Document J8paJvXBmz44zEBmJ7Eg94GK

Lab0ratoyRep-t-Revision1 Analytical Reportof Data for PFOS Dietary LC50 Studiyn Northern Bobwhite (Sera and Liver) Laboratory Report No.FACT-TOX-129 (U2723) Testing Laboratory ~~~ ~ 3M Environmental Technology& Safety Services 3M Environmental Laboratory FluorineAnalybcal 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: (65 1) 778-601 8 FAX:(651) 778-6176 Revision Date: March 19,2001 1 Introduction , i This study was designed to quantitatively assess perfluorooctane sulfonate (PFCO8SF, l7S0iC, AS # 279539-3) levels in the sera and liver of the northern bobwh(Citeohus virginianus) exposed to PFOS through diet. Detailsof dosing conditions, clinical observations and other toxicity data were generated by the in-Me testing facdity (WildMe International) and are not included in this report. Subsets ofgrtohuepseoigfhttest animalswere sacrificedon day 8 and day22, post-dose. Sera and liver collected fiom the sacrificed animals were sent to th3eM Environmental Laboratory and analyzed quantitatively for PFOS. Tissueasnifimoamls that died before the scheduled sacrifice date were collected and aanlasoly;zresdults are included itnhis report. Elucidation of data trends was complicabtyedthe sporadicnature of the PFOS levels in sera. Within a particular dosegroup there was dramatic variation between individuaanlimals. The relative standard deviation of PFOS sera levels within a dgorsoeup ranged fiomgreaterthan20% to over200%. By comparison, theresults of PFOS leveilns the livers of the bobwhites were far more consiwsittehnitn a particular dose leve(lgroup RSD between 10-55%) In general, both sera and liver levoeflPsFOS increased with dose level, although due to antoimaanlimal variability,thistrend is not pronounced in the sera. With the exceptiongroufp 4 anm i asl,average levelsof PFOS in the seroaf the bobwhites generally increased or stayed the same betwe8enandadyay 22. In all dose groups, the group average for PFOS concentration in liver decreased dramatically between8 adnady day 22. Livers were collected fiom bobwhites thatbedtiwedeen day1and day8, prior to the scheduled sacrificeIn. every case, PFOS levels in the livers of these birds w1e1re0pogv/egr wetweight. Although rigorous quality control measures 3aMndEnvironmental Laboratory Standard Operating Procedures were followed, the acquisitoiofnthisdata was not necessarily collected according to applicable Good Laboratory Practices. Data presented ihsetrhee highest quality data availablethaist time. No data was collected for any analytes otthhearnPFOS. It is recommended that bobwhite quail be avoindefudture studres because thsemall amount of tissue available causes analytical challenges that make verificMatiocnult. The Environmental Laboratory receive6d5 liver, 34 blood, and34 serum specimens,frozen, on 05/19/99. Upon specimen receipt3Ma chain of custody was initiated. The Environmental Laboratory receive3d5 liver, 35 blood, 35 serum, and 6 bile specimens,hzen, w 06/29/99. Upon specimen receipt 3aM chain of custody was initiated. The Environmental Laboratory receive3d0 pooled bobwhite quail serum tubes, fiozoenn,06/30/99. Upon specimen receipt 3aM chain of custodywas initiated. A copy of each form can be found in an Attachment to