Document BROGBNEqxwQQ2r9ad6egzpggk

25) OECD 205-OPPTS 850.2200, Analysis of fluorochemicals in Bobwhite quail samples, E02-0659 (from study 454-1 12) e- __-. E02-0659 Analyticaf Data Summary: 3M Environmental Lab SAN ITlZED Anatybcal Report DEC 0 9 2003 Quantitative Analysis of Fluorochemicals in Bobwhite Quail Samples Obtained from Wildlife International, Ltd Amendment I- September 8,2003 3M Study No./3M Laboratory L l M S No. E02-0659 r Testing Laboratory ExtractiodAnalyses 3M Environmental Technology & Safety Services 3M Environmental Laboratow I 1 Building 2-3E-09,935Bush Avenue I I St. Paul, MN 55106 Laboratory Contact Bldg. 2-3E-09 P.O. Box 3331 St. Paul, MN 55133-3331 Phone: FAX: Requester 3M Environmental Technology and Safety Services Bldg. 2-3E-09 P.O. Box 3331 St. Paul, MN 55133-3331 Page 1of8 E02-0659 Analytical Data Sininmoly: 3M Environmental Lab 1 Amendment This amendment was written to update "Table 1:Target ions Monitored in 3M Laboratory Analyses" to "Table 3: Target Ions Monitored in 3M Laboratory Analyses", to update the tables following Table 3 to the correct numbers, and to change the analyte listed in the header of Table 5 to PFBS. 2 Introduction The purpose of the study is to determine the presence and concentration of pemuorobutanesulfonate(PFBS) in bobwhite liver and sera samples (collectedfrom Day 8 of an acute dietary LC50 study, from birds exposed to 5620 or 10000 mglkg in the diet) collected by Wildlife International, Ltd (E00-1429). Analyses of liver and sera samples were completed by the 3M Environmental Laboratory under study number E02-0659,and the results of these analyses are presented in this report. This study was not intended to be performed under compliance with the Good Laboratory Practice (GLP) Standard. 3 Sample Receipt The 3M Environmental Laboratory received liver and sera specimens on 18 June 2002 and 03 July 2002. All samples were received frozen, in good condition, and were transferred to frozen storage at -20Cf 10C upon arrival. 1 Table I . Bobwhite Liver Samples Received I Sample ~~~ 5454-1 12-79 Timepoint Day 8 Dietary Exposure Concentration uglg ~~ 5.620 53454-1 12-80 5454-112-81 S454-112-82 S454-112-83 954-112-84 S454-112-85 Day 8 10,000 S454-112-86 S454-112-87 5454-112-88 5454-112-89 354-112-90 Page 2 of8 E02-0659 Analytical Data Summaly: 3M Environmental Lab Table 2. Bobwhite Sera Samples Received Sample Timepoint S454-112-126 S454-112-127 S454-112-129 954-112-130 S454-112-131 S454-112-132 9 5 4 - 112-133 s454-112-134 5454-112-135 Day 8 Day 8 1 Dietary Exposure Concentration ug/g 5,620 10,000 4 Holdinq Times No known holding times are associated with these samples. 5 Methods - Preparatory and Analytical Preparatory/AnalyficaI Methods ETS-8-231.I,"Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices".This method was used to preparelanalyzeliver and sera samples. PFBS was extracted from approximately 1.O mL of homogenized liver or approximately 1.O mL of diluted sera using C18 solid phase extraction (SPE) cartridges. The compounds were eluted from the C18 cartridge using 2.0 mL of methanol. Quantitative analyses were performed by monitoring a single product ion selected from a primary ion characteristic of a particular fluorochernical using HPLC-ESIMSIMS. For example, molecular ion 299, selected as the primary ion for PFBS (C4F9S03-)analysis, was fragmented further to produce ion 99 (FS03-). The characteristic product ion 99 was monitored for quantitative analysis. The method was modified as approximately 0.16 g were weighed from each of the six liver samples, combined together, and homogenized using 9.0 mL of water (labeled as 5454-112- Cornp(79-84)and S454-112-Comp(85-90). For the sera samples, approximately0.02 - 0.3 mL were measured and