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BACK TO MAIN Analytic Report Analysis o f PFOS in Test Organisms from the 96-Hour Static Acute Toxicity Test with the Freshwater M ussel Laboratory Report No. W2078 FACT Project No. FACT-TOX132 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 Requester Susan Beach 3M Company ' Environmental Technology & Safety Services Bldg 2-3E-09 I St. Paul, MN 55133 P a g e lo f BACK TO MAIN M ENVIRONMENTAL LABORA TORY ANALYTICAL REPORTNO. FACT-TOXI32 This study was designed to quantitatively assess perfluorooctane sulfonate (PFOS, C8F17S03-, CAS # 2795 39-3) levels in freshwater mussels after a 96-hour exposure. Details o f exposure conditions, clinical observations and other toxicity data were generated by the in-life testing facility (W ildlife International) and are not included in this rep o rt A nalytical results o f the test animals representing 5 dosage groups and 1 control group are presented here. In each dose group, there were 20 mussels exposed to PFOS. W ithin each group, 10 m ussels were exposed in one o f two chambers labeled A and B. Six o f the twenty test animals per group (3 from chamber A and 3 from chamber B) were analyzed for PFOS. PFOS w as identified in all test animals analyzed, and most o f the matrix mussels and control mussels. M atrix m ussels were sacrificed upon arrival at the in-life testing facility, prior to the start o f the study. Control m ussels were housed near die test animals in w ater to which no PFOS was added. PFOS levels w ere determined to increase with each increase in dose lev el Refer to attachment D for results o f individual mussel analysis. - Based on m atrix spike analysis studies, data generated in support o f this study should be considered to be semi-quantitative w ith a m argin o f error o f +/-35%. N o data was collected for analytes other than PFOS. A lthough rigorous quality control m easures and 3M Environmental Laboratory Standard Operating Procedures w ere followed, the acquisition o f this data was not necessarily collected according to applicable G ood Laboratory Practices. D ata presented here is the highest quality data available at this time. In-phase testing was conducted according to applicable Good Laboratory Practices. The conduct o f the in-life phase is the sole responsibility o f W ildlife International Ltd. See attachment C for die in-life protocoL M ussel specimens were collected on 06/01/99 and sent to die Environmental Laboratory on 06/07/99. The 3M Environm ental Lab received, on 06/08/99, a total o f 105 frozen mussel specimens. Twenty specimens w ere from group 1,20 from group 2 ,2 0 from group 3 ,2 0 from group 4 ,18 from group 5 ,2 from group 6, and 5 m atrix mussels. U pon specim en receipt, a chain o f custody was initiated and a copy o f this form can be found in Attachment C. There are no holding tim e criteria associated with this analysis. k- m ram Preparatory and Analytical Methods FA CT-M -1.1, Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis U sing HPLC-Electrospray/M ass Spectrometry Spectrometry with thefollowing exception:Due to die tim e constraints and lack o f control material, standard curves were prepared in m ethanol instead o f from m ussel tissue. 