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Report No . FACT G e n -021, G e n -024, G e n -030, G e n -033 LRN--W2491, W2845, W3197, E00-1386
Analytical Study Initiation
GEN021: 08/25/99 GEN024: 10/12/99 GEN030: 12/13/99 GEN033: 03/14/00
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3M Environmental Laboratory Report No. Gen-021, Gen-024, Gen-030, Gen-033
Table of Contents
Analytical Study Personnel and Contributors....................................................................................1 Introduction......................................................................................................................................... 2
Purpose.......................................................................................................................................... 2 Test and Control Article.................................................................................................................2 Sample Collection and Analysis....................................................................................................3 Sample Receipt and Maintenance.....................................................................................................3 Chemical Characterization................................................................................................................ 3 Procurement.................................................................................................................................. 3 Method Summaries.... ;...................................................................................................................... 3
Preparatory and Analytical Methods....................................................................................... 4 Analytical Equipment............................................................................................................... 5 Data Summary, Analyses, and Results.................................................................... ......................... 6 Summary of Quality Control Analyses Results............................................................................ 6 Summary of Sample Results........................................................................................................7 Data Quality Objectives......................................................................................................................8 Statement of Conclusion.................... :.............................................................................................. 9 References..........................................................................................................................................9 Attachments........................................................................................................................... ............ 10 Report Signature................................................................................................................................ 10
List of Tables
Table 1. Description of Samples, by Study........................................................................................2 Table 2. Procurement Information for Reference Materials in the Analysis of Environmental
Samples................................................................................................................................ 3 Table 3. Ions Monitored in the Analyses of Extracts of Groundwater................................................ 6 Table 4. Range of LOQs for Sera, by Study.......................................................................................8 Table 5. Range of LOQs for Liver and Other Tissues, by Study....................................................... 8
Proprietary and Confidential
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3M Environmental Laboratory Report No. G en-021, Gen-024, Gen-030, Gen-033
Analytical Study Personnel and Contributors
Analytical Chemistry Laboratories
3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory Fluorine Analytical Chemistry Team (FACT) 2-3E-09 935 Bush Avenue St. Paul, MN 55106
Kris Hansen, Ph.D., PAI Harold Johnson, Analytical Chemist Lisa A. Clemen, Analytical Chemist Mark Ellefson, Analytical Chemist
Sponsor
3M EJ&SS Buildfrig 2-3E-09 St. Paul, MN 55133
Dale Bacon, Sponsor Representative
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3M Environmental Laboratory Report No. G en-021, Gen-024, Gen-030, Gen-033
Introduction
Purpose
The purpose of this composite report is to provide a summary of the analytical data collected for 3M studies Gen-021, Gen-024, Gen-030, and Gen-033. All of the samples included in these studies are tissue samples collected from fish, birds, mammals, and amphibians; Dr. John Geisy of Michigan State University has supplied all samples to 3M. These analyses have been conducted to support studies designed by Dr. Geisy.
The target analytes for these four studies were perfluorooctane sulfonate (PFOS; CAS# 2795-393), perfluorooctanesulfonyiamide (PFOSA; CAS# 754-91-6), perfluorooctanoate (PFOA or POAA; CAS# 3825-26-1), and perfluorohexane sulfonate (PFHS; no CAS# available).
Due to the variety of matrices analyzed (with respect to both species and tissues), and due to evolving analytical methods, some analytical data quality objectives, such as the limit of quantitation (LOQ) were quite variable. A summary of the achieved LOQ (by specie, tissue and study number) is presented in Table 2 of this report. The stated data quality is based on results of data collection quality controls, sample prep quality controls, and recovery of target analytes from prepared matrix spike samples. More specific data quality objectives and parameters for these analytical studies are outlined later in this report.
Test and Control Article
The test articles for each study consisted of various tissues from various species and are listed below, in Table 1. For all studies, the control article consisted of rabbit sera and rabbit liver, as appropriate. Rabbit tissues were chosen as the control articles because previous studies have indicated very low levels of endogenous fluorochemicals in these matrices. Samples of the control articles were provided by the 3M Environmental Laboratory.
This report does not include details for the collection of the test articles; these details should be obtained from Dr. Geisy.
Table 1. Description of Samples, by Study
Study Number G en -021 G e n -024 G e n -0 30
G e n -0 33
S era/P lasma/B lood
C o rm o ra n t B lood, C a s p ia n S e a l B lood, S e a O tte r B lood
A lbatross sera, A lbatross plasm a, C orm orant plasm a, H erring Gull P lasm a, Bald E agle plasm a,
C orm orant blood, H erring G ull blood
L iver C a lifo rn ia S e a L ion, E le p h a n t S e a l,
H a rb o r S e a l, G o z z i, M in k , R iv e r O tte r, S e a O tte r, T u rtle
Loon, Brown Pelican, A lbatross
N orthern Fu r S eal blood (juvenile, sub adult, adult), Polar B ear blood, S tellar
S ea Lion blood
N orthern Fur S eal, Polar B ear, M ink, M a p T u rtle , T e rrap in , T u n a , G reen
Frog, C hinook S alm on, Lake W h itefts h , Brow n T ro u t
N one Subm itted
M ink, Baikal S eal, G an g e s D olphin, C orm orant (adult and ju ven ile),
B ottlenose D olphin, S triped D olphin, W ed d ell S eal, S w ordfish, T u n a, B lacktailed G u ll
O ther
S ea O tte r B rain, S e a O tte r K idney
A lbatross kidney, C orm orant yolk, G ull y o lk
C a rp body, Frog m uscle, Frog body, G reen Frog eg g s, Lake W h ite fis h eg gs, Brown T ro u t eg g s, C a rp m u scle, C hinook S alm on m uscle, Lake W h ite fis h m uscle,
Brow n T ro u t m uscle
N on e S ubm itted
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3M Environmental Laboratory Report No. G en-021, Gen-024, Gen-030, G en-033
Following analysis, extracts generated from these samples have been retained in cold storage.
Sample Collection and Analysis Tissue samples were submitted to the Environmental Laboratory- Fluorine Analytical Chemistry Team by Kurunthachalan Kannan of Michigan State University. Details of the sample receipt are documented on the chain of custody forms located in appendices of this report.
Sample Receipt and Maintenance
Samples were received in the Environmental Lab cold or frozen on the following dates: Gen-021 (8/24/99), Gen-024 (10/11/99), Gen-030 (12/13/99), and Gen-033 (3/13/00). Sample receipt, identification, and chain of custody information are located in the study folder for each report; the folders are located in the 3M archives.
The sample extracts will be maintained in cold storage at the 3M Environmental Laboratory until the quality of preparation no longer affords preservation.
Chemical Characterization
The target analytes characterized in the samples include PFOS, PFOSA, PFOA, and PFHS. Procurement details of the reference standards used for analysis are summarized below.
Procurement
Table 2. Procurement Information for Reference Materials in the Analysis of Environmental Samples
R e f e r e n c e M a t e r ia l P F O S (potassium salt)
PFOSA P F H S (potassium salt) P F O A (am m onium salt)
Lot N umber
171 G en-021: L-2353; all others: L -157 09
N B 116638-16 Gen-024: 245; all others: com m ercial
S ource 3M IC P /P C P Division
3M Specialty Chem icals (R. Buckanin)
3M Specialty Chem icals (G . M oore) Gen-024: 3M Specialty Chem icals;
all others: Aldrich
Full chemical characterization studies, including purity and stability determination, have not been completed at this time. Upon completion of these studies, a report will be archived in the 3M Environmental Lab.
Method Summaries
Following is a brief description of the methods used during this analytical study by the 3M Environmental Laboratory. Copies of the actual methods used for these studies are located in attachment H.
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3M Environmental Laboratory Report No. G en-021, Gen-024, Gen-030, Gen-033
Preparatoryand Analytical Methods
ETS-8-004.1, "Extraction of PFOS or Other Fluorochemical Compounds from Serum for Analysis using HPLC-Electrospray Mass Spectrometry" with some modifications, described below.
Because the matrices were so variable and sample size extremely limited, it was not possible to prepare extracted standard curves. All extracts were evaluated versus unextracted standard curves. When sample size permitted, two matrix spikes were prepared in each tissue sample from each specie tested to provide some level of extraction efficiency determination.
For some samples, less than 1 mL of sample was available. For these samples, the available volume was extracted according to the method with the exception that the final volume of extraction solvent was adjusted to match the volume of the initial sample.
This method was used for the extraction of sera, plasma, and whole blood samples.
ETS-8-005.1, "Analysis of PFOS or Other Fluorochemical Compounds in Serum Extracts Using HPLC-Electrospray Mass Spectrometry" with some modifications, described below.
Because the matrices were so variable and sample size extremely limited, it was not possible to prepare extracted standard curves. All extracts were evaluated versus unextracted standard curves; as a result, all sample concentrations were adjusted by a factor of 1.25 to adjust for the removal of 4/5 of the MTBE from the extract. The factor is unnecessary when an extracted curve is used for evaluation.
ETS-8-006, "Analysis of PFOS or Other Fluorochemical Compounds in Liver Extracts using HPLC-Electrospray Mass Spectrometry" with some modifications, described below.
Because the matrices were so variable and sample size extremely limited, it was not possible to prepare extracted standard curves. All extracts were evaluated versus unextracted standard curves. When sample size permitted, two matrix spikes were prepared in each tissue sample from each specie tested to provide some level of extraction efficiency determination.
For some samples, less than 1 g of sample (as called for in the method) was available. For these samples, the available mass of tissue was extracted according to the method.
Samples of kidney, brain, egg, and muscle were extracted by this method.
ETS-8-007, "Extraction of PFOS or Other Fluorochemical Compounds from Liver for Analysis using HPLC-Electrospray Mass Spectrometry' with some modifications, described below.
Because the matrices were so variable and sample size extremely limited, it was not possible to prepare extracted standard curves. All extracts were evaluated versus unextracted standard curves; as a result, all sample concentrations were adjusted by a factor of 1.25 to adjust for the removal of 4/5 of the MTBE from the extract. The factor is unnecessary when an extracted curve is used for evaluation.
For Gen-030 and Gen-033 only: Due to the lack of excess test material for method development, all samples determined to contain greater than 0.015 pg/g of PFOS were subject to an additional PFOS verification process. Each sample was analyzed separately with respect to the 499 -> 99 transition and the 499 -> 80 transitions. The quantitative results
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obtained from each transition analysis were compared. When these results agreed to with 30%, the identity of PFOS was confirmed (see Reference 1). Those samples where the identity of PFOS could not be confirmed are noted in the data table.
In Gen-021, no PFHS standard was available. In these samples, qualitative determination of PFHS was conducted based on reasonable retention time and a known PFHS transition (399
99).
Specific instrumental parameters are available in appendix l-L of this report, stored in the 3M Environmental Lab archives.
Analytical Equipment
For HPLC-Electrospray Tandem Mass Spectrometry: Liquid Chromatograph: Hewlett-Packard Series 1100 Liquid Chromatograph system Analytical column:
1x30 mm C18 BetasllTM Column temperature: 30 degrees C Cycle Time: 10 minutes
Mobile phase components: ' Component A: 2mM ammonium acetate Component B: Methyl alcohol
Flow rate: 300 pL/min
Injection volume: 10 pL Solvent Gradient:
Time (min) 0 1 5.5 7.5 8 10
%B 10 10 95 95 10 10
For Detection: Mass Spectrometer: Micromass API/Mass Spectrometer Quattro Ultima Triple Quadrapole system or Micromass API/Mass Spectrometer Quattro II Triple Quadrapole system Acquisition Mode: MRM (refer to Table 3) Software: Mass LynxTM 3.3 Mode: Electrospray Negative Source Block Temperature: 125-150C Source: Z-spray
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Table 3. Ions Monitored in the Analyses of Extracts of Groundwater
T ar g et A nalyte
P r im a r y Io n ( a m u )
P r o d u c t io n (a m u )
PFOS
499.0
80, 99*
PFOSA
498
78
PFOA PFHS
i I
413 399
169 99
* Indicates the ion used for quantitation
Refer to the analytical methods and equipment logs found in the raw data for details on the actual analytical equipment settings used in the present study. These settings may have varied somewhat during actual data collection. However, slight variations in the instrument settings will not adversely affect the quality of the data. Exact settings during all phases of data collection are recorded and presented in the appendix of this report.
Data Summary, Analyses, and Results
Summary of Quality Control Analyses Results
Standard Curves: The coefficient of determination (r2) for all 1/X weighted curves bracketing useable data was > 0.982. High or low curve points may have been excluded to provide a better fit over the linear range appropriate to the data. High or low curve points were deactivated if the calculated concentration varied from the theoretical concentration by more than 30%. Acceptable data was evaluated versus a standard curve containing at least 5 points. All actions are acceptable and are documented in specific data sets. All standard curves used to evaluate quantitative data were acceptable.
C ontinuing Calibration Verifications: On average, one calibration check is analyzed for every five samples. Acceptable data is bracketed by calibration checks quantitated to be within 30% of the theoretical value, evaluated at least every ten samples. All quantitative data is bracketed by acceptable calibration checks, as required.
Blanks: Extraction blanks were compliant if no target analyte was detected above the limit of quantitation (LOQ) for a specific analyte. In this study, extraction blanks were often higher than low curve points. Because analyte levels in the blank are used to determine the LOQ, by default, all blanks were determined to be below the limit of quantitation for the compounds of interest.
Internal Standards: Internal standard response was monitored in Gen-030 and Gen-033 only. Internal standard response was required to be within 50% of the theoretical value. If samples showed an internal standard response that deviated more than 50%, the samples were reanalyzed. If the deviant IS response was confirmed, the analyte data was reported, but noted in the data table.
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Summary of Sample Results GEN-021:
PFOS was detected in at least one sample from the following matrices: California Sea Lion liver, Harbor Seal liver, Gozzi liver, Mink liver, River Otter liver, Turtle liver, Cormorant blood, Otter blood, and Caspian Seal blood.
PFOSA was tentatively identified in at least one sample from the following matrices: California Sea Lion liver, River Otter liver, Sea Otter liver, Sea Otter brain, and Otter blood.
PFOA was tentatively identified in at least one sample from the following matrices: California Sea Lion liver and Caspian Seal blood.
PFHS was tentatively identified in at least one sample from the following matrices: California Sea Lion liver, Gozzi liver, Mink liver, River Otter liver, Sea Otter liver, Turtle liver, Cormorant blood, Caspian Seal blood, and Otter blood.
GEN-024:
PFOS was detected identified in at least one sample from the following matrices: Albatross plasma, Albatross sera, Cormorant plasma, Cormorant blood, Herring Gull plasma, Herring Gull blood, Bald Eagle plasma, Loon liver, Albatross liver, Brown ' Pelican liver, Albatross kidney, Cormorant yolk, and Gull yolk.
PFOSA was tentatively identified in at least one sample from the following matrices: Cormorant blood, Bald Eagle plasma, Loon liver, Brown Pelican liver, and Albatross liver.
PFOA was tentatively identified in at least one sample from the following matrices: Cormorant blood, Albatross liver, Cormorant yolk, and Gull yolk.
PFHS was tentatively identified in at least one sample from the following matrices: Herring Gull plasma and Bald Eagle plasma, Loon liver, Albatross liver, Brown Pelican liver, Albatross kidney, Cormorant yolk, and Gull yolk.
GEN-030:
PFOS was detected in at least one sample from the following matrices: Polar Bear blood, Polar Bear liver, Mink liver, Northern Fur Seal liver, Map Turtle liver, Tuna liver, Green Frog liver, Chinook Salmon liver, Lake Whitefish liver, Brown Trout liver, Whole Carp, Frog muscle, Lake Whitefish eggs, Brown Trout eggs, Carp muscle, Chinook Salmon muscle, Lake Whitefish muscle, and Brown Trout muscle.
PFOSA was tentatively identified in at least one sample from the following matrices: Mink liver.
PFOA was not tentatively identified in any sample analyzed.
PFHS was not tentatively identified in any sample analyzed.
GEN-033:
PFOS was detected in at least one sample from the following matrices: Mink liver, Baikal Seal liver, Cormorant liver, Bottle Nosed Dolphin liver, Ganges Dolphin liver, Striped Dolphin liver, Swordfish Liver, Tuna liver, and Black Tailed Gull liver.
PFOSA was tentatively identified in at least one sample from the following matrices: Mink liver, Cormorant liver, and Bottle Nosed Dolphin liver.
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PFOA was tentatively identified in at least one sample from the following matrices: Cormorant liver.
PFHS was tentatively identified in at least one sample from the following matrices: Mink liver, Striped Dolphin liver, and Swordfish Liver.
Appendices contain data summary tables.
Data Quality Objectives
No circumstances existed during the present study that would have affected the quality or integrity of the data. The data quality objectives (DQOs) followed during the present are indicated below.
Linearity: The coefficient of determination (r2) of the standard curve was equal to or greater than 0.985 with at least 5 active points using a linear regression curve with 1/x weighting.
Instrument Quantitation Lim it (IQL): The IQL is equal to the lowest acceptable standard in the calibration curve (acceptable standard is defined as a standard within 30% of the theoretical value). As this value is not useful in consideration of the sample data, the IQL was not specifically determined or stated for every study.
Limits o f Quantitation (LOQ): The LOQ is equal to the lowest acceptable standard in the calibration curve (defined as a standard within 30% of the theoretical value), and is at least two times the analyte peak area detected in the extraction blanks. The LOQ may vary due to the amount of sample available for analysis (particularly for samples extracted according to ETS-8-006) or to day-to-day variations in the analytical system. The ranges of LOQs for various tissues are listed in Table 4 (sera, plasma, and blood) and Table 5 (liver, kidney, muscle, egg, and brain).
Table 4. Range of LOQs for Sera, by Study
A nalyte
G en-021
PFOS
0.0116 pg/mL
PFOSA
0.00625pg/mL
PFHS
NA
PFOA
0.00599 pg/mL
GEN024 0.00116 pg/mL 0.00626 pg/mL 0.00114 pg/mL 0.0299 pg/rnL
G en-030
0.0029-0.0579 pg/mL 0.000625 pg/mL 0.00114 pg/mL
0.00240-0.00958 pg/mL
G en-033
NA NA NA NA
Table 5. Range of LOQs for Liver and Other Tissues, by Study
A nalyte
G en-021
GEN024
G en-030
PFOS
0.0348 pg/g
0.0348 pg/g
0.00696-0.0696 pg/g
PFOSA
0.0375 pg/g
0.00750 pg/g
0.0188 pg/g
PFHS
NA
0.00683 pg/g
0.00683-0.0342 pg/g
PFOA
0.0359 pg/g
0.180 pg/g
0.0180-0.0719 pg/g
NA = not applicable
G en-033 0 .0 0 6 9 6 -0.0694pg/g
0.0376 pg/g 0.00683 pg/g 0.00719-0.0718 pg/g
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Duplicate/acceptable precision (extraction): Spikes conducted on samples of control tissues were reproducible to within 15%
Quality Control Response: A continuing calibration verification (CCV) was analyzed every 5-10 samples. Acceptable CCV response was within 30% of the theoretical value. No more than 10 samples were analyzed between acceptable CCVs.
Spike/acceptable recoveries: Due to the number of different matrices analyzed, there was great variability in spike recoveries. For any given matrix (specie and tissue), spike recoveries within 70-130% of the expected concentration indicate quantitative data (gopd to 30%); spike recoveries between 50-150% indicate semi-quantitative data for that matrix (good to 50%). Spike recoveries outside of this range indicate that sample data should be used for qualitative purposes only. Due to sample limitations, matrix spike studies were not conducted for all matrices. For PFOS analyses, sample data that is not supported by matrix spike studies should be considered for qualitative purposes only. Since no identity verification experiments were performed for PFOA, PFHS, and PFOSA, for these analytes, all analyses that are not supported by matrix spike studies should be considered to provide unconfirmed qualitative data only.
Use o f Internal Standards: Tetrahydro-perfluorooctane sulfonate (THPFOS) was spiked into the extracts post-extraction and used as an internal standard for samples in Gen-030 and Gen-033. For all samples in these studies, THPFOS levels were monitored to verify the analytical soundness of the data. THPFOS levels that were determined to be deviant from expected values by more than 50% were reanalyzed. If the deviant THPFOS levels were confirmed, analyte levels were reported but are noted in the results table.
Use of confirm atory methods: Given the selectivity of the analytical tool used (HPLCESMSMS) 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 of an analyte-specific parent ion.
Assuming spike recovery studies form a suitable indication of endogenous analyte recovery, matrix spike studies have been used as an indicator of data quality (see above). The validity of this assumption has not been verified by other techniques.
Statement of Conclusion
Under the conditions of the present studies, the presence of fluorochemicals was observed in the quantitative analysis of a selection of environmental matrices.
References
1) "Acceptance Criteria for Ultratrace HPLC-Tandem Mass Spectrometry. Quantitative and Qualitative Determination of Sulfonylurea Herbicides in Soil"; Li, L.Y.; Campbell, D.A.; Bennet, P.K.; Henion, J.; Anal. Chem., 68 (19), 3397-3404, 1996
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Attachments
Attachment A: Gen-021 Sera/Plasma/Blood Results Attachment B: Gen-021 Liver/Miscellaneous Results Attachment C: Gen-024 Sera/Plasma/Blood Results Attachment D: Gen-024 Liver/Miscellaneous Results Attachment E: Gen-030 Sera/Plasma/Blood Results Attachment F: Gen-030 Liver/Miscellaneous Results Attachment G: Gen-033 Liver Results Attachment H: Analytical Methods Attachment I - L (additional bound document available in the 3M Environmental Lab
archives): Analytical Details for Gen-021, Gen-024, Gen-030, and Gen-033
Report S ignature
____ l b - - - ______________________________________ S~/ IQ /P O
Kristen J. Hansen, Ph.D., Principal Analytical Investigator
Date
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FACT-GEN-021
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates o f Extraction/Analyst: Dates o f Analysis/Analyst: Date o f Data Reduction/Analyst:
Sample Data BLOOD
G roup Dose
Sample #
Method Blk
H 20 Blk-1 8/25/99
H 2 0 Blk-2 8/25/99
H 20 Blk-3 8/25/99 H 20 Blk-4 8/25/99
Caspian Seal Blood
W2491-40.J 53 W 2491-41J 11
W2491-42.J 46
W2491-43.J 17
W 2491-44J 13
W2491-45J 8
W 2491-46.J 12
W 2491-47.J9
W2491-48.J 14
W 2 4 9 M 9 .J 18 t W2491-50.J 52
"
W2491-5 U 55
.V- W2491-52.J 10
W2491-53,J 15
No curve analyzed for PFHS, PFDS. PFHS based on PFOS response.
No PFOS qualitative confirmation performed. Identifications are preliminary. *
Deviant Surrogate levels are not noted and w ere not confirmed.
Date Entered/By:
08/27/99,09/01/99, 12/30/99,01/20/00,02/14/00 LAC
Date Verified/By:
02/22/00 MEE
GEN021 Various Matrices from MSU None Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Amelia 062498 Masslynx 3.2 See list to right See Attachments See Attachments See Attachments 08/25/99 MCH/KK/SAL/SEE 08/26/99,08/27/99,08/28/99, 12/09/99 MEE/IAS 08/27/99, 08/30/99, 08/31/99, 12/10/99, 01/20/00 MEE/IAS
Concentration of PFOS ug/m L
Mean PFOS ug/m L
< L O Q ( 0.0116) <LO Q ( 0.0116) < L O Q (0 .0 1 1 6 ) <LO Q ( 0.0116)
<LOQ
0.0180 < L O Q ( 0.0116) < L O Q ( 0.0116)
0.0166 < L O Q ( 0.0116)
0.0131 <LOQ ( 0.0116) < L O Q ( 0.0116) <LO Q ( 0.0116) <LOQ (0.0116) < L O Q (O .O H 6 )
< L O Q (O .O tl6) < L O Q ( 0.0116) < L O Q (0.0116)
<LOQ - 3 Outliers
LOQ =*Limit o f Quantitation NA =*N ot Applicable
RSD = Relative Standard Deviation ND =Not Detected
D = Detected
RSD Std. Dev.
NA NA
NA NA
Concentration of PFOSA ug/mL '
M ean PFOSA ag/m L
RSD S td. Dev.
<LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625)
<LOQ
NA NA
<LO Q ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625) <LOQ ( 0.00625)
<LOQ ( 0.00625) <LO Q ( 0.00625) <LOQ ( 0.00625)
<LOQ
NA NA
PFOS = Perfluorooctanesulfonate PFOSA * Perfluorooctanesulfonamide
POAA = Perfluorooctanoate PFHS = Perfluorohexanesulfonate
ETS-8-5.1 Excel Version 5/95
G E N -0 2 1-scra.xls
000015
5/10/00
FACT-GEN-021
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates o f Extraction/Analyst: Dates o f Analysis/Analyst: Date o f Data Reduction/Analyst:
Sample Data BLOOD
G roup Dose
Sample #
Method Blk
H 2 0 Blk-1 8/25/99
H 20 BIk-2 8/25/99
H 20B lk-3 8/25/99
H 20 Blk-4 8/25/99
Caspian Seal Blood
W 2491-40J 53
W 2491-41J 11 W2491-42,J.46
W 2491-43J 17
W 2491-44J 13
W 2491-45J 8
W 2491-46J 12
W 2491-47J 9
W 2491-48J 14
W 2491-49J 18
W 2491-50J 52
' ' . W 2491-51J55 A W 2491-52J 10
W 2491-53J 15
No curve analyzed for PFHS, PFDS. PFHS based on PFOS response.
No PFOS qualitative confirmation performed. Identifications are preliminary.
Deviant Surrogate levels are not noted and were not confirmed.
Date Entered/By:
08/27/99, 09/01/99, 12/30/99,01/20/00,02/14/00 LAC
Date Verified/ By:
02/22/00 MEE
G EN 021 Various Matrices from MSU None Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Amelia 062498 Masstynx 3.2 See list to right See Attachments See Attachments See Attachments 08/25/99 MCH/KK/SAL/SEE 08/26/99, 08/27/99,08/28/99, 12/09/99 MEE/IAS 08/27/99, 08/30/99, 08/31/99, 12/10/99, 01/20/00 MEE/IAS
Concentration of POAA ug/m L
0.00629 <LOQ ( 0.00599) <LOQ ( 0.00599) <LOQ ( 0.00599)
<LOQ ( 0.00599) 0.00759
<LOQ ( 0.00599) <LOQ ( 0.00599) < L O Q ( 0.00599) <LOQ ( 0.00599) <LOQ ( 0.00599) <LOQ ( 0.00599)
0.00728 <LOQ ( 0.00599)
0.0108 0.0234 <LOQ ( 0.00599) <LOQ ( 0.00599)
Mean POAA ug/m L
RSD Std. Dev.
Concentration of PFHS ug/m L
Mean PFHS u g/m L
ND
<LOQ - 1 Outlier
NA NA
ND ND ND
ND
D
D
ND
D
ND
ND
ND
ND
ND
ND
ND
<LOQ - 4 Outliers
NA NA
ND ND ND ND - 3 Outliers
LOQ " Limit o f Quantitation
PFOS - Perfhiorooctanesulfonate
NA - Not Applicable
PFOSA * Perfiuorooctanesulfonamide
RSD - Relative Standard Deviation POAA - Perfiuorooctanoate
ND = Not Detected D = Detected
PFHS = Perfiuorohexanesulfonate
RSD Std. Dev.
NA NA
NA NA
ETS-8-5.1 Excel Version 5/95
GEN-021-sera.xls
000016
5/10/00
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Softwarc/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst: Dates of Analysis/Analyst: Date of Data Reduction/Analyst
Sample Data
BLOOD
QC
Group Dose
Sample #
Method Blk
H20 Blk-l 8/25/99
H 20 BIk-2 8/25/99
H20 Blk-3 8/25/99
H20 Blk-4 8/25/99
Caspian Seal Blood
W2491-40.J 53-MS
W 2491-4U U -M S
W2491-42.J 46-MS
W2491-43.J 17-MS
W2491-44.J 13-MS
W2491-45J 8-MS
W2491-464 12-MS W2491-47J 9-MS
W2491-48J 14-MS
W2491-494 18-MS W2491-50J 52-MS
W2491-514 55-MS
W2491-52.J 10-MS
' V.
W2491-53.J 15-MS
No curve analyzed for PFHS, PFQS. PFHS based on PFOS response.
^ No PFOS qualitative confirmation performed Identifications are preliminary.
Devient Surrogate levels are not noted and were not confirmed.
Date Entered/By:
02/16/00, 02/17/00 LAC
Date Verified/By:
02/22/00 MEE
FACT-GEN-021
GEN021 Various Matrices from MSU None Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Amelia 062498 Masslynx 3.2, 3.3 08/28/99, 12/09/99 IAS 08/28/99, 12/09/99 IAS 08/30/99, 12/10/99 IAS See Attachments 08/25/99 MCH/KK/SAL/SEE 08/28/99, 12/09/99 IAS 08/30/99, 12/10/99 IAS
Concentration of PFOS
ug/mL or V* Ree
NA NA NA NA 164% 115%
101% 2431%
67% 103% 150% 103% 103% 73% 65% 17% 12% 17%
Mean PFOS ug/mL or % Rec
NA
RSD Std. Dev.