this report. There is no holdmg time criteria associated with these samplesLfoCrESMSMS analysis. Analytical methods ETS-84.1, Extraction of PotassiumPeffluorooctanesulfonateor Other FluorochemicaClompounds from Senun for Analysis Using HPLC-ElectrosprayMass Spectromewtirtyh the following exceptions: Due tothe small volume of bobwhite sera available, standard curves were preipnamreedthanol insteadof from bobwhite sera. Four matrix spikes were prepared fiom the pooled qbuoabiwl sheirtuem received on 06/30/99.PFOS lot171 was usedas reference material. FACT-M- 1.1, Extractioonf Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds fiom Liver for Analysis Using HPLC-ElectrosprayMass Spectrometry rhwefiothZZoowing exception: The extraction was performed following ETS-8-6th.i0s;method had beenvddated but not approvedat the time of extraction. PFOSlot 171 was usedas reference material. ETS-8-6.0, Extraction of PotassiuPmerfluorooctanesulfonateor Other Fluorochemical Compoundfisom Liver for Analysis Using I-IPLC-EIectrospray/Mas SpectromweitthrythefoZZowing erception: Due tothe s d amount of bobwhite liver available, standard curves were prinepmareetdhanol instead of fiom bobwhite liver.In some cases, liver samples wveerrey limited andtwo or more samples htaodbe combined for extraction. These combinations are indicattheed riensults. Twenty-twomatrix spikes were prepared h m the bobwhite liver. PFOS lot 171 was aussreedference material. ETS-8-5.1, Analysis of PotassiuPmerfluorooctanesulfonateor Other Fluorochemicailns Serum Extracts Using HPLC-ElectrosprayMass spectrometry. FACT-M-2.1, Analysisof Potassium Peduorooctanesulfonate or Other Fluorochemicals on Liver Extracts Using HPLC-Electrosprayhiass Spectrometwryith the followingexception: The analysis was performed following ETS-8-7.0, which had been validated but not aatpthperovtiemde of extraction.As described in the method, all results are reported in otferpmgsof analyte pergram of liver (wet weight). ETS-8-7.0, Analysisof PotassiumPerfluorooctanesulfonateor Other Fluorochemicals on Liver Extracts Using HPLC-ElectrosprayhiassSpectrometry. Sample preparation-HPLC/ESMSMS: ion-pairing extraction Analyte is extracted from a sample matrix iwonitphairing 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 anisaltyratensferred into a non-polar organic solvent (methyl-tertbutyl ether), dried, and reconstituted in methanol for MS analysis. HPLC/ESMSMS In HPLC, an aliquot of extract is injected and passed through a reverse phase liquid chromatographic column. Based on the afinityof the analyte for thsetationary phase in the column relative to the liquid mobile phase, the analyitseretained for a characteristic amouofntime. For example,in a standard solutionPFOSmay elute a8t.0 minutes. Retention times between a standard PFOS solution and the analyte extractedfkom tissuein thisanalysis were matched on tHhePLC system towithin 1%. Following HPLC separation, ESMSMS provides a rapid and accurate means for analyzing a owfide range organic compounds, includmg fluorochemicaEllse.ctrospray, an ionization technique used primarily for detection of molecular ioniss,generally operated at relatively mild temperatures. Molecules are ionized, and aprimary ion, characteristic of the