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/M ass Spectrometry with thefollowing exception: Page2of6 BACK TO MAIN 3M ENVIRONMENTAL LABORATORY ANALYTICAL REPORT NO. FACT-TOX132 Due to the time constraints and lack o f control material, standard curves were prepared in m ethanol instead o f from m ussel tissue. PFOS lot 171 was used as reference material. FACT-M -2.1, Analysis o f Fluorochem icals in Liver Extracts Using HPLC-Electrospray/M ass Spectrom etry ETS-8-7.0, Analysis o f Potassium Perfluorooctanesulfonate or Other Fluorochem icals in Liver Extracts U sing HPLC-Electrospray/M ass Spectrometry The characterization o f the reference m aterial (PFOS, lot #171), w ith respect to purity, stability, and hom ogeneity, was not complete on the date this report was issued. Once the characterization is com plete, a copy o f die report will on file in the 3M Environm ental Lab archives. Sample preparation - ion-pairing extraction On July 0 1,1999,1.0 mL o f each m ussel homogenate (six specimens per group) w ere extracted following m ethod FACT-M -1.1 which utilizes an ion pairing reagent [tetrabutyl ammonium hydrogen sulfate (TBA)] in a pH -controlled environm ent The cationic reagent selectively targets anionic fluorochemicals. Once the anion-TBA pair is formed, the analyte is transferred into a non-polar organic solvent, dried, and reconstituted in 1.0 mL o f m ethanol for M S analysis. Sam ples o f the test matrix from Group 4 animals were spiked w ith PFOS and extracted according to FACT-M -1.1 o n Ju ly O l, 1999. Samples o f test m atrix from the control group were spiked and extracted follow ing ETS-8-6.0 on July 22,1999 and A ugust 2,1999. A lthough two different extraction m ethod num bers were used for these specimens, die extractions were perform ed as described in m ethod ETS-8-6.0. For example, the July 01,1999 extraction follow ing m ethod FA CT-M -1.1 used methyl tert butyl ether solvent as stated in m ethod ETS-8-6.0, instead o f ethyl acetate as stated in FACT-M -1.1. HPLC/ESMSMS: for detailed quanitative work On July 07,1999 die m ussel extracts w ere analyzed using tandem electrospray m ass spectrometry (ESM SM S), analytical system "Soup020199" according to FACT-M -2.1, w ith die following exception. Because large amounts o f blank m atrix were not available, analyte levels were evaluated versus a solvent curve instead o f versus a curve extracted from matrix. Extra m atrix spike studies were conducted in order to validate this action. Results o f these m atrix spike studies were good (extraction efficiency greater than 65% ), therefore, these sample results are considered to be acceptable. In this analysis, a 10 pL aliquot o f extract is injected and passed through a reverse phase liquid chrom atographic column. Based on die affinity o f the analyte for the stationary phase in die colum n relative to the liquid mobile phase, die analyte is retained for a characteristic am ount o f tim e. For example, in a standard solution PFOS m ay elute at 5.0 minutes. Retention tim es betw een a standard PFOS solution and the analyte extracted from m ussel in this analysis were m atched on die HPLC system to w ithin 2%. Follow ing HPLC separation, ESMSMS provides a rapid and accurate means for analyzing a w ide range o f organic compounds, including fluorochemicals. Electrospray, an ionization technique used prim arily for detection o f molecular ions, is generally operated at relatively m ild temperatures. M olecules are ionized, and a prim ary ion, characteristic o f die analyte, is selected. This ion is bom barded w ith high-energy gas; subsequent collisions create sm aller secondary ionic fragments unique to the prim ary ion, w hich are detected. For example, for PFOS (C*FnSOi") analysis, ion 499 is selected as the characteristic prim ary ion. This ion is fragm ented into other ions such as 99 amu (corresponding to FSOj"), and 230 amu (C3F6SO3O. Each o f these secondary