NA NA
* outlier excluded
58%
84% 48%
LOQ = Limit of Quantitation
N A 88Not Applicable
RSD * Relative Standard Deviation
ND Not Detected
D = Detected
NS Not Spiked
Concentration
Mean
of PFOSA
PFOSA
ug/mL or % Rec
ug/mL or % Rec
NA
NA
NA
NA NA
106%
98%
73% 1991%
50%
79%
116%
85%
90%
47%
49%
10%
10% * outlier excluded
12% 63%
PFOS = Perfluorooctancsulfonate
PFOSA = Perfluorooctanesulfonamide
POAA ~ Perfluorooctanoate
PFHS = Perfluorohexanesulfonate
RSD Std. Dev.
NA NA
58% 37%
ETS-8-5.1 Excel Version 5/95
GEN-021-sera.xls
000017
5/10/00
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst Dates of Analysis/Analyst: Date of Data Reduction/Analyst:
Sample Data
BLOOD
QC
Group Dose
Sample #
Method Blk
H 20 Blk-1 8/25/99
H 20 Blk-2 8/25/99
H 20 Blk-3 8/25/99
H 20 Blk-4 8/25/99
Caspian Seal Blood
W2491-40J 53-MS
W 2491-4U 11-MS
W2491-42J 46-MS
W2491-434 17-MS
W2491-44.J 13-MS
W2491-45,J 8-MS
W2491-464 12-MS
W2491-47J9-MS W2491-484 14-MS
W2491-49.J 18-MS
W2491-50J 52-MS
W2491-51J 55-MS
W2491-52.J 10-MS
W2491-53J 15-MS
No curve analyzed for PFHS, PFDS PFHS based on PFOS response.
NoVFOS qualitative confirmation performed. Identifications are preliminary.
Devient Surrogate levels are not noted and were not confirmed.
Date Entered/By:
02/16/00,02/17/00 LAC
Date Verified/ By:
02/22/00 MEE
FACT-GEN-021
GEN021 Various Matrices from MSU None Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Amelia 062498 Masslynx 3.2,3.3 08/28/99,12/09/99 IAS 08/28/99,12/09/99 IAS 08/30/99, 12/10/99 IAS See Attachments 08/25/99 MCH/KK/SAL/SEE 08/28/99, 12/09/99 IAS 08/30/99, 12/10/99 IAS
Concentration of POAA
ug/mL or Ree NA NA NA NA 157% 115% 100%
2056% 89% 108% 174% 111% 112% 91% 78% 14% 19% 19%
Mean POAA ug/mL or % Rec
RSD Std. Dev.
NA NA NA
* outlier excluded 91%
54% 50%
LOQ Limit o f Quantitation
NA = Not Applicable
RSD = Relative Standard Deviation
ND = Not Detected
D = Detected
NS =*Not Spiked
Concentration of PFHS
ug/mL or % Rec
NS NS NS NS
NS NS NS NS NS NS NS NS NS NS NS NS NS NS
Mean PFHS ug/mL or % Rec
NS
RSD Std. Dev.
NS NS
NS NS NS PFOS = Perfluorooctanesulfonate PFOSA = Perfluorooctanesulfonamidc POAA = Perfluorooctanoate PFHS = Perfluorohexanesulfonate
ETS-8-5.1 Excel Version 5/95
GEN-021-sera.xts
000018
5/10/00
FACT-GEN-021
Study: Product Numbei(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Softwarc/Version: Date of Extraction/Analyst: Date of Analysis/'Analyst: Date of Data Reduction/Analyst:
Sample Data
GEN021 Various Matrices from MSU None Various Matrices . ETS-8-6.0 & ETS-8-7.0 using unextracted curves Amelia 062498 Masslynx 3.2 08/25/99 MCH/KK/SAUSEE 08/26/99,08/27/99, 08/28/99, 12/09/99 MEE/IAS 08/27/99, 08/30/99, 08/31/99, 12/10/99, 01/20/00 MEE/IAS
LIVER/W HOLE BLOOD Group Dose
Sample#
Method Blk
H 20B tk-l 8/25/99
H20 BUc>2 8/25/99
H20 Bfic-3 8/25/99
H20 BDc-4 8/25/99
California Sea Lion Liver
W2491-3.CSL 3448
W2491-4.CSL 3395
W2491-6.CSL 3020
W2491-7.CSL 2169
W2491-9.CSL 2839
W2491-10.CSL 2367
Elephant Seal Liver
W249l-2ES 1500
W249l-5.ES 1552
W 249l-lt,ES 808
W24$1-I2,ES 772
W2491-14,ES 782
Harbor Seal Liver
W2491-8.HS
W2491-13.HS 1191
W2491-15.HS 1199
Gozzl Liver
W2491-1.NFS 100
Mink Liver
W2491-34JM030 USFWS W2491-35J31146 USFWS
W2491-36,D1158 USFWS
River O tter Liver A
W2491-29JIAG 066 W2491-30,RAG 028 W2491-31.RAG 148
W2491-32.RAG 230 W2491-33.RAG 237
Sea Otter Liver
W249M6.SO 12593-001
W2491-19.SO 11494-001
W2491-22.SO 11940-001
W2491-24,SO 11309-001
W2491-25.SO 12797-001
W249I-26.SO 13110-001
W2491-27.SO 12679-001 W2491-28,SO 12707-001
Turtle Liver
W2491-37Jdal< Turtle (-2,8),Turtle Liver W249L-38,Male Turtle (2,12),Turtle Liver
W2491-39,Female Turtle (-3,9),Turtle Liver
Sea O tter Brain
W249M8.SO 12593-001,Sea Otter Brain W2491-21,SO 11494-001,Sea Otter Brain
Sea Otter Kidney
W2491-17.SO 12593-OOl.Sea Otter Kidney W2491-20.SO 11494-001,Sea Otter Kidney
W2491-23.SO 11940-001,Sea Otter Kidney
Whole Blood
W2491-54,Cortnorant DCCO L Charity W2491-55,Cormorant DCCO Hym Island, Lake Sup
W249I-56,Otter DCCO Great Lakes
PFDS/PFHS - no curve analyzed PFHS based on PFOS response.
No PFOS qualitative confirmation performed. Identifications are preliminary.
Devient Surrogate levels are not noted and were not confirmed.
Date Entercd/Analyst: Date Verified/Analyst:
08/27/99,09/01/99, 12/30/99,01/20/00,02/14/00 LAC 02/22/00 MEE
Concentration
Mean
of PFOS
PFOS
UR/Ro r % Ree. <LOQ ( 0.0348)
tit
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ
0.0384
0.0494
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ - 2 Outliers
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348) <LOQ ( 0.0348)
<LOQ
<LOQ ( 0.0348)
<LOQ ( 0.0348)
0.0571
<LOQ - 1 Outlier
0.133
0.133
4.85
2.41
0.587
2.62
0.279
0.994
0.189
0.0336
0.151
0.329
<LOQ ( 0.0348)
<LOQ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ ( 0.0348)
<LOQ
0.239
0.358
0.099
0.232
<LOQ 0.0348)
<LOQ 0.0348)
<LOQ
<LOQ 0.0348)
<LOQ 0.0348)
<LOQ 0.0348)
<LOQ
0.190
0.0422 0.0392
0.0904
LOQ 3 Limit o f Quantitation
RSD 3 Relative Standard Deviation
NA 3 Not Applicable
ND 3 Not Detected
D 3 Detected
RSD Std. Dev.
<LOQ <LOQ
NA 0.00773
NA NA NA NA NA 81.7 2.14
116 0.382
NA NA 56.0 0.130 NA NA NA NA 95.2 0.0861
Concentration of PFOSA
ug/R or % Ree. <LOQ ( 0.0375)
Mean PFOSA
UR/R
RSD Std. Dev.
<LOQ( 0.0375)
<LOQ ( 0.0375) <LOQ ( 0.0375)
0.0443
<LOQ
NA NA
<LOQ ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375) <LOQ ( 0.0375)
<LOQ - I Outlier
NA NA
<LOQ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375) <LOQ ( 0.0375)
<LOQ
NA NA
<LOQ ( 0.0375)
<LOQ ( 0.0375)
NA
<LOQ ( 0.0375)
<LOQ
NA
<LOQ ( 0.0375)
<LOQ
NA
<LOQ ( 0.0375)
<LOQ( 0.0375) <LOQ ( 0.0375)
<LOQ
NA NA
0.0371
0.0448
0.0716
<LOQ ( 0.0375) 0.0393
0.0482 - 1 Outlier
33.1 0.0160
0.0806
<LOQ ( 0.0375)
<LOQ ( 0.0375)
<LOQ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375)
<LOQ ( 0.0375) <LOQ ( 0.0375)
NA <LOQ -1 Outlier . NA
<LOQ ( 0.0375)
<LOQ ( 0.0375)
NA
<LOQ ( 0.0375)
*LOQ
NA
0.0664
NA
<LOQ ( 0.0375)
NA NA
<LOQ 0.0375)
<LOQ 0.0375)
NA
<LOQ 0.0375)
<LOQ
NA
<LOQ 0.0375)
<LOQ( 0.0375) 0.112
NA <LOQ - l Outlier ... NA
PFOS 3 Perfluorooctanesulfonate
PFOSA 3 Perfluorooctanesulfonamide
POAA 3 Periluorooctanoate
PFHS 3 Perfluorohexanesulfonate
ETS-8-7.0 Excel Version 5/95
GEN-021-liver.xls
000019
5/10/00 ` 6:07 AM
FACT-GEN-021
Study: Product NumbcrfTest 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
GEN021 Various Matrices from MSU None Various Matrices ETS-8-6.0 & ETS-8-7.0 using unextracted curves
BEST COPY AVAILABLE
Amelia 062498
Masslynx 3.2
08/25/99 MCH/KK/SAL/SEE
08/26/99, 08/27/99, 08/28/99, 12/09/99 MEE/IAS
08/27/99, 08/30/99, 08/31/99, 12/10/99, 01/20/00 MEE/IAS
LIVERAVHOLE BLOOD Group Dose
Sample U
Method Blk
H20BUC-1 8/25/99 H 20 Blk-2 8/25/99
H20 Blk-3 8/25/99
H20 BUc-4 8/25/99
California Sea Lion Liver
W2491-3.CSL 3448
W2491-4.CSL 3395
W2491-6.CSL 3020
W2491-7.CSL 2169
W2491-9.CSL 2839
W2491-10.CSL 2367
Elephant Seal Liver
W2491-2JES 1500 W249t-5.ES 1552
W 2491-UJS808
W 249l-llES772
W 249M4,ES 782
Harbor Seal Liver
W2491-8.HS W2491-13,HS 1191
W2491-15.HS 1199
Gozzi Liver
W2491-1.NFS 100
Mink Liver
W2491-34.D1030 USFWS W2491-35.D1146 USFWS
W2491-36.D1158 USFWS
River Otter Liver
,>
W2491-29,RAG 066
1 W2491-30JIAG 028
W2491-31JIAG 148 W2491-32.RAG 230
W2491-3J,RAG 237
Sea Otter Liver
W249M 6.SO 12593-001 W2491-19,SO 11494-001
W2491-22,SO 11940-001 W2491-24,SO 11309-001
W2491-25,SO 12797-001
W2491-26.SO 13110-001
W2491-27.SO 12679-001
W2491-28,SO 12707-001
Turtle Liver
W249l-37,Male Turtle (-2,8),Turtle Liver W2491-38,Male Turtle (2,12XTurtle Liver W2491-39,Female Turtle (-3,9),Turtle Liver
Sea Otter Brain
W2491-18.SO 12593-001,Sea Otter Brain W2491-21,SO 11494-001,Sea Otter Brain
Sea O tter Kidney
W2491-17.SO 12593-OOl.Sea Otter Kidney W2491-20.SO 11494-001,Sea Otter Kidney
W2491-23.SO U940-00I.Sea Otter Kidney
Whole Blood
W2491-54,Cormorant DCCO L Charity
W2491-55,Cormorant DCCO Hym Island, Lake Sup W2491-56,Otter DCCO Great Lakes
PFDS/PFHS * no curve analyzed PFHS based on PFOS response.
No PFOS qualitative confirmation performed. Identifications are preliminary.
Devient Surrogate levels are not noted and were not confirmed.
Date Entered/Analyst: Date Verified/Analyst
08/27/99, 09/01/99, 12/30/99, 0I/2CV00, 02/14/00 LAC 02/22/00 MEE
Concentration or POAA
ug/g o r % Ree. 0.00602
<LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359)
0.0409 <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0,0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ( 0.0359) <LOQ( 0.0359) <LOQ ( 0.0359) <LOQ( 0.0359) <LOQ( 0.0359) <LOQ( 0.0359) <LOQ 0.0359) <LOQ 0.0359) <LOQ( 0.0359) <LOQ 0.0359) <LOQ 0.0359) <LOQ 0.0359) <LOQ 0.0359)
Mean POAA
RSD Std. Dev.
<LOQ -1 Outlier
NA NA
<LOQ -1 Outlier
NA NA
<LOQ
<LOQ <LOQ
<LOQ
NA NA
NA NA NA
NA NA
<LOQ
NA NA
<LOQ
NA NA
<LOQ <LOQ
NA NA NA NA
<LOQ
NA NA
NA
<LOQ
NA
LOQ =Limit o f Quantitation
RSD - Relative Standard Deviation
NA = Not Applicable
ND = Not Detected
D = Detected
Concentration of PFHS
ug/g o r % Ree. ND ND ND ND ND ND ND ND D ND ND ND ND ND ND ND ND ND
D D ND ND D D D ND D
ND D ND ND D ND ND ND ND D D ND ND ND ND ND ND D D
Mean PFHS ug/g
ND ,
RSD Std. Dev.
NA NA .
ND -1 Outlier
NA NA
ND
ND D
ND -1 Outlier
NA NA
NA NA NA
NA NA
D -1 Outlier
NA NA --
ND - 2 Outliers
NA NA
D - 1 Outlier ND
NA NA NA NA
NA ND NA
NA
D - 1 Outlier
... NA
PFOS - Perfluorooctanesulfonate
PFOSA **Perfluorooctanesulfooamide
POAA = Perfluorooctanoate
PFHS " Perfluorohexanesulfonate
BTS-8-7.0 Excel Version 5/95
GEN-021-liver.xls
000020
5/10/00 6:07 AM
FACT-GEN-021
Study: Product NumbeifTest Substance): Matrix: Melhod/Revision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reduction/Analyst:
Sample Data
LIVERAVHOLE BLOOD_______________________________QC
Group Dose
Sample #
Method Blk
H20Blk-l 8/25/99
H20 Blk-2 8/25/99
H20 Blk-3 8/25/99
H20 Blk-4 8/25/99
California Sea Lion Liver
W2491-3.CSL 3448-MS
W249M.CSL 3395-MS
W2491-6,CSL 3020-MS
W2491-7.CSL 2169-MS
W2491-9,CSL 2839-MS
W2491-10.CSL 2367-MS
Elephant Seal Liver
W2491-2.ES 1500-MS
W2491-5S 1552-MS
W2491-lt,ES 808-MS
W2491-12JES 772-MS
W2491-14.ES 782-MS
Harbor Seal Liver
W249I-8.HS-MS
W2491-13.HS 1191-MS
W2491-15.HS 1199-MS
Gozzi Liver
W249MJ4FS 100-MS
Mink Liver
W2491-34J ) 1030 USFWS-MS
W2491-35.D1146 USFWS-MS
v j W2491-36J31158 USFWS-MS
' River O tter Liver
W2491-29JLAG 066-MS ,
W2491-30.RAG 028-MS
W2491-31JLAG 148-MS
W2491-32JLAG 230-MS
W2491-33JLAG 237-MS
Sea Otter Liver
W 249M 630 12593-001-MS
W2491-19.SO 11494-001-MS
W2491-22.SO 11940-001-MS
W2491-24.SO 11309-001-MS
W2491-25.SO 12797-001-MS
W2491-26.SO 13110-001-MS
W2491-27.SO 12679-001-MS
W2491-28.SO 12707-001-MS
Turtle Liver
W249l-37,MaIe Turtle (-2,8)-MS
W2491-38,Male Turtle (2,12)-MS
W2491-39fem ale Turtle (-3,9)-MS
Sea Otter Brain
W2491-18.SO 12593-OOt-MS
W2491-21 ,SO.11494-001 -MS
Sea Otto* Kidney
W2491-1730 12593-001-MS
W2491-20.SO 1I494-00I-MS
W2491-23.SO U940-001-MS
Whole Blood
W2491 -54,Cormorant DCCO L Charity-MS
W2491-55,Cormorant DCCO Hym Island, Lake Sup-MS
W2491-56,Otter DCCO Great Lakes-MS
PFDS/PFHS no curve analyzed, PFHS based on PFOS response.
No PFOS qualitative confirmation performed. Identifications are preliminary.
NR * Not reported, appears the spike wasn't detectable from endogenous levels.
Date Entered/Analyst Date Verified/Analyst
02/16/00,02/17/00 LAC 20/22/00 MEE
GEN021 Various Matrices from MSU None Various Matrices ETS-8-6.0 & ETS-8-7.0 using unextracted curves Amelia 062498 Masslynx 3.2,3.3 08/25/99 MCH/KK/SAL/SEE 08/28/99,12/09/99 IAS 08/30/99,12/10/99 IAS
mumbest copy
Concentration
Mean
of PFOS
PFOS
ug/g or % Rec
Recovery
NA
NA
NA NA NA
91%
78%
69%
45%
31%
64% 63%
105%
77%
51%
44%
33% 62%
35%
81%
50% 55%
57% NA
NR
NR
NR NR
NR
NR
48%
38%
37% 41%
42%
61%
44%
27%
34%
69%
36%
65% 47%
37%
35%
51% 41%
42%
46% 44%
61%
75%
24% 53%
63%
69%
115%
82%
LOQ Limit of Quantitt on
RSD - Relative Standard Deviation
NA Not Applicable
NS Not spiked
RSD Std. Dev.
NA NA
35% 22%
47% 29% 42% 23% NA NA NA
15% 6%
33% 16% 20% 8% 7% 3% 49% 26% 34% 28%
Concentration of PFOSA
ug/g or % Rec NA
Mean PFOSA
tic/g
NA
NA NA -
NA
66%
52%
48%
30%
15%
43% 42%
78%
65%
35%
18%
29% 45%
42%
79%
42% 54%
63% NA
NR
NR
NR NR
NR
NR
47%
34%
29% 36%
11%
50%
35%
16%
22%
44%
26%
33% 30%
42%
5t%
39% 44%
-3%
27% 12%
66%
57%
14% 46%
65%
64%
104%
77%
PFOS Perfluorooctanesulfonate PFOSA ~ Perfluorooctanesulfonamide
POAA = Perfluorooctanoate
PFHS * Periluorohexanesulfonate
RSD Std. Dev.
NA ' NA
42% 18%
56% 25% 39% 21% NA NA NA
26% 9%
45% 13% 15% 6% 171% 21% 61% 28% 29% 23%
ETS-8-7.0 Excel Version 5/95
GEN-02t-liver.xls
000021
5/10/00 6:07 AM
FACT-GEN-021
Study: Product NumberfTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Date of ExtractionAnalyst: Date of Analysis/Analyst: Date of Data Reduction/Analyst:
Sample Data
LIVER/WHOLE BLOOD
Group Dose
__________ QC
Sample #
Method Blk
H20 Blk-\ 8/25/99 H20 Btk-2 8/25/99
H20 Blk-3 8/25/99 H20 Blk-4 8/25/99
California Sea Lion Liver
W2491-3.CSL 3448-MS W2491-4.CSL 3395-MS
W2491-6.CSL 3020-MS
W2491-7.CSL 2169-MS W2491-9,CSL 2839-MS
W2491-10.CSL 2367-MS
Elephant Seal Liver
W2491-2JES 1500-MS JY2491-5,ES 1552-MS
W2491-11JES 808-MS W2491-12.ES 772-MS W2491-14.ES 782-MS
Harbor Seal Liver
W2491-8JHS-MS
W2491-13.HS 1191-MS W2491-15S 1199-MS
Gozzl Liver
W2491-1.NFS 100-MS
Mink Liver
W2491-34J31030 USFWS-MS W2491-35J)1 146 USFWS-MS
W2491-36.D1158 USFWS-MS
'-V River Otter Liver
W2491-29.RAG 066-MS , W2491-30.RAG 028-MS W249I-31JRAG 148-MS
W2491-32.RAG 230-MS
W249I-33JRAG 237-MS
Sea Otter Liver
W2491-16.SO 12593-001-MS W2491-19.SO H494-001-MS W2491-22.SO 11940-001-MS
W2491-24,SO 11309-001-MS
W2491-25.SO 12797-001-MS W2491-26.SO 13110-001-MS W2491-27.SO 12679-001-MS W2491-28.SO 12707-001-MS
Turtle Liver
W2491-37,Male Turtle (-2,8)-MS W2491-38,Malc Turtle (2,12)-MS
W2491-39Jemale Turtle (-3,9)-MS
Sea Otter Brain
W2491-18.SO 12593-001-MS W2491-21,SO 11494-001-MS
Sea Otter Kidney
W2491-17.SO 12593-001-MS
W2491-20.SO 11494-001-MS W2491-23.SO 11940-001-MS
Whole Blood
W2491-54,Cormorant DCCO L Charity-MS W2491-55,Cormorant DCCO Hym Island, Lake Sup-MS
W2491-56,Otter DCCO Great Lakes-MS
PFDS/PFHS no curve analyzed, PFHS based on PFOS response.
No PFOS qualitative confirmation performed Identifications are preliminary.
NR * Not reported, appears the spike wasn't delectable from endogenous levels.
Date Entercd/Analyst: Date Verified/Analyst
02/16/00,02/17/00 LAC 20/22/00 MEE
GEN021 Various Matrices from MSU None Various Matrices ETS-8-6.0 & ETS-8-7.0 using unextracted curves Amelia 062498 Masslynx 3.2,3.3 08/25/99 MCH/KK/SAL/SEE 08/28/99, 12/09/99 IAS 08/30/99, 12/10/99 IAS
i
W ttM
Concentration of POAA
u |/( or % Ree NA NA NA NA 109% 85% 68% 45% 20% 64%
100% 80% 44% 31% 36%
48% 94% 58%
91% NR NR NR 21% 46% 69% 52% 70%
47% 64% 47% 22% 30% 83% 44% 64%
48% 71% 42% 41% 50%
94% 95% 9%
118% 122% 123%
Mean POAA US/
RSD Std. Dev.
Concentratimi of PFHS
ug/g or % Ree.
Mean PFHS U/
RSD Std. Dev.
NS
NA NA NA
NS NS NS
NA NS 'NA
NS
NS
NS
47% 65% 31%
NS NS NA NS NS NA
NS
NS
52% 58% 30%
NS NS NA NS NS i NA
37% 67% 25%
NS NS NA NS NS NA
NA NA
NS NS NA
NS
NA NS
NA
NR NA
NS NS NA
NS
NS
NS 39% NS
NA
52% 20%
NS NS NA
NS
NS
NS
NS
NS
39% 50% 20%
NS NS * NA NS NS NA
29% 54% 15%
NS NS NA NS NS NA
13% 46% 6%
NS NA NS NS NA
NS
75% 66% 49%
NS NA NS NS NA
NS
121%
2% 3%
NS NA NS NS NA
LOQ * Limit of Quantitation
PFOS * Periluorooclanesulfonate
RSD = Relative Standard Deviation PFOSA Periluorooctanesulfonamide
NA = Not Applicable
POAA * Periluorooctanoate
NS * Not spiked
PFHS =*Periluorohexanesulfonate
ETS-8-7.0 Excel Version 5/95
GEN-021-Kver.xls
000022
5/10/00 6:07 AM
FACT-GEN-024
Study: Product Numbcr^Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst: Dates of Analysis/Analyst: Date of Data Reduction/Analyst:
Sample Data SERA
Group Dose
Sample#
Method Blk
Birdl0129-WBlk-5-J
Matrix Blk
NE
NE
MS/MSD 250 ppb
Bird 040-Alb sera-MS-250ppb-5-l Bird 040-Alb scra-MSD-250ppb-5-l
Bird 060-E plasma-MS-250 ppb-S-2
Bird 060-E plasma-MSD-250 ppb-5-2
Bird 054-Cblood-MS-250 ppb-5 * Bird 054-Cblood-MSD-250 ppb-5
Albatross
Bird 034-AJbatross Chick sera Bird 03S-Albatross plasma
Bird 036-Albatross s tr i
Bird 037-Albatross plasma
Bird 038-Albatross sen
Bird 039-Albatross sen
Bird 040-Albatross sen
Bird 041-Albatross sen
Bird 042-Albatross sen
Bird 043-Albatross sen
Bird 044-Albatross sen
Bird 045-Albatross sen
Bird 046-Albatross plasma
Comorant
Bird 047-Comorant plasma Bird 048-Comonnt plasma
Bird 049-Comorant plasma
Bird 050-Comonnt blood
Bird 051-Comorant blood
Bird 052-Comonnt blood
Bird 053-Comonnt blood
Bird 054-Comorantblood
Bird 055-Comonnt blood
Herring
Bird 056-HcTring Gull plasma
Bird 057-Herring Gull plasma
Bird 058-Hefring Gull blood Bird 059-Herring Gull blood
Bald Eagle
Bird 060-Bald Eagle plasma
Bird 061-Bald Eagle plasma
Bird 062-Bald Eagle plasma
Bird 063-Bald Eagle plasma
Bird 064-Bald Eagle plasma
Bird 065-Bald Eagle plasma Bird 066-Bald Eagle plasma
Bird 067-Bald Eagle plasma
Bird 068-Bald Eagle plasma Bird 069-Bald Eagle plasma
Bird 070-Bald Eagle plasma
Bird 071-Bald Eagle plasma
Bird 072-Bald Eagle plasma
Bird 073-Bald Eagle plasma
Bird 074-Bald Eagle plasma
Bird 075-Bald Eagle plasma
Bird 076-Bald Eagle plasma
Bird 077-Bald Eagle plasma
Bird 078-Bald Eagle plasma
Bird 079-Bald Eagle plasma
Bird 080-Bald Eagle plasma
Bird 081-Bald Eagle plasma
Bird 082-Bald Eagle plasma Bird 083-Bald Eagle plasma
Bird 084-Bald Eagle plasma Bird 085-Bald Eagle plasma
No PFOS qualitative confirmation performed. Identifications are preliminaiy.