diluted 1/20 with water. From each of three diluted sera samples, 0.33 mL was removed for the first composite (labeled S454-112-Comp(126-129). From each of six diluted sera samples, 0.16 mL was removed for the second composite (labeled S454-112Comp(130-135). Note: As standards were prepared for this study, they were correctedfor purity. Page 3 of 8 E02-0659 Analytical Data Summary: 3M Environmental Lab Analytical Equipment The following are representative of the actual settings used during the analytical phase of this study. liquid Chromatograph: Hewlett-Packard' Series I100 Liquid Chromatograph system Analytical column: Keystone@BetasiP CIB 2 x 50 mm (5 pm) Column temperature: 30C Mobile phase components: ComponentA: 2mM ammonium acetate Component B: methanol Flow rate: 300 pUmin Injectionvolume: 10 V L Solvent Gradient: 10.0 minutes . Time %B (minutes) 0.0 10% 1 .o 10% 5.5 95% 7.5 95% 8.0 10% Mass Spectrometer: Micromass@AfVMass Spectrometer Quattro HTYTriple Quadrupole system Software: Mass Lynx" 3.4 Cone Voltage: 15-60 V CollisionGas Energy: I C 4 0 eV Mode: ElectrosprayNegative Source Block Temperature: 150C klOC Electrode: Z-spray Analysis Type: Multiple ReactionMonitoring (MRM) Table 3. Target Ions Monitored in 3M Laboratory Analyses Target Analyte Primary Ion (AMU) Product Ion (AMU) Retention Time I PFBS . I 299 I 99 I -5.8min. I Liver samples were homogenized beginning on 08 October 2002. Sera samples were diluted beginning on 14 October 2002. All were extracted beginning on 14 October 2002 and analyzed beginning on 17 October 2002 following method ETS-8-231.1 using high-performance liquid chrornatography-electrospray/tandemmass spectrometry (HPLC-ES/MS/MS) in the multiple reaction mode versus extracted matrix curves. PFBS levels were quantitated by external calibration. Page 4 of8 E02-0659 Analytical Data Surnrnaly: 3M Environmental Lab 6 Data Quality Objectives The following data quality objectives were indicated in the method performance section of ETS8-231.1, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices". The data quality objectives were followed loosely in this analysis. Practiced procedures are adequate to support the stated data quality for this data set. System Suitability: System suitability will be determined prior to the start and at the completion of each analytical run. Prior to the calibration curve and after the last sample of the run three (3) mid-level unextracted calibration standards will be analyzed. As applicable, the peak area precision, retention time precision, resolution, and peak asymmetry will be monitored at the beginning and the end of the run separately. The peak area precision must be equal to or less than 5.0% RSD, the precision of the retention time must be equal to or less than 2.5% RSD, the resolution must be > 2.0, and the peak asymmetry (fronting or tailing) must be 0.5<AF<2.0, where AF is the.asymmetryfactor. Calibration: One calibration curve will be prepared from extracted matrix standards, in the same matrix as the samples. It will consist of 8 levels. The curve will be plotted by quadratic regression, weighted Ilx, and not forced through the origin, using MassLynx or other suitable software. The coefficient of determination (P) equal to or greater than 0.990, All active calibration curve points must be within 25% of the theoretical value with the exception of the LOQ point, which may deviate up to 30%. Calibration standards with peak areas less than two times the curve matrix blank will be deactivated to disqualify a data range that may be affected by background levels of the analyte. A valid calibration curve must contain at least 6 active points above and including the LOQ Blanks: Water and matrix blanks must have a peak area 5 % the