Document 8RQrdQK1Nmw0BjkYrzqG9mggK

Report EO1-1256 PFOS: A Reproductiun Study with the Mallard Study Title A nalytical Phase Report for PFO S : A Reproduction Study w ith th e Mallard Data Requirement FIFR A Subdivision E, Section 7 1-4 O E C D Guideline 206 Author Lisa A. Stevenson Analytical Phase Initiation Date February 19, 2002 Analytical Phase Completion Date At Signing Performing Laboratory 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Environmental Laboratory Study # E01-1256 Wildlife International Ltd: 4 5 4 -1 0 9 Total Number of Pages 216 3M Environmental Laboratory Page 1 o '> 0 6 2 Report EO1-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard GLP C o m p lia n c e S ta tem en t Study Title: Analytical Phase Report for PFOS: A Reproduction Study with the Mallard Study Identification Number: E01-1256 This analytical phase was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards with the exceptions listed below: Exceptions to GLP compliance: Automated data collection systems are not validated. The hardcopy printouts are considered as the original raw data. The stability of the samples has not been determined. Not all data generated were recorded directly or promptly, or initiated and dated on date of entry. J _isa A. Stevenson, Principal Analytical Investigator (2. A. Beach, Sponsor Representative _0[ / al t. Date ol I 3 Dat F n v ir r u n m o n ta l I ahrralr>r\r 3M Environmental Laboratory D o n a O r *f O f Page 2 C01063 Report EOI-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard Q u a l ity A s s u r a n c e S tatem ent Study Title: Analytical Phase Report for PFOS: A Reproduction Study with the Mallard Study Identification Number: E01-1256 The analytical phase was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings were reported to the study director and laboratory management. In s p e c t io n D a t e s 02/22/02 04/03/02-04/08/02,04/17/02 05/13/02-05/15/02 P hase In-phase Data Final Report D ate R ep o r te d to M anagement S tu d y D ir ec to r 02/22/02 02/22/02 04/08/02,05/21/02 04/08/02,05/21/02 05/16/02 05/16/02 i/C^ Quality Assurance Representative 3M Environmental Laboratory 3M Environmental Laboratory Paae 3 of 216 Page 3 vf J Report EOI-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard Table of C ontents GLP Compliance Statement.................................................................................................................. 2 Quality Assurance Statem ent............................................................................................................... 3 List of Tables.......................................................................................................................................... 5 Summary................................................................................................................................................ 7 Purpose...................................................................................................................................................8 Reference Substances...........................................................................................................................8 Specimen Receipt and Storage............................................................................................................. 9 Method Summary................................................................................... ............................................... 9 Preparatory M ethod............................................................................................................................... 9 Sample Receipt, Preparation and Analysis...........................................................................................10 Analytical Method.................................................................................................................................... 11 Analytical Results.................................................................................................................................... 12 Data Summary....................................................................................................................................... 14 Statement of Conclusion........................................................................................................................21 Statistical Methods................................................................................................................................. 23 Statement of Data Quality......................................................................................................................23 References............................................................................................................................................. 24 List of Attachments................................................................................................................................. 24 Signature P a g e ...................................................................................................................................... 25 Attachment A: Extraction and Analytical Method, ETS-8-231.1, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices".................................................... 26 Attachment B: Data Summary Tables................................................................................................... 