* Appears to not have been spiked. LAC 10/19/99
Date Entcred/By: Date Verified/ By:
10/18/99, 10/20/99 LAC 12/30/99 MMH 2/17/00 rranh
GEN024 MSU Environmental Samples Various Various ETS-8-4.1 A ETS-8-5.1 using unextracted curves Amelia 062498 Masslynx 3.3 See Attachments See Attachments See Attachments See Attachments 10/12/99 SAL/KK 10/15/99, 10/19/99 IAS/MMH, 12/13/99 IAS 10/18/99, 10/20/99 HOJ, 12/14/99 MMH
Copy m u m
Concentration
orPFOS
og/mL or % Ree
<LOQ (0.00116 uR/mL) NE NE 81% 96% 11% 23% 84% -46%
0.0393 0.0256 0.00755 0.0196 0.00348 0.00677 0.00621 0.00430 0.00402 0.00546 0.00897 0.00657 0.00889
0.431 0.242 0.110 0.0725 0.218 0.153 <LOQ (0.00116 ug/mL) 0.144 0.273
0.453 0.277 0.0790 0.0664
0.449 0.316 0.185 0.167 0.0402 0.0147 0.122, 0.0865 2.57 2.35 0.0324 0.0908 0.217 0.168 0.163 0.0822 0.0238 0.0573 <LOQ (0.00116 ug/mL) 0.286 0.281 0.649 0.335 0.0718 0.0381 0.118
Mean
RSD
Concentration
PFOS
Std. Dev.
of PFHS
ug/mL
RPD-MS/MSD
ug/mL or % Ree
<LOQ
<LOQ
<LOQ (0.00114 ug/mL)
NE
NE NE
NE
68%
89% 17%
81%
59%
17% 69%
65%
76%
19% 678%
14%
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
Plasma
47.0 <LOQ (0.00114 ug/mL)
0.0180
0.00847
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
Sen 0.00926
115 0.0t07
<LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL) 61.8 <LOQ (0.00114 ug/mL)
0.261
0.161
<LOQ (0.00114 uR/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
44.5 <LOQ (0.00114 ug/mL)
0.172 - 1Outlier
0.0764
<LOQ (0.00114 ug/mL)
34.1 0.00243
0.365
0.125
<LOQ (0.00114 UR/mL)
12.2 <LOQ (0.00114 ug/mL)
0.0727
0.0088S
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
0.00148
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
0.00131
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
0.00617
<LOQ (0.00114 ug/mL)
<LOQ (0.00114 ug/mL)
0.356 - 1Outlier
183 0.6S1
<LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL)
NE * Not extracted
NA " Not Applicable
NS * Not Spiked
LOQ Limit of Quantitation
RSD s Relative Standard Deviation
RPD = Relative Percent Difference
Mean PFHS ug/mL <LOQ
NE <
75%
62%
45%
RSD Std. Dev. RPD-MS/MSD
<LOQ
NE
17%
11%
139%
Plasma <LOQ
Sen <LOQ
<LOQ
<LOQ NA
<LOQ
NA NA
NA NA
NA NA
NA NA NA NA NA . NA
NA
<LOQ - 3 Outliers
NA
PFS - Perfluorooctanesulfonate
PFHS Perfluorohexansesulfonate
POAA * Perfluorooctanoate
PFOSA " Perfluorooctanesulfonamide
ETS-8-5.1 Excel Version 5/95
GEN-024-sen.xla
000023
5/10/00
FACT-GEN-024
Study: Product NumbcrfTcst Substance): Matrix: Method/Rcvision: Analytical Equipment System Number Instrument Sofhvare/Version: Filename: R-Squared Value: Slope: Y-Intercept: Dates of Extraction/Analyst: Dates of Analysis/Analyst: Date of Data Reduction/Analyst:
Sample Data SERA
Group Dose
Sample #
Method Blk
Birdl0129-WBlk-5-l
Matrix Blk
NE
NE
MS/MSD
Bird 040-Alb scra-MS-250ppb-5-t
250 ppb
Bird 040-Alb sera-MSD-250 ppb-5-1
Bird 060-E plasma-MS-250 ppb-5-2
Bird 060-E plasma-MSD-250 ppb-5-2
Bird 054-C blood-MS-250 ppb-5 * Bird 054-C blood-MSD-250 ppb-5
Albatross
Bird 034-Albatross Chick sere Bird 035-Albatross plasma
Bird 036-Albatross sera
Bird 037-Albatross plasma
Bird 038-Albatross sera
Bird 039-Albatross sere
Bird 040-Albatross sen
Bird 041-Albatross sen
Bird 042-Albatross sera
Bird 043-Albatross sere
Bird 044-Albatross sen
Bird 045-Albatross sera
v Bird 046-Albatross plasma
'V Comorant
Bird 047-Comorant plasma
Bird 048-Comorant plasma
Bird 049-Comorant plasma
Bird 050-Comorant blood
Bird 051-Comorant blood
Bird 052-Comorant blood
Bird 053-Comorant blood
Bird 054-Comorant blood
Bird 055-Comonmt blood
Herring
Bird 056-Herring Gull plasma
Bird 057-Herring Gull plasma
Bird 058-Herring Gull blood
Bird 059-Herring Gull blood
Bald Eagle
Bird 060-Bald Eagle plasma
Bird 061-Bald Eagle plasma Bird 062-Bald Eagle plasma
Bird 063-Bald Eagle plasma
Bird 064-Bald Eagle plasma
Bird 065-Bald Eagle plasma
Bird 066-Bald Eagle plasma
Bird 067-Bald Eagle plasma
Bird 063-Bald Eagle plasma
Bird 069-Bald Eagle plasma
Bird 070-Bald Eagle plasma
Bird 071-Bald Eagle plasma
Bird 072-Bald Eagle plasma
Bird 073-Bald Eagle plasma
Bird 074-Bald Eagle plasma
Bird 075-Bald Eagle plasma
Bird 076-Bald Eagle plasma
Bird 077-Bald Eagle plasma
Bird 078-Bald Eagle plasma
Bird 079-Bald Eagle plasma
Bird 080-Bald Eagle plasma
Bird 081-Bald Eagte plasma
Bird 082-Bald Eagte plasma Bird 083-Bald Eagle plasma
Bird 084-Bald Eagle plasma
Bird 085-Bald Eagle plasma
No PFOS qualitative confirmation performed. Identifications are preliminary.
* Appears to not have been spiked. LAC 10/19/99
Date Entered/By: Date Verified/ By:
10/18/99, 10/20/99 LAC 12/30/99 MMH 2/17/00 mmh
GEN024 MSU Environmental Samples Various Various ETS-8-4.1&ETS-8-5.1 using unextracted curves Amelia 062498 Masslynx3.3 See Attachments See Attachments See Attachments See Attachments I(yi2/99 SAL/KK 10/15/99, 10/19/99 IAS/MMH, 12/13/99 IAS l<yi8/99, 10/20/99 HOJ, 12/14/99 MMH
BEST copy AVAILABLE
Concentration of POAA
ug/mL or % Ree <LOQ (0.0299 ug/mL)
NE NE 89*/. 105% 77% 84% 90% 23%
<LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL)
<LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL)
0.0489
<LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL)
<LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) , <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL) <LOQ (0.0299 ug/mL)
Mean POAA ug/mL <LOQ
NE
97%
80%
56%
RSD Std. Dev. RPD-MS/MSD
<LOQ
NE
17%
9%
119%
Plasma <LOQ
Sere <LOQ
<LOQ
<LOQ -1 Outlier <LOO <LOQ
NA NA
NA NA
NA NA
NA NA NA NA NA NA
NA
<LOQ
NA
NE * Not extracted
NA * Not Applicable
NS * Not Spiked
LOQ * Limit of Quantitation
RSD " Relative Standard Deviation
RPD Relative Percent Difference
Concentration
Mean
or PFOSA ug/mL or % Ree
PFOSA ug/mL
<LOQ (0.00625 ug/mL)
<LOQ
NE
NE NE 70%
80% 75% 59%
65% 62% 77%
14% 45%
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
Plasma
<LOQ (0.00625 ug/mL)
<LOQ
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
Sera
<LOQ (0.00625 ug/mL)
<LOQ
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL) <LOQ (0.00625 ug/mL)
<LOQ
0.014S
0.00871
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
0.0426
0.0219 3 Outliers
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
0.0751
0.0996
<LOQ (0.00625 ug'mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ (0.00625 ug/mL)
<LOQ - 2 Outliers
PFOS = Perfluorooctaneaulfonate
PFHS = Periluorohexansesulfonate
POAA - Periluorooctanoate
PFOSA = Perfluorooctanesulfonamide
RSD SOL Dev. RPD-MS/MSD
<LOQ NE 14% 9% 140%
NA NA NA NA NA ' NA
82.7 r 0.0181
NA NA NA NA
NA NA
ETS-8-5.1 Excel Version 5/95
GEN-024-sera.xls
000024
5/10/00
FACT-GEN-024
Study; Product NumbcifTcst Substance): Matrix: Method/Rcvision:
Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reducdon/Analyst:
Sample Data
LIVER, KIDNEY, YOLK
Dose
Sample #
Method Blk
QC 250 ng/g
Bird10129-wblk-5-1 Bird10129-wb0c-6-1 BirdOOl-Loon Lvr-MS BirdOOl-Loon Lvr-MSD
Bird023-Albatross Kdny-MS Bird023-Aibatross Kdny-MSD
Bird030-Cormorant Yolk-MS Bird030-Cormorant YoDc-MSD
Liver
BirdOOl-Looo
BmK)02-Loon Bird003-Loon
Bird004-Loon^ Bird005-Looo Btrd006-Looo
Bird007-Looo
Bird008-Loon
Liver
Bird009-Brown Pelican BirdOlO-Brown Pelican
Liver
\ ,
BirdOU-Albatross BirdO12-Albatross BirdOl 3-AIbatross BirdOl 4-AIbalrcss
BirdOl 5-AIbatross BirdOl 6-Albatross Bird020-Albatross
Bird021-Albatross Bird022-Albatross
Kidney
BirdOl 7-Albatross BirdOl 8-Albatross Bird019-Albatross
Bird023-Albatross
Bird024-Albatross
Bird025-Albatross Bird026-Albatross
Yolk
Bird027-Comorant
Bird028-Comorant Bird029-Comorant Btrd030-Comorant
Yolk
Bird03l-Gull Bird032-Gull Bird033-Gull
No PFOS qualitative confirmation performed. Identifications are preliminary.
LOQ * Limit of Quantitation RSD = Relative Standard Deviation
RPD * Relative Percent Difference
Date Entered/Analyst:
10/23/99 LAC, 12/1/99 GML, 12/30/99 MMH
Date Verified/Analyst:
2/17/00 MMH
GEN024 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0 using unextracted curves Amelia 062498 Masslynx 3.3 10/12/99 SAL/KK 10/14/99, 12/13/99 HOJ/IAS 10/15/99,11/15/99,12/14/99 HOJ/MMH
Concentration of PFOS
ug/g or % Ree <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g)
28% 28% 71% 87% 121% 134% 0.345 0.689 0.185 0.199 0.202 0.105 <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) 0.0460 0.294 0.617 <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) <LOQ (0.0348 ug/g) 0.134 0.317 0.254 0.146 <LOQ (0.0348 ug/g) 0.0541
NA Not Applicable
Mean PFOS
RSD Std. Dev. MS/MSD RPD
Concentration of PFHS
ug/g or % Ree
<LOQ 28% 79%
<LOQ (0.0683 ug/g) NA <LOQ (0.0683 ug/g)
54% 3% 65%
63% 20% 77%
128%
10%
71% 75%
<LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
0.287-2 Outliers
73.6 0.212
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
0.170
103 0.175
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ -1 Outlier
NA
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ
<LOQ (0.0683 ug/g) NA <LOQ (0.0683 ug/g)
0.235 -1 Outlier
39.5 0.0930
<LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g) <LOQ (0.0683 ug/g)
<LOQ (0.0683 ug/g)
64.8 <LOQ (0.0683 ug/g)
0.0999 - 1 Outlier
0.0648
<LOQ (0.0683 ug/g)
PFOS = Pcrfluorooctanesulfonate
PFHS - Perfluorohexanesulfonate POAA Periluorooctanoate
PFOSA * Pefluorooctane sulfonamide
Mean PFHS
U |/g
<LOQ 60% 70% 73%
<LOQ <LOQ
<LOQ
<LOQ <LOQ <LOQ
RSD Std. Dev. MS/MSD RPD
NA 18% 20% 5%
NA NA
NA -
NA . NA NA
ETS-8-7.0 Excel Version 5/95
GEN-024-liver.xls
000025
5/10/00 6:07 AM
FACT-GEN-024
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 Reducrion/Analyst:
Sample Data
LIVER, KIDNEY, YOLK
Group Dose
Sample#
Method Blk
Birdl0129-wbIk-5-l
Birdl0 129-wbDc-6-1
QC BirdOOl-Loon Lvr-MS
250 ng/g
BirdOOl-Loon Lvr-MSD
Bird023-AIbatross Kdny-MS
Bird023-AIbatross Kdny-MSD
Bird030-Cormorant Yolk-MS
Bird030-Cormorant Yolk-MSD
Liver
BirdOOl-Loon
Bird002-Loon
Bird003-Loon
Bird004-Loon Bird005-Loon
Bird006-Loon
Bird007-Loon
Bird008-Loon
Liver
Bird009-Brown Pelican
BirdOlO-Brown Pelican
Liver
BirdOl 1-Albatross BirdO12-Albatross
Bird013-Albatross
Bird014-Albatross
A,. BirdOl 5-AJbatross BirdOl 6-Albatross
Bird020-Albatross
Bird021 -Albatross
Bird022-Albatross
Kidney
BirdOl7-Albatross BirdOl 8-Albatross
BirdOl 9-Albatross
Bird023-Albatross
Bird024-Albatross
Bird025-AIbatross
Bird026-Albatross
Yolk
Bird027-Comorant
Bird028-Comorant
Bird029-Comorant
Bird030-Comorant
Yolk
Bird031-Gull
Bird032-Gull
Bird033-Gull
No PFOS qualitative confirmation performed. Identifications are preliminary.
LOQ * Limit of Quantitation
RSD = Relative Standard Deviation
RPD = Relative Percent Difierenee
Date Entered/Analyst:
10/23/99 LAC, 12/1/99 GML, 12/30/99 MMH
Date Verified/Analyst:
2/17/00 MMH
GEN024 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0 using unextracted curves Amelia 062498 Masslynx 3.3 10/12/99 SAL/KK 10/14/99,12/13/99 HOJ/IAS 10/15/99, 11/15/99,12/14/99 HOJ/MMH
ttSCOPYmiLABLt
Concentration of POAA
ugIt or % Ree. <LOQ (0.180 ug/g) <LOQ (0.180 ur/r)
101% 108% 113% 121% 105% 130%
<LOQ (0.180 ug/g) <LOQ(0.180ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g)
0.182 <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g) <LOQ (0.180 ug/g)
0.245 <LOQ (0.180 ug/g)
0.192 0.197 <LOQ (0.180 ug/g) 0.196
NA Not Applicable
Mean POAA <LOQ 104% 117% 117%
<LOQ < LOQ
< LOQ -1 Outlier
< LOQ 0.218- 2 outliers 0.196 -1 Outlier
RSD Std. Dev. MS/MSD RPD
NA 6% 7% 22%
NA NA NA NA
NA NA
NA NA
17.2 0.0374 0.528 0.00104
Concentration
Mean
of PFOSA ug/g or % Ree <LOQ (0.00750 ug/g)
PFOSA tig's
<LOQ (0.00750 ug/g) 53%
<LOQ
51% - 52% 56%
58% 57%
62%
72% 67%
0.0153
<LOQ (0.00750 ug/g)
0.0147
0.0262
0.0213
0.0242
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g) 0.0204 - 3 Outliers
<LOQ (0.00750 ug/g)
0.178
0.178-1 Outlier
0.527
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g) <LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ (0.00750 ug/g)
<LOQ
PFOS * Perfiuorooctancsulfonate
PFHS * Perfluorohexanesulfonate
POAA = Perfluorooctanoate
PFOSA - Pcfiuorooctane sulfonamide
RSD Std. Dev. MS/MSD RPD
NA 4% 5% 15%
.25.5 0.00520
NA
NA
' NA
NA NA
ETS-8-7.0 Excel Version 5/95
GEN-024-liver.xls
000026
5/10/00 6:07 AM
FACT-GEN-030
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept: Oates of Extraction/Analyst Dates of Analysis/Analyst:
Date o f Data Reduction/Analyst:
Sample Data BLOOD
Group Dose
Sample#
Method Blk
MSU 12129-H20blk unfiltered 5-3
MSU12 129-H20blk filtered 5-4
Matrix Blk
HMB12129-blood blk-S-1
HMB12129-blood blk-5-2
HMB 12129-blood blk-5-3
HMB 12129-blood blk-5-4
HMB 12129-blood blk-5-5
HMB12129-blood Wk-5-6*
HMB 12129-blood blk-5-7
HMB 12129-blood blk-5-8
HMB12129-blood bIk-5-9
QC PBB-6255-250MS-5-1-2 PBB-6255-250MSD-5-1 -2
FSB-S009-250ppb MS-5-1-2
FSB-S009-250ppb MSD-5-1-2
SSB-SSL49-250ppb MS-5-1-2
SSB-SSL49-250ppb MSD-5-1-2
HMB-FE52189-250 MS-5-1-2 HMB-FE52189-250 MSD-5-1-2
Blood
P205
Northern Fur Seal
P206
Pups
P207
P208 P209
P210
P211
P212
P215
P217
P219
P220
P221
P222
P223 P224
P226
P229
P230
Blood Northern Fur Seal
Adult Females
M104 M105 M106 M107
M112
M115
MI16
M118
M119
M122
Blood Northern Fur Seal
S001 S002
Subadult Males
S003
S006 S007
S008
S009
Blood
P298B
Northern Fur -Seal
P236A 368C P406
P411
98 CUKB 3*
98CUKB7
NA2 EDTA
98 CUKB 9
* Surrogate >30% deviation, not confirmed
I = May need to rerun all samples for PFOSA, interfrent present in both analyses.
** PFOS NOT confirmed; MS transitions variation > 30%
Date Entered/By:
01/21/00 LAC
GEN030 MSU Environmental Samples Various Various Blood ETS-8-4.1 & ETS-8-S. 1 using unextracted curves Soup020199 Masslynx 3.3 See Attachments See Attachments See Attachments See Attachments 12/14/99 SAL/SRP/KK 01/06/00, 01/07/00 MMH/IAS
01/07/00, 01/10/00IAS/MMH
Concentration of PFOS
ug/mL or % Ree. <LOQ (0.00290 ug/mL) <LOQ (0.00290 ug/mL)
0.0253 0.0248 0.0262 0.0243 <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) 74% 91%
-6% -5% 77% 65% -1% 77% <LOQ (0.00579 ug/mL) <LOQ(0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0 00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0 00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL)
Mean PFOS ug/mL
<LOQ
RSD Std. Dev. MS/MSD RPD
NA NA
Concentration of PFHS
ug/mL or % Ree.
<LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL)
Mean PFHS ug/mL
<LOQ
<LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL)
<LOQ (0 00114 ug/mL)
<LOQ (0.00114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL) NA <LOQ (0.0114 ug/mL)
<LOQ - 4 outliers
0.000790
<LOQ (0.0114 ug/mL) 76%
<LOQ
82% 21%
77% 76%
-6% 25%
-1% -1% -1%
71% 17%
83% 77% 80%
38% 208%
1% 74% 37%
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0 0114 ug/mL) <LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ
NA <LOQ (0.0U4 ug/mL)
NA
<LOQ (0.0114 ug/mL)
<LOQ
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
NA <LOQ (0.0114 ug/mL)
<LOQ
NA
<LOQ (0.0114 ug/mL)
<LOQ
<LOQ (0.0114 ug/mL)
<LOQ (0.0U4 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
NA <L0Q (0.0114 ug/mL)
<L0Q
NA
<L0Q (0.0114 ug/mL)
<LOQ
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ (0.0114 ug/mL)
<LOQ
<LOQ (0.0114 ug/mL)
NA <LOQ (0.0114 ug/mL)
NA
<LOQ (0.0114 ug/mL)
<LOQ
NE Not Extracted
PFOS = Perfluorooctanesulfonate
E 3 Lost during extraction PFOSA Pefluorooctane sulfonamide
NA =Not Applicable
PFHS 3 Perfluorohexanesulfonate
LOQ 3 Limit of Quantitation POAA * Perfluorooctanoate
RSD StiL Dev. MS/MSD RPD
NA NA
NA NA 2% 20% 9% 196%
--
--
NA NA
NA NA
NA NA
NA NA
ETS-8-5.1 Excel Version 5/95
GEN-030-sera.xls
000027
5/10/00 6:29 AM
FACT-GEN-030
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number: Instrument Software/Veision: Filename: R-Squared Value: Slope: Y-Intercept: Dates o f Extraction/Analyst: Dates of Analysis/Analyst:
Date of Data Reduction/Analyst:
Sample Data BLOOD
Group Dose
Sample #
Method Bile
MSU 12129-H20blk unfiltered 5-3
MSU12129-H20b!k filtered 5-4
Matrix BIk
HMB12 129-blood blk-5-1
HMB12129-blood Wk-5-2
HMB 12129-blood bIk-5-3
HMB 12129-blood bIk-5-4
HMB 12129-blood blk-5-5
HMB 12129-blood blk-5-6*
HMB 12129-Hood bIk-5-7
HMB 12129-blood blk-5-8
HMBl2129-blood bllc-5-9
QC PBB-6255-250MS-5-1 -2
PBB-6255-250MSD-5-1-2
FSB-S009-250ppb MS-5-1-2
FSB-S009-250ppb MSD-5-1-2
SSB-SSL49-250ppb MS-5-1-2
SSR-SSL49-250ppb MSD-5-1-2
' "v
HMB-FE52189-250 MS-5-1-2
HMB-FE52189-250 MSD-5-1-2
Blood
P205
Northern Fur Seal
P206
Pups
P207
P208
P209
P210
P211
P212
P215
P217
P219
P220
P221
P222
P223
P224
P226
P229
P230
Blood
M104
Northern Fur Seal
M105
Adult Females
M106 M107
M112
M115
Ml 16
Ml 18
Ml 19
M122
Blood
S001
Northern Fur Seal Subadult Males
S002 S003
S006
S007
S008
S009
Blood
P29SB
Northern Fur Seal
P236A
368C
P406
P411
98CUKB3
98CUKB7
NA2EDTA
98 CURB 9
* Surrogate >50% deviation, not confirmed
I * May need to rerun all samples for PFOSA, interfrent present in both analyses.
** PFOS NOT confirmed; MS transitions variation > 30%
Date Entered/By:
01/21/00 LAC
GEN030 MSU Environmental Samples Various Various Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Soup020199 Masslynx 3.3 See Attachments See Attachments See Attachments See Attachments 12/M/99 SAUSRP/KK 01/06/00, 01/07/00 MMH/IAS
01/07/00,01/10/00 ia s/m m h
ft S
T
CO
py
a v a il a b l e
Concentration of POAA
ug/mL or % Ree<LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/fflL) <LOQ (0.00240 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL)
76% 75% 0% 1% 76% 71% 1% 70%
<LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL)
LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL)
LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL) LOQ (0.00958 ug/mL)
Mean POAA ng/mL
<LOQ
RSD Std. Dev. MS/MSD RPD
NA NA
<LOQ 76% 1% 74% 35%
NA NA 1% 36% 7% 195%
<LOQ
NA NA
<LOQ
NA NA
<LOQ
NA NA
NA
<LOQ
NA
NE Not Extracted
E * Lost during extraction
NA Not Applicable
LOQ Limit o f Quantitation
Concentration
Mean
of PFOSA
PFOSA
ng/mL or % Ree
ng/mL
<LOQ (0.000625 ug/mL)
<LOQ (0.000625 ug/mL) <LOQ
0.00195
0.00190
0.00225
0.00189
I
I
I
I
II
50%
47% 49%
I
II
I
1I
I
II
I
I
I
I
I
I
I
1
I
I
1
I
I
I
I
I
l
I
II
I
I
1
I
1
I
I
I
I
II
I
I
I
I
I
I
I1
I
I
I
I
I
I
I
II PFOS *' Perfluorooctanesulfonate
PFOSA =" Pefluorooctane sulfonamide
PFHS Perfluorohexanesulfonate
POAA - Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA 6% NA NA NA
`*
NA NA
-
-
NA NA
NA NA
NA NA
ETS-8-5.1 Excel Version 5/95
GEN-030-scra.xls
000028
5/10/00 6:29 AM
FACT-GEN-030
Study:
GEN030 MSU Environmental Samples
Product Number(Test Substance):
Various
Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Version: Filename: R-Squared Value:
Various Blood ETS-8-4.1 & ETS-8-5.1 using unextracted curves Soup020199
Masslynx 3.3 See Attachments See Attachments
^ S r c o p YAVAILABi[
Slope:
See Attachments
Y-Intercept:
See Attachments
Dates of Extraction/Analyst: Dates of Analysis/Analyst: Date of Data Reduction/Analyst:
12/14/99 SAL/SRP/KKK 01/06/00, 01/07/00 MMH/IAS 01/07/00, 01/10/00 IAS/MMH
Sample Data
BLOOD
Group Dose
Sample #
PFOS Verified
Concentration of PFOS
ug/mL o r % Ree.
Mean PFOS ug/mL
RSD Std. Dev. MS/MSD RPD
Concentration of PFHS
ug/mL or % Ree.
Mean PFHS ug/mL
Method Bile Matrix Blk
QC
MSU 12129-H20blk unfiltered 5-3 MSU12129-H20blk filtered 5-4 HMB12129-blood blk-5-1 HMB12129-blood blk-5-2 HMB 12129-blood blk-5-3 HMB 12129-blood blk-5-4 HMB 12129-blood blk-5-5 HMB 12129-blood blk-5-6* HMB 12129-blood blk-5-7 HMB 12 L29-blood blk-5-8 HMB12129-blood blk-5-9 PBB-6255-250MS-5-1-2 PBB-6255-250MSD-5-1-2
NA
NA
NA
NA NA NA NA NA NA
NA
NA NA NA
<LOQ (0.00290 ug/mL) <LOQ (0.00290 ug/mL)
0.0253 0.0248 0.0262 0.0243 <LOQ (0.0579 ug/mL) <LOQ (0.0579 ug/mL) <LOQ (0.0579 ug/mL) <LOQ (0.0579 ug/mL) <LOQ (0.0579 ug/mL) 74% 91%
<LOQ
<LOQ - 4 outliers 82%
NA NA
NA 0.000790
21%
<LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL) <LOQ (0 00114 ug/mL) <LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL) <LOQ (0.00114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL)
76% 77%
<LOQ
<LOQ 76%
FSB-S009-250ppb MS-5-1-2 FSB-S009-250ppb MSD-5-1-2 SSB-SSL49-250ppb MS-5-1-2 SSB-SSL49-250ppb MSD-5-1-2 HMB-FE52189-250 MS-5-1-2 HMB-FE52189-250 MSD-5-1-2
NA NA NA
NA
NA
NA
-6% -5% 77% 65% -1% 77%
-6% 25% 71% 17% 38% 208%
-1%
-1% -1 % 83% 77% 80% 1% 74% 37%
Blood Polar Bear
6255 (Heparin) 20436 (NA2 EDTA) 20467 (NA2 EDTA)
20468 (EDTA) 20470 20472 20473*
20474 (NA2 EDTA) 20475 (NA2 EDTA) 20476 (NA2 EDTA)
20477 20485 (Heparin) 20486 (Heparin) 20487 (Heparin)
X 0.0518
<LOQ (0.00114 ug/mL)
X 0.0381
<LOQ (0.00114 ug/mL)
X 0.0358
<LOQ (0.00114 ug/mL)
X 0.0315
<LOQ (0.00114 ug/mL)
X 0.0281
<LOQ (0.00114 ug/mL)
X 0.0327
<LOQ (0.00114 ug/mL)
X 0.0309
<LOQ (0.00114 ug/mL)
X 0.0256
<LOQ(0 00114 ug/mL)
X <LOQ (0.00290 ug/mL)
<LOQ (0.00114 ug/mL)
X 0.0345
<LOQ (0.00114 ug/mL)
X 0.0272
<LOQ (0.00114 ug/mL)
X 0.0429
<LOQ (0.00114 ug/mL)
X 0.0316
0.214
<LOQ (0 00114 ug/mL)
X
<LOQ (0.00290 ug/mL) 0.0342 - 2 outliers
0.00731
<LOQ (0.00114 ug/mL)
<LOQ
Blood Stellar Sealion
SSL49 (7.2 mg K2 EDTA) SSL50 (7.2 mg K2 EDTA) SSL51 (7.2 mg K2 EDTA) SSL52 (7.2 mg K2 EDTA) SSL53 (7.2 mg K2 EDTA)
NA NA NA NA NA
<LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL) <LOQ (0.00579 ug/mL)
<LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL)
SSL54 (7.2 mg K2 EDTA) SSL55 (5.4 mg K2 EDTA) SSL56 (5.4 mg K2 EDTA)** SSL57 (5.4 mg K2 EDTA) SSL58 (5.4 mg K2 EDTA)*
N A <LOQ (0.00579 ug/mL) NA <LOQ (0.00579 ug/mL) NA <LOQ (0.00579 ug/mL) N A <LOQ (0.00579 ug/mL) NA <LOQ (0.00579 ug/mL)
<LOQ (0 0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL) <LOQ (0.0114 ug/mL)
SSL59 (5.4 mg K2 EDTA) SSL60 (5.4 mg K2 EDTA)*
NA <LOQ (0.00579 ug/mL) NA <LOQ (0.00579 ug/mL)
<LOQ
NA <LOQ (0.0114 ug/mL)
NA
<LOQ (0.0114 ug/mL)
<LOQ
I " May need to rerun all samples for PFOSA, interfrent present in both analyses.