analyitse,selected. This ion is bombarded withhgh-energy gas; subsequentcohsions create smaller secondary ionihcpents unique to the primary ion, whaicreh detected. For example, for PFO(SC8F17SOia)nalysis, ion499 is selectedas the characteristicprimary ion. This ion is hgmented into other ions suchas 80 amu (corresponding toSO;), and 230 amu(CZ6S03-). Each of these secondaryhgments is detected and can be used to differentiate PfrFoOmSother compounds that might have the same characterist4ic99 m u primary ion, but different chemical compositions and secondary ionhgmentation patterns. 5 Analysis 5.1 Calibration A mid-level calibration check standairsdanalyzed every5- 10samples to monitor instrumentadlrift. Calibration check standardwsere compliantif instrumental response was withi+n/- 30% of the expected value. Quantitationof the target analytes was basoend linearregression analysis (weightedl/x) of two unextracted methanol curves bracketing geraocuhp of samples. Quantitation of each analyte was baosend the responseof one or more specific daugihontesrusing the multiple response-monitoring mofdtehe instrumeXlL 3M lot #217 (purity 86.9%) of PFOS was used for caliimtion standardAs.s purity informationwas not availableat the timeof data collection, reported results have not been corrected 5.2 Extraction and MethodBlanks At least two extraction blanks, utilizing wastesrurrogate matrix, were extmcted with each boaftch samples. Additionally,two method blanks, utilizing control sample matrix (ifavailable), were extracted along with each batch of samanpdlescurves. Blanks were complianiftno target d y t e was detected above the limit of detection for a specific analyte. In this study, all blanks were compliant. 5.3 Surrogates Tetra-hyclroperfluorooctanesulfonatewas usedas a surrogate itnhisanalysis. Surrogate response was monitored to confirm gross instrumental failure, ifnecessary. Surrogate response was acceptable for all analyses. 5.4 Matrix Spikes Over the course of the analysis, twenty-six matrixwespreikpersepared in the bobwhite liver matrAixl.l of the matrix spikes were recovered at gretahtaenr 50% of the expected value witthwo exceptions. The two 50 nglg matrix spikes thawt ere preparedon September24,1999 with the day3-7 liver samples showed analyte recoveryof approximately350% of the expected value. These samples amnadtrix spikeswere re- extractedon October08,1999. The re-extracted matrix spike recoveries were grethataenr 50% (104%- 11OYO)t,herefore day 3-7data are reported fiom extractiondate October 08,1999- referto extraction sheet for animalnumbers. Because the bobwhite sera wseorelimited, only foumratrix spike samples wereprepared All matrix spikes showed analyte recovery of greate9r0%thoanf the expected value. 5.5 Laboratory Control Samples Laboratory Conml Samples were not a component of this study. 5.6 Sample Related Comments Samples were not analyzfeodr any residuaol r potential metabolite othetrhanPFOS. Other matrices submitted withthisstudy will be analyzedas needed upon written requesEtromthe requestor.In the event of W e r analysis,an additionalreport will be generated. 