46 Attachment C: Sample Chromatograms, Calibration Information, and Instrument Information....... 55 Attachment D: Sample Preparation Sheets..........................................................................................144 Attachment E: Study Protocol, Protocol Amendments, and Deviations............................................. 178 Attachment F: Example Calculations.................................................................................................... 216 .TM Fnvirnnmfintel l.ahorafnrv 3M Environmental Laboratory Paae 4 of 216 Page 4 001065 Report E01-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard L is t o f Ta b les Table 1. Summary Table, C8F17S 0 3` Concentration In Seraand Liver.............................................7 Table 2. Summary Table, C8F17S 03' Concentration In Seraand Liver with Outliers Removed.... 8 Table 3. Reference Substances............................................................................................................ 8 Table 4. SPE Extraction Summary....................................................................................................... 9 Table 5. Sample Analysis....................................................................................................................... 10 Table 6. Serum QC Summary Table..................................................................................................... 13 Table 7. Serum Matrix Spike Summary Table...................................................................................... 13 Table 8. Liver QC Summary T a b le........................................................................................................13 Table 9. Liver Matrix Spike Summary Table......................................................................................... 13 Table 10. C8F17S 0 3' Data Summary, Female Mallard Sera (Adult and Offspring)............................. 14 Table 11. C8F17S 0 3' Data Summary, Male Mallard Sera (Adult and Offspring)................................. 16 Table 12. C8F17S 0 3' Data Summary, Female Mallard Liver (Adult and Offspring)............................. 18 Table 13. C8F17S 0 3' Data Summary, Male Mallard Liver (Adult and Offspring)................................. 20 Table 14. Summary Table, CaF17S 0 3' Concentration in Sera and Liver............................................. 22 Table 15. Summary Table, C8Fi7S 0 3' Concentration in Sera and Liver with Outliers Removed..... 23 3M Environmental Laboratory 3M Environmental Laboratory Page 5 of 216 Page 5 001066 Report EO1-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard St u d y In fo rm atio n Study Director Sean P. Gallagher Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 Sponsor 3M Environmental Technology and Safety Services 3M Environmental Laboratory Building 2-3E-09 St. Paul, MN 55106 Susan A. Beach, Sponsor Representative Analytical Chemistry Laboratory 3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory (3M Lab) Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Lisa Stevenson, Principal Analytical Investigator Study Personnel Sean P. Gallagher, Wildlife International LTD., Study Director Joann B. Beavers, Wildlife International LTD., Laboratory Management William K. Reagen, 3M Environmental Laboratory Management Susan A. Beach, Sponsor Representative Harold O. Johnson, III, Original Principal Analytical Investigator Lisa A. Stevenson, Principal Analytical Investigator Kent R. Lindstrom, Senior Research Chemist Ognjenka Krupljanin, Analytical Chemist Marlene Heying, Analytical Chemist Experimental Dates Sample Analysis Initiation: 19 February 2002 Sample Analysis Completion: 11 April 2002 Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The analytical reference standard reserve samples, as well as the samples pertaining to the analytical phase of this study, are archived at the 3M Environmental Laboratory according to 40 CFR Part 792. 3M Fnvirnnrrwsntal I ahnratorv 3MEnvironmental Laboratory Paae 6 of 216 Page 6 Report E01-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard S ummary The concentration of the C8Fi7S 0 3' anion of perfluorooctane sulfonate, potassium salt (PFOS, C8F17S 0 3'K+) was determined in the liver and sera of adult mallards and their offspring in both a control group and a 10ppm dosed group from Wildlife feeding study #454-109. The following tables summarize the CgF17S 0 3' concentrations found in the liver and sera of study samples. Extracted-matrix PFOS standards are used to establish the HPLC-ES/MS/MS instrument calibration curve for C8F17S 0 3`. Concentrations of C8F17S 0 3' are reported after correction for the purity and the potassium contribution to the mass of the PFOS reference substance. Table 1 summarizes all study sample data. Table 2 summarizes study data after removing certain results that have been statistically determined to be outliers utilizing the Dixon Q-Test. Table 1. Summary Table, CbFi 7S 03' Concentration in Sera and Liver S a :.