* Surrogate >50% deviation confirmed
Surrogate >50% deviation, not confirmed
** PFOS NOT confirmed; MS transitions variation > 30%
Date Entered/By:
1/13/00, 01/21/00 MMH/LAC
Date Verified/ By:
02/10/00 kjh
N E " Not Extracted E " Lost during extraction NA * Not Applicable NV = Not Verified LOQ111Limit of quantitation
PFOS 0 Perfiuorooctanesulfonate PFOSA * Pefluorooctane sulfonamide PFHS **Periluorohexanesulfonate POAA = Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD . na
NA
NA NA 2% 20% 9% 196%
NA NA
NA NA
ETS-8-5.1 Excel Version 5/95
GEN-030-sera,xls
000029
5/9/00 5:37 PM
FACT-GEN-030
Study:
GENQ30 MSU Environmental Samples
Product NumberfTest Substance):
Various
Matrix: Method/Revision:
Various Blood ETS-8-4.1 & ETS-8-5.1 using unextracted-curves
Analytical Equipment System Number:
Soup020199
Instrument Software/Version: Filename: R-Squared Value: Slope: Y-Intercept:
Masslynx 3.3 See Attachments See Attachments See Attachments See Attachments
BEST COPY AVAILABLE
Dates of Extraction/Analyst:
12/14/99 SAUSRP/KKK
Dates of Analysis/Analyst:
01/06/00, 01/07/00 MMH/IAS
Date of Data Reduction/Analyst
01/07/00, 01/10/00 IAS/MMH
Sample Data
BLOOD
Group Dose
Method Blk
Matrix Blk
Sample M
MSU!2129-H20blk unfiltered 5-3 MSU 12129-H20blk filtered 5-4 HMB 12129-blood blk-5-1 HMB12129-blood blk-5-2 HMB 12129-blood blk-5-3
Concentration of POAA
ng/mL or % Ree
<LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL)
Mean POAA ng/mL
<LOQ
RSD Std. Dev. MS/MSD RPD
NA NA
Concentration of PFOSA
ug/mL or % Ree. <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL)
Mean PFOSA ug/mL
<LOQ
RSD Std. Dev. MS/MSD RPD
.NA NA
HMB 12129-blood bIk-5-4 HMBI 2129-Wood blk-5-5 HMB12129-blood blk-5-6*
<LOQ (0.00240 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL)
<LOQ (0.000625 ug/mL) I 1
HMB12129-blood bIk-5-7
<LOQ (0.00958 ug/mL)
I
HMB 12129-blood blk-5-
<LOQ (0.00958 ug/mL)
NA !
NA
QC
S
HMB12129-blood blk-5-9 PBB-6255-250MS-5-1-2 PBB-6255-250MSD-5-1-2 FSB-S009-250ppb MS-5-1-2 FSB-S009-250ppb MSD-5-1-2 SSB-SSL49-250ppb MS-5-1-2 SSB-SSL49-250ppb MSD-5-1-2 HMB-FE52189-250 MS-5-1-2
<LOQ (0.00958 ug/mL) 76% 75% 0% 1% 76% 71% 1%
<L0Q 76% 1% 74%
NA 1% 36% 7%
I I NA 50% 47% 49% 6%
1 I I NA I I 1 NA I
HMB-FE52189-250 MSD-5-1-2
70%
35%
195%
I I NA
Blood Polar Bear
6255 (Heparin) 20436 (NA2 EDTA)
<LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL)
<LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL)
20467 (NA2 EDTA)
<LOQ (0.00240 ug/mL)
<LOQ (0.000625 ug/mL)
20468 (EDTA) 20470 20472 20473*
20474 (NA2 EDTA)
<LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL) <LOQ (0 00240 ug/mL)
<LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL)
20475 (NA2 EDTA)
<LOQ (0.00240 ug/mL)
<LOQ (0.000625 ug/mL)
20476 (NA2 EDTA)
<LOQ (0.00240 ug/mL)
<LOQ (0.000625 ug/mL)
20477 20485 (Heparin)
<LOQ (0.00240 ug/mL) <LOQ (0.00240 ug/mL)
<LOQ (0.000625 ug/mL) <LOQ (0.000625 ug/mL)
20486 (Heparin)
<LOQ (0.00240 ug/mL)
NA <LOQ (0.000625 ug/mL)
NA
20487 (Heparin)
<LOQ (0.00240 ug/mL)
<LOQ
NA
<LOQ (0.000625 ug/mL)
<LOQ
NA
Blood Stellar Sealion
SSL49 (7.2 mg K2 EDTA) SSL50 (7.2 mg K2 EDTA) SSL51 (7.2 mg K2 EDTA)
<LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL)
I I I
SSL52 (7.2 mg K2 EDTA)
<LOQ (0.00958 ug/mL)
I
SSL53 (7.2 mg K2 EDTA)
<LOQ (0.00958 ug/mL)
I
SSL54 (7.2 mg K2 EDTA)
<LOQ (0.00958 ug/mL)
I
SSL55 (5 4 mg K2 EDTA)
<LOQ (0.00958 ug/mL)
I
SSL56 (5.4 mg K2 EDTA)**
<LOQ (0.00958 ug/mL)
I
SSL57 (5.4 mg K2 EDTA)
<LOQ (0.00958 ug/mL)
I
SSL58 (5.4 mg K2 EDTA)* SSL59 (5.4 mg K2 EDTA) SSL60 (5.4 mg K2 EDTA)*
<LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL) <LOQ (0.00958 ug/mL)
I = May need to rerun, all samples forPFOSA, interfrent present in both analyse
<LOQ
NA NA
I
I NA I I NA
PFOS = Perfluorooctanesulfonate
** Surrogate >50% deviation confirmed
NE ="Not Extracted
PFOSA * Pefluorooctane sulfonamide
* Surrogate >50% deviation, not confirmed
** PFOS NOT confirmed; MS transitions variation > 30%
Date Entered/By:
1/13/00,01/21/00 MMH/LAC
Date Verified/ By:
02/10/00 kjh
E <BLost during extraction NA * Not Applicable N V * Not Verified LOQ = Limit of quantitation
PFHS = Perfluorohexanesulfonate POAA = Pcrfluorooctanoate
ETS-8-5.1 Excel Version 5/95
GEN-030-sera.xls
000030
5/9/00 5:37 PM
FACT-GEN-030 MSU Environmental Samples
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
CEN030 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0 Soup 020199, Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 01/10/00 MMH 01/12/00 IAS
B (s r COPYAVAILABLE
MUSCLE
Dose
Sample #
PFOS Verified
Method Blk Unfiltered water
Method Blk Filtered water
Matrix Blk
MSU 12149-H20unfil 5-1* MSU 12149-H20unfil 5-2*
MSU12149-H20fil 5-1* MSU12149-H20fil 5-2* MSU 12149-Fishblk 5-1
NA NA NA NA NA
Fish Liver
MSU 12149-Fishblk 5-2 MSU12149-Fishblk 5-3*
NA NA
QC 250 ng/g
CPM-B1N7-MS 5-1-2 CPM-B2N6-MS 5-1-2 CSM-1999030-03-01-MS 5-1-2 CSM-1999030-03-01-MSD 5-1-2 LWM-1999029-16-MS 5-1-2 LWM-1999029-11 -MS 5-1-3
"
NA NA NA NA NA NA
BTM- 1999040-09-MS 5-1-2
NA
BTM-1999040-10-MS 5-1-2
NA
Muscle Carp
B1N1 B1N2 B1N7
X X X
B1N10
X
B2N2 B2N6
X X
B2N8 B2N10
X X
Carpi*
X
Carp2*
X
Muscle Chinook Salmon
1999030-01 1999030-02
NA NA
1999030-02-01 1999030-02-04
X X
1999030-03-01 1999030-03-04
X X
Muscle
1999029-11*
X
Lake Whitefish
1999029-12* 1999029-13 1999029-14 1999029-16
X X X X
Muscle Brown Trout
1999040-01 1999040-02 **
1999040-03 1999040-04 1999040-05 1999040-06 1999040-07 1999040-08 1999040-09 1999040-10
X X** NA NA
X X NA X NA NA
* High (>50%) surrogate deviations
** PFOS NOT confirmed; MS transitions variation > 30% Date Entered/Analyst: 01/24/00, 01/25/00, 01/28/00 LAC
Date Verified/Analyst: 02/10/00 kjh
Concentration of PFOS
ng/g or % Rec. 0.0163
<LOQ ( 0.00696) <LOQ ( 0.00696)
0.0257 0.00948 0.00985 0.00454
59% 31% 160% 127% 67% 122% 148% 166% 0.101 0.0784 0.0905 0.0878 0.105 0.0894 0.0596 0.0838 0.297 0.243 0.189 0.126 <LOQ ( 0.00696) 0.113 0.0514 0.0573 0.168 0.130 0.0967 0.0983 0.1659
<LOQ ( 0.00696) <LOQ ( 0.00696) <LOQ ( 0.00696) <LOQ ( 0.00696) <LOQ ( 0.00696) <LOQ ( 0.00696) <LOQ ( 0.00696)
0.0460 <LOQ { 0.00696) <LOQ ( 0.00696)
Mean PFOS
Q g/g
NA NA
0.00796 45% 143% 95% 157%
RSD Std. Dev. MS/MSD RPD
NA NA NA NA
37.2 0.00296
63%
23%
59%
11%
0.124 0.107-1 outlier
0.132
64.2 0.0793
0.524 0.0562
26,4 0.0348
<LOQ -1 outlier
NA NA
NE = Not Extracted
E * Lost during extraction
NA Not Applicable
LOQ = Limit o f Quantitation
X = Verified PFOS concentration
Concentration of PFHS
ug/g or % Rec. <LOQ ( 0.0342)
Mean PFHS ug/g
<LOQ ( 0.0342)
<LOQ
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
60%
45% 148%
53%
183% 129%
166%
130% 144%
130%
168%
156%
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
<L0Q ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ ( 0.0342)
<LOQ
PFOS - Perfluorooctanesulfonate
PFOSA * Pefluorooctane sulfonamide
PFHS = Perfluorohexanesulfonate
POAA Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD
NA NA NA NA NA 28% 21% 0% 15%
NA NA
1
NA NA -
N j^
NA-:
NA NA
ETS-8-7.0 Excel Version 5/95
GEN-030-ltver.xls
000031
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
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
GEN030 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0 Soup 020199, Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 01/10/00 MMH 01/12/00 IAS
B iSTM P Y AVAILABLE
MUSCLE
Group Dose
Sample#
Method Blk Unfiltered water
Method Blk Filtered water
Matrix Blk Fish Liver
QC 250 ng/g
Muscle Carp
v' f
Muscle Chinook Salmon
Muscle Lake Whitefish
Muscle Brown Trout
MSU 12149-H20unfil 5-1* MSU 12149-H20unfil 5-2*
MSU12149-H20fil 5-1* MSU12149-H20fil 5-2* MSU12149-Fishblk 5-1 MSU12!49-Fishblk 5-2 MSU12149-Fishblk 5-3* CPM-B1N7-MS 5-1-2 CPM-B2N6-MS 5-1-2 CSM-1999030-03-01-MS 5-1-2 CSM-1999030-05-01-MSD 5-1-2 LWM-1999029-16-MS 5-1-2 LWM-1999029-11-MS 5-1-3 BTM-1999040-09-MS 5-1-2 BTM-1999040-10-MS 5-1-2
BINI BIN2 BIN7 B1NI0 B2N2 B2N6 B2N8 B2N10 Carpi* Carp2* 1999030-01 1999030-02 1999030-02-01 1999030-02-04 1999030-03-01 1999030-03-04 1999029-11* 1999029-12* 1999029-13 1999029-14 1999029-16 1999040-01 1999040-02** 1999040-03 1999040-04 1999040-05 1999040-06 1999040-07 1999040-08 1999040-09 1999040-10
kPFOS NOT confirmed; MS transitions variation > 30% Date Entered/Analyst: 01/24/00, 01/25/00, 01/28/00 LAC Date Verified/Analyst: 02/10/00 kjh
Concentration of POAA
ug/g or % Ree. <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359)
6% 39% 139% 178% 146% 105% 147% 152% <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ { 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ< 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0 0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ ( 0.0359)
Mean POAA
<LOQ <LOQ
<LOQ 23% 158% 125% 150%
RSD Std. Dev. MS/MSD RPD
NA NA NA NA
NA NA
148%
25%
33%
3%
<LOQ <LOQ <LOQ
NA NA
NA NA
NA NA
<LOQ
NA NA NE " Not Extracted E = Lost during extraction NA = Not Applicable LOQ Limit of Quantitation X Verified PFOS concentration
Concentration of PFOSA
*g/g o r % R ee <LOQ( 0.0188)
Mean PFOSA
Ug/i
RSD Std. Dev. MS/MSD RPD
NA
<LOQ( 0.0188) <LOQ( 0.0188) <LOQ( 0.0188) <LOQ( 0.0188)
<LOQ <LOQ
NA NA NA
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA NA
11%
13% 12% 19% 77%
69% 73% 11% 61%
76% 68% 22% . 80%
87% 83%
8%
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ{ 0.0188) <LOQ{ 0.0188)
<LOQ
NA NA
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<l o q ( o .o m )
<LOQ( 0.0188)
NA
<LOQ( 0.0188) <LOQ
NA
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ{ 0.0188)
<LOQ( 0.0188)
NA
<LOQ( 0.0188) <LOQ
NA
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0 0188)
<LOQ ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
NA
<LOQ( 0.0188) <LOQ
NA
PFOS = Perfluorooctanesulfonate
PFOSA = Pefiuorooctane sulfonamide
PFHS *Periluorohexanesulfonate
POAA * Perfluorooctanoate
ETS-8-7.0 Excel Version 5/95
GEN-030-liver.xls
000032
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study: Product Number(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Venion: Date of Extraction/Analyst: Date of Analysis/Analyse Date of Data Reduction/Analyst:
Sample Data
GEN030 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0 Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 01/10/00 MMH 01/11/00 IAS
MUSCLE and EGGS
Group Dose
Sample #
Method Blk Water
Matrix Blk Egg
MSU 12159-H20Blk-unfiltered-5-9 MSUl2159-H20Blk-filtered-5-9
EggBlk-5-I EggBlk-5-2 EggBUc-5-3
QC FGW-ALSWCR-TD2 6/25/98-MS-S-1-2
250 ng/g
FGW-ALSWCR-TD2 6/25/98-MSD-5-1-2
FGE-AL118-HP26/25/98-MS-5-1-2
FGE-AL118-HP26/25/98-MSD-5-1-2
LWE-19999029-13-MS-5-1*2
LWE-1999902913-MSD-5-1-2
BTE-19999040-01 -MS-5-1-2 BTE-19999040-01-MSD-5-1-2
Carp
Diet 1(Carpi)
Diet 2 (Caip6)
Diet 3 (Carp4)
Diet 4 (Carp3)
Frog Muscle Wholebody
AL-118-YOY 08/26/98 ** ALSWCR-TD 06/03/98 *
ALSWCR-TD2 06/25/99 *
KZCKDM-JUV 08/26/98 *
K2CKDM-TD 06/05/98 **
K2CKDM-TD-2 06/25/98 *
V SJ0002-TD-2 06/05/98 **
Green Frog Eggs
1 SJ0002-TD-2 06/25/98 AL-118-HP89 06/25/98 AL-118-HP94 06/25/98
SJ0001 06/03/98**
SJ0001 06/05/98
Lake Whltensh Eggs
1999029-13
1999029-14
Brown Trout Eggs
1999040-01
1999040-04
1999040-06
* High (>50%) surrogate deviations
** PFOS NOT confirmed; MS transitions variation > 30*/*
Date Entered/Analyst:
01/21/00,01/24/00 LAC
Date Verified/Analyst:
02/10/00kjh
PFOS Verified
NA NA NA NA NA X X X X X X X X X** X X X X** X** X** X** X** X** X** X** X X X** X X X X X X
Concentration of PFOS
ug/t or % Ree. <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174)
44% 47% 13% 15% 146% 104% 121% 167%
<LOQ (0.0174) <LOQ (0.0174)
0.0267 0.0278
0.00243 <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174)
0.0216 <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174)
<LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174) <LOQ (0.0174)
0.145 0.381
0.0749 0.0675 0.0488
Mean PFOS _______" t ' e <LOO <LOQ 46% 14% 125% 144%
0.0272 - 2 outliers
<LOQ - 2 outliers
<LOQ 0.263 0.0637
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA
Concentration of PFHS
ug/g or % Ree. <LOQ (0.0342) <LOQ (0.0342) <LOQ (0.0342) <LOQ (0.0342) <LOQ (0.0342)
16% 8% 16%
10% 8% 12%
30% 33% 35%
57% 32% 57%
<LOQ (0.0342) <LOQ (0.0342) NA <LOQ (0.0342) NA <LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342) NA <LOQ (0.0342) NA <LOQ (0.0342)
<LOQ (0.0342)
<LOQ (0.0342) NA <LOQ (0.0342) NA <LOQ (0.0342)
63.5 0.167
<LOQ (0.0342) <LOQ (0.0342)
21.1 0.0134
<LOQ (0.0342) <LOQ (0.0342) <LOQ (0.0342)
NE * Not Extracted E * Lost during extraction NA9 Not Applicable LOQ9 Limit of Quantitation X 9 Verified PFOS concentration
Mean PFHS u/g <LOQ
<LOQ 16% 11% 33% 57%
<LOQ
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA
3%
13%
16%
1%
NA . NA
<LOQ
NA NA
<LOQ <LOQ
NA NA
NA NA
-- NA
<LOQ
NA
PFOS * PwflurvrvytnnrsMlff>ate
PFOSA * Pefluorooctane sulfonamide
PFHS * Perfluorohexanesulfonate
POAA = Perfluorooctanoate
ETS-8-7.0 Excel Version 5/95
GEN-030-1iver.xls
000033
5/IQ/00 6:46 AM
FACT-GEN-030 MSU Environmental Samples
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
GEN030 MSU Environmental Samples Various Various ETS-8-6.0 A ETS-8-7.0 Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 01/10/00 MMH Oim/OO IAS
COPYAVJHl m i
MUSCLE and EGGS
Group Dose
Method Blk Water
Matrix Blk EgS
QC 250 ng/g
Sample 0
MSUI2!59-H20BDc*unfiltered-5-9 MSU12159-H20Btk-filtered-5-9
EggBlk-5-1 EggBlk-5-2 EggBflc-5-3 FGW-ALSWCR-TD2 6/25/98-MS-5-1-2 FGW-ALSWCR-TD2 6/25/98-MSD-S-1-2 FGE-AL118-HP26/25/98-MS-5-1-2
Concentration of POAA
ug/g or % Rec. <LOQ ( 0.0359) <LOO ( 0.0359) <LOQ ( 0.0359) <LOQ ( Q.0359) <LOQ ( 0.0359)
34% 33% 22%
Mean POAA
<LOQ
<LOQ 34%
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA
4%
Concentration of PFOSA
ug/g or % Rec <LOQ( 0.0188) <LOQ( 0.0188) <LOQ( 0.0188) <LOQ(0.Q188) <LOQ( 0.0188)
49% 49% 46%
Mean PFOSA
ug/g <LOQ
<LOQ
49%
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA
1%
FGE-AL118-HP26/25/98-MSD-5-1-2
26% 24% 15%
54%
50%
16%
LWE-19999029-U-MS-5-1-2
90%
68%
LWE-19999029-13-MSD-5-1-2 BTE-19999040-01-MS-5-1-2
83% 86% 9% 98%
64% 104%
66%
6%
BTE-19999040-01-MSD-5-1-2
112%
105%
13%
108%
106%
4%
Carp
Diet 1 (6arp5)
<LOQ ( 0.0359)
<LOQ( 0.0188)
Diet 2 (Carp6) Diet 3 (Caip4) Diet 4 (Caip3)
<LOQ ( 0.0359) <LOQ ( 0.0359) <LOQ( 0.0359)
<LOQ
NA NA
<LOQ( 0.0188) <LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA NA
Frog Muscle
AL-118-YOV 08/26/98**
<LOQ( 0.0180)
<LOQ( 0.0188)
Wholebody
ALSWCR-TD 06/03/98 *
<LOQ( 0.0180)
<LOQ( 0.0188)
ALSWCR-TD2 06/25/99
<LOQ( 0.0180)
<LOQ( 0.0188)
KZCKDM-JUV 08/26/98 **
<LOQ( 0.0180)
<LOQ(0.0188)
KZCKDM-TD 06/05/98
<LOQ( 0.0180)
<LOQ( 0.0188)
K2CKDM-TD-2 06/25/98
<LOQ(0.0180)
<LOQ ( 0.0188)
SJ0002-TD-2 06/05/98 *
<LOQ( 0.0180)
NA <LOQ( O.OI88)
NA
A
SJ0002-TD-2 06/25/98 *
<LOQ( 0.0180) <LOQ
NA
<LOQ( 0.0188)
<LOQ
NA
Green Frog Eggs
AL-118-HP89 06/25/98
<LOQ ( 0.0180)
<LOQ( 0.0188)
AL-U8-HP94 06/25/98
<LOQ( 0.0180)
<LOQ ( 0.0188)
SJ0001 06/03/98** SJ0001 06/05/98
<LOQ( 0.0180) <LOQ( 0.0180)
<LOQ
NA NA
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA NA
Lake Whiteflsh Eggs
1999029-13 1999029-14
<LOQ( 0.0180) <LOQ( 0.0180)
<LOQ
NA NA
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA NA
Brown Trout Eggs
1999040-01
<LOQ( 0.0180)
<LOQ( 0.0188)
1999040-04 1999040-06
<LOQ ( 0.0180) <LOQ ( 0.0180)
<LOQ
NA NA
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA NA
* High (>50%) surrogate deviations
NE = Not Extracted
PFOS = Perfluorooctanesulfonate
** PFOS NOT confirmed; MS transitions variation > 30%
E = Lost during extraction
PFOSA = Pefluorooctane sulfonamide
Date Entcred/Analyst:
01/21/00,01/24/00 LAC
NA = Not Applicable
PFHS Perfluorohexanesulfonate
Date Verified/AnalysC
02/10/00 kjh
LOQ Limit of Quantitation
POAA Perfluorooctanoate
X - Verified PFOS concentration
ETS-8-7.0 Excel Version 5/95
GEN-03O-fiver.xls
000034
5/10/00 6:46 AM
FACT-GEN-030 MSU Environmental Samples
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
GEN030 MSU Environmental Samples Various Various ETS-8-60 & ETS-8-7.0 Soup 020199, Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 12/17/99, 12/20/99, 12/28/99, 12/29/99,01/03/00,01/06/00 IAS/MMH 12/20/99,12/21/99,12/22/99,12/30/99,01/03/00, 01/05/00,01/07/00 MMH/IAS
COPYM ahable
Miscellaneous Liver
Group Dose
Sample #
Method Blk Unfiltered water
MSU 12129-H20Blk-unfiltercd-5-1 MSU 12129-H20B1k-unfiltered-5-2 MSU 12129-H20Blk-unfiItered-5-3 MSU 12129-H20Blk-unfiltered-5-4 MSU12129-H20Blk-unfiltcred-5-5
MSU 12129-H20Blk-unfiltered-5-6 MSU12129-H20Blk-unfiltered-5-7 MSU12129-H20BIk-unfiltered-5-8
Method Blk Filtered water
MSU12129-H20BIk-filtered-5-1 MSU12129-H20Blk-filtered-5-2 MSU 12129-H20Blk-filtered-5-3 MSU12129-H20Blk-filtered-5-4
MSU12129-H20Blk-fiUered-5-S MSU12 129-H20Blk-filtered-5-6
MSU! 2129-H20Blk-filtered-5-7
MSU 12129-H20Blk-filtered-5-8
Matrix Blk Fish Liver
FSH12 129-LvrBlk-5-1 FSH12129-LvrBlk-5-2
Matrix Blk Rabbit Liver
A
FSH12129-LvTBlk-5-3 RBL12129-LvrBlk-5-1 RBL12129-LvrBIk-5-2 RBL12129-LvrBlk-5-3 RBL12129-LvrBIk-5-4
RBL12129-LvrBlk-5-5*
RBL12129-LvrBlk-5-6*
QC 250 ng/g
Mink Liver, D530, MS-5-1-1 Mink Liver, D530, MSD-5-1-2* CSL-1999030-03-0 l-MS-5-1 -2* CS L -1999030-03-01-M SD -5-1-2 *
LWL-1999029-12-MS-5-1-2*
LWL-1999029-12-MSD-5-1-2* BTL-1999040-01-MS-5-1-2* BTL-1999040-01 -MSD-5-1-2*
TNL-TU54-MS-5-1-2 TNL-TU54-MSD-5-1-2
FSL-P295-MS-5-1 FSL-P295-MSD-5-1 PBL-980390LB-MS-5-1
PBL-980390LB-MSD-5-2* GFL-KZCKDM-D1-MS-5-1-2 GFL-KZCKDM-D1-MSD-5-1-
TTL-LCPTR99503C-MS-5-1-2* TTL-LCPTR99503C-MSD-5-1-2
MTL-10Vancleave98-MS-5-1 -2 MTL-I0Vancleave98-MSD-5-l-2
Liver Chinook Salmon
Liver Lake Whitefish
1999030-01 1999030-02 1999030-02-01 1999030-02-04 1999030-03-01 1999030-03-04
1999029-U 1999029-12
1999029-13 1999029-14 1999029-16
Liver
1999040-01
Brown Trout
1999040-02 1999040-03 1999040-04 1999040-05 1999040-06 1999040-07 1999040-08 1999040-09 1999040-10
* High (>50%) surrogate deviations
Date Entered/Analyst: Date Verified/Analyst:
12/22/99, 12/28/99,12/29/99,12/30/99, 01/12/00,01/17/00,01/18/00 LAC 0
PFOS Verified
NA NA NA NA NA NA NA NA NA *NA NA v NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA X X NA NA NA NA
NA NA X X NA NA NA NA NA NA NA NA NA NA NA
.
Concentration of PFOS
ug/g or % Ree. NA NA
<LOQ (0.0347) <LOQ (0.0347)
E <LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696)
NA NA E <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696) 0.0305 0.0331 0.0170 <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0696) <LOQ (0 0696) <LOQ (0.0696) <LOQ (0.0696)
145% 538% 196% 140% 321% 287% 138% 132% 68%
E 91% 86% 18% 179% 93% 105% 90% 100% 79% 89%
0.109 0.169 0.0328 0.126 0.173 0.0405
0.0679 0.0812 0.0738 0.0329 0.0778 <LOQ( 0.0174) <LOQ( 0.0174) <LOQ( 0.0174) <LOQ( 0 0174) 0.0255 <LOQ( 0.0174) <LOQ( 0.0174) <LOQ( 0.0174) <LOQ( 0.0174) <LOQ( 0.0174)
Mean
PFOS vtflt
RSD Std. Dev. MS/MSD RPD
<LOQ
NA NA
<LOQ 0.0269
<LOQ 342% 168% 304% 135% 68% 88% 99% 99% 95% 84%
0.108
0.0667
NA NA 32.1 0.00862
NA NA It 5% 33% 11% 5% NA 6% 163% 12% 11% 11%
56.1 0.0608
29.3 0.0195
<LOQ - 1 outlier
NE - Not Extracted . E = Lost during extraction NA * Not Applicable LOQ Limit of Quantitation X - Verified PFOS concentration
NA NA
Concentration
Mean
of PFHS
PFHS
ng/g or % Ree. NA
ng/g
NA
<LOQ (0.00683)
<LOQ (0 00683)
E
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ
NA
NA
E
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0 0171)
<LOQ
61%
50% 55%
116%
112%
114%
57%
68% 63%
81%
84% 83%
55%
E 55%
97%
84% 90%
70%
76% 73%
82%
89% 86%
66%
72% 69%
74%
84% 79%
<LOQ( 0.0171)
<LO Q( 0.0171)
<LOQ( 0.0171)
<LO Q( 0.0171)
<LOQ{ 0.0171)
<LOQ( 0.0171)
<LOQ
<LO Q( 0.0171)
<LO Q( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ
<LOQ( 0.0171)
<LO Q( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ
PFOS Perfluorooctanesulfonate
PFOSA Pefluorooctane sulfonamide
PFHS = Pcrfluorohexanesulfonatc
POAA " Perfluorooctanoate
RSD Std. Dev. -- Mj-S--/-M---S--D---R--P--D-------
NA NA
NA NA NA NA
NA NA 21% 4% 18% 4% NA 15% 9% 9% 8% 7. 13%
NA NA
NA NA
- ...
NA NA
ETS-8-7.0 Excel Version 5/95
GEN-030-liver.xls
000035
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study: Product Number(Tcst 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
GEN030 MSU Environmental Samples Various Various ETS-8-6.0 A ETS-8-7.0 Soup 020199, Amelia 062498 Masslynx 3.3 12/12/99 SAL/KK/SRP/CSH 12/17/99, 12/20/99, 12/28/99,12/29/99,01/03/00,01/06/00 IAS/MMH 12/20/99, 12/21/99, 12/22/99,12/30/99, 01/03/00, 01/05/00,01/07/00 MMH/IAS
Miscellaneous Liver
Group Dose
Sample U
Method Blk
MSU 12129-H20Blk-unfiltercd-5-1
Unfiltered water
MSU 12129-H20Blk-unfiltered-5-2
MSU 12129-H20Blk-unfiltered-5-3
MSU 12129-H20BIk-unfiItered-5-4
MSU 12129-H20Blk-unfiltered-5-5
MSU 12129-H20Blk-unfiItered-5-6
MSU12129-H20Blk-unfiltered-5-7
MSU 12129-H20Blk-unfiltered-5-8
Method Btk
MSU12129-H20Blk-filtered-5-1
Filtered water
MSU12129-H20Blk-fi1tered-5-2
MSU 12129-H20Blk-filtered-5-3
MSU 12129-H20Blk-filtered-5-4
MSU 12129-H20BIk-filtered-5-5
MSU 12129-H20Blk-fiItered-5-6
MSU 12129-H20B\k-fiItered-5-7
MSU 12129-H20Blk-filtcred-5-8
Matrix Blk
FSH12129-LvrBlk-5-1
Fish Liver
FSH12129-LvrBlk-5-2
FSH12129-LvrBlk-5-3
Matrix Blk .