6 Data Summary I ' "' .,:a 5 1 Elucidation of data trends was complicated by the sporadic naturoef the PFOS levels insera. Withm a particular dose groutphere was dramatic variation between individuanlimals. The relative standard deviationof PFOS sera levels withina dose group ranged &om gretahtaenr 20%to over200%. By comparison,the results of PFOS levels in the loivfetrhse bobwhites were far more consistwenitthin a particular dose leve(lp u p RSD between1045%) In general, both sera and liver levels increased with dose level, although due to animal to animal variability, thls trend is not pronounced in the sera. With the exception of g4roaunipmals,average levelsof PFOS in the sera of the bobwhtes generally increased or stayed the same between8daanyd day22. In all dosegroups, the group average for PFOS levels in liver decreased dramatically betwee8nadnadyday22. Livers were collectefdrombobwhites that died between da1yand day7, prior to the scheduled sacrifice.In every case, PFOS levels in the livers of these birds were over 110 pg/g. No circumstances existed during the present study that wouldahdavvmeely affected theq d t y or integrity of the data. The following data quality objectives were followed during the study: Linearity - The coefficient of determination (r'2) of the standard curve was equal to or greater than 0.985 using l/x weighting 0 Limits of quantitation- PFOS in serais 0.010 pg/mL; PFOS in liveris approximately0.070 pg/g 0 Acceptable duplicate precision- <30%,assessed by analysisof the MS/MSDs. 0 Acceptable spike recoveries-sera: 70% to 130%;liver: 50% to 150% 0 Use of c o n f i i t o r y methods - Given the selectivity of the analytical tool used (I-IPLC-ESMSMS) and lack of a viable alternative for analynsoisc,onfirmatory methods were used. 0 Demonstration of specificity - Specificitywasdemonstratedbychromatographicretentiontime (matched to standards towithin 3%) and the response of at least one characteristic product ion arising fiom collisions of an analyte-specific pareniot n. The characterization of the reference material (PFOS,lot#171), with respect to purity, stability,and homogeneity, was notcomplete on thedate this report wasissued.Oncethecharacterization is complete, a copoyf the reportwilon file in the3M Environmental Lab archives. Given the parameterlsistedabove and assuming spike recoveries form a suitable mdoicfaetinodnogenous analyte recovery, sera data are quantitativ+e/-t3o0 % and liver dataare quantitative to+A50 %. The validity of these assumptionshas not been verified by other techniques.aIgfreater degreeof accuracyis required, additionalmatrix should be providedso samples can be evaluated versus a standard curve extrfaiocmtedthe same matrix as the samples. It is recommended that bobwhite quail be avoided in fskttudrees because the small amount of tissue available causes analytical challenges. 0 Attachment A: Table of results of liver analysis 0 Attachment B: Table of results of sera analysis 0 Attachment C: SampleReceipt 0 Attachment D: In-life protocol from Wildlife Int., LTD 0 Attachment E:Extraction andanalytical data Lisa ClemenD, Aadtevanced Chemist Kris Hdken, PhD., hcp ialAnaIlnytviecsatDligaatteor DaRtepresentaQtiUvuenailtity Assurance 0 3 / 2 3 / 01 - - Lisa Clemen,DAadtevanced Chemist a1AnalyticaDl Iantveestigator ~L/Ol/OQ 6*7--00 Date LaboraforyReport 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 