fragments is detected and can be used to differentiate PFOS from other compounds that Page3of6 BACK TO MAIN I 3MENVIRONMENTAL LABORATORY ANALYTICAL REPORT NO. FACT-TOXI32 I m ight have die same characteristic 499 amu prim ary ion, but different chemical compositions and secondary ion fragm entation patterns. ] A fter analysis on July 07,1999, it w as determ ined that dilutions were required for extracts from group 2-6 test anim als. D ilutions w ere prepared and analyzed on July 09,1999 using analytical system "Madeline 040198" according to m ethod FACT-M -2.1. The diluted samples were w ithin the calibration range. 5.1 Calibration For quantitative determ inations, a m id-level, unextracted m ethanol calibration curve verification (CCV) standard was analyzed every 5 to 10 sam ples to m onitor instrum ental d rift Calibration curve verification standards are compliant if instrum ental response is w ithin +/- 30% o f die expected value. In this study, all CCV standards were com pliant Q uantitation o f the target analytes is based on linear regression analysis (weighted 1/x) o f two curves prepared in methanol, bracketing each group o f samples. Q uantitation o f each analyte is based on the response o f one or more specific daughter ions using the m ultiple response-monitoring mode o f the instrum ent A nalyses on 08/02/99 and 08/03/99 contained poor calibration curve response; these data were not reported. 5.2 Blanks Four extraction blanks, utilizing w ater as surrogate matrix, were extracted w ith each batch o f samples. B lanks are compliant if no target analyte is detected above the lim it o f quantitation for a specific analyte. In this study, all blanks w ere com pliant . 5.3 Surrogates Tetra-hydro perfluorooctane sulfonate is used as a surrogate in this analysis. Surrogate response is m onitored to confirm gross instrum ental failure, if necessary. Surrogate response results were acceptable. 5.4 Matrix Spikes Four m atrix spikes at 100 ppb and four m atrix spikes at 1.0 ppm were extracted and analyzed utilizing all target analytes for w hich quantitative data is presented. Sam ples o f the test m atrix from Group 4 animals were spiked w ith PFOS and extracted according to FACT-M -1.1 on July 01,1999. Spike recoveries were erratic, probably due to exceptionally high endogenous levels o f PFOS; the m atrix spike studies were repeated on July 22,1999 and August 02,1999. Spike recoveries from the July 22,1999 extraction at 100 ppb and 1.0 ppm were within analytical acceptance criteria o f 65-130% . A s the m atrix spike data collected July 22,1999 was acceptable, data collected from analyses o f the A ugust 2,1999 extraction were not included in this report 5.5 Laboratory Control Samples Laboratoiy Control Samples are not a com ponent o f this study. 5.6 Sample Related Comments The level ofPFO S in sam ples o f m ussel tissue was determ ined by comparison w ith a standard curve prepared in solvent Typically, PFOS levels are determined versus a curve extracted from tissue in order to m ore accurately assess any m atrix effects. Extra matrix spike studies were conducted in order to validate this action. Results o f these m atrix spike studies were good (extraction efficiency greater than 65%), therefore these sample results are considered to be acceptable. . Page4of6 BACK TO MAIN 3MENVIRONMENTAL LABORATORY ANALYTICAL REPORT NO. FACT-TOXI32 PFOS was detected in the test animals analyzed. Refer to Table 1 for a summary o f average PFO S levels in mussels by dose group. Please refer to the attached spreadsheets for individual test animal