RBL12129-LvrBlk-S-l
4Rabbit Liver
RBL12129-LvrBlk-5'2 RBL 12129-LvrBlk-5-3
RBL 12129-LvrBlk-5-4
RBL12129-LvrBlk-5-5*
RBL12129-LvrBlk-5-6*
QC 250 ng/g
Mink Liver, D530, MS-5-1-1 Mink Liver, D530, MSD-5-1-2*
CSL-1999030-03-01-MS-5-1-2*
C S L -1999030-03-01-M SD-5-1-2 *
L W L -1999029-12-MS-5-1 -2*
LWL-1999029-12-MSD-5-1-2*
BTL-1999040-01 -MS-5-1-2*
BTL-1999040-01 -MSD-5-1-2*
TNL-TU54-MS-5-1-2
TNL-TU54-MSD-5-1-2
FSL-P295-MS-5-1
FSL-P295-MSD-5-1
PBL-980390LB-MS-5-1
PBL-980390LB-MSD-5-2* .
GFL-KZCKDM-D1 -MS-5-1-2
GFL-K2CKDM-D1-MSD-5-1-
TTL-LCPTR99503C-MS-5-1-2*
TTL-LCPTR99503C-MSD-5-1 -2
MTL-10Vancleave98-MS-5-1 -2
MTL- 10VancIeave98-MSD-5-1-2
Liver
1999030-01
Chinook Salmon
1999030-02
1999030-02-01
1999030-02-04
1999030-03-01
1999030-03-04
Liver
1999029-11
Lake Whitefish
1999029-12
1999029-13
1999029-14
1999029-16
Liver
1999040-01
Brown Trout
1999040-02
1999040-03
1999040-04
1999040-05
1999040-06
1999040-07
1999040-08
1999040-09
1999040-10
* High (>50%) surrogate deviations
Date Entered/Analyst: Date Verified/Analyst:
12/22/99,12/28/99,12/29/99,12/30/99, 01/12/00,01/17/00,01/18/00 LAC 0
Concentration of POAA
ug/g o r % R ee NA NA
<LOQ (0.0719) <LOQ (0.0719)
E <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
NA NA E <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
86% 84% 131% 111% 133% 145% 127% 120% 84%
E 72% 80% 85% 129% 82% 103% 73% 64% 100% 75% <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
<LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
Mean POAA
nt/t
RSD Std. Dev. MS/MSD RPD
<LOQ
NA NA
<LOQ <LOQ
NA NA
NA NA
<LOQ 85% 121% 139% 124% 84% 76% 107% 92% 69% 88%
NA NA 3% 17% 9% 6% NA 10% 41% 23% 13% 29%
<LOQ <LOQ
NA NA
NA NA
NA
<LOQ
NA
NE Not Extracted
E Lost during extraction
NA " Not Applicable
LOQ = Limit of Quantitation
X Verified PFOS concentration
Concentration
Mean
of PFOSA ug/g or % R ee
NA
PFOSA ng/g
NA
<LOQ(0.0188)
<LOQ( 0.0188) E
<LOQ( 0.0188)
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
NA
NA
E
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ
<LOQ( 0.0188)
<LOQ ( 0.0188)
<LOQ( 0.0188)
<LOQ
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188) <LOQ( 0.0188)
<LOQ
43%
81% 62% 64%
52% 58% 77%
83% 80% 94%
92% 93%
71%
E 71%
87%
82% 85% 71%
65% 68%
82%
93% 88% 78%
71% 74% 74%
74% 74%
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ
<LOQ( 0.0188)
<LOQ ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ
PFOS = Perfluorooctanesulfonate
PFOSA = Pefluorooctane sulfonamide
PFHS - Perfluorohexanesulfonate
POAA = Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA NA NA
NA NA 60% 21% 7% 3% NA 7% 9% 12% 10% 0%
NA NA
NA NA
NA NA
ETS-8-7.0 Excel Version 5/95
GEN-030-liver.xls
000036
i
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study; Product NumberfTc* Substance): Matrix: Melhod/Revision: Analytical Equipment System Number lastnunenl Software/Vcraion: Date o f Extractton/Analyst: Data o f Analysis/Analyst Date o f Data Reduction'Analyst:
Sample Data
Miscellaneous Liver
Group Doee
Sample
Method Bik
MSU12129-H20BUc-unlittered-5-l
Unaltered water
MSUJ2129-H20Blk-*filtered-5-2 M S U 1 2 l294i2 0B lk -onfih ered-5-3 MSUl 2129-H20BDc-mfittend-S'4 MSU 12129-H20BUt-unfihered-3-5 MSU12129-H20BDc-aafiMerad-5'6 MSU 2129-H2OB0c-uafihered-5-7
Method Bik Filtered water
Matrix Bik Fish Liver Matrix Bik Rabbit Liver
MSUI2129-H20Btk-anfiHerad-5-8 MSUl2129-H20Btk-fihe*ed-S-l M S U I2 129*H 20Btk-0tered-J* 2 MSU12129-H20Blk-Ghered-5-3 MSU12 129-H20BIk-fQtoed-5-4 MSU12129-H20BIk-Dtered-5-5 MSUl 2129-H20BIk-fita*ed-5-6 MSU12129-H20BJk-5h*fe-5-7 MSUl 2129-H2QBIk-fillen>d-5-8 FSH12l29-LvrBk-5-l FSH12129-Lvifilk-5-2 FSH12129-LvrBJk-J-3 RBL12I29-LvrfiIk-5-1
RBL121294.viBIk-5-2 ^ RBL12129-LvrBIk-S-3 RBL12129-LvrBft-5-4 RBLI2l29-LviBlk-5-5* RBL12t29-LvrBlk-5-6*
QC Mink Liver, DS30, MS-5-1-1
250 aft/g
Mink Liver. D530. MSD-5-1-2* CSL-1999030-03-01-MS-5-1-2* CSL-19990304)3-0 l-MSD-5-1 -2*
LWL-1999029-12-MS-5-1 -2* LWL-1999029-12-MSD-S-l -2*
BTL-l 999040411-MS-5-1-2*
BTL-19990404)1 -MSD-5-1-2*
TNL-TU54-MS-5-1-2 TNL-TU54-MSD-5-1-2
FSL-P295-MS-5-1
FSL-P295-MSD-5-1
PBL-980390LB-MS-5-1 PBL-980390LB-MSD-5-2*
GFL-KZCKDM-DI-MS-5-1-2 GFL-KZCKDM-D1-MSD-5-1-
TTL-LCPTR99503C-MS-5-1-2*
TTL-LCPTR99503C-MSD-5-I-2 MTL-10V anc)eav98 -MS-5-1 -2 MTL-10Vockave98-MSD-5-l-2
Liver Northern Fur Seel
P283 P285**
P295 97 CU 02
98 CKB 02 98 C U R B 03 98 CURB 07 98 CURB 09 98 CURB 10 98CURB tl 98 CURB 12 98 CU KB 13
98 CURB 14 98 CURB 15
Liver Polar Bear
970012 970201 980341 692-PLBR43033
980I27LB
980387LB 980390LB** 980563LA
980565LB 990112LB 990592LA
990594LB
990598LB 990600LB
Liver Mink
990610LB 990652LC 990658LB
D0530 D0566* 0590 D06I8 D0630*
D0684* D1000
D1024 D1050 D1092 D ll 10* D ll 34
D1150X* DU94 D1198 D1244
D124S D1660*
* High (>J0H) surrogate deviations continued. ** positive analyte coofinnation was not achieved, used 499 --> 99 transition.
Dale Entered/Analjnt Date Verified/Analyst:
1202/99,12/28/99,12 0 9 /9 9 ,12/30/99, 01/12/00, 01/17/00,01/111X1,01/19/00, 01/2000 LAC 0
GENQ30 MSU Environmental Samples Various
Various
ETS-8-6.0 A ETS-B-7.0
Soup 020199, A m lia 062498
Messlynx 3.3
12/12/99 SAL/KK/SRP/CSH
12/17/99, 12/20/99, 12/28/99, 12/29/99, 01/03/00, 01/05/00, 01/06/00, 01/08/00 1AS/MMH
12/20/99, 12/21/99,12/22/99, 12/30/99, 01/03/00, 01/05/00.01/06/00,01/07/00,01/11/00 MMH/IAS
M
um
Verified
NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA
NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA X NA NA NA NA NA NA NA NA NA NA NA NA
X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X
C oncentra don
ef nos
u r t tr%Ree. NA NA
<LOQ (0.0347) <LOQ (0.0347)
E <L0Q (0.0696) <LOQ (0.0696) <LOO (0.0696)
NA NA E <LOQ (0.0347)
<LOQ (0.0347) <LOQ (0.0696) <LOQ (0.0696)
<LOO (0.0696) 0.0305
0.0331
0.0170 <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0696)
<LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696)
145% 538% 258% 202%
351% 317%
138%
132% 68%
E 111% 106% 77%
238% 101%
112% 92%
102% 108% 118% <LOQ (0.0347)
0.t22 0.0547
NE <L0Q (0.0347)
<LOQ (0 0347) <LOQ (0.0347) <LOQ (0.0347)
<LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0347)
<LOQ (0.0347) <LOQ (0.0347)
0.456 0.301
0.678 0.471 0.539
0.221 0.209
0.438
0.328 0.175 0.313 0.356 0.436
,
0.224 0.295
0.235 0.282
0.974 2.68
2.38 0.974 175
3.50
3.42 113
3.28
3.22 3.35 3.67 1.96 IBS 182
3.42 1.21 3.68
Meen
pros
r t ______
<LOO
<LOQ 0.0269
<LOQ 342% 230% 334% 135% 68% 108% 158% 107% 97% 113%
<LOQ - 2 outliers
0.350
2.63
rsd Std. Dev. MS/MSD RPD
Concentrado o f PFHS
a r t o r % Ree. NA
NA <LOQ (0.00683)
<LOQ (0.00683)
E <LOQ (0.0171)
NA <LOQ (0.0171)
NA <LOO (0.0171) NA NA
E
<LOQ (0.00683) <LOQ (0.00683) <LOQ (0.0171) NA <LOQ (0.0171) NA <LOO (0.0171) <LOQ (0.0171) 32.1 <LOQ (0.0171)
0.00862
<LOO (0.0171) <LOQ (0.00683) <LOQ (0.00683)
<L0Q (0.0171)
<LOQ (0.0171)
NA <LOQ (0.0171)
NA <LOQ (0.0171)
61%
115%
50% 116%
24% 112% 57%
10% 68%
81% 5% 84%
NA 5% 102%
55% E
97% 84% 70% 76%
82% 11% 89%
66% 10% 72%
74% 8% 84%
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.00683) NE
<LOQ (0.00683)
0.0587 .
<LOQ (0.00683) <LOQ (0.00683)
<LOQ (0.00683) <LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.00683) NA <LOQ (0.00683) NA 0.0858
<LOQ (0.00683)
<LQQ (0.00683) <LOQ (0.00683) <LOQ (0.00683) <LOQ (0.00683)
<LOQ (0 00683)
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.00683) <LOQ (0.00683) <LOQ (0.00683)
<LOQ (0.00683) <LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.00683)
38.6 0.135
<LOQ (0.00683) <LOQ (0.00683)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171) <LOQ (0.0171)
<LOQ (0.0171) <LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
< L 0 Q (0 .D I7 |)
<LOQ (0.0171)
<LOQ (0.0171)
<LOQ (0.0171)
33.0 0.919
<LOQ (0.0171) <LOQ (0.0171) <LOQ (0.0171)
N E Not Extracted
E " Lost during extraction
NA - Not Applicable LOQ " Limit o f Quantitation X - VeriBed PFOS ooncentratioo
M esa PFHS
RSD Std. Dev. MS/MSD RPD
<LOO
NA NA
<LOQ
<LOQ
<LOQ 55% 114% 63% 83% 55% 90% 73% 86% 69% 79%
NA NA NA NA
NA NA 21% 4% 18% 4% NA 15% 9% 9% 8% 13%
<LOQ- 2 outliers
NA NA
<LOQ
NA NA
NA
<LOQ
NA
PFfts - Perflurm w -- .irTM.*
PFOSA - Pefluorooctane enlfonan
PFHS " Perfluorohexanesuifonate
POAA - Periluorooctanoete
ETS-8-7.0 Excel Version 5/95
GEN -030-liver,x li
000037
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study; Product NumberfTest Substance): Matrix: Method/Revision: Analytical Equipment System Number [oatnuneut Software/Version: Date o f Extraction/Analyst: Date o f Amiyaia'Analyst: Date o f Data Reduction/Analyst:
Sample Data
Miscellaneous Liver
Group Dose
Sarape*
Method BUt Unfiltered water
MSU12129-H20fUk-tmfiterad-3-] MSU 12129-H20eft-nfiherod-3-2 M S U 1 2 l2 9 -H 2 0 6 R - ifh e re d -S -3 MSU12129-H20BSc-mifi]tend-3-4
MSU12129-H20B*-nnfilterod-5-S MSU12129-H2O0fo-anfiherad-5-6 MSU12129-H20B-unEltered-3-7
Method Blk Filtered water
MSUI2l29-H20Bflt-aafi]tered-S-8 MSUI2l294O0Blk-fiRerod-5-l MSU12l29-H2Ceft-fillered-3-2 M S U I2l2 9-H 20G & -filure d-3-3 M S U 1 2 l2 9 4 O 0 C * -fih e re d -5 -4
MSU12l29-H20CHk-fihcrod-5-5 MSU12l29-H2CBfc-filtered-S-6 MSU12t29-H20Bi-Ghered-5-7
Matrix Btk Fish Liver
Matrix Blk Rabbit Liver
M SU 12l294C 0k-fiterod-S - F S H l2 l2 9 -L v tB lk -5 -l FSH12129-LnBDc-3-2 FSH12 129-LvrBIk-S-3 RBL12129-LvrBtk-5-l RBLl2l-LwBlk-5-2 RBL12129-LvrBlk-5'3 RBL12129-LvrBIk-5-4
RBL12129-LiBlk-5-S* RBL12129-Lwfilk-5-6*
QC 250 ng/g
Mink Uver. D530, MS-5-1-1 Mink Liver. DS30. MSD-5-1-2* CSL-1999030-03-01- M S -5 -I-2* CSL-1999030-03-01-MSD-5-1-2* LWL-1999029-12-MS-5-1-2* LWL-1999029-12-MSD-5-1-2*
BTL-1999040-01-MS-5-1-2" BTL-1999040-01-MSD-5-1 -2*
TNL-TUS4-MS-5-1 -2 TNL-TU54-MSD-S-1-2
FSL-P295-MS-5*1
FSL-P29S-MSD-5-1 PBL-9S0390LB-MS-5-1
PBL-980390LB-MSD-5-2* QFL-KZCKDM-D1-MS-5-1-2 GFL-KZCXDM-DI-MSD-5-TTL-LCPTR99303C-MS-S-1 -2
TTL-LCPTR99503C-MSD-5-1 -2
MTL-10Vauckafw98-MS-5-I -2 MTL-IOVaneeave98-MSD-5-1-2
Liver Northern Fur Seal
P283 P33** P295
97 CU 02
98 CU KB 02 98 CU KB 03 98 CU KB 07
98 CU KB 09 * CU KB 10
9 CU KB 11 98 CU KB 12 98 CU KB 13 98 CU KB 14
98 CU KB 13
Liver Polar Bear
970012 970201
980341 692-PLBR-0033
980I27LB 980387LB
980390LB** 980S63LA 980563LB
990112LB 99Q592LA
990594LB 990398LB
99000LB
990610LB
Liver Mink
9906S2LC 990658LB
D0530
D0S66* D0390
D06I8 D0630*
D0684* D1000 D I 024
D I 050 D I 092 DI1I0* DI 134 DIIS0X* D1194
DI 198 D1244
D1248 D I 660*
High (>30%) surrogate davialions aoofinncd. * positive aoalyte confirmation was not achieved, mad 499 --> 99 transition. Date Entered/Analyst: 12/22/99.12/28/99, 12/29W9,12/3Q/99,
0 1 /t2 0 0 ,01/17/00,01/1100,0l/1900,01/20/00 LAC
Date Verified/Analyst: 0
GENO30 MSU Environmental Samples Various Various ETS-8-6.0 & ETS-8-7.0
BEST COPYAVAILABLE
Soup 020199, Amalia 06249S
Maaalyux 3.3
12/12A SAL/KK/SRP/CSH
12/17/99, 12/20/99, 12/28/99, 12/29/99, 01/03/00, 01/05/00, 01/06/00,01/08A 1AS/MMH
12/20/99, 12/21/99, 12/22/99.12/30/99,01/03/00, 01/03/00, 01/0600, 01/07/00,01/11/00 MMH/IAS
Cenceatratioa e f POAA
a*/* o r % Ree. NA NA
<LOQ (0.0719) < L O Q (0 .0 7 I9 )
E < O Q (0.0719) <LOQ (0.0719) <LOQ (0.0719)
NA NA E <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOO (0.07191 <LOQ (0.0719) <LOQ (0.0719) <LOO (0.0719) < O Q (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <UOQ (0.0719) <LOQ (0.0719)
86% 84% 131% 111% 133% 143% 127% 120% 84%
E 72% 80% 85% 129% 82% 103% 73% 64% 100% 75%
<LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
NE <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0 0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
<LOQ (0.0719) <LOQ (0.0719) < O Q (0.0719) <LOQ (0.0719) < O Q (0.0719) < O Q (0.0719) <LOQ(0.0719) < O Q (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ(0.0719) <LOQ (0.0719) <LOQ (0.0719) < O Q (0.0719) <LOQ (0.0719) <LOQ (0.0719)
<LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) < O Q (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) < O Q (0.0719)
Mean POAA
<LOO <LOO <LOQ 83% 121% 139% 124% 84% 76% 107% 92% 69% 88%
<LOQ
<LOQ
<OQ
RSD Std. Dev. MS/MSDRPD
Conesatratfoa of PFOSA
ug/g o r % Ree.
NA NA <LOQ( 0.0188) <LOQ( 0.0188) E <LOQ( 0.0188)
NA <LOQ(O.OI38) NA < .00(0.0188)
NA
NA E <LOQ( 0.0188) <LOQ( 0.0188)
<LOQ( 0.0188) NA <LOQ( 0.0188) NA <LOO( 0.0188)
<LOQ( 0.0188) NA <LOQ ( 0.0188) NA < 0 0 ( 0 .0 1 8 8 )
< O Q ( 0.0188) < O Q ( 0.0188)
< O Q ( 0.0188) <LOQ( 0.0188)
NA <LOQ( 0.0188) NA < O Q (0.0188)
43%
3% 81% 33%
17% 41%
67% 9% 73%
83% 6% 81%
59% NA E
76% 10% 71%
60% 41% 34%
71% 23% 82%
67%
13% 59%
63% 29% 63%
< O Q ( 0.0188) < O Q ( 0.0188) < O Q ( 0.0188)
NE <LOQ( 0.0188)
< O Q ( 0.0188) <LOQ( 0.0188) < O Q ( 0.0138) <LOQ( 0.0138)
<O Q (0.0188) < O Q ( 0.0188)
< O Q ( 0.0188) NA <LOQ( 0.0188) NA < O Q ( 0.0188)
<LOQ( 0.0188) <LOQ( 0.0188) < O Q ( 0.0188)
< O Q ( 0.0188) <LOQ(OOI88) <LOQ{0 0188)
< L O Q (O .O t8 8)
<LOQ(O.OI88) < O Q (O .O l8 8 )
<LOQ(0.0188) <LOQ( 0 0188)
<LOQ( 0.0188) <LOQ( 0.0188) < O Q ( 0.0188)
< O Q (O .O I8 8 ) NA <LOQ( 0.0188) NA < O Q ( 0.0188)
0.0334
0.0266 0.0209 0.0581
0.0703
0.0612 0.0294 < O Q (0.0188) < L O Q (0 .0 1 8 f) <LOQ (0.0181) 0.0367 <LOQ (0.0188) 0.0340
0.0833 0.0242
< O Q (0.0188) NA 0.0345 NA 0.0621
NE " N o t Extracted
E " Lost during extraction NA ~ Not Applicable LOQ " Linai o f Quantitation
X " Verified PFOS concentration
M esa PFOSA
U t/|
RSD Std. Dev. MS/MSDRPD
<LOO
<LOQ <LOO
<LOQ 62% 47% 70% 82% 59% 73% 57% 77% 63% 63%
NA NA
>
NA NA NA NA
NA NA 60% 26% 8% 3% NA 8% 11% 14% 12% 1%
<LOQ
NA NA
<LOQ
NA NA
0.5
0.0442 - 5 outliers
0.0201
PFOS - Perfluorooctanesulfonate
PFOSA " Pefluorooctane sulfonamide
PFHS " Periluorobexanesulfonate
POAA Periluorooctanoate
ETS-8-7 0 Excel Veraion 3/93
OEN-030-bvwjda
000038
3/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study: Product NumbcrtTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Versioo: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reduction/Analyst: Sample Data
GEN030 MSU Environmental Samples Various
M ST COPYAVAILABLE
Various
ETS-S-6.0 St ETS-8-7.0
Soup 020199, Amelia 062498
Masslynx 3.3
12/12/99 SAL/KJC/SRP/CSH
12/17/99, 12/20/99, 12/28/99,12/29/99, 01/03/00, 01/06/00, 02/01/00 IAS/MMH/MEE
12/20/99, 12/21/99, 12/22/99, 12/30/99, 01/03/00, 01/05/00, 01/07/00, 02/03/00 MMH/1AS/MEE
Miscellaneous Liver Group Dose
S am ple#
Method Blk Unfiltered water
Method Blk Filtered water
Matrix Blk Fish Liver Matrix Blk Rabbit Liver A
QC 250 ng/g
Liver Map Turtle
MSUl2129-H20Blk-unfiItered-5-l MSU 12129-H20Blk-unfiltered-5-2 MSU 12129-H20Blk-unffltered-5-3 MSU12129-H20BIk-unfiltered-5-4 MSU 12129-H20Blk-\mfiltered-5-S MSU 12129-H20Blk-unfiltered-5-6 MSU 12129-H20Blk-unfiltered-5-7 MSU12129-H20BUc-unfiltered-5-8
MSU 12129-H20Blk-filtered-5-1 MSU 12129-H20Blk-filtered-5*2 MSU12129-H20Blk-filtered-5-3 MSU12129-H20Blk-filtered-5-4 MSU12129-H20Blk-filtered-5-5 MSUl2129-H20Blk-filtered-5-6 MSU12129-H20Blk-filtered*5-7 MSU 12l29-H20Bflc-fi1tered-5-8
FSH12129-LvtBDc-5-1 FSH12129-LvrBIk-5-2 FSH 12129-LvrBlk-5-3 RBL12129-LvrBlk-5-l RBL12129-LvrBlk-5-2 RBL 12129-Lvrfilk-5-3 RBL 12129-LvrBlk-5-4 RBLI2129-LvTBlk-5-5* RBLI2129-LvTBlk-5-6*
Mink Liver, D530, MS-5-1-1 Mink Liver, D530, MSD-5-1-2* CSL-1999030-03-01 -MS-5-1-2* CSL-1999030-03-01-MSD-5-1-2* LWL-1999029-12-MS-5-1-2* LWL-1999029-12-MSD-5-1-2*
BTL-1999040-01-MS-S-1-2* BTL-1999040-01-MSD-S-1-2*
TNL-TU54-MS-5-1-2 TN L-TU54-M SD-5 -1 -2
FSL-P295-MS-S-1 FSL-P295-MSD-5-1 PBL-980390LB-MS-5-1 PBL-980390LB-MSD-5-2* GFL-KZCKDM-D1-MS-5-1-2 GF L-KZC KDM-D1-MSD-5 -1TTL-LCPTR99503C-MS-5-1-2* TTL-LCPTR99503C-MSD-5-1-2 MTL-10Vancleave98-MS-5-l-2 MTL-I0Vancleave98-MSD-5-l-2
F, #10, Vancleave 98 F, #09, Vancleave 98 F, #02, Leeksville 98 F, #06. Leeksville 99
"
Uver Terrapin
Uver Tuna
Liver Green Frog * High (>50%) surrogate deviations
F, (89,8912) LCPTR 9503C LCPTR 9504C LCPTR 9505C
TU25* TU34* TU4I* TU48* TU49 TU54* TU58 TU63* TU66 TU84 TU88 TU90
KZCKDM-D1 KZCKDM-D2
Pool o f 4 SJ0001
Date Entered/Analyst: Date Verified/Analyst:
12/22/99, 12/28/99, 12/29/99, 12/30/99, 01/12/00, 01/17/00, 01/18/00, 02/04/00 LAC
PFOS Verified
NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA X
X X X NA X NA X X NA X NA NA X NA X
Concentration of PFOS
at / t o r % Rec. NA NA
<LOQ (0.0347) <LOQ (0.0347)
E <LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696)
NA NA E <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0696) <LOQ (0.0696) <100(0.0696) 0.0305 0.0331 0.0170 <LOQ (0.0347) <LOQ (0.0347) <LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696) <LOQ (0.0696)
. 145% 538% 196% 140% 321% 287% 138% 132% 68% E 91% 86% 18% 179% 93% 105% 90% 100% 79% 89% 0.0801 0.0514 0.0739 0.0394 0.703
ai7 9 <LOQ ( 0.0347) <LOQ( 0.0347) <LOQ ( 0.0347)
<LOQ ( 0.0696) <LOQ ( 0.0696) <LOQ (0.0696) <LOQ ( 0.0696) <LOQ (0.0696) <LOQ ( 0.0696) <LOQ 0.0696) <LOQ 0.0696)
0.00698 <LOQ 0.0696) <LOQ ( 0.00696) <LOQ 0.0696) <LOQ 0.0347)
0.285 <LOQ 0.0347) <LOQ 0.0347)
Mean PFOS
______ "S li______
RSD Std. Dev. MS/MSD RPD
Concentration of PFHS
u r/r or % Rec NA
NA
<LOQ (0.00683)
<LOQ (0.00683)
E
<LOQ (0.0171)
NA <LOQ (0.0171)
<LOQ
NA <LOQ (0.0171)
NA
NA
E
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.0171)
NA <LOQ (0.0171)
<LOQ
NA <LOQ (0.0171)
<LOQ (0.0171)
32.1 <LOQ (0.0171)
0.0269
0.00862
<LOQ (0.0171)
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ (0.0171)
<LOQ (0.0171)
NA <LOQ (0.0171)
<L0Q
NA <LOQ (0.0171)
61%
342%
115%
50%
116%
168%
33%
112%
304%
11%
57% 68%
81%
135%
5%
84%
55%
68% NA
E
97%
88% 6%
84%
70% 99% 163% 76%
82% 99% 12% 89%
66% 95% 11% 72%
74% 84% 11% 84%
<LOQ ( 0.00683)
<LOQ ( 0.00683)
<LOQ ( Q.00683)
<LOQ ( 0.00683)
<LOQ ( 0.00683)
<LOQ ( 0.00683)
<LOQ ( 0.00683)
0.730
<LOQ ( 0.00683)
0.188 - 3 outliers
0.257
<LOQ( 0.00683)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
<LOQ( 0.0171)
NA <LOQ( 0.0171)
<L0Q -1 outlier
NA
<LOQ( 0.0171)
<LOQ (0.00683)
<LOQ (0.00683)
<LOQ -1 outlier
NA NA
<LOQ (0.00683) <LOQ (0.00683)
NE = Not Extracted
E = Lost during extraction
NA = Not Applicable
LOQ Limit o f Quantitation
X Verified PFOS concentration
Mean PFHS US'S
RSD Std. Dev. MS/MSD RPD
<LOQ
NA NA
<LOQ
<LOQ
<LOQ 55% 114% 63% 83% 55% 90% 73% 86% 69% 79%
NA NA NA NA
NA NA 21% 4%
l 78% 4% NA 15% 9% 9% 8% 13%
<LOQ
NA NA
i`
<LOQ
NA NA
NA
<LOQ
NA
PFO S Periluorooctanesulfonate
PFOSA = Pefluorooctane sulfonamide
PFHS * Perfluorohexanesulfonatc
POAA - Periluorooctanoate
ETS-8-7.0 Excel Version 5/95
GEN-030-liver.xls
000039
5/9/00 4:46 PM
FACT-GEN-030 MSU Environmental Samples
Study: Product Number(Test Substance): Matrix: Method/Rcvision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst: Date of Analysis/Analyst Date of Data Reduction/Analyst: Sample Data
GEN030 MSU Environmental Samples Various Various ETS-8-6.0 A ETS-8-7.0 Soup 020199, Amelia 002498 Masslynx 3J 12/12/99 SAL/KK/SRP/CSH 12/17/99, 12/20/99, 12/28/99, 12/29/99,01/03/00,01/06/00,02/01/00 IAS/MMH/MEE 12/20/99, 12/21/99, 12/22/99, 12/30/99, 01/03/00, 01/05/00, 01/07/00, 02/03/00 MMH/1AS/MEE
F
Miscellaneous Liver Group Dose
Sam ple#
Method Blk Unfiltered water
Method Blk Filtered water
Matrix Blk Fish Liver Matrix Blk Rabbit Liver
QC 250 ng/g
Liver Map Turtle
Liver Terrapin
Liver Tuna
Liver Green Frog * High (>50%) surrogate deviations
MSUI2129-H20BIk-unfiltered-5-l M SU 12129-H20Blk-unfil tered-5-2 MSU12 !29-H20Blk-unfiltered-5'3 MSU 12129-H20Blk-unfiltered*5-4 MSU 12! 29-H20Blk-unfiltered-5-5 MSU 12129-H20Blk-unfiltered-5-6 MSU 12129-H20Blk-unfiltered-5-7 MSU 12129-H20B flc-unfiltered-5-8
MSUI2129-H20Blk-filtered-5-l MSU12 129-H20BQc-filtered-5-2 MSU 12129-H20Blk-filtered-5-3 MSU12129-H20Blk-fHtered-5-4 MSU 12129-H20Blk-filtered-5-5 MSU 12129-H20BQb-61tered-5-6 MSU 12129-H20BIk-filtered-5-7 MSU 12129-H20Blk-fiItered-5-8
FSHl2129-LvrBlk-5-l FSH12129-LvrBflc-5-2 FSHl2129-LvrBDc-5-3 RBL12l29-LvrB1k-5-l RBL121 29-LvtBIk-5-2 RBL12 129-LvrBlk-5-3 RBL12 129-LvrBIk-5-4 RBLl2129-LvrBIk-5-5* RBL12129-LvrBUc-5-6*
Mink Liver, D530, MS-5-M Mink Liver, D530, MSD-5-1-2* CSL-1999030-03-01-MS-5-1-2* CSL-1999030-03-01-MSD-5-1-2* LWL-1999029-12-MS-5-1-2* LWL-1999029-12-MSD-5-1-2*
BTL-1999040-01-MS-5-1-2* BTL-1999040-01-MSD-S-1-2*
TNL-TU54-MS-5-1-2 TNL-TU54-MSD-5-1-2
FSL-P295-MS-5-1 FSL-P295 -MSD-S-1 PBL-980390LB-MS-5-1 PBL-980390LB-MSD-5-2* GFL-RZCKDM-Dt-MS-5-1-2 GFL-K2CKDM-D l -MSD-5-1TTL-LCPTR99503C-MS-5-1-2* TTL-LCPTR99503C-MSD-5-I-2 MTL-I0Vancleave98-MS-5-l-2 MTL-10Vancleave98-MSD-5-l-2
F, #10, Vancleave 98 F. #09, Vancleave 98 F, #02, Leeksville 98 F. #06, Leeksville 99
M .(-l) F, (89, 8912) LCPTR 9503C LCPTR 9504C LCPTR 9505C
TU25* TU34* TU41* TU48* TU49 TU54* TU58 TU63* TU66 TU84 TU88 TU90
KZCKDM-D1 KZCKDM-D2
Pool o f 4 SJ0001
Date Entered/Analyst: Date Verified/Analyst:
12/22/99, 12/28/99, 12/29/99, 12/30/99, 01/12/00,01/17/00,01/18/00,02/04/00 LAC
Concentration of POAA
ug/g o r % Ree. NA NA
<LCX3 (0.0719) <LOQ (0.0719)
E <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
NA NA E <LOQ (0.0719) <LOQ (0.0719) <LOQ (a0719) <LOQ(a0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) 86% 84% 131% 111% 133% 145% 127% 120% 84% E 72% 80% 85% 129% 82% 103% 73% 64% 100% 75%
<LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
<LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719) <LOQ (0.0719)
Mean POAA
ti*
<LOQ
<LOO <LOO
<LOQ 85% 121% 139% 124% 84% 76% 107% 92% 69% 88%
<LOQ
<LOQ <LOQ
RSD Concentration
Std. Dev. MS/MSD RPD
of PFOSA ug/g or % Ree.