Day3-7 Day8 Day22 FACT-TOX-129 Study: Product Number(Tcs1 Substance): Mamx: Melhod/Revision: Analytical Equipment System Number: Instrument SohvarcNenion: Date of Extnction/Analyst: Date of AnalysislAnalyst Date of 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 10/21/99.01118/00 I A S / " H 10/22/99.01/19/00 1 A S I " H -BOBWHITE LIVER DAY 3-7 Group Sample # Dose - I Method Blk BWL10089-HZ0 Blk-11 I - I I BWL10089-HZ0 Blk-12 -QC R5304p2p-0b-Days-SoPpb-MS1 I I ~~ I QC-IoOOppb I R342-0-Day8-50ppbMSD-11 B439-0-Day8-lppm-MS-12 - I I B439-0-Day8-lppm-MSD-I2 Crow1 I ~3420~~8 I 0.0 mgkg food -- Croup5 146 mgkg food B4390Day8 G355-ihy7 Croup6 I 293 mgYk4g1f7o,od R352, B450-DayS R353-Day6 G35G8-3D5a7y. 7 - I Group7 I 586 mgkg food Y41B64D4a9y, 7 AL354-Day3 i R354-Day4 R3B154255.29Y-5D740ay15. Y418. B451. G360-Day6 - I I GrOUD8 I I 1171 mgkg food G359, AL355-Day7 -56. . R1335378..5Y2419-Dav3 B45A3L. 357. G361-Dai4 8454, G362-Day4 I PFOS = Perfluorooctanesulfonate R356, Y420-Day4 Limit of Quantitation (LOQ):PFOS is approximately0.070 ug/g NA = noatpplicable G1 Day 8 data isincluded to support QC NE = not exhacted EnDteart0e1d/B18y/:00, 01/26/00 LAC Date Verified/ By: 04/12/00 KJH PFOS Cale. Cone. ng/g 0.00 0.00 68.7 64.6 980 1034 229 383 I10527 248693 123521 NE .NE I79 235 111299 131788 122934 124333 Concentration of PFOS .uUg or % Ree. <LOQ <LOQ 110% 104% 82% 86% 0.229 0.383 111 249 202 153 124 NE NE I26 235 111 134 132 I 123 124 Filename: R-Squared Value: Slope: Y-Intercept Mean PMS UgIg or %Ree CLOQ Io x 84% 0.306 Ill 182 I57 I 128 See Attachments See Attachments See Attachments See Attachments RSD (Ye) Std. Dev.(uvg) 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. I Excel Version 5/95 TOX129 liver-3.xls 3/8/01 9:40 AM FACT-TOX-129 Study: Product Number(Test Substance): Matrix: Methomevision: Analytical Equipment System Number: Instrument SoRwareNenion: Date of ExhactiodAnalyst: Date of AnalysislAnalyst: Date of Data ReductiodAnalyst: Sample Data 454-103 Bobwhite Quail LC50 PFOS Bobwhite Quail Liver FACT-M- I . I and FACT-M-2.I Amelia 062498 Masslynx 3.2 06/30/99SAH 07/02/99. 07/22/99.09/01/99 MEUPTF 07/20/99, 07/23/99.09/02/99 DRB/MEEQTF Filename: R-Squared Value: Slope: Y-Intercept: See Attachments See Attachments See Attachments See Attachments BOBWHITE LIVER DAY 8 Group Dose Sample # Method Blk QC - PPb QC - SOWb QC- l w p p b Croup1 0.0 mgkg food BWL-Hz0 Blk-6130-1 BWL-Hz0 Blk-6/30-2 , BWL-HZ0 Blk-U30-3 BWL-Hz0 Blk-6l30-4 G347-MSD-I Y405-MS2 Y405-MSD-2 G346-0-MSll-1 G348-0-MSll-2 G347-0-MS11-3 Y405-0-MSll-4 G346-0-MSll-1 G348-0-MS11-2 G347-0-MS11-3 Y405-0-MS11-4 B438lAL341 Y405 8440 Y406 Y407 R341 R342 B439 G346R346 Day6 G348lAL342 Group2 18.3 mgkg food B442lAL345 R340 G350N409 Group3 36.6 mgrkg food Y411 B444P.347 AL347lG352 Group4 73.2 mgkg food R348/B446 G354lAL349 Y413 Croup5 I46 mgkg food Group 6 293mgkg food PFOS = Perfluomtanesulfonate AL3SIN415 G356/B448 AL353-Day8 Limit of Quantitation (LOQ): PFOS is approximately0.070 ugig NA = not applicable NE =not exhactcd EDnatteeredBy: 0712 1/99, 07123199.09/03/99 LAC DVaetreifBieyd:/ 08/09/99 GML. o 4 / 1 z m KJH PFOS Calc. Conc. Concentradon of PFOS nglg uglg o r @/e Rcc. 0.00 <LOQ 0.00 <LOQ 0.00 <LOQ 0.00 <LOQ 977 G347-MS1 82% 972 81% 920 76% 872 47.2 79% 72.1 120% 42.8 71% 51.7 86% 84 I 70.9 955 79% 680 57% 804 67% 0.00 G347 34.7 0.00 86.2 AL343 <LOQ <LOQ <LOQ 0.0862 39.1 <LOQ 0.00 <LOQ 60.6 <LOQ 9.72 <LOQ 16.5 <LOQ 0.00 <LOQ 147 0.147 0.00 22780 22.8 <LOQ 15691 15.7 17080 17.1 28909 28.9 20463 20.5 28.0 28049 43414 43.4 48741 48.7 39880 39.9 65404 65.4 75.3 75293 NE NE Mean PFOS uglg o r %Ree <LOQ 81% 74% 72% 89% 68% <LOQ 18.5 25.8 44.0 70.3 NA RSD (%) Std. Dcv.