peak area of the lowest acceptable standard (LOQ). Surrogate Standards: Surrogates will be added to all samples prior to the start of the preparatory procedure. The surrogate to be used is nonadecafluorodecanoicacid (PFDA) and it will be used to determine method performance. Surrogates standards must have a percent deviation c 50%. Limits of Quantitation (LOQ):The LOQ is equal to the lowest acceptable standard in the calibration curve. Matrix Spikes: Matrix spikes are not a component of this study. Quality Control (QC) Samples: Three (3) quality control samples (QC) will be prepared for each matrix during the course of the study. QC samples will consist of one sample at each of three levels of analyte. The levels listed below may be used: Low level. 3X to 5X the LLOQ, Mid-level:equivalentto a point near the middle of the calibration curve, High level: 80% of the ULOQ Two QC sample levels are analyzed afler every tenth sample injection starting afler the last calibration standard injection, with a minimum of three QC per analysis. Solvent blanks are not considered samples but may be included as such for determining when QC samples will be analyzed. Page 5 of 8 E02-0659 Analytical Data Sumrnav: 3M Environmental Lab Each QC is expected to show an accuracy of 75-125% of expected. A minimum of 2/3 of all QC samples analyzed within an analytical run must meet this criteria, and a minimum of 1/2 of the QC samples at each level within an analytical run must meet this criteria. If not, the set must either be re-analyzedor re-extracted 7 Analysis System Suitability: System suitability was determined prior to the start and at the completion of each analytical run. The peak area precision was less than 5.0% RSD (0.5 - 3.0%), the precision of the retention time was less than 2.5% RSD (0.0 - 0.34%). Calibration: Quantitation of the target analyte was based on quadratic regression fit analysis weighted I l x of a single extracted curve for each group of liver or sera samples. Points on the curve which weren't within +/- 25% criteria or low curve points with peak areas less than 2 times that of the matrix blanks were deactivated. Quantitation of each analyte was based on the response of one specific product ion using the multiple reaction-monitoring mode of the instrument. Extracted calibration standards were preparedto run, undiluted, at approximately 10 ng/g or n g h L - 1000 ng/g or ng/mL. The coefficient of determination (P) of each standard curve was a.990. Demonstration of Specificity: Specificity for analyte identification was demonstrated by chromatographicretentiontime (-5.8 minutes) and mass spectral daughter ion characterization. Blanks: Kandiyohiwater was used for all water blanks. Bobwhiteliver and Bobwhite sera were used as matrix blanks. All blank peak areas were 5 X the peak area of the limit of quantitation for the compounds of interest. Surrogate Standards: PFDA surrogate standard was spiked into all samples during the course of this study. The percent difference of each surrogate standard was c 50%. Limits of Quantitation (LOQ): The LOQ was equal to the lowest acceptable standard in the calibration curve (defined as a standard within *30% of the theoretical value), and had a peak area 2 times greater than the analyte peak area detected in the average of the surrogate matrix blanks. Because low levels of the target analytes are ubiquitous in the laboratory, it IS imperative that these criteria for LOQ determination be observed. The approximate LOQ for each analyte (reported as <L.OQ) in each matrix is listed on the results summary tables. Matrix Spikes: Matrix spikes were not a component of this study. Quality Contra1(QC) Samples: Three (3) quality controlsamples (QC) were prepared for each matrix