NA
NA
<LOQ( 0.0188)
<LOQ( 0.0188)
E
<LOQ( 0.0188)
NA <LOQ( 0.0188)
NA <LOQ ( 0.0188)
NA
NA
E
<LOQ( 0.0168)
<LOQ( 0.0188)
<LOQ( 0.0188)
NA <LOQ ( 0.0188)
NA <LOQ( 0.0188)
<LOQ( 0.0188)
NA <LOQ( 0.0188) NA <LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
NA <LOQ ( 0.0188)
NA <LOQ( 0.0188)
43%
3% 81%
64%
17% 52%
77%
9% 83%
94%
6% 92%
71%
NA E
87%
10% 82%
71%
41% 65%
82%
23% 93%
78%
13% 71%
74%
29% 74%
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
NA <LOQ ( 0.0188)
NA <LOQ ( 0.0188)
<LOQ ( 0.0188)
<LOQ ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ ( 0.018B)
<LOQ(O.Q188)
<LOQ 0.0188)
<LOQ( 0.0188)
NA <LOQ( 0.0188)
NA <LOQ( 0.0188)
<LOQ( 0.0188)
<LOQ( 0.0188)
NA <LOQ(0.0188)
NA <LOQ( 0.0188)
NE - Not Extracted
E " Lost during extraction
NA 3 Not Applicable
LOQ " Limit o f Quantitation
X * Verified PFOS concentration
Mean PFOSA
ag/g
RSD Std. Dev. MS/MSD RPD
<LOQ
NA NA
<LOQ
<LOQ
<LOQ 62% 58% 80% 93% 71% 85% 68% 88% 74% 74%
NA NA NA NA
NA NA 60% 21% 7% 3% NA 7% 9% 12% 10% 0%
<LOQ
NA NA
<LOQ
NA NA
<LOQ
NA NA
PFOS " Pofluorooctanesulfonate
PFOSA " Pefluorooctane sulfonamide
PFHS 3 Perfluorohexanesulfonate
POAA ' Perfluorooctanoate
ETS-8-7.0 Excel Version 5/95
CEN-03O-liver.xls
000040
5/9/00 4:46 PM
Study: Product NumbeifTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Venion: Date o f Extraction/Analyst: Date o f Analysis/Analyst: Date of Data Reduction/Analyst: Sample Data Various Livers
Group Dose Mink Liver
Baikal Seal Liver
Ganges Dolphin Liver LOQ =Limit of Quantitation NA = Not Applicable
Sample#
C23 (45) 06(44) C27 (49) C33 (46) 05(47) C37 (48) C42 (43) C44 (54) D10 (50) F15 (55) F19 (56) F2I (57) F24 (58) P0h(60) P03 (64) P09 (61) SII (37) S15 (41) S18 (40) S19 (59) S25 (39) S30 (36) S35 (42) S39 (38) T01 (51) T04 (53) T03 (52) V12 (62) V03 (65) V08 (63) J08 (81) J09 (87) J10 (86) J12 (89) J19 (84) J20 (88) J24 (82) J27 (83) J36 (85) J37 (80) R04 (69) R13 (78) R14 (74) R16 (71) R29 (72) R42 (66) R43 (73) R45 (79) R46 (77) R47 (70) R54 (67) R55 (75) R 57(76) R64 (68) L04 (91) L05 (90)
Date Entered/Analyst: Date Verified/Analyst:
03/28/00,04/0S/00,04/07/00,05/07/00 MMH/LAC
FACT-GEN-033
BEST COPY AVAILABLE
GEN033, MSU - Liver Samples
NA
Various livers - Unextracted Curves ETS-8-6.0 and ETS-8-7.0
Filename: R-Squared Value:
Davey 070799, Amelia 062498 Masslynx 3.3 03/14/00 SAL/CSH/KKK
Slope: Y-Intercept:
03/16/00, 03/17/00, 3/19/00, 03/20/00, 03/210, 03/29/00, 04/07/00 IAS/MMH
03/20/00, 03/22/00, 03/23/00, 03/24/00, 04/04/00, 04/11/00 IAS/MMH
See Attachments See Attachments See Attachments See Attachments
Concentration of PFOS
ug/g or % Ree 0.801 0.443 0.145 0.435 0.355 0.833 0.420 0.237 1.67 0.548 0.783 1.03 0.868 2.16 4.80 0.841 0.902 1.27 1.99 2.68 0.509 0.186 0.0933 0.317 0.633 1.35 0.565 4.87 1.52 3.65 0.0127 0.0228
<LOQ (0.0347 ug/g) 0.0141 0.00931 0.0154
<LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g)
0.0146 <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g)
0.0100 <LOQ (0.0347 ug/g)
0.00808 0.00795 0.0138 0.0156 0.00848 0.00778 <LOQ (0.0347 ug/g) <LOQ (0.0347 ug/g) 0.0133 0.0158 0.00786
<LOQ (0.0347 ug/g) 0.0813
Mean PFOS
1.23
0.0123 NA
RSD Std. Dev. MS/MSD RPD
103 1.27
33.7 0.00416
NA NA
Concentration
Mean
of POAA ug/g or */ Ree < LOQ (0.0359 ug/g)
POAA ug/g
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.03S9 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
<LOQ
< LOQ (0.0359 ug/g)
< LOQ (0.07IB ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0718 ug/g)
< LOQ (0.0359 ug/g)
< LOQ (0.0359 ug/g)
<LOQ
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ
PFOS Periluorooctanesul fonate
PFOSA = Perfluorooctanesulfonamide
PFHS 9 Periluorohexane sulfonate
POAA = Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD -
NA NA
NA NA NA NA
\
i
ETS-8-7.0 Excel Version 5/95
GEN-033-Irvcr.xls
000041
5/i/O O 4:49 PM
Study: Product NumbeifTest Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Vcnion: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reduction/Analyst: Sample Data Various Livers
Groop Dose Mink Liver
t V-
Baikal Seal Liver
Ganges Dolphin Liver LOQ = Lindt of Quantitation NA = Not Applicable
Sam ple#
C23 (45) C26 (44) C27 (49) C33 (46) C3S (47) C37 (48) C42 (43) C44 (54) D10 (50) F15 (55) F 19 (56) F21 (57) F24 (58) POI (60) P03 (64) P09 (61) S U (37) SIS (41) S 18 (40) S19 (59) S25 (39) S30(36) S35 (42) S39 (38) T01 (51) T04 (53) T03 (52) V12 (62) V03 (65) V08 (63) J08 (81) J09 (87) J10 (86) J12 (89) J19 (84) J20 (88) J24 (82) J27 (83) J36 (85) J3 7 (80) R04 (69) RI3 (78) R14 (74) R16 (71) R29 (72) R42 (66) R43 (73) R4S (79) R46 (77) R47 (70) R54 (67) R5S (75) R57 (76) R64 (68) L04 (91) L05 (90)
Date Entered/Analyst: Date Verified/Analyst:
03/28/00, 04/05/00, 04/07/00,05/07/00 MMH/LAC
FACT-GEN-033
GEN033, MSU - Liver Samples
BESTCflPy AVAILABLE
NA
Various livera - Unextncted Curves
Filename:
See Below
ETS-8-6.0 and ETS-8-7.0
R-Squared Value: See Attachments
Davey 070799, Amelia 062498
Slope:
See Attachments
Masslynx 3.3
Y-Intercept
See Attachments
03/14/00 SAL/CSH/KKK
03/16/00, 03/17/00, 3/19/00, 03/20/00,03/21/00, 03/29/00,04/07/00 IAS/MMH
03/20/00, 03/22/00,03/23/00,03/24/00,04/04/00,04/11/00 IAS/MMH
Concentration of PFOSA
ug/g or % Ree
0.0383 < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
0.0828 < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
0.0579 < LOQ (0.0376 ug/g)
0.5S1 0.590 < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) 0.0414 0.132 < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) 0.345 0.0594 0.0586
< LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) <LO Q (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
< LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
Mean PFOSA
t / i
<LOQ -1 0 Outliers
<LOQ <LOQ
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA NA NA
Concentration
Mean
of PFHS
PFHS
ug/g o r % Ree
uj/g
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
0.00833
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
0.0315
0.0852
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
0.0102
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
0.0104
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
<LOQ - 5 Outliers
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
<LOQ
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g)
<LOQ
PPOS * Perfluorooctanesulfonate
PPOSA " Perfluorooctancsulfonamide
PFHS * Perfluorohexane sulfonate
POAA Perfluorooctanoate
RSD Std. Dev. MS/MSD RPD
`'
NA NA
,
NA NA NA NA
ETS-8-7.0 Excel Version 5/95
G E N -033-liver.xls
000042
5/9/00 4:49 PM
FACT-GEN-033
Study; Product Number(Test Substance); Matrix; Metho<yRevision: Analytical Equipment System Number Instrument Soflware/Version: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Reduction/Analyst: Sample Data
Various Livers Group Dose
Sample
Comorant Liver Female, Adult
Comorant Liver Female, Juvenile Comorant Liver Male, Juvenile Bottlenose Dolphin Liver
Striped Dolphin Liver Weddell Seal Liver Swordfish Liver
Tunafiah Liver
,
Blacktailed Gull Liver
NO PFOS conflrraation performed. LOQ = Limit ofQuantitation NA Not Applicable
<F(1W\A 29F (5XF.A** 30F(6)JA 32F (7XFA 34F(9XF.AM 42F (lIXFwA
7F(2*FJ 9F(3XFJ 3F(10LFJ
22F(4)M J 33F (8XMJ 4F02XMJ
TI5/91 (13) Tt8/9I (17) T161 (14) TtAI 1(16) Ttxnns) 9CQD2 (18) SCVl (19) SCP03 (20) 9CP04(2I)
WS1 (22) S23 (23) S24(24) S25 (25) S32(27) S48 (26)
Tl (33) T1LFI56(3S)
T2 (29) TIS(28) T17(3I) T20(32) T23 (30) T25 (34) BHG01 (100)
BHG02 (101) BHC03 (102) BHG04 (103) BHG05 (104) BTG9305 (92) BTG9306 (99) BTG9310 (96) BTG931I (98)
BTG9312 (97) BTG940I (93) BTGbongstol (94) BTGoandol (95) HRG04 (106) HRG09(105)
Date Entered/Analyst: Date Verified/Analyst:
03/28/00, 04/05/00.0407/00,05A>7A MMH/LAC
GEN033, MSU - Liver Samples NA Various livers unextracted curves
Ksl c iln mFilename:
ETS-8-6.0 and ETS-8-7.0
R-Squared Value:
Davty 070799, Amelia 062498
Slope:
See Attachments
Masslynx3.3
Y-Intercept:
See Attachments
03/14/00 SAL/CSH/KKK
03/16/00,03/17/00,3/19/00,03/20/00, 03/21/00,03/29/00.04/07/00 tAS/MMH
03/20/00,03/22/00,03/23/00,03/24/00,04/04/00,04/11/00IAS/MMH
su
Concentration of PFOS
uf/tor% R ec
0.0432 0.0565 0.0485 0.0977 0.0335 0.150 0.0913 0.468 0.0499
0.0433 0.0316 0.0337
0.181 0.296 0.169 0.425 <LOQ (0.00696 ug/g)
0.161 0.0891 0.0944 0.0647
< LOO (0.0347 ug/g)
< LOQ (0.00696 ug/g) < LOQ (0.00696 ug/g)
0.00774 < LOQ (0.00696 ug/g)
0.0133
< LOQ (0.0347 ug/g) 0.0433 0.0874 0.0568 0.0491 0.0207 0.0560 0.0250
0.292 0.260 0.148 0.503 0.271 0.0881 0.107 0.215 0.143 0.0705 0.126 0.0737 0.0707 0.116 0.0939
Mean PFOS
ug/g
BSD Std. Dev. MS/MSDRPD
0.0715 0.203 0.0362
61.9 0.0442
113 0.230
17.2 0.00623
0.268 -One Outlier
0.102 <LOQ
44.5 0.119
40.2 0.0410
NA
<LOQ - Two Outliers
NA NA
0.0483 - One Outlier
46.2 0.0223
0.172
69.2 0.119
PFOS *PerQuorooctainsulfonate
POAA Perfluoroocatdoate
PFOSA = Perfluorooctanesulfonamide
PFHS - Perfiuorohexane sulfonate
Concentration of POAA
ug/gor % Rec
0.0414 0.102 0.0506 0.0897 0.0297 0.143 0.0841 0.444 0.0467
0.0454 0.0303 0.0293
<LOQ (0.0359 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0718 ug/g)
<LGQ (0.0718 ug/g) <LOQ (0.0359 ug/g) <LOO (0.0359 ug/g) <LOO (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g)
<LOQ (0.0359 ug/g) <LOQ (0.0718 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.07IB ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0718 ug/g)
<LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOO (0.0359 ub/b) <LOQ (0.0359 ua/s) <LOQ (0.0359 ug/g) cLOQ (0.0359 ug/g) <LOQ (0 0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0 0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g) <LOQ (0.0359 ug/g)
Mean POAA ug/g
0.0761 0.192 0.0350 <LOQ <LOQ <LOQ <LOQ
<LOQ
<LOQ
BSD Std. Dev. MS/MSDRPD
56.9 0.0432 , 115 0.219
25.7 0.00899
NA NA NA NA NA
NA NA
NA NA
NA NA
ETS-8-7.0 Excel Versioo 5/95
GEN-0334tver.xls
000043
5/9/00 4:49 PM
FACT-GEN-033
Study: Product Numbei(Test Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Softwaie/Vereton: Date o f ExtractkHv'Analyst Date o f Analysis/Analyst: Dale o f Data {Induction/Analyst: Sample Data
Various Livers Group Dose
Sample#
Comorant Liver Female, Adult
Comonnt Liver Female, Juvenile Cemerant Liver Male, Juvenile Bottlesose Delphin Liver
Striped Delphin Liver Weddell Seal Liver Swordfish Liver
Tunafisb Liver
Blacktailed Gull Liver
NO PFOS confirmation performed. LOQ - Limit of Quantitation NA * Not Applicable
6F(lWsA 29F (5)JLA" 30F (6)J,A 32F (7)J,A 34F (9)^,A " 42F(11)JFA
7F(2XFJ 9F (3 )J J 36F (1 0 W 22F (4)Jv4J 33F (8)JVU 44F(12XMJ
T15/91 (13) T18/91 (17) Tt61 (14) TtAll (16) TCT2 (15)
SCP02 (18) SCVl (19) SCP03 (20) SCP04 (21)
^
WS1 (22)
S 2 3 (23) S24 (24) S25 (25) S32 (27) S48 (26)
T1 (33) TILF156(35)
T2 (29) TI5 (28) T17 (31) T20(32) T23 (30) T25 (34) BHGOl (100) BHG02 (101) BHG03 (102) BHG04 (103) BHG05 (104) BTG9305 (92) BTG9306 (99) BTG9310 (96)
BTG9311 (98) BTG9312 (97) BTG9401 (93) BTGhongdol (94) BTGnandol (95) HRG04 (106) HRG09(105)
,
Date Entered/Analyst: Date Verified/Analyst
03/28/00,04/05/00.04/07/00,05/07/00 MMH/LAC
GEN033, MSU - Liver Samples
NA
Various liven - unextncted curves
Filename:
ETS-8-6.0 and ETS-8-7.0
R-Squared Value:
Davey 070799, Amelia 062498
Slope:
Masslynx 3.3
Y-Inlercept:
03/14/00 SAL/CSH/KKK
03/16/00,03/17/00,3/19/00,03/20/00,03/21/00,03/29/00,04/07/00 IA&MMH
03/20/00,03/22/00,03/23/00,03/24/00, 04/04/00,04/11/00IAS/MMH
See Below See Attachments See Attachments See Attachments
Concentration f PFOSA
ug/g er % Rec
<LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g)
0.0883 <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g)
0.224 0.358 0.129 0.129 0.115
<LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g)
<LOQ (0.0376 ug/g)
<LOQ (0.0376 ug/g) <LOQ <0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOO (0.0376 ug/g)
<LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g) <LOQ (0.0376 ug/g)
Mean PFOSA
ug/g
<LOQ <LOQ - One Outlier
<LOQ 0.191 <LOQ - One Outlier <LOQ <LO0
<LO0
<LOQ
RSD Std. Dev. MS/MSD RPD
Concentration of PFHS
ug/g or % Rec
< LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g)
54.1 0.103
< LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) NA < LOQ (0.006B3 ug/g) NA 0.0270
NA < LOQ (0.00683 ug/g)
0.00954
< LOQ (0.00683 ug/g)
< LOQ (0 00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g)
< LOQ (0 00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0 00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0 00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0 00683 ug/g)
< LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g) < LOQ (0 00683 ug/g)
< LOQ (0.00683 ug/g)
< LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0 00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g) NA < LOQ (0.00683 ug/g)
PFOS **Perfluorooctanesulfonale POAA - Perfluoroocantoate PFOSA **Perfluorooctanesulfonamide PFHS * Perfluorohexane sulfonate
Mean PFHS ug/g
RSD Std. Dev. MS/MSD RPD
<LOQ <LOQ <LOQ
NA NA
NA NA
NA NA
<LOQ
<LOQ - One Outlier <LOQ
NA NA
NA NA NA
<LOQ - One Outlier
NA NA
<LOQ
NA NA
<LOQ
NA NA
ETS-8-7.0 Excel Version 5/95
G E N -0 3 3 4 iv e r.x ls
000044
5/9/00 4 49 PM
Study: Product Number(Tcst Substance): Matrix: Method/Revision: Analytical Equipment System Number Instrument Software/Versicm: Date of Extraction/Analyst: Date of Analysis/Analyst: Date of Data Redaction/Analyst
Sample Data
Various Livers Group Dose
Sample#
Method Blk
Matrix Blk
QC 250 ppb
1
LOQ - Limit of Quantitation NA " Not Applicable
RBL03140-H20 Blk-5-1 RBL03140-H20 Blk-5-2
RBL0314O-H2O Blk-5-3 RBL03140-H20 Blk-5-4 RBL03140-H2O Blk-5-5 RBL03140-H20 Blk-5-6 RBL03140-H2O Blk-5-7 RBL03140-H20 BUt-5-8 RBL03140-H2O Blk-5-9 RBL03140-H20 BIk-5-10 RBL03140-H2O BIk-5-ll RBL03140-H20 Btk-5-12 RBL03I40-H2O Blk-5-13 RBL03140-Liver Blk-S-1
RBL03140-Livcr Blk-5-2 RBL03140-Liver Blk-5-3 RBL03140-Livcr Blk-5-4 RBL03140-Livcr'BIk-5-S RBL03140-Liver Btk-5-6 RBL03140-Liver Blk-5-7 RBL03140-Livcr BIk-5-8 RJBL03140-Liver Blk-5-9
RBL03140-Liver Blk-5-10 RBL03140-Livcr Blk-5-11 RBL03140-Liver Blk-5-12 RBL03140-Livcr Blk-5-13
RBL03140-MS-1 pph-J-1* RBL03140-MSD-1 ppb-5-2* RBL03140-MS-250 ppb-5-1 RBL03140-MSD-250 pob-5-2
44F(12)-MS
44F(12)-MSD Tt8/91(l7)-MS Tt8/9I (17>MSD SCVl (19)-MS SCVl (19VMSD WS1 (221-MS
WS1 (22VMSD S24 (24V-MSD T15 (28)-MS T15 (28VMSD TILF156 (35>-MS T1LF156 (35VMSD S19 (59)-MS S19C59VMSD
P9 (61)-MS P9 (6l)-MSD R47 (70)-MS R47 (70VMSD L4 (91)-MS L4 (91)-MSD BTG9305 (92)-MS BTGhongdol (94)-MS BTGhongdol (94VMSD
Date Entered/Analyst: 03/28/00, 04/05/00, 04/07/00, 05/07/00 MMH/LAC Date Verifled/Analyst
FACT-GEN-033
BEST COPY AVAILABLE
GEN033, MSU - Liver Samples
NA
Various liven - oncxtncted curves ETS-8-6.0 and ETS-B-7.0
Filename: R-Squared Value:
Davey 070799, Amelia 062498
Slope:
Masslynx 3.3
Y-Intercept:
03/14/00 SAL/CSH/KKK
03/16/00, 03/17/0L 3/19/00,03/20/00, 03/21/00, 03/29/00, 04/07/00 1AS/MMH
03/20/00, 03/22/00.03/23/00, 03/24/00. 04/04/00, 04/11/00IAS/MMH
See Attachments See Attachments See Attachments See Attachments
Conccutntt--
of PFOS ug/g or % Rec
< LOQ (0.00696 ug/g) < LOQ (0.00696 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0347 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0347 ug/g) < LOQ (0.0347 ug/g) < LOQ (00347 ug/g) < LOQ (0.0347 ug/g)
< LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g)
O.OOS83 < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0694 ug/g) < LOQ (0.0347 ug/g) < LOQ (0.0347 ug/g) < LOQ (0.0347 ug/g)
104% 114% 120% 133% ' 98% 107% 131% 140% 66% 93% 81% 73% 92% 84% 103% 93% 72% 326% 234% 231% 142% 68% 69% 69% 187% 143% 122% 120%
Mean
RSD Coucentratioa
PFOS rt
Std. Dev. MS/MSD RPD
of POAA ag/g or % Rec
<LOO (0.0359 ue/s)
<LOQ (0.0359 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.00719 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.00719 ug/g)
<LOQ (0.0359 ug/g)
<LOQ
NA <LOQ (0.0359 ug/g) NA <LOQ (0.0359 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.07IBug/g)
<LOQ (0.0359 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0718 ng/g)
<LOQ <0.0718 ug/g)
<LOQ (0.0718 ug/g)
<LOQ (0.0359 ug/g)
NA <LOQ (0.0359 ug/g)
<LOQ - One Outlier
NA
<LOQ (0.0359 ug/g)
112%
109%
9%
106%
127%
10%
128% 128%
103%
9%
106% 109%
59%
135%
6%
64%
56%
79% 35%
76%
77% 10%
63% 62%
NA NA
101%
93%
94% 21%
105%
92%
82% 25%
80%
280%
33%
126% 154%
102%
187%
48%
112%
69% 2%
70% 68%
73%
128% NA
92% NA
165% 155%
121%
2%
137% 156%
PFOS " Pcrfiuorooctanesulfocale
POAA - Pcrflnoroocantoatc
PFOSA * Perfluorooctanesulfonamide
PFHS " Perfluorobexane sulfonate
Mean POAA g/g
RSD Std. Dev. MS/MSD RPD
<LOQ
NA NA
<LOQ 109% 128% 107% 61% 66% 62% ' NA 99% 86% 140% 107% 69% 119% NA 146%
NA NA
6% 0% 2% 9%
30% 2% NA 12% 15%
20% 9% 3% 78% NA 13%
ETS-8-7.0 Excel Version 5/95
GEN-033-livcr.xls
000045
5/9/00 4:49 PM
FACT-GEN-033
Study: Product Numbcr(Test Substance): Matrix: Metbod/Revision: Analytical Equipment System Number Instrument Software/Version: Date of Extraction/Analyst: Date of AnalysisAnalyst: Date of Data Reduction/Analyst:
Sample Data
Various Livers Group Dose
Sample #
Method Blit
Matrix Blk
QC 250 ppb
iv
LOQ = Limit of Quantitation NA " Not Applicable
RBL03140-H20 BUc-5-l RBL0314O-H2O Blk-5-2 RBL0314O-H2O Blk-5-3 RBL03140-H20 Blk-5-4 RBL0314O-H2O Blk-5-5 RBL03140-H20 Blk-5-6 RBL03140-H2O Blk-5-7 RBL0314O-H2O Blk-S-8 RBL0314O-H2O Blk-5-9 RBL03I40-H20 Blk-5-10 RBL0314O-H2O Blk-S-11
RBL03I4O-H2O Blk-S-12 RBL03140-H20 Blk-5-13
RBL03140-Liver Blk-5-1 RBL03340-Liver B0c-S-2 RBL03140-Liver Blk-5-3 RBL03140-Liver Blk-5-4 RBL03140-LiverBtk-5-5 RBL03140-Lives Blk-5-6
RBL03140-Liver Btk-$-7 RBL03140-Lives BIk-5-8 RBL03140-Lives BUc-5-9 RBL03140-Lives Blk-5-10 RBL03140-Liver BIk-5-11 RBL03140-Lives BIk-5-12 RBL03140-Lives Blk-5-13
RBL03140-MS-1 ppb-5-1* RBL03140-MSD-I ppb-5-2* RBL03140-MS-250 ppb-5-l RBL03140-MSD-250 ppb-5-2
44F (12)*MS 44FU2VMSD Tt8/9l (17>MS Tt8/91 (17)-MSD SCV1 (19)-MS SCV1 (19VMSD WS1 (22>MS WS1 (22VMSD S24 (24VMSD T15 (28)-MS TI5 (28)-MSD T1LF156(35)-MS TILF156 (35VMSD S19 (59)-MS S19(59>MSD
P9(61)-MS P9 (61J-MSD R47 (70)-MS R47 (70)-MSD L4(91)-MS L4 <9I)-MSD BTG9305 (92VMS BTGbongdol (94)-MS BTGhnngdoI (94)-MSD
Dale Entered/Analyst: 03/28/00, 04/05/00, 04/07/00, 05/07/00 MMH/LAC Date Verified/Analyst:
GEN033, MSU Liver Samples
NA
Various liven unextracted carves
Piletume:
See Below
ETS-8-6.0 and ETS-8-7.0
R-Squared Value:See Attachments
Davey 070799, Amelia 062498
Slope:
See Attachments
Masstynx3.3
Y-lntercept:
See Attachments
03/14/00 SAL/CSH/KKK
03/16/00, 03/17/00, 3/19/00, 03/20/00, 03/21/00, 03/29/00,04/07/00 IAS/MMH
03/20/00, 03/22/00, 03/23/00,03/24/00, 04/04/00, 04/11/00 IAS/MMH
Concentration ofPFOSA
i | / | or % Rec
< LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (00376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
< LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
< LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g) < LOQ (0.0376 ug/g)
66% 67% 114% 84% 94% 89% 146% 191% 53% 87% 69% 69% 98% 66% 95% 86% 66% 140% 2% 73% 68% 68% 78% 50% 119% 70% 62% 63%
Mean PFOSA
l/t
<LOQ
<LOQ 67% 99% 92% 168% 70% 69% NA 81% 76% 71% 70% 73% 85% NA 63%
RSD Std. Dev. MS/MSD RPD
Concentration of PFHS
og/g or % Rec
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ <0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g) NA <LOQ (0.00683 ug/g)
NA <LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ng/g)
<LOQ (0.00683 ng/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ng/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ng/g)
<LOQ (0.00683 ug/g)
<LOQ (0.00683 ng/g)
<LOQ (0.00683 ug/g)
NA <LOQ (0.00683 ug/g) NA <LOQ (0.00683 ug/g)
87%
2% 57%
61%
30% 48%
84%
5% 87%
65%
26% 65%
54%
48% 64%
59%
0% 60%
NA 76% 67%
36% 102%
88%
26% 73% 97%
195%
-1%
52% 7% 48%
55%
14% 58%
58%
82% 145%
NA 31%
18% 2% 22%
PFOS = Perfluorooctanesnlfonatc
POAA " Perfluoroocantoate
PPOSA " Perfluoroocunesulfboamide
PFHS Perfluorobexane sulfonate
Mean PFHS f/<
<LOQ
<LOQ 72% 54% 86% 65% 59% 60% NA 84% 80% 48% 50% 57% 102% NA 20%
RSD Std. Dev. MS/MSD RPD
NA NA
NA NA 42% 25% 3% 1% 17% 1% NA 40% 19% 203% 6% 5% 86% NA 20%
ETS-8-7.0 Excel Version 5/95
GEN-033-liver.xls
000046
5/9/00 4:49 PM
3M Environm ental Laboratory
M ethod
Extraction of Potassium Perfluorooctanesulfonate or O ther Fluorochemical compounds from Serum for A nalysis U sing HPLC-
Electrospray/M ass Spectrometry
M ethod N um ber: ETS-8-4.1
A doption D ate: 03/01/99
Author: Lisa Clemen, Glenn Langenburg V,
Approved By:
Revision Date:
Laboratory Manager
Date
Group Leader
Date
Technical Reviewer
Date
1.0 Scope and A p p lic a tio n ____________________________________________________
1.1 Scope: This method is for the extraction o f potassium perfluorooctanesulfonate (PFOS) or other fluorochemical compounds from serum.