results. Assuming spike recovery studies form a suitable indication o f endogenous analyte recovery, data are quantitiative to 35 %. The validity o f this assumption has not been verified by other techniques. The estim ated lim it o f quantitation (LOQ) for PFOS is 25 ppb (ng/mL) for the July 09,1999 analysis and 5.0 ppb (ng/mL) for the rem aining analyses (July 07,1999 and August 03,1999). The LOD and LOQ values are stated as ng/mL since the extracts were evaluated versus a solvent curve, not an extracted mussel curve. This report reflects die m ost accurate data available at this time. T able 1 A nalysis R esults-M ussel FRESHW ATER MUSSEL G roup/ A verage Cone. D osage PFO S in M ussel M atrix ug/g 0.0808 G roup 1 A: <0.0174pg/g C o n tro l B: 0.0502 Group 2 5.3 ppm A: 3.41 B: 3.96 Group 3 12 ppm A: 4.83 B: 5.61 Group 4 20 ppm A: 8.07 B: 6.58 Group 5 41 ppm A : 11.2 B: 12.5 Group 6 79 ppm A: 91.2 B: 86.4 M atrix = M ussel specimens which were sacrificed upon arrival o f all mussel specimens at die in-life testing facility and not exposed to control or dosed water. A = Specimens stored in Cham ber A. B = Specimens stored in Chamber B. Specimens and extracts w ill be retained for a minimum o f one year for future evaluation, if necessary. Page 5 o f6 BACK TO MAIN 3MENVIRONMENTAL LABORATORY ANALYTICAL REPORTNO. FACT-TOX132 8 .1 A ttachm ent A: Analytical Report Addendum 1 8 .2 A ttach m en t!): W ildlife International Protocol 8 .3 A ttachm ent C: Extraction and Analytical Methods 8 .4 A ttachm ent D: Chain o f custody form(s) 8 .3 A ttachm ent E: Complete Analytical Results 8 .6 A ttachm ent F; Extraction and Analysis Information 'h ' 'tifyijttifoy* * - a L,,iisa Clemen, Advanceed Chemist L , h -- Kris Hansen, P hD ., Specialist u la ilh b D ate Q (* / 3 /& 0 D ate 6> u c OLnoo Page 6 o f6 BACK TO MAIN 3MENVIRONMENTAL LABORATORY ANALYTICAL REPORT NO. FACT-TOX132 AnalyticalReport Analysis o f PFOS in Test Organisms from the 96-Hour Static Acute Toxicity Test with the Freshwater M ussel Laboratory Report No. W2078 FACT Project No. FACT-TOX132 Attachment A Analytical Report Addendum 1 BACK TO MAIN 3M ENVIRONMENTAL LABORA TORY ANALYTICAL REPORTNO. FACT-TOXI32 ADDENDUM I AnalyticalReportAddendum 1 Analysis o f PFOS in Test Organisms from the 96-Hour Static Acute Toxicity Test with the Freshwater M ussel Laboratory Report No. W2078 FACT Project No. FACT-TOX132 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 .B ox 3331 St. Paul, M N 55133-3331 Phone: (651)778-6018 FAX: (651) 778-6176 P agelof4 I I 1 BACK TO MAIN 3M ENVIRONMENTAL LABORA TORY ANALYTICAL REPORTNO. FACT-TOX132 ADDENDUM I s 3 a a s e :;* i . - ^ f e This study addendum was designed to quantitatively assess perfluorooctane sulfonate (PFOS, C8F17S03-, CAS # 2795-39-3) levels in unexposed, freshwater "matrix" mussels. M atrix mussels were sacrificed upon arrival at die in-life testing facility, prior to die start o f die study. In the original study, the samples o f m atrix mussels were determined to contain an average o f0.0808 ug/g o f PFOS. This level o f PFOS in unexposed mussels was unexpected, so further analyses were conducted. The results o f die reanalysis, reported in this addendum indicate that PFOS levels in the samples o f matrix m ussels are less than 0.0174 pg/g. It is likely that the original samples o f mussel m atrix that were analyzed were contaminated during sample prep. 