during the course of the study at approximately 25 ppb, 250 ppb, and 750 ppb. Ail were within +/-25% criteria (range: 3-25%). Laboratory Control Samples: Laboratory Control Samples were not a component of this study. Page 6 of 8 E02-0659 Analytical Data Sumrnaly: 3M Environmental Lab Sample Related Comments: It is not possible to verify true recovery of endogenous analyte from liverlsera without radiolabeled reference material. The only measurement of accuracy available at this time, QC samples, indicate that these data can be reported with a stated accuracy of +/-25%. These data were corrected for purity of the target analytes. Available purity information was included in the raw data. 8 Data Summary PFBS was detected in the Bobwhite liver and sera samples. Refer to Tables 4 and 5 for more information. I Table 4. PFBS Data Summary of Bobwhite Liver Samples Sample Timepoint Dietary Exposure Concentration ug/g PFBS Liver Concentration ug/g 1 Mean ug/g S454-112-Cornp(79-84>1 Day 8 5,620 1.29 1.36 S454-112-Cornp(79-84)-2 1.43 S454-112-Comp(85-90)-1 10,000 1.40 1.34 S454-112-Cornp(8590)-2 1.27 I Table 5. PFBS Data Summary of Bobwhite Sera Samples Sample Timepoint Dietary Exposure 954-112-Comp(126-129>1 S454-112-Cornp(126-129)-2 3 5 4 - 112-Comp(l30-135)-1 S454-11Z-Cornp(l30-135)-2 Day 8 5,620 10,000 PFBS Sera 2.55 2.78 1.30 1.22 I 2.67 1.26 Example Calculation: AR (ng/mL) x DF x d curve (mUmL) x VR ImL) Curve x FV (mL) in Curve x = (VglmL) 8 sample (mUmL) VR (mL) Matrix N (mL) in Matrix 1000 ng AR = Analytical result from MassLynx summary DF = Dilutionfactor MA = Matrix amount 3 curve = MA of tissuelfluidcalibration curve, assumed to be 1 g or 1 mU5 mL water d sample = MA of tissuehluid sample L g or mL of sample/5 mL water) VR = Volume removed for extraction FV = Final volume Page 7 of 8 E02-0659 Analytical Data Sumrnaly: 3M Environmental Lab Example calculationfor sample S454-112-Comp(l26-129)-1: 101.13 ng/mL x 25 x 1 mu5 mL x 1.0 mL x 2.0 mL x 1.0 uq = 2.55 pg/mL 1 mU5mL 0.99mL 2.0mL l000ng Refer to Attachment A for data summary tables. 9 Data / Sample Retention All original raw data and analytical report will be archived at the 3M Environmental Laboratory. The analytical reference standard reserve sample and remaining specimens pertaining to the analytical phase of this study will be archived at the 3M Environmental Laboratory according to applicable standard operating procedures. 70 Attachments 10.1 Attachment A: Data Summary Tables 77 Signatures 0 9/os lu3 Date 1 -- d Laboratory Management Date ,lualityAssurance Representative Date' Page 8 of 8 E02-0659 Analytical Data Summary: 3M Environmental Lab ~ Anafytical Report Quantitative Analysis of Fluorochemicals in Bobwhite Quail Samples Obtained from Wildlife International, Ltd Amendment 1 - September 8,2003 3M Study No./3M Laboratory LlMS No. E02-0659 Attachment A Data Summary Tables Various Tissues E02-0659 E02.0659 NA Bobuhilc Liver versus Bobuhtc Liver crlracled cunFdenvne ETS-a-2II.I Amclid161498 MasrLynx 3 4 10/14mz RWW R-Squucd Value: Slope: Y-lnlcrccpl. 10/17102 LAS IOR3lOl LAS See Anachmmu Scc Anachmcnls See Anrchmenu Scc And"ncna 11/13/02 LAS 0 ETS-I-211I. hlicroroll Excel 2wO E02-0659 Study. Producl Nurnber(Tcii Subrtmce). Matrix. MelhodlRcvlrion Andricd Equipment System Numbu. Instrument SoRwucNvsim Date of Emaenon/Andys~ Date of AndyrdAndyst D a l e of Data Rcductidhdyrt Sample Data Various Tissues E024659 NA BobwhiteS c r i versu Bobwhtc Sera criraclcd cg~lcnamc ETS-8-231 I R-Squared Value Amclia062498 Slope MarrLyn. 3 4 10/14m2 RWW Y-hterscp1 I WI 7102 LAS 1 o n i m 2 LAS SCK Anwhmcnts See Anachments See Anachments See Attachments BWS101402-Sera Blank-3 11/13102 LAS 0 ETS-8.23 I. I Microsoh Exccl ZOO0 sera E02-0659-8obhtc liver-rcndr