1.2 A pplicable compounds: Fluorochemical surfactants or other fluorinated compounds.
1.3 M atrices: Rabbit, rat, bovine, monkey, and human serum or other fluids as designated in the validation report.
Word 6/95
ETS-8-4.1 Extraction o f PFOS from Serum
000047
Page 1 of 14
2.0 Summary of Method
2.1 This method describes the procedure for extracting potassium perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants from serum, or other fluids, using an ion pairing reagent and methyl-terf-butyl ether (MtBE). In this method, seven fluorochemicals were extracted: PFOS, PFOSA, PFOSAA, EtFOSE-OH, PFOSEA, M556, and surrogate standard (see 3.0 Definitions). An ion pairing reagent is added to the sample and the analyte ion pair is partitioned into MtBE. The MtBE extract is removed and put onto a nitrogen evaporator until dry. Each extract is reconstituted in 1.0 mL o f methanol, then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials.
2.2 These sample extracts are analyzed following method ETS-8-5.1 or other appropriate method.
3.0 De fin itio n s _______________________________________________________________
3.1 PFOS: perfluorooctanesulfonate (anion o f potassium salt) CgFnSOs'
3.2 PFOSA: perfluorooctane sulfonylamide C8F17SO2NH2
3.3 PFOSAA: perfluorooctane sulfonylamido (ethyl)acetate C8Fi7S02N(CH2CH3)CH2C02'
3.4 EtFOSE-OH: 2(N-ethylperfluorooctane sulfonamido)-ethyl alcohol C8F17S02N(CH2CH3)CH2CH20 H
3.5 PFOSEA: perfluorooctane sulfonyl ethylamide C8F i7S02N(CH2CH3)H
3.6 M556: C gF nSO zN ^C H jC O O H )
3.7 Surrogate standard: 1H-1H-2H-2H perfluorooctane sulfonic acid
4.0 W arnings and Cautio ns______________ 4.1 H ealth and safety w arnings
4.1.1 Use universal precautions, especially laboratory coats, goggles, and gloves when handling animal tissue, which may contain pathogens.
5.0 INTERFERENCES_____________________________________________________________ 5.1 There are no interferences known at this time.
6.0 Eq uipm ent________________________________________________________________ 6.1 The following equipment is used while performing this method. Equivalent equipment is
acceptable. 6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.1.2 Centrifuge, Mistral 1000 or IEC 6.1.3 Shaker, Eberbach or VWR 6.1.4 Nitrogen evaporator, Organomation
ETS-8-4.1 Extraction of PFOS from Serum
Page 2 o f 14
000048
6.1.5 Balance ( 0.100 g)
7.0 Supplies and M a ter ia ls_______________________________________________ _ 7.1 Gloves 7.2 Eppendorf or disposable pipettes 7.3 Nalgene bottles, capable o f holding 250 mL and 1 L 7.4 Volumetric flasks, glass, type A 7.5 I-CHEM vials, glass, 40 mL glass 7.6 Centrifuge tubes, polypropylene, 15 mL 7.7 Labels 7.8 Oxford Dispenser --3.0 to 10.0 mL 7.9 Syringes, capable o f measuring 5 pL to 50 pL 7.10 Graduated pipettes 7.11 Syringes, disposable plastic, 3 cc 7.12 Syringe filters, nylon, 0.2 pm, 25 mm 7.13 Timer 7.14 Crimp cap autovials and caps 7.15 Crimpers Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with
Milli-QTM water. Rinse syringes a minimum o f 9 times with methanol, 3 rinses from 3 separate vials.
8.0 Reagents and Standards___________________________________________________ 8.1 Type I reagent grade water, Milli-QTM or equivalent; all water used in this method should
be Milli-QTM water and may be provided by a Milli-Q TOC PlusTM system 8.2 Sodium hydroxide (NaOH), J.T Baker or equivalent 8.3 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent 8.4 Sodium carbonate (Na2C03), J.T. Baker or equivalent 8.5 Sodium bicarbonate (NaHCOs), J.T. Baker or equivalent 8.6 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 8.7 Methanol, Omnisolv, glass distilled or HPLC grade 8.8 Serum or blood, frozen from supplier 8.9 Fluorochemical standards
8.9.1 PFOS (3M Specialty Chemical Division), molecular weight = 538 8.9.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499 8.9.3 PFOSAA (3M Specialty Chemical Division), molecular weight = 585
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8.9.4 EtFOSE-OH (3M Specialty Chemical Division), molecular weight = 570
8.9.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527
8.9.6 M556 (3M Specialty Chemical Division), molecular weight = 557
8.9.7 Surrogate standard: 4-H, perfluorooctane sulfonic acid (1-H,1-H, 2-H, 2-H C8F13SO3H) molecular weight = 428
8.9.8 Other fluorochemicals, as appropriate
8.10 R eagent preparation
N O TE: When preparing larger volumes than listed in reagent, standard, or surrogate preparation, adjust accordingly.
8.10.1 10 N sodium hydroxide (NaOH): Weigh approximately 200 g NaOH. Pour into a 1000 mL beaker containing 500 mL Milli-Q water, mix until all solids are dissolved. Store in a 1 L Nalgene bottle.
8.10.2 1 N sodium hydroxide (NaOH): Dilute 10 N NaOH 1:10. M easure 10 mL o f 10 N NaOH solution into a 100 mL volumetric flask and dilute to volume using MilliQTM water. Store in a 125 mL Nalgene bottle.
8.10.3 0.5 M tetrabutylammonium hydrogen sulfate (TBA): Weigh approximately 169 g o f TBA into a 1 L volumetric containing 500 mL Milli-Q water. Adjust to pH 10 using approximately 44 to 54 mL o f 10 N NaOH (While adding the last mL o f NaOH, add slowly because the pH changes abruptly). Dilute to volume w ith MilliQTM water. Store in a 1 L Nalgene bottle.
8.10.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1 N NaOH solution.
8.10.4 0.25 M sodium carbonate/sodium bicarbonate buffer (Na2C03/N aH C 0 3): W eigh approximately 26.5 g o f sodium carbonate (Na2C03) and 21.0 g o f sodium bicarbonate (N aH C 03) into a 1 L volumetric flask and bring to volume with MilliQTM water. Store in a 1 L Nalgene bottle.
8.11 S tandards preparation
8.11.1 Prepare PFOS standards for the standard curve.
8.11.2 Prepare other fluorochemical standards, as appropriate. Multicomponent fluorochemical standards are acceptable (for example, one working standard solution containing 1.00 ppm PFOS, 1.02 ppm PFOSA, 0.987 ppm PFOSAA, and 1.10 ppm EtFOSE-OH.)
8.11.3 Weigh approximately 100 mg o f PFOS into a 100 mL volumetric flask and record the actual weight.
8.11.4 Bring to volume with methanol for a stock standard of approximately 1000 ppm (pg/mL).
8.11.5 Dilute the stock solution with methanol for a working standard 1 solution of approximately 50 ppm.
8.11.6 Dilute working standard 1 with methanol for a working standard 2 solution of approx. 5.0 ppm.
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OGOQSO
8.11.7 Dilute working standard 1 with methanol for a working standard 3 solution of approx. 0.50 ppm.
8.12 Surrogate stock standard preparation
8.12.1 Weigh approximately 50-60 mg o f surrogate standard 1-H,1-H, 2-H, 2-H, C8F13SO3H into a 50 mL volumetric flask and record the actual weight.
8.12.2 Bring to volume with methanol for a surrogate stock of approximately 1000-1200 ppm.
8.12.3 Prepare a surrogate working standard. Transfer approximately 1 mL of surrogate stock to a 10 mL volumetric flask and bring to volume with methanol for a working standard o f 100 ppm. Record the actual volume transferred.
9.0 Sample Handling________________________________________ ___________________
9.1 All samples are received frozen and must be kept frozen until the extraction is performed.
9.2 Allow samples to thaw to room temperature prior to extraction.
10.0 Quality Control_________________________________________________________
10.1 Solvent B lanks, M ethod blanks an d m a trix blanks
10.1.1 An aliquot o f 1.0 mL methanol is used as a solvent blank.
10.1.2 Extract two 1.0 mL aliquots o f Milli-QTM water following this procedure and use as method blanks.
10.1.3 Extract two 1.0 mL aliquots o f the serum following this procedure and use as matrix blanks. See 11.1.4,
10.2 M atrix spikes
10.2.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine the accuracy o f the extraction.
10.2.2 Prepare each spike using a sample chosen by the analyst, usually the control matrix received with each sample set.
10.2.3 Expected concentrations will fall in the mid-range o f the initial calibration curve. Additional spikes may be included and may fall in the low-range o f the initial calibration curve.
10.2.4 Prepare one matrix spike and matrix spike duplicate per 40 samples, with a minimum o f 2 matrix spikes per batch.
10.3 C ontinuing calibration checks
10.3.1 Prepare continuing calibration check samples to ensure the accuracy o f the initial calibration curve.
10.3.2 Prepare, at a minimum, one continuing check per group o f 10 samples. For example, if a sample set = 34, four checks are prepared and extracted.
10.3.3 Prepare each continuing calibration check from the same matrix used to prepare the initial curve.
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10.3.4 The expected concentrations will fall within the mid-range o f the initial calibration curve. Additional spikes may be included that fall in the low-range o f the initial calibration curve. This is necessary if the analyst must quantitate using only the low end o f the calibration curve (for example, 5 ppb - 100 ppb, rather than 5 ppb - 1000 ppb).
11.0 Ca l ib r a t io n and St a n d a r d iza t io n _____________________________
11.1 Prepare matrix calibration standards
11.1.1 Transfer 1 mL o f serum to a 15 mL centrifuge tube.
11.1.2 If most sample volumes are less than 1.0 mL, extract standards with matrix volumes equal to the sample volumes. Do not extract less than 0.50 mL o f matrix. Record each sample volume on the extraction sheet.
11.1.3 While preparing a total o f twenty aliquots in 15 mL centrifuge tubes, mix or shake between aliquots.
11.1.4 Two 1 mL aliquots, or other appropriate volume, serve as matrix blanks. Typically use the standard concentrations and spiking amounts listed in Table 1, at the end o f this section, to spike, in duplicate, two standard curves, for a total o f eighteen standards, two matrix blanks, and two method blanks.
11.1.5 Refer to validation report ETS-8-4.0 & ETS-8-5.0-V-1, which lists the working ranges and the Linear Calibration Range (LCR) for calibration curves.
11.1.6 Use Attachment D' as an aid in calculating the concentrations o f the working standards. See Section 13.0 to calculate actual concentrations o f PFOS in calibration standards.
11.2 To each standard, blank, or continuing check, add appropriate amount o f surrogate working standard for the concentration to fall within the calibration curve range 5 ppb 1000 ppb.
11.3 Extract spiked matrix standards following 12.6-12.16 o f this method. Use these standards to establish each initial curve on the mass spectrometer.
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Table 1
Approximate spiking amounts for standards and spikes
Using 1.0 mL of m atrix
Working standard
pL Approx, final cone, o f
(approx, cone.)
analyte in matrix
- - Blank
0.500 ppm
10 0.005 ppm
0.500 ppm
20 0.010 ppm
5.00 ppm
5 0.025 ppm
5.00 ppm
10 0.050 ppm
5.00 ppm
20 0.100 ppm
50.0 ppm
5 0.250 ppm
50.0 ppm
10 0.500 ppm
50.0 ppm
15 0.750 ppm
50.0 ppm
20 1.00 ppm
12.0 Procedure________________
12.1 Obtain frozen samples and allow to thaw at room temperature or in a lukewarm waterbath.
12.2 Vortex mix for 15 seconds, then transfer 1.0 mL or other appropriate volume to a 15 mL polypropylene centrifuge tube.
12.3 Return unused samples to freezer after extraction amounts have been removed.
12.4 Record the initial volume on the extraction worksheet.
12.5 Label the tube with the study number, sample ID, date and analyst initials. See attached worksheet for documenting the remaining steps.
12.6 Spike all samples, including blanks and standards, ready for extraction with surrogate standard as described in 11.2.
12.7 Spike each matrix with the appropriate amount o f standard as described in 11.1, or Table 1 in that section, for the calibration curve standards. Also prepare matrix spikes and continuing calibration standards.
12.8 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds.
12.9 Check to ensure the 0.5 M TBA reagent is at pH 10. If not, adjust accordingly.
12.10 To each sample, add 1 mL 0.5 M TBA and 2 mL o f 0.25M sodium carbonate/sodium bicarbonate buffer.
12.11 Using an Oxford Dispenser, add 5 mL methyl-fe/7-butyl ether.
12.12 Cap each sample and put on the shaker at a setting o f 300 rpm, for 20 minutes.
12.13 Centrifuge for 20 to 25 minutes at a setting o f 3500 rpm, or until layers are well separated.
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12.14 Label a fresh 15 mL centrifuge tube with the same information as in 12.5. 12.15 Remove 4.0 mL of the organic layer to this clean 15 mL centrifuge tube. 12.16 Put each sample on the analytical nitrogen evaporator until dry, approximately 1 to 2
hours. 12.17 Add 1.0 mL o f methanol to each centrifuge tube using a graduated pipette. 12.18 Vortex mix for 30 seconds. 12.19 Attach a 0.2 pm nylon mesh filter to a 3 cc syringe and transfer the sample to this syringe.
Filter into a 1.5 mL glass autovial or low-volume autovial when necessary. 12.20 Label the autovial with the study number, animal number and gender, sample timepoint,
matrix, final solvent, extraction date, and analyst(s) performing the extraction. 12.21 Cap and store extracts at room temperature or at approximately 4 C until analysis. 12.22 Complete the extraction worksheet, attached to this document, and tape in the study
notebook or include in study binder, as appropriate.
13.0 Da t a A n alysis and Ca lc u la tio n s__________________________________________ 13.1 C alculations
13.1.1 Calculate actual concentrations o f PFOS, or other applicable fluorochemical, in calibration standards using the following equation: mL o f standard x concentration o f standard Cue /mLf___________________= mL o f standard + mL o f surrogate standard + initial matrix volume (mL)
Final Concentration (pg/mL) o f PFOS in matrix
14.0 M ethod Performance____________ ____________________ ___________________ 14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to M DL report
for specific MDL and limit o f quantitation (LOQ) values (see A ttachm ents B an d C).
14.2 The following quality control samples are extracted with each batch o f samples to evaluate the quality o f the extraction and analysis. 14.2.1 Method blanks and matrix blanks. 14.2.2 Matrix spike and matrix spike duplicate samples to determine accuracy and precision of the extraction. 14.2.3 Continuing calibration check samples to determine the continued accuracy o f the initial calibration curve.
14.3 Refer to section 14 o f ETS-8-5.1 for method performance criteria.
15.0 Po llu tio n Pr even tio n and W aste M anag em ent ____________________
15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory.
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16.0 Records
16.1 Complete the extraction worksheet attached to this method, and tape in the study notebook or include in the 3-ring study binder, as appropriate.
17.0 Attachments____________________________________________________ _ 17.1 Attachment A, Extraction worksheet 17.2 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Calibration standard concentration worksheet
18.0 References_______________________________________________________________
18.1 The validation report associated with this method is ETS-8-4.0 & 5.0-V -l.
18.2 FACT-M-3.1, "Analysis o f Serum or Other Fluid Extracts for Fluorochemicals using HPLC-Electrospray Mass Spectrometry"
19.0 Affected Documents______________________________________________________
19.1 T S-8-5.1, "Analysis o f Serum or Other Fluid Extracts for Fluorochemicals using HPLCElectrospray Mass Spectrometry"
20.0 Revisions____________
Revision Number
1
Reason For Revision
Section 12.21 Changed to include sample storage at room temperature.
Section 12.13 Added the shaker speed.
Section 12.17 Final volume is 1.0 mL; not adjusted for initial volumes less than 1.0 mL.
Revision
Date
04/02/99
ETS-8-4.1 Extraction o f PFOS from Serum
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4.1.2 When handling samples or solvents wear appropriate protective gloves, eyewear, and clothing.
4.2 Cautions:
4.2.1 Operate the solvent pumps below a back pressure o f 400 bar (5800 psi). If the back pressure exceeds 400 bar, the HP 1100 will initiate automatic shutdown.
4.2.2 Do not run solvent pumps to dryness.
5.0 I nterferences_____________________________________________________________
5.1 To minimize interferences when analyzing samples, Teflon shall not be used for sample storage or any part o f instrumentation that comes in contact with the sample or extract.
6.0 Eq uipm ent_____________________________________________________
6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations.
6.1.1 6.1.2
Micromass Quattro II triple quadrupole Mass Spectrometer equipped with an electrospray ionization source.
HP 1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler
7.0 Supplies and M ate r ia ls ____________________________________________________
7.1 Supplies 7.1.1 High purity grade air regulated to approximately 100 psi (house air system) 7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data 7.1.3 Capped autovials or capped 15 ml centrifuge tubes
8.0 Reagents and Standards___________________________________________________ 8.1 Reagents
8.1.1 Methanol, HPLC grade or equivalent
8.1.2 Milli-QTM water (ASTM type I), all w ater used in this method should be ATSM type I, or equivalent, and be provided by a Milli-Q TOC Plus system or other vendor
8.1.3 Ammonium acetate, reagent grade or equivalent
8.1.3.1 When preparing different amounts than those listed, adjust accordingly.
8.1.3.2
2.0 mM ammonium acetate solution: Weigh approximately 0.300 g ammonium acetate. Pour into a 2000 mL volumetric container containing 2000 mL Milli-QTM water, mix until all solids are dissolved. Store at room temperature.
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8.2 Standards
8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. Refer to ETS-8-6.0.
9.0 Sa m p le H a n d lin g ______________________________________________ ___ 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples
are stored in capped autovials or capped 15 ml centrifuge tubes until analysis.
9.2 If analysis will be delayed, extracted standards and samples may be stored at room temperature, or refrigerated at approximately 4 C, until analysis can be performed.
10.0 Qu a l it y Control_________________________________________________________ 10.1 M ethod B lanks a n d M atrix B lanks
10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each batch to determine contamination or carryover.
10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve.
10.2 M atrix Spikes
10.2.1 Matrix spikes are prepared and analyzed to determine the matrix effect on the recovery efficiency. -
10.2.2 Matrix spike duplicates are prepared and analyzed to measure the precision and the recovery for each analyte.
10.2.3 Analyze a matrix spike and matrix spike duplicate per forty samples. With a minimum o f 2 spikes per batch.
10.2.4 Matrix spike and matrix spike duplicate concentrations will fall in the mid-range o f the initial calibration curve. Additional spike concentrations may fall in the lowrange o f the initial calibration curve.
10.3 C ontinuing C alibration Checks
10.3.1 Continuing calibration verifications are analyzed to verify the continued accuracy of the calibration curve.
10.3.2 Analyze a mid-range calibration standard every tenth sample, with a minimum o f one per batch.
11.0 Ca l ib r a t io n and St a n d a r d iza t io n _________________________________________ 11.1 Analyze the extracted matrix standards prior to and following each set o f sample extracts.
The average o f two standard curves will be plotted by linear regression (y = mx + b), weighted 1/x, not forced through the origin, using MassLynx or other suitable software.
11.2 If the curve does not meet requirements perform routine maintenance or reextract the standard curve (if necessary) and reanalyze.
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11.3 For purposes o f accuracy when quantitating low levels o f analyte, it may be necessary to use the low end o f the calibration curve rather than the full range o f the standard curve. Example: when attempting to quantitate approximately 10 ppb o f analyte, generate a calibration curve consisting o f the standards from 5 ppb to 100 ppb rather than the full range o f the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting o f high concentration standards.
12.0 Procedures______________________________________________________ _ _ _ _ _
12.1 A cquisition Set up
12.1.1 Set up the sample list.
12.1.1.1 Assign a sample list filename using MO-DAY-last digit o f year-increasing letter o f the alphabet starting with a
12.1.1.2 Assign a method (MS file) for acquiring 12.1.1.3 Assign an HPLC program (Inlet file) 12.1.1.4 Type in sample descriptions and vial position numbers
12.1.2 To create a method click on method in the Acquisition control panel then mass spectrometer headings and select SIR (Single Ion Recording) or MRM (Multiple
' Reaction Monitoring). Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected along with the SIRs. Save acquisition method. If MS/MS instruments are employed, additional product ion fragmentation information may be collected. Refer to Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM
12.1.3 Typically the analytical batch run sequence begins and ends with a set o f extracted matrix standards.
12.1.4 Samples are analyzed with a continuing calibration verification injected standard after every tenth sample. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered samples but may be included as such.
12.2 Using the A utosam pler
12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1,
12.2.2 Set-up the HP1100/autosampler at the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook:
12.2.2.1 Sample size = 10 pL injection
12.2.2.2 Inject/sample = 1
12.2.2.3 Cycle time = 9 minutes
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12.2.2.4 Solvent ramp conditions
Time
MeOH
0.00 min. 1.0 min. 4.5 min. 6.5 min. 7.0 min. 9.0 mi.
40% 40% 95% 95% 40% 40%
2.0 mM Ammonium acetate
60% 60% 5% 5% 60% 60%
12.2.2.5 Press the "Start" button.
12.3 Instrument Set-up -
12.3.1 Refer to ETS-9-24.0, "Operation and Maintenance o f the Micromass Quattro II Triple Quadrupole Mass Spectrometer Fitted with an Atmospheric Pressure Ionization Source," for more details.
12.3.2 Check the solvent level in reservoirs and refill if necessary.
12.3.3 Check the stainless steel Capillary at the end o f the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary.
12.3.4 Turn on the nitrogen.
12.3.5 Open the tune page. Clicks on operate to initiate source block and desolvation heaters.
12.3.6 Open the Inlet Editor.
12.3.6.1 Set HPLC pump to "On" 12.3.6.2 Set the flow to 10 - 500 uL/min or as appropriate 12.3.6.3 Observe droplets coming out o f the tip o f the probe. A fine mist should be
expelled with no nitrogen leaking around the tip of the probe. Readjust the tip o f the probe if no mist is observed 12.3.6.4 Allow to equilibrate for approximately 10 minutes.
12.3.7 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response:
12.3.7.1 Drying gas 250-400 liters/hour 12.3.7.2 ESI nebulizing gas 10-15 liters/hour 12.3.7.3 HPLC constant flow mode flow rate 10 - 500 pL/min 12.3.7.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the
HPLC is operating correctly.)
12.3.7.5 Source block temperature 150
12.3.7.6 Desolvation temperature 250
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12.3.8 Print the tune page, with its parameters, and store it in the study binder with a copy taped into the instrument log.
12.3.9 Click on start button in the Acquisition Control Panel (this may vary among MassLynx versions, refer to appropriate MassLynx U ser's Guide). Ensure start and end sample number includes all samples to be analyzed.
13.0 Da t a A n alysis and Calc u la tio n s__________________________________________ 13.1 Calculations:
13.1.4 Calculate matrix spike percent recoveries using the following equation:
% Recovery =
Observed Result - Background Result x 100 Expected Result
13.1.5 Calculate percent difference using the following equation:
% Difference = Expected Cone. - Calculated Cone, x 100 Expected Cone.
13.1.6 Calculate actual concentrations in matrix (pg/g):
fng o f PFOS calc, from std. Curve x Dilution Factor) (Initial Weight o f Liver fal Final Volume (mL)
x 1 ue 1000 ng
14.0 Method Performance____________________________________________________ 14.1 M ethod Detection Limit (MDL) and Limit o f Quantitation (LOQ) are method, analyte, and
matrix specific. Refer to ETS-8-6.0, Attachment B for a listing o f current validated MDL and LOQ values.
14.2 Solvent Blanks, Method Blanks and Matrix Blanks
14.2.1 Solvent blanks, method blanks, and matrix blanks must be below the lowest standard in the calibration curve.
14.3 Calibration Curves
14.3.1 The r2 value for the calibration must be 0.980 or better.
14.4 Matrix Spikes
14.4.1 Matrix spike percent recoveries must be within 30% o f the spiked concentration.
14.5 Continuing Calibration Verification
14.5.1 Continuing calibration verification percent recoveries must be within 30% o f the spiked concentration.
14.6 I f criteria listed in the method performance section are not met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook.
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14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text o f the report.
15.0 Po llu t io n Preventio n and W aste M anag em ent___________________________ _ 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass
pipette waste is disposed in broken glass containers located in the laboratory.
16.0 Records_________________________________________________________________ 16.1 Each page generated for a study must have the following information included either in the
header or hand written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst.
16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument runlog.
16.3 Plot the calibration curve by linear regression, weighted 1/x, then print these graphs and store in the study folder.
16.4 Print data integration summary, integration method, and chromatograms from MassLynx and store in the study folder.
16.5 Summarize data using suitable software (Excel 5.0+) and store in the study folder, refer to A ttachm ent A for an example o f a summary spreadsheet.
16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location o f backup electronic data.