(ug/g) MSlMSD 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. I Excel Version 5/95 TOX129 liver-3,xls 3/8/01 9:OlAM FACT-TOX-129 Study: Product Number(Test Substance): Matrix: McthodRcvision: Analytical Equipment SystemNumber: Instrumcnt S o h a r e N e n i o n : Date of ExbactiodAnalyst: Date of AnalysidAnalyst: Date of Data ReductiodAnalyst: Sample Data BOBWHITE LIVER DAY 22 - I Group I Dose Method Blk I - QC - 1.00 ppm - - Sample X BWL07209-H20 BIk-9-1 BWL07209-HZO Blk-9-2 BWL07209-H20 Blk-9-3 BWL07209-HE2l0k-94 B435-MS9-1 B435-MSD-9-1 (3343-MS9-2 0.0 mgkg food B43S G343 R344 Sample spilled lost-3 mL Y402 AL339 B436 G344 R338 Y403 AL340 8437 G345 R339 Y404 Group2 AL344 18.3 mgkg f w d B441 G349 R345 Y408 Group3 AL346 36.6 mgkg food 8443 G351 R343 Y410 Group4 AL348 73.2 mgkg food B445 G353 Group5 I46 mgkg fwd Group6 293 mglkg food I R348 Y412 AL35O B447 R349 Y414 AL352 FOS = Perfluorooetanesulfonate Limit ofQuantitation (LOQ): PFOS is approximately0.070 uglg NA = not applicable Date EnteredlBy: Date Verified/ By: 07121/99.07123/99 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.I Soup 020199 Masslynx 3.2 07/20/99MCH 07/22/99. 07/23/99 DRB 07/23/99.07/26/99 DRB Filename: R-Squattd Value: Slope: Y-Intercept: See Attachments See Attachments See Attachments See Attachments PFOS Calc. Cone. nvg 0.00 0.00 O.Oo0 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 92 1 2215 4324 11905 10253 10755 12618 33391 17719 23 I67 29026 39326 Concentration of PFOS ugle or % Rec. Mean PFOS ug/g or %Rec RSD (Ye) Std. Dev.(ug/g) M m S D RPD <LoQ <LoQ <LoQ <LOQ <LOQ NA 85% 78% 1 82% 1 8% 1 95% 91% 93% 5% 113% 1 0.164 0.266 0.136 0.153 0.141 0.265 0.285 0.235 0.2 15 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 I 43.4 0.2 15 0.093 24.8 3.25 0.806 53.4 2.92 1.56 33.0 10.0 3.29 26.5 25.8 6.84 NA NA FACT-M-2.1 Excel Version 5/95 TOX129 liver-3.xls 3/8/01 9:OlAM LabomvRePrt 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 of Results - Sera Analysis Day8 e Day22 e Results of MS/MSD Analysis FACT-TOX-I29 Study Fmdun NumtufTat Suhtancc) Matrix. MahodlRcvislm Analytical EquipmentSyttml Number Insbumcnt SomVueNmion~ Filename: R-Squared Value Slope. . Y-lntercepl: Dues of ExUaclicmIAndyR. D u n of AdyritlAndyst: Date of Data RodunionlAnalyn: Sample Data I O B W H m SERUM Day 8 GOUP Dor Sample # Mahod Blk h i m i x Blk Group1 0.0 m& food <LOQ Group2 18.3mgkg fmd H20 BIk-62899-I 1-1 H20 BL-62899-I 1-2 H20 Blk-62899-11-3 H20 Blk-62899-114 RBS Blk-062899-11-1 RBS BIk-062899-I 1-2 RBS Blk-062899-11-3 RBS Blk-062899-1I 4 G347 B438 Y405 AL343 B440 Y406 Y407 R341 R342 AL342 8439 G346 G348 AL341 B442 R340 G350 PFOS Dielay LC50 Study with Nonhm Bobwhitu PFOS BobwhiteS a w n ETS-84 I and ETS-8-5 I Amelia 062498.Soup 020199 Masslynx 3.2 S a AtUchmenU See Attachments S a Atochmmu See Anvhmen~ 06/28/99.08/24/99 