-p i in g E v f.u r Male Control Group N l.V R P t OF S era Samples AVC GAGE C ,F ,:S O j' Conc. S era R ange of R esults Nil ' > K C.F i R S a . h t , <100 ng/mL (14 of 18) 18 NA 20 107-1400 ng/mL range (4 of 18) A verage: C .T Y S O i C o n c . L iv e r <50ng/g Range of R e s u lts NA Female Control Group Male 10ppm Group Female 10ppm Group <100 ng/mL (15 of 20) 20 NA 20 166-2680 ng/mL range (5 of 20) 20 89700 ng/mL 11300-210000 ng/mL 20 20 20300 ng/mL 6350-91400 ng/mL 20 <50ng/g NA 60900 ng/g 24200-125000 ng/g 10800 ng/g 2480-30700 ng/g Male Offspring <100 ng/mL (2 of 3) 3 NA 3 <50 ng/g NA Control Group 103 ng/mL (1o f3) Female Offspring 7 NA <100 ng/mL (7 of 7) 7 <50 ng/g NA Control Group Male Offspring 10ppm Group 3 4410 ng/mL 3670 - 5450 ng/mL 3 3170 ng/g 22004640 ng/g Female Offspring 10ppm Group 7 4960 ng/mL 3420 - 6760 ng/rnL 7 3620 ng/g 2240-5930 ng/g For individual sample values see Tables 10 to 13 and Attachment B. 0114 C r ttiic A n m n M ^ I I i* ih A m tn n f 3M Environmental Laboratory D a a o 7 rf 0 4 ft Page 7 001068 Report E01-126: Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard Table 2. Summary Table, C8F17SO3' Concentration in Sera and Liver with Outliers Removed Sampling, Even' N: r.'.Bi v of Sera Samples A v t rage: C;iF t;SO ;" Conc. Sera Male Control Group 18* R ango of R esults * Number or L iv e r Samples 20* A verage C .F,;SO f Conc. Lever * Range of R e s u lts * Female Control Group 17 NA <100 ng/mL (15of 17) 166-172 ng/mL range (2 of 17) Male 10ppm Group 18 87300 ng/mL 68700-108000 ng/mL 20* 20* * * * * Female 10ppm Group 19 16600 ng/mL 635042800 ng/mL 20* * * Male Offspring Control Group Female Offspring Control Group 3* 7* * * * 3* * * 7* * Male Offspring 10ppm Group 3* * * 3* * * Female Offspring 10ppm Group 7* * * r| * * * No outliers removed from this data set. Refer to Table 1 for results. For individual sample values see Tables 10 to 13 and Attachment 6. Purpose The purpose of the analytical phase of this study Is to quantify CbF^S C V in the liver and sera of both control and dosed mallards and in their offspring. R eference S ubstances Table 3. Reference Substances R eference S ubstance Source Expiration Date Storage Conditions 3M Laboratory Identification Number Physical Description Purity PFOS 3M Internal 08/31/2006 Frozen -20C +/- 10C SD-018, Lot #217 White Crystalline Powder 86.9% PFDA Oakwood Products, Inc. 12/01/2010 Frozen -20"C +/- 10"C SD-036 White Solid 98% PFDA is nonadecafluorodecanoic acid, or Perfluorodecanoic add, CioF,90 2H and is used solely to monitor for gross instrument failure and not as an internal standard for quantitation. osa I . k . , 3M Environmental Laboratory o rone Page 8 Report E01-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard S p e c im e n R e c e ip t a n d S to r a g e Mallard serum and liver samples were obtained from the sacrifice of the test animals upon the conclusion of the in-life phase of the study (#454-109) conducted at Wildlife International, Ltd. and sent to 3M Environmental Laboratory for analysis. The specimens were received at 3M Environmental Laboratory, assigned a laboratory tracking (LIMS) number (E01-1256, E01-1326, or E01-1367), and were stored at -20C +/- 10C until prepared for analysis. Method Summary The determination of C8F17S 0 3' concentration in the liver and sera was performed according to 3M Environmental Laboratory method ETS-8-231.1, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices". The sera samples were prepared by measuring a volume of serum and diluting with water in a 1/20 ratio. The liver samples were prepared by measuring a weight of liver and diluting with water to a 1/10 ratio prior to homogenizing. Five mL of acetonitrile was added to 1.0mL of the diluted sera or 1.OmL of the homogenate of liver and was shaken for 20 minutes. These samples were then centrifuged for 10 minutes; the supernatant was decanted into 40 mL of water and passed through a conditioned solid phase extraction (SPE) column. After drying the column, 2.0 mL of methanol was passed through the column to elute the analyte. The samples were analyzed for C8F17S 0 3 using HPLC-ES/MS/MS. Preparatory M ethod ETS-8-231.1 "Solid Phase Extraction and Analysis of Fluorochemical Components from Biological matrices." This method describes the extraction of target analytes from biological matrices, such as sera and liver, using solid phase extraction (SPE). Sera samples were diluted with Kandiyohi (commercially bottled) water 1/20, before extraction. Liver samples were diluted 1/10, approximately 1g of liver and 9ml of Kandiyohi water, and then homogenized. A 1.0mL aliquot of the diluted sera or homogenate of liver was extracted by adding 5.0mL of acetonitrile, shaking for 20 minutes, and centrifuging for 10 minutes. The supernatant was transferred to a clean tube, diluted with 40 ml of Kandiyohi water, and filtered through a conditioned C18 (SPE) column. Analytes were eluted off the column, and analyzed by high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ES/MS/MS). Table 4. SPE Extraction Summary S tep S am ple P r ep a r /oicm 1 Measure volume of sera or weigh portion of liver 2 I Dilute sera (1/20) and liver (1/10) with water 3 Mix sera/water or homogenize liver/water 4 Aliquot 1.0ml of diluted/homogenized sample and add 5.0ml of ACN. Spike samples accordingly. Shake for 20 minutes, centrifuge for 10 minutes. 5 Dilute supernatant with 40 ml of water 6 Filter sample through conditioned SPE C18 column. 7 Elute target analytes from the column with 2.0mL methanol (MeOH). 3M Environmental Laboratory 3M Environmental Laboratory Paoe 9 of 216 Page 9 0 0 *0 7 0 Report EO1-1256 Report E01-1256 PFOS: A Reproduction Study with the Mallard PFOS: A Reproduction Study with the Mallard S a m p le R ec e ipt, P r eparatio n a n d A na lysis 3ME v r nment l Labor t r Pa e 10 C 'v*.071