17.0 T ables. Diag r am s. Flo w charts, and V a l id a t io n Da t a ______________________ 17.1 Attachment A: ETS-8-7.0 Data summary spreadsheet
18.0 References_____________________________________________________________ 18.1 FACT-M -2.1, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical
Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry" 18.2 ETS-9-24.0, "Operation and Maintenance o f the Micromass Atmospheric Pressure
Ionization/Mass Spectrometer Quattro II triple quadrupole Systems" 18.3 The validation report associated with this method is ETS-8-6.0 & 7.0-V-l
19.0 A ffected Documents____________________________________________________ 19.1 ETS-8-6.0, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical
Compounds from Liver or Fluid for Analysis Using HPLC-Electrospray/Mass Spectrometry"
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20.0 Revisions____________________________
Revision Number
Reason For Revision
Revision Date
ETS-8-7.0
Analysis of Liver Extract Using ES/MS
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Page 9 o f 10
Laboratory Study #
Study: Test Material: Matrix/Final Solvent: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y Intercept: Date o f Extraction/Analyst: Date of Analysis/Analyst:
Group Sample# Concentration Dose ng/g
ii'
Initial Wt. g
Dilution Factor
*
Final Cone, ug/g
Slope: Taken from linear regression equation. Group/Dose: Taken from the study folder. Sample#: Taken from the study folder.
Concentration (ng/g): Taken from the MassLynx integration summary. Initial Wt. (g): Taken from the study folder. Dilution Factor: Taken from the study folder. Final Cone, (ug/g): Calculated by dividing the initial volume from the concentration
Attachment A: Summary Spreadsheet -
ETS-8-7.0
Analysis of Liver Extract Using ES/MS
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Page 10 o f 10
3M Environm ental Laboratory
M ethod
Extraction of Potassium Perfluorooctanesulfonate or other Fluorochemical Compounds from Liver for A nalysis using HPLC-
Electrospray/M ass Spectrometry
M ethod N um ber: ETS-8-6.0
Author: Lisa Clemen, Robert Wynne Approved By:
Adoption Date: Revision Date:
Laboratory Manager
Date
Group Leader
Date
Technical Reviewer
Date
1.0 Scope and A p p lic a t io n ___________________________________________________ 1.1 Scope: This method is for the extraction o f potassium perfluorooctanesulfonate (PFOS) or
other fluorochemical compounds from liver.
1.2 A pplicable C om pounds: Fluorochemical surfactants or other fluorinated compounds.
1.3 M atrices: Rabbit, rat, bovine, and monkey livers or other tissues as designated in the validation report.
Word 6.0/95
- ETS-8-6.0 Extraction of PFOS from Liver
Page 1 o f 14
000064
2.0 Summary of Method
2.1 This method describes the procedure for extracting potassium perfluorooctanesulfonate (PFOS) or other fluorochemical surfactants from liver, or other tissues, using an ion pairing reagent and methyl-te/7-butyl ether (MtBE). In this method, seven fluorochemicals can be extracted: PFOS, PFOSA, PFOSAA, EtFOSE-OH, PFOSEA, M556, and surrogate standard. An ion pairing reagent is added to the sample and the analyte ion pair is partitioned into MtBE. The M tBE extract is transferred to a centrifuge tube and put onto a nitrogen evaporator until dry. Each extract is reconstituted in 1.0 mL methanol then filtered through a 3 cc plastic syringe attached to a 0.2 pm nylon filter into glass autovials.
2.2 These sample extracts are analyzed following method ETS-8-7.0 or other appropriate methods.
3.0 Definitions_______________________________________________________________ 3.1 PFOS: perfluorooctanesulfonate (anion o f potassium salt) CgFnS03 3.2 PFOSA: perfluorooctane sulfonylamide C8F17SO2NH2 3.3 PFOSAA: perfluorooctane sulfonylamido (ethyl)acetate C8Fi7S02N(CH2CH3)CH2C02 3.4 EtFOSE-OH: 2(N-ethylperfluorooctane sulfonamido)-ethyl alcohol
C 8Fi7S02N (C H 2CH 3)C H 2C H 2 0 H 3.5 PFOSEA: perfluorooctane sulfonyl ethylamide CgFnS02N(CH2CH3)H 3.6 M556: C8F17S02N(H)(CH2C 0 0 H ) 3.7 Surrogate standard: 1H-1H-2H-2H perfluorooctane sulfonic acid
4.0 W arnings and Cautions________________________________________________________ 4.1 H ealth an d Safety W arnings:
4.1.1 Use universal precautions, especially laboratory coats, goggles, and gloves when handling animal tissue, which may contain pathogens.
5.0 Interferences____________________________________________________________ 5.1 There are no interferences known at this time.
6.0 Equipment________________________________________________________________
6.1 The following equipment is used while performing this method. Equivalent equipment is acceptable.
6.1.1 6.1.2 6.1.3 6.1.4
Ultra-Turrax T25 Grinder for grinding liver samples Vortex mixer, VWR, Vortex Genie 2 Centrifuge, Mistral 1000 or IEC Shaker, Eberbach or VWR
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6.1.5 Nitrogen Evaporator, Organomation 6.1.6 Balance (sensitivity to 0.100 g)
7.0 Supplies and M ater ials_________________________________________________ _ 7.1 Gloves 7.2 Dissecting scalpels 7.3 Eppendorf or disposable pipettes 7.4 Nalgene bottles, capable o f holding 250 mL and 1 L 7.5 Volumetric flasks, glass, type A 7.6 I-CHEM vials, 40 mL glass 7.7 Plastic sampule vials, Wheaton, 6 mL (or appropriate size) 7.8 Centrifuge tubes, polypropylene, 15 mL 7.9 Labels 7.10 OxfordD ispensor- 3 .0 to 10.0ml 7.11 Syringes, capable of measuring 5 pL to 50 pL 7.12 Graduated pipettes 7.13 Syringes, disposable plastic, 3 cc 7.14 Syringe filters, nylon, 0.2 pm, 25 mm 7.15 Timer 7.16 Crimp cap autovials and caps 7.17 Crimpers Note: Prior to using glassware and bottles, rinse 3 times with methanol and 3 times with Milli-
QTM water. Rinse syringes a minimum o f 9 times with methanol, 3 rinses from 3 separate vials.
8.0 Reagents and Standards_________________________________________________ 8.1 Type I reagent grade water, Milli-QTM or equivalent; all water used in this method should
be Milli-Q water and be provided by a Milli-Q TOC PlusTM system 8.2 Sodium hydroxide (NaOH), J.T Baker or equivalent 8.3 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent 8.4 Sodium carbonate (Na2C03), J.T. Baker or equivalent 8.5 Sodium bicarbonate (N aH C 03), J.T. Baker or equivalent 8.6 Methyl-terf-butyl ether, Omnisolv, glass distilled or HPLC grade 8.7 Methanol, Omnisolv, glass distilled or HPLC grade 8.8 Liver, frozen from supplier 8.9 Dry ice from supplier 8.10 Fluorochemical standards
8.10.1 PFOS (3M Specialty Chemical Division), molecular weight = 538
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8.11 8.12
8.10.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499
8.10.3 PFOSAA (3M Specialty Chemical Division), molecular weight = 585
8.10.4 EtFOSE-OH (3M Specialty Chemical Division), molecular weight = 570
8.10.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527
8.10.6 M556 (3M Specialty Chemical Division), molecular weight = 557
8.10.7 Surrogate standard: 4-H, perfluorooctane sulfonic acid (1-H,1-H, 2-H, 2-H CgFnSOsH) molecular weight = 428
8.10.8 Other fluorochemicals, as appropriate
Reagent preparation
N O TE: When preparing larger volumes than listed in reagent, standard, or surrogate preparation, adjust accordingly.
8.11.1 10 N sodium hydroxide (NaOH): Weigh approximately 200 g NaOH. Pour into a 1000 mL beaker containing 500 mL Milli-QTM water, mix until all solids are dissolved. Store in a 1 L Nalgene bottle.
8.11.2 1 N sodium hydroxide (NaOH): Dilute 10 N NaOH 1:10. Measure 10 mL o f 10 N NaOH solution into a 100 mL volumetric flask and dilute to volume using Milli-QTM water. Store in a 125 mL Nalgene bottle.
8.11.3 0.5 M tetrabutylammonium hydrogen sulfate (TBA): Weigh approximately 169 g o f TBA into a 1 L'volumetric containing 500 mL Milli-QTM water. Adjust to pH 10 using approximately 44 to 54 mL o f 10 N NaOH (While adding the last mL o f NaOH add slowly because the pH changes abruptly). Dilute to volume with Milli-QTM water. Store in a 1 L Nalgene bottle. .
8.11.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using 1 N NaOH solution.
8.11.4 0.25 M sodium carbonate/sodium bicarbonate buffer (Na2C03/NaHC03): Weigh approximately 26.5 g o f sodium carbonate (Na2C03) and 21.0 g o f sodium bicarbonate (N aH C 03) into a 1 L volumetric flask and bring to volume with MilliQTM water. Store in a 1 L Nalgene bottle.
Standards preparation
8.12.1 Prepare PFOS standards for the standard curve.
8.12.2 Prepare other fluorochemical standards, as appropriate. Multicomponent fluorochemical standards are acceptable (for example, one working standard solution containing 1.00 ppm PFOS, 1.02 ppm PFOSA, 0.987 ppm PFOSAA, and 1.10 ppm EtFOSE-OH.)
8.12.3 Weigh approximately 100 mg o f PFOS into a 100 mL volumetric flask and record the actual weight.
8.12.4 Bring to volume with methanol for a stock standard o f approximately 1000 ppm (pg/mL).
8.12.5 Dilute the stock solution with methanol for a working standard 1 solution o f approximately 50 ppm.
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8.12.6 Dilute the stock solution with methanol for a working standard 2 solution o f approx. 5.0 ppm.
8.12.7 Dilute the stock solution with methanol for a working standard 3 solution o f approx. 0.50 ppm.
8.13 Surrogate stock standard preparation
8.13.1 Weigh approximately 50-60 mg o f surrogate standard 1-H .l-H , 2-H, 2-H, C8F13SO3H into a 50 ml volumetric flask and record the actual weight.
8.13.2 Bring to volume with methanol for a surrogate stock o f approximately 1000-1200 ppm.
8.13.3 Prepare a surrogate working standard. Transfer approximately 1.0 ml o f surrogate stock to a 10 ml volumetric flask and bring to volume with methanol for a working standard o f 10-20 ppm. Record the actual volume transferred.
9.0 Sample Handling________________________________ __________________________ 9.1 All samples are received frozen and must be kept frozen until the extraction is performed.
10.0 Q u a l it y C o n t r o l ______________________________________________________________ 10.1 Matrix blanks and method blanks
10.1.1 An aliquot o f 1.0 mL methanol is used as a solvent blank. 10.1.2 Extract two 1.0 mL aliquots o f Milli-QTM water following this procedure and use
as method blanks.
10.1.3 Extract two 1.0 mL aliquots o f liver homogenate following this procedure and use as matrix blanks. Refer to 11.1.6,
10.2 Matrix spikes
10.2.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine the accuracy o f the extraction.
10.2.2 Prepare each spike using a sample chosen by the analyst, usually a control liver received with each sample set.
10.2.3 Expected concentrations will fall in the mid-range o f the initial calibration curve. Additional spikes may be included and may fall in the low-range o f the initial calibration curve.
10.2.4 Prepare one matrix spike and matrix spike duplicate per 40 samples, with a minimum o f 2 matrix spikes per batch.
10.3 Continuing calibration verifications 10.3.1 Prepare continuing calibration verification samples to ensure the accuracy o f the initial calibration curve. 10.3.2 Prepare, at a minimum, one continuing calibration verification sample per group o f 10 samples. For example, if a sample set = 34, four verifications are prepared and extracted.
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10.3.3 Prepare each continuing calibration verification from the same matrix used to prepare the initial curve.
10.3.4 The expected concentrations will fall within the mid-range o f the initial calibration curve. Additional spikes may be included that fall in the low-range o f the initial calibration curve. This is necessary if the analyst must quantitate using only the low end of the calibration curve (for example, 5 ppb - 100 ppb, rather than 5 ppb 1000 ppb).
11.0 Calibration and Standardization___________________________________ ______ 11.1 Prepare matrix calibration standards
11.1.1 Weigh approximately 40 g o f liver into a 250 mL Nalgene bottle containing 200 mLs Milli-QTM water. Grind to a homogeneous solution.
11.1.2 I f 40 g is not available, use appropriate amounts o f liver and w ater to ensure a 1:5 ratio.
11.1.3 Refer to 13.0 to calculate the actual density o f liver homogenate and the concentration o f solid liver tissue dispersed in 1.0 mL o f homogenate solution.
11.1.5 Add 1 mL o f homogenate to a 15 mL centrifuge tube. Re-suspend solution by shaking between aliquots while preparing a total o f eighteen 1 mL aliquots of homogeneous solution in 15 mL centrifuge tubes.
11.1.6 Two 1 mL aliquots, or other appropriate volume, serve as matrix blanks.
11.1.7 Typically use the standard concentrations and spiking amounts listed in Table 1, at the end o f this section, to spike, in duplicate, tw o standard curves, for a total o f eighteen samples, two matrix blanks, and two method blanks.
11.1.8 Refer to validation reports ETS-8-6.0 and ETS-8-7.0-V-1 or Attachment B,
which lists the working ranges and the Linear Calibration Range (LCR) for calibration curves..
11.1.9 Use Attachment C as an aid in calculating the concentrations o f the working standards. Refer to 13.0 to calculate actual concentrations o f PFOS in calibration standards.
11.2 To each working standard, blank, or continuing verification, add appropriate amount o f surrogate working standard for the concentration to fall within the calibration curve range 5 ppb - lOOOppb.
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11.3 Extract spiked liver homogenates following 12.14-12.25 o f this method. Use these standards to establish each initial curve on the mass spectrometer.
Table 1 Approximate Spiking Amounts for Calibration Standards
Working Standard (Approx. Cone.)
-
0.50 ppm 0.50 ppm 0.50 ppm 0.50 ppm 0.50 ppm 5.0 ppm 5.0 ppm 5.0 ppm 50 ppm
III Approx, final cone, of PFOS in liver
- Blank 2 0.005 ppm 4 0.010 ppm 10 0.025 ppm 20 0.050 ppm 40 0.100 ppm 10 0.250 ppm 20 0.500 ppm 30 0.750 ppm 4 1.00 ppm
12.0 Procedure_______________________________________________________________ 12.1 Obtain frozen liver samples.
12.2 Cut approximately 1 g o f liver using a dissecting scalpel. This part o f the procedure is best performed quickly, not allowing the liver to thaw.
12.3 Weigh the sample directly into a tared plastic sampule vial.
12.4 Record the liver weight in the study notebook.
12.5 Return unused liver portions to freezer.
12.6 Add 2.5 mLs o f water to sampule vial.
12.7 Grind the sample. Put the grinder probe in the sample and grind for about 2 minutes, or until the sample is homogeneous.
12.8 Rinse the probe into the sample with 2.5 mLs water using a pipette.
12.9 Take the grinder apart and clean it with methanol after each sample. Refer to AMDT-EP22.
12.10 Cap the sample and vortex for 15 seconds. Label the sampule vial with the study number, weight, liver ID, date and analyst initials.
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12.11 Pipette 1.0 mL, or other appropriate volume, o f homogenate into a 15 mL polypropylene centrifuge tube. Label the centrifuge tube with the identical information as the sampule vial. Refer to attached worksheet for documenting the remaining steps.
12.12 Pipette tw o 1 mL aliquots o f Milli-QTM water to centrifuge tubes. These will serve as method blanks.
12.13 Spike all samples, including blanks and standards ready for extraction with surrogate standard as described in section 11.2.
12.14 Spike each matirx with the appropriate amount o f standard as described in 11.1, or Table 1 o f that section, for the calibration curve standards. Also prepare matrix spikes and continuing calibration standards.
12.15 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration samples for 15 seconds.
12.16 Check to ensure 0.5 M TBA reagent is at pH 10. I f not, adjust accordingly.
12.17 To each sample, add 1 mL 0.5 M TBA and 2 mL o f the 0.25 M sodium carbonate/sodium bicarbonate buffer.
12.18 Using an Oxford Dispenser, add 5 mL methyl-fe/7-butyl ether.
12.19 Cap each sample and put on the shaker at a setting o f 300 rpm, for 20 minutes.
12.20 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm, or until layers are well separated.
12.21 Label a fresh 15 mL centrifuge tube with the same information as in 12.10.
12.22 Remove 4.0 mL o f the organic layer to the fresh 15 mL centrifuge tube.
12.23 Put each sample on the analytical nitrogen evaporator until dry, approximately 1 to 2 hours.
12.24 Add 1.0 mL to each centrifuge tube using a graduated pipette.
12.25 Vortex mix for 30 seconds.
12.26 Attach a 0.2 pm nylon mesh filter to a 3 cc syringe and transfer the sample to this syringe. Filter into a 1.5 mL glass autovial or low-volume autovial when necessary.
12.27 Label the autovial with the study number, animal number and gender, sample timepoint, matrix, final solvent, extraction date, and analyst(s) performing the extraction.
12.28 Cap and store extracts at room temperature or at approximately 4 C until analysis.
12.29 Complete the extraction worksheet, attached to this document, and tape in study notebook or include in study binder, as appropriate.
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13.0 Data Analysis and Calculations______ ____________________________________ 13.1 C alculations:
13.1.1 Calculate the average density o f the liver homogenate by recording each mass o f ten separate 1.0 mL aliquots o f homogenate. Average density (mg/mL) = Average mass (me) o f the aliquots
1.0 mL aliquot
13.1.2 Calculate the amount o f liver (mg) per 1.0 mL homogenate (or concentration o f dispersed solid tissue per mL o f homogenate suspension) using the following equation:
g o f Liver x. Average density* o f homogenate (mg/mLl (g of Liver + g of Water)
* refer to 13.1.1 for details.
13.1.3 Calculate actual concentrations o f PFOS and other fluorochemicals in calibration standards using the following equation:
uL o f Standard x Concentration (ug /m D = Final Concentration (gg/g or mg/kg)
mg L iv e r/1 mL homogenate*
o f PFOS in Liver
*refer to 13.1.2 for details.
14.0 Method Performance______________________________________ __________'
14.1 The method detection limit (MDL) is analyte and matrix specific. Refer to MDL report for specific MDL and limit of.quantitation (LOQ) values (refer to A ttachm ents B and C).
14.2 The following quality control samples are extracted with each batch o f samples to evaluate the quality o f the extraction and analysis.
14.2.1 Method blanks and matrix blanks.
14.2.2 Matrix spike and matrix spike duplicate samples to determine accuracy and precision of the extraction.
14.2.3 Continuing calibration verification samples to determine the continued accuracy of the initial calibration curve.
14.3 Refer to section 14 o f ETS-8-7.0 for method performance criteria.
15.0 Pollution Prevention and Waste Management____________________________
15.1 Sample waste is disposed in biohazard containers, flammable solvent waste is disposed in high BTU containers, and used glass pipette waste is disposed in broken glass containers located in the laboratory.
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16.0 Records
16.1 Complete the extraction worksheet attached to this method, and tape in the study notebook or include in the 3-ring study binder, as appropriate.
17.0 Tables. Diag r a m s . Flo w char ts, and V a l id a t io n Da t a ______________________ 17.1 Attachment A, Extraction worksheet 17.2 Attachment B, MDL/LOQ values and summary 17.3 Attachment C, Calibration standard calculation and concentration worksheet
18.0 References______________________________________________________________ 18.1 The validation report associated with this method is ETS-8-6.0 & 7.0-V -l.
18.2 AMDT-EP-22, "Routine Maintenance o f Ultra-Turrax T-25"
18.3 FACT-M -1.1, "Extraction o f PFOS or Other Anionic Fluorochemical Surfactants from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry"
19.0 Affcted Documents______ ,______________________________________________
19.1 ETS-8-7.0, "Analysis o f Potassium Perfluorooctanesulfonate or other Fluorochemicals in Liver Extracts using HPLC-Electrospray Mass Spectrometry"
20.0 Revisio ns
Revision Number.
Reason For Revision
Revision Date
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Page 10 o f 14
BESTCOPYAVAILABLE
Study #
M atrix Box # W k/Day
D ate Spiked/Analyst ccv MS MSD
Surrogate Std
approx, ppm
actual
#
ppm
F C M ix Std
approx. 0.5 ppm
actual
ppm
#
F C M ix Std approx. 5 ppm actual ppm
#
F C M ix Std
approx. 50 ppm
actual
ppm
#
Com m ents
--
--
* --
--
--
--
--
--
--
--
--
--
--
--
- -
Blank
Liver Homogenate: Std #
Liver amount =
Liver Extraction Method Soike surrogate and Standard mix. Vortex 15 sec. Pinette 1 mL of Liver Solution Pipette 1 mL of t0.5 M TBA, pH 10. pH = Pipette 2 mL of 0.25 Na2CO3/0.25M NaHCOj Buffer
...................
Std. #
Std. #
Dispense 5ml of Methyl-t-Butyl Ether
TN-A-
Shake 20 min. Centrifuse 20-25 min. Remove a 4 mL aliauot of organic laver Put on Nitrogen Evanorator to drvness Add 1.0 mL of Methanol
Shaker Sneed Centrifuge Sneed
Evaporator Temnerature TN-A-
Vortex 30 sec.
Filter using a 3cc B-D svringe with a 0.2um SRI filter into autosamnle vial Cont. Cal. Verifications used the same matrix as for the standard curve.
. . . -
g
Date & Initials
Attachment A: Extraction Worksheet
. ETS-8-6.0 Extraction o f PFOS from Liver
Page 11 o f 14
000074
MDL/LOQ values for rabbit liver
Compound MDL LOQ Linear Calibration Range (LCR)
(ppb) (PP*>) Approximate concentrations to be used for preparing the
Standard Calibration Curve
PFOS
8.45 26.9 30 ppb - 1200 ppb
PFOSA
3.50 11.1 12 ppb - 1200 ppb
PFOSAA
24.6 78.3 30 ppb - 1200 ppb
EtFOSE-OH 108
345 60 ppb - 900 ppb*
M556
82.3 262 60 ppb - 1200 ppb
PFOSEA
33.9 108 30 ppb- 1200 ppb
MDL/LOQ values in rat, bovine, and monkey liver were not statistically determined. Two curves in each o f these matrices were extracted and analyzed with the rabbit liver curves to determine equivalence. Responses in the rat, bovine, and monkey liver curves were equivalent to the rabbit responses, therefore, their M DL and LOQ will be assumed to be equivalent to those values as determined for the rabbit liver.
Refer to LOQ Summary and MDL study in ETS-8-6.0 & 7.0-V-l for further information
* EtFOSE-OH estimates only for MDL and LOQ. Did not meet criteria for validation.
Compound: PFOS _____________ ___________________________ ______
Prepared 1 Range of
LCR from
Range of
LCR from
Range of
Liver
range of 1 average
ave curve
low std
low std
high std
matrix
standards 1 curve
curve
curve
curve
(ppb) (ng/mL) | (ppb) (ng/mL) (ppb) (ng/mL) (ppb) (ng/mL) (ppb) (ng/mL) (ppb) (ng/mL)
Rabbit
6 .1 9 -12 3 7 12 - 1200
12 - 1200
6-300
12-300
60 - 1200
L C R from high Std curve
(ppb) (ng/mL)
60 - 1200
Compound: PFOSA
Prepared
Liver
range of
matrix
standards
(ppb) (ng/mL)
Range of average
curve
(ppb) (ng/mL)
Rabbit
6 .1 9 -12 3 7 12 -1200
LCR from ave curve
(ppb) (ng/m L ).
Range of low std curve
(ppb) (ng/mL)
LCR from low std curve
. (PPb) (ng/mL)
12 * 1200
12 - 300
12 * 300
Range of high std
curve
(ppb) (ng/mL)
60 - 1200
LOR from high shl curve
(ppb) (ng/mL)
60 -1200
Compound: PFOSAA
Prepared
Range of
Liver
range of
average
matrix
standards
curve
(ppb) (ng/mL) (ppb) (ng/mL)
Rabbit
6.16 - 1232 12 - 1200
LCR from ave curve
(ppb) (ng/mL)
30-1200
Range of low std
c u rv e (ppb) (ng/mL)
30 - 900
LCR from low std
c u rv e (ppb) (ng/mL)
60 - 900
Range of high std
curve
(ppb) (ng/mL)
N/A
L C R from high std curve
(ppb) (ng/mL)
N/A
Attachment B: MDL/LOQ Values
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Page 12 o f 14
Compound: EtFOSE-OH
Prepared
Range of
Liver
range of
average
matrix
standards
curve
(ppb) (ng/mL) (ppb) (ng/mL)
Rabbit
6.17-1235 31 -900
LCRfrom ave curve
(ppb) (ng/mL)
31 -900
Range of low std curve
(ppb) (ng/mL)
N/A
LCR from low std curve
(ppb) (ng/mL)
N/A
Range of high std
curve
(ppb) (ng/mL)
L C R from h i g h std curve
(ppb) (ng/mL)
N/A f j l l l l l l J
Compound: PFOSEA
Prepared
Range of
Liver
range of
average
matrix
standards
curve
(ppb) (ng/mL) (ppb) (ng/mL)
Rabbit
6.17-1235 31-1200
LCRfrom ave curve
(ppb) (ng/mL)
31-1200
Range of low std curve
(ppb) (ng/mL)
N/A
LCR from low std curve
(ppb) (ng/mL)
N/A
Range of high std
curve
(ppb) (ng/mL)
N/A
LCR from high std
curve
(ppb) (ng/mL)
illllilll
Compound: M556
Prepared
Liver
range of
matrix
standards
(ppb) (ng/mL)
Rabbit
6 .1 7 -1 2 3 5
Range o f average
curve
(ppb) (ng/mL)
31-1200
LCRfrom ave curve
(ppb) (ng/mL)
60-1200
Range of low std curve
(ppb) (ng/mL)
N/A
L C R from low std curve
(ppb) (ng/mL)
N/A
Range of high std
curve
(ppb) (ng/mL)
N/A
L C R from high std curve
(ppb) (ng/mL)
N/A
Attachment B: MDL/LOQ Values
. ETS-8-6.0 Extraction of PFOS from Liver
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000076
Ion Pair Standard Curves - Tissue
Prep date(s): Analyte(s): Sample matrix:
Method/revision: Target analyte(s): FC mix std approx. 0.500 ppm: FC mix std approx. 5.00 ppm: FC mix std approx. 50.0 ppm: Surrogate std approx. 100 ppm:
Standard number: Equipment number: Final solvent and TN: Blank liver/identifier:
Actual concentrations of standards in the FC mix
PFOS PFOSA PFOSAA EtFOSE PFOSEA
Std cone Std cone Std cone Std cone Std cone
ug/mL ug/mL
ug/mL
ug/mL
ug/mL
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500 ' * 0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
0.500
5.00 5.00 5.00 5.00 5.00
5.00 5.00 5.00 5.00 5.00
5.00 5.00 5.00 5.00 5.00
50.0 50.0 50.0 50.0 50.0
M556 Std cone ug/mL
0.500 0.500 0.500 0.500 0.500 5.00 5.00 5.00 50.0
Std cone ug/mL
All Am't spiked
mL 0.002 0.004 0.010 0.020 0.040 0.010 0.020 0.030 0.004
A ll Density
g 0.167 0.167 0.167 0.167 0.167 0.167 0.167 0.167 0.167
Calculated concentrations of standards in the sample matrix
PFOS
PFO SA PFO SAA EtFOSE PFOSEA M556
Final
Final Final cone Final
Final
Final
Std cone
cone
cone
ng/g
cone
cone
cone
ng/g
ng/g
ng/g
ng/g
ng/g
ng/g
5.99 5.99 5.99 5.99 5.99 5.99
12.0 12.0 12.0 12.0 12.0 12.0
29.9 29.9 29.9 29.9 29.9 29.9
59.9 59.9 59.9 59.9 59.9 59.9
120 120 120 120 120 120
299 299 299 299 299 299
599 599 599 599 599 599
898 898 898 898 898 898
1198
1198
1198
1198
1198
1198
Surrogate Std cone ng/mL
100
S u rro g ate Final cone
ng/mL 0.500
All Am't spiked mL 0.005
Validated ranges - approximate concentrations
L iv e r
PFOS
PFOSA
PFOSAA
Rabbit
5-1000 ppb
5-1000 ppb
5-1000 ppb
Bovine
Estim ates only, use rabbit values.
Rat Estim ates only, use rabbit values.
Monkey
Estim ates only, use rabbit values.
EtFO SE-O H 5-1000 ppb
POAA 5-1000 ppb
PFOSEA 5-1000 ppb
Attachment C: Standard Calculations
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3M Environm ental Laboratory
M ethod
A nalysis of P otassium Perfluorooctanesulfonate or Other Fluorochemicals in Serum Extracts U sing H PLC-Electrospray/M ass Spectrometry
M ethod N um ber: ETS-8-5.1
Author: Lisa Clemen, Robert Wynne
A** t
Approved By:
Adoption Date: 03/01/99 Revision Date:
Laboratory Manager
Date
Group Leader
Date
Technical Reviewer
Date
1.0 Scope a n d Ap p lic a t io n
1.1 Scope: This method describes the analysis o f serum extracts for fluorochemical surfactants using HPLC-electrospray/mass spectrometry.
1.2 A pplicable C om pounds: Fluorochemical surfactants or other fluorinated compounds, or other ionizable compounds.