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 G ood Laboratory Practices. Data presented here is the highest quality data available at this time. M ussel specimens were collected on 06/01/99 and sent to the Environmental Laboratory on 06/07/99. The 3M Environmental Lab received, on 06/08/99, a total o f 105 frozen mussel specimens. One hundred and five specimens were received; 5 o f these were matrix mussels. Upon specimen receipt, a chain o f custody was initiated and a copy o f this form can be found in Attachment C o f the final analytical report (TOX132). There are no holding time criteria associated w ith this analysis. Preparatory and Analytical Methods Please refer to the analogous section in the full analytical report tv ` ,, , ^ av ^ Tm jgkl^. 14.1 Calibration For quantitative determinations, a mid-level, unextracted methanol calibration curve verification (CCV) standard was analyzed every 5 to 10 samples to monitor instrumental drift. Calibration curve verification standards are compliant if instrumental response is within +/- 30% o f the expected value. In this study, all CCV standards were com pliant Q uantitation o f die target analytes is based on linear regression analysis (weighted 1/x) o f two curves prepared in methanol, bracketing each group o f samples. Quantitation o f each analyte is based on the P a g e2 o f4 BACK TO MAIN 3M ENVIRONMENTAL LABORATORY ANALYTICAL REPORTNO. FACT-TOXI32 ADDENDUM I response o f one or more specific daughter ions using the multiple response-monitoring m ode o f the instrum ent 14.2 Blanks Two extraction blanks, utilizing w ater as surrogate matrix, were extracted w ith each batch o f samples. Blanks are compliant if no target analyte is detected above die lim it o f quantitation for a specific analyte. In this study, all blanks were com pliant 14.3 Surrogates Tetra-hydro perfluorooctane sulfonate is used as a surrogate in this analysis. Surrogate response is m onitored to confirm gross instrumental failure, if necessary. Surrogate response results w ere acceptable. 14.4 Matrix Spikes Two matrix spikes at 50 ppb were extracted and analyzed for PFOS. The m atrix spikes show ed recoveries o f 119% and 75%, within the acceptable range o f +/-30%. 14.5 Laboratory Control Samples Laboratory Control Samples are not a component o f this study. 14.6 Sample Related Comments The level o fPFOS in samples o f mussel tissue was determined by comparison w ith a standard curve prepared in solvent Typically, PFOS levels are determined versus a curve extracted from tissue in order to m ore accurately assess any matrix effects. Results o f m atrix spike studies conducted in the original study and described in the final analytical report validate the choice o f using an unextracted curve. w The results o f the reanalysis, reported in this addendum indicate that PFOS levels in die samples o f m atrix mussels are less than 0.0174 pg/g. It is likely that the original samples o f m ussel m atrix that were analyzed were contaminated during sample prep. Assuming spike recovery studies form a suitable indication o f endogenous analyte recovery, data are quantitative to 35 %. The validity o f this assumption has not been verified by other techniques. Specimens and extracts w ill be retained for a minimum o f one year for future evaluation, if necessary. P a g e3 o f4 1 I BACK TO MAIN 3M ENVIRONMENTAL LABORA TORY ANALYTICAL REPORTNO. FACT-TOX132 ADDENDUM 1 17.1 None. ,>Yfjf ( ft i t ! 