SEWRP 07/23/99,07/26/99.08/24199.09/01/99 DRB/MEF/PTF 07/26/99,07/27/99,09/02/99, DRBiMEWTF Concatradon of?FOS uL/mL <LOQ <uQ <toQ <LoQ <LoQ <toQ nOQ aOQ <LoQ <LoQ aOQ <LoQ <LoQ aoQ <LOQ <LOQ a@? <LOQ <LOQ <LOQ <LOQ aOQ 0.136 Man PFOS uL/mL RSD Y. Std. Dcv. (uL/mL) <LOQ NA NA AL345 croup3 Y411 36.6 mgkg fmd B444 R347 AL347 G352 Group4 R348 73.2 mgkg food B446 G354 9.38 AL349 Y413 21.4 Group5 146 mgkg food AL35I 25.2 Y415 I Group6 4I 3.6 I 293mgkgfood G356 8448 AW53 imit of QuMtilation(LOPQF)O: S 0.010 @mL <LOQ 0.200 3.51 8.41 0.0542 <LOQ NA 57.7 76.3 1 I4 21.6 5.18 <LOQ 1 outlier NA 129 304 3.92 75.6 41.2 31.2 I19 49.4 41.6 - -I NA I NA NA Nu Amlicable PFOS Pcrfluorartmesulfautc Date EntaedlBy: Due Verified/ By 07/26199.07/27/99.09103/99 LAC 08/10199 GML, 04/1UOO KJH ETS-8-5 I Excel Venion 5 0195 TOX129 sera-3 XIS 31810 I FACT-TOX-I29 study Produn Nwnbe~iTaSt ubrunce) Mauix: MnhodlRcvisim: AnalyticalEquipmentSyrtm N w n k lnmwnmt Softwdmicm. Filename: R-Squad Value Slop: Y-lntercep: Dam of E x a u t i d A n d y s t : . Datcl of AnalysidAnalyst: Due of Dau RcducticdAnalysc Sample Data tBOBWHITE SEI UM Day22 Croup Sample I Method BIk Manix BIk H20Blk-71499-11-1 HZ0 BIk-71499-11-2 H20 Blk-71499-11-3 H20 Blk-71499-11-4 RBS BIk-071499-11-1 RBS BIk473499-11-2 RBS Bk-071499-11-3 RBS Blk471499-I 1-4 Au38 0.0 m& food B435 843 R344 Y402 u339 B436 G344 R338 Y403 AU40 B437 G345 R339 Y4M Group2 18.3mgkg food Au44 B441 G349 R345 Y408 Cronp.7 AW46 36.6 mgkg food B443 r73.2mgkg food I 4 6 mgkg food G351 R343 Y410 AW48 8445 G353 Iu49 Y412 hum B447 rum Y414 293 @g food Limit of Quvltiudm --52 OQ): PFOS 0 010 ugh PFOS Diemy LC50 Studywith Nonhm Bobwhitn PFOS Bobwhite Serum ETS-8-4.1 and ETS-8-5. I Soup 020199, Amelia 062498 Murlynx 3.2 Sce Attachments Sce Attachmmn Sce Anschmmts Sce Atrvhmmn 07/14199.08124l99 SEE 07l1~.07/20199,08/25/99 G"EEISAH 07/19199.07/21/99.09/02/99 DRBlMEE 0.0326 0.0284 0.0874 0. I25 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 7.27 9.31 16.7 16.6 69.6 18.8 60.0 36.5 27.0 38.4 1 I2 0.115-TwoOutlim 217 0.251 21.7 5.89 1.28 57.3 4.17 93.6 26.2 24.5 34.5 40.5 14.0 UA NA -PFOS P d u a m n a n a u l f a u t c bEntacd/By: Date Verified/ By 07/26/99,07I27199.09/03/99 LAC 08/1(y99 GML. 04/IYOO KM ETS-8-5 I Excel Vmion 5 0/95 TOXI29 sera-3 XIS 3/8/01 FACT-TOX-I 29 StUdy. PFOS Diary LC50 Study with Nachcm Bobwhita Roduct Number(Tat SubaI~cc): PFOS Matrix: Bobwhite S a m McfhodlRcvirim: ETS-8-4.I and ETS-8-5.I Andytiul Equipnmt Spanfrlumbcr Amelia 062498 InsmMmt SoftwuJvarion: Maulynx 3.3 FilrnmK: See Anrhmenu R - S q d Value: S a Anrhmcno Slope: SaAdmenu . Y-lnlerecp: Ducs of ExmnionlAnalyn: Dues of AndyridArulyn: S a Au- I lnU99 SEE I1/30/99 HOJ Duc of DmR c d u a i a J A r u l y n : 12/3/99GML Sample Data BOBWHITE SERUM MS/MSD's Group Dose Smpk # Conamtloll o t ?QOS Mun ?QOS RSD K Std. Dcv. H rJmL or H Rer nJmL or HRec M&od Blk H20BIk-11249-11-1 <I.oQ <LOQ NA Mlaix Blk BWS Blk-11249-11-1 CLOQ - BWS Bk-11349-11-1 <LoQ <LOQ NA QC 50 ppb BWSll249-MS-I 1-1 96% BWS11249-MSBll-I 104% - - 250ppb BWSll249-MS-11-1 91% BWSl1249-MSD-11-1 103% 97K 12% Limit of Quatiution (LOQ): PFOS 0.010 ug/mL NA N a Appriulbc DucEnmedBy: I y y 9 9 G M L DucVmfiedlBr: WlMo KIH ETS-S4.I Eaccl Vcnion 5 0195 TOX129 sera-3.xlr