125 0.0303 0.0338 0.0293 <LOQ NA R338 Y403 AJL340 B437 0.0478 0.0393 0.945 <LOQ Group! 18.3 mg/kg food Group! 36.6 mg/kg food G345 R339 Y404 AL344 B44I G349 R345 Y408 AL346 B443 G351 R343 Y410 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 0.115-Two Outliers 217 0.251 21.7 5.89 1.28 57.3 7.27 4.17 Group4 AU48 9.31 73.2 mg/kg food B445 16.7 G3S3 16.6 R349 69.6 93.6 Y412 18.8 26.2 24 5 Group! 146 mg/kg food Group* 293 mg/kg food AJL350 B447 A350 Y414 AL352 60.0 365 27.0 38.4 112 34.5 40.5 M.0 NA NA Limit of Quantitation (l^Q)7PFOS 0010 ug/mL NA * Not Applicable PFOS - Ferfluoroocisncmlfonatc Dale Eniercd/By: 07/26/99.07/27/99 ,09/03/99 LAC Date Verified/ By: 08/10/99 GML, 04/11*00 KJH E7S-8-3 1 Excel Version 50/95 TOXI29 scra-3 ill 3/1/01 4 7 ;J FACT-TOX-1I9 Study: Product Number<Test Sufananee): PFOS Dietary 1X50 Study with Northern Bobwhitea PFOS Matrix: Bobwhne Scram Mcthod/Rcvision: ETS-8-4. I and ETS-8-5. t Analytical Equipment SystemNumber Amelia 062498 Instrument Software/Verticxi: Maaslyiu 3.3 Filename: See Attachments R-Squared Value: See Attachments Slope: See Attachments YImerccpt: D ata of Extraction/Analyst: See Attachments ] 1/24/99 SEE Dates of Analysia/Anafyst: Date of Data Reduction/Analyst: 11/30/99 HOJ 12/3/99 GML Sample Data BOBWHITE SERUM MS/MSD's Group Dose Sam ple# C eaeeatratiM ef PFOS C/m LarH Rec M ean PFOS ag/mL er % Rcc r sd % S id . Dev. % Method Blk Matrix Blk H 20Blk-l 1249-11-1 BWS Btk-l1249-1M BWS Blk-I1249*11*1 <LOO <LOQ <LOQ <LOO <LOQ NA NA QC - 30 ppb BWSl 1249-MS-l1-1 96% BWS1I249-MSD-II-1 104% 2 P I* BWSl 1249-MS-l M 91% BWS11249-MSD-1M 103% 97% 12% Limit of Quantttatxm (LOQ): PFOS" 0.010 ug/mL N A -N o t Applicalbe Date Eatered/By: 12/3/99 GML Date Verified/By: 04/12/00 KJH ETS-I*5.| Excel Version 3 0/95 TOXI29 sera-3 xk 3/8/01 600679a.