1 f i ______ L isa Clemen, Advanced Chemist U * rL K ris Hansen, PhD ., Principal A nalytical Investigator Date O L /6 9 /o 0 Date I I I I Page4of4 FACT-TOX-132 BACK TO MAIN Study: Product Number(Test Substance): Matrix: Method/Revision: 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 96 Hour Static Acute Toxicity Study with the Freshwater Mussel PFOS Freshwater Mussel Filename: ETS-8-6.0, ETS-8-7.0 R-Squared Value: Amelia 062498 Slope: Masslynx 3.2 Y-Intercept: 09/14/99 RWW 09/19/99 MEE 09/20/99 MEE See Attachments See Attachments See Attachments See Attachments MUSSEL Group Dose Sample # Method Blk H 20 Blk-1 H 20 Blk-2 QC - 100 ppb, 500 ppb 1A-10-100 ppb-1 36417 1A-10-100 ppb-2 lA-10-500ppb-l lA-10-500ppb-2 Matrix MATRIX-1-1 clean mussels MATRIX-1-2 MATRIX-1-3 MATRIX-1-4 MATRIX-1-5 Group 1 1A-6 Control 1A-10 IB-3 IB-6 PFOS = Perfluorooctancsulfonate Limit o f Quantitation (LOQ): PFOS = 0.0347 ug/g Date Entered/By: Date Verified/ By: 09/21/99 LAC 12/13/99 GML Concentration of PFOS ng/g <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) 105% 110% 99% 90% <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) 0.0663 NA: not applicable Mean PFOS ng/g <LOQ 108% 95% <LOQ <LOQ NA RSD Std. Dev. MS/MSDRPD NA NA 5% 10% NA ' NA NA NA NA FACT-M-2.1 Excel Version 5/95 TOX-132-Mussel.xls 5/24/00 5:27 PM FACT-TOX-132 BACK TO MAIN Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst: Date o f Analysis/Analyst: Date of Data Reduction/Analyst: Sam ple D ata . 96 Hour Static Acute Toxicity Study with the Freshwater Mussel PFOS Freshwater Mussel Filename: ETS-8-6.0, ETS-8-7.0 R-Squared Value: Amelia 062498 Slope: Masslynx 3.2 Y-Intereept: 9/22/99 MCH 09/24/99 IAS 09/27/99 IAS See Attachments See Attachments See Attachments See Attachments MUSSEL Group Dose Sample # Method Blk H 20 Blk-9 H 20 Blk-10 QC - 50 ppb MATRIX-1-3-MS-9 e x t 9/22/99 MATRDC-1-3-MSD-10 M atrix MATRIX-1-1 clean mussels MATRIX-1-2 MATRIX-1-3 MATRIX-1-4 MATRIX-1-5 PFOS = Perfluorooctanesulfonate Limit of Quantitation (LOQ): PFOS - 0.0174 ug/g Concentration of PFOS ug/g <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 119% 75% <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) NA: not applicable Mean PFOS ug/g <LOQ 97% <LOQ RSD Std. Dev. MS/MSDRPD NA , 45% NA NA Date Entered/By: Date Verified/By: 10/06/99 LAC 12/13/99 GML FACT-M-2.1 Excel Version 5/95 TOX-132-MusseI.xls 5/24/00 5:27 PM BACK TO MAIN 3M ENVIRONMENTAL LABORA TORY ANALYTICAL REPORTNO. FACT-TOXI32 ADDENDUM I AnalyticalReport Analysis o f PFOS in Test Organisms from the 96-Hour Static Acute Toxicity Test with the Freshwater M ussel Laboratory Report No. W2078 FACT Project No. FACT-TOX132 Attachment E Complete Analytical Results FACT-TOX-132 Study: Product Numbcr^Tcit Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst Date of Analysis/Analyst Date of Data Reduction/Anal yst Sample Data 96 Hour Static Acute Toxicity Study with the Freshwater Mussel PFOS Freshwater Mussel FACT-M-1.1, FACT-M-2.I, ETS-8-6.0, ETS-8-7.0 Soup 020199, Madeline 040198 Masslynx 3.2 07/01/99.07/22/99, 08/02/99 SRP/RWW 07/07/99, 07/09/99,08/02/99,08/03/99 DRB/GML/SAH 07/09/99, 07/30/99,08/04/99 DRB/GML Filename: R-Squared Value: Slope: Y-Intercept See Attachments See Attachments See Attachments See Attachments MUSSEL Group Pose Sample # Method Blk H20 Blk-1 H20 Blk-2 H20 Blk-3 H20 BIk-4 Precision Check 4B Blk-1-1 4B Blk-1-2 4B Blk-1-3 4B Btk-t-4 Matrix Blk lA-10-Bflc-l 07/22/99 QC - 100 ppb, 1.0 ppm 07/22/99 lA-10-BIk-2 1A-10-100 ppb-1 1A-10-100 ppb-2 IA-UM00 ppb-3 1A-1(V100ppb-4 1A-10-I.0ppm-1 1A-10-I.0 ppm-2 1A-10-I.0ppnv3 tA-lO-l.Oppm-4 Matrix Blk 08/02/99 lA-10-Blk-l lA-10-BIk-2 QC - 1.0 ppm, 10 ppm 08/02/99 1A-10-1.0 ppm-MS-1 1A-10-1.0 ppm-MSD-1 1A -10-1.0 ppm-MS-2 1A-10-1.0 ppm-MSD-2 1A-10-10 ppm-MS-1 1A-10-10 ppm-MSD-1 1A-10-10 ppm-MS-2 1A -10-10 ppm-MSD-2 G roupl 1A-6 Control 1A-9 IA-10 IB-1 IB-3 IB-6 Groop2 2A-4 5.3 ppm 2A-5 2A-7 2B-6 2B-7 . 2B-10 Group3 12 ppm 3A-7 3A-8 3A-9 3B-5 3B-7 3B-10 Group* 4A-5 20 ppm 4A-8 4A -10 4B-1 4B-2 4B-6 Group5 5A-7 41 ppm 5A-8 5A-9 5B-4 5B-7 5B-9 Group6 6A-1 79 ppm 6B-1 M atrix MATRIX-1-1 clean mussels MATRIX-1-4 MATRIX-1-5 PFOS - Pcrfluorooctanesulfooate Limit of Quantitation (LOQ): 0.0867 ug/g (070999Files) Limit of Quantitation (LOQ): 0.0174 ug/g (070799 and 080399 Files) Concentration of PFOS 8/8 <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 7.89 7.68 8.76 7.40 <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 102% 91% 121% 103% 89% 83% 92% 96% <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 83% 74% 80% 85% 89% 89% 77% 87% <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 0.0227 <LOQ (0.0174 ug/g) 0.0778 3.87 3.70 2.67 3.25 3.15 5.47 4.85 5.76 3.88 5.11 5.75 5.97 6.91 10.2 7.07 8.06 4.79 6.88 13.1 10.9 9.55 14.0 14.5 8.92 91.2 86.4 0.213 0.0434 0.0251 NA Not applicable _ Mean PFOS rt RSD Std. Dev. MS/MSD RPD <LOQ NA 7.93 <LOQ 7.38 0.585 NA 104% 8% 12% 90% <LOQ 4% 5% NA 4% 80% 5% 5% 86% 6% <LOQ NA 0.0502 77.5 0.0389 19.1 3.41 0.654 33.2 3.96 1.31 19.5 4.83 0.940 8.00 5.61 0.449 23.2 8.07 1.87 25.2 6 58 1.66 16.1 11.2 1.79 24.8 12.5 3.10 NA 88.8 - 0.0537 ' 0.0937 * 110 0.103 Date Entered/By: Date Vcrified/ By: 07/09/99,07/30/99,08/04/99 LAC 08/05/99 GML BACK TO MAIN FACT-M-2.1 Excel Version 5/95 TOX-132-Mussel,xls 5/24/00 5:27 PM I I I i I I I I 1 I 1 I I I a a a a FACT-TOX-132 BACK TO MAIN Study: Product Number(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 96 Hour Static Acute Toxicity Study with the Freshwater Mussel PFOS Freshwater Mussel Filename: ETS-8-6.0, ETS-8-7.0 R-Squared Value: Amelia 062498 Slope: Masslynx 3-2 Y-Intercept: 09/14/99 RWW 09/19/99 MEE 09/20/99 MEE See Attachments See Attachments See Attachments See Attachments MUSSEL Group Dose Sample # Method Blk H 20 Blk-1 H 20 BIk-2 QC - 100 ppb, 500 ppb 1A-10-100 ppb-1 36417 1A-10-100 ppb-2 lA-10-500ppb-l lA-10-500ppb-2 Matrix MATRIX-1-1 clean mussels MATRIX-1-2 MATRIX-1-3 MATRIX-1-4 MATRIX-1-5 Group 1 1A-6 Control 1A-10 IB-3 IB-6 PFOS = Perfluorooctanesulfonate Limit of Quantitation (LOQ): PFOS = 0.0347 ug/g Concentration of PFOS ug/g <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) 105% 110% 99% 90% <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) 0.0663 NA: not applicable Mean PFOS ug/g <LOQ 108% 95% <LOQ <LOQ NA RSD Std. Dev. MS/MSDRPD NA NA 5% 10% NA NA NA NA NA Date Entered/By: Date Verified/ By: 09/21/99 LAC 12/13/99 GML FACT-M-2.1 Excel Version 5/95 TOX-132-MusselJds 5/24/00 5:27 PM FACT-TOX-132 BACK TO MAIN Study: Product Number(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: Sam ple D ata . 96 Hour Static Acute Toxicity Study with the Freshwater Mussel PFOS Freshwater Mussel Filename: ETS-8-6.0, ETS-8-7.0 R-Squared Value: Amelia 062498 Slope: Masslynx 3.2 Y-Intercept: 9/22/99 MCH 09/24/99 IAS 09/27/99 IAS See Attachments See Attachments See Attachments See Attachments MUSSEL Group Dose Sample # Method Blk H 20 Blk-9 H 20 Blk-10 QC - 50 ppb MATRIX-1-3-MS-9 e x t 9/22/99 MATRIX-1-3-MSD-10 M atrix MATRIX-1-1 clean mussels MATRIX-1-2 MATRIX-1-3 MATRIX-1-4 MATRIX-1-5 PFOS = Periluorooctanesulfonate Limit of Quantitation (LOQ): PFOS = 0.0174 ug/g Concentration of PFOS ug/g <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) 119% 75% <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) <LOQ (0.0174 ug/g) NA: not applicable Mean PFOS " g/g <LOQ 97% <LOQ RSD Std. Dev. MS/MSD RPD NA 45% NA NA Date Entered/By: Date Verified/ By: 10/06/99 LAC 12/13/99 GML I I 8 8 8 8 FACT-M-2.1 Excel Version 5/95 TOX-132-Mussel.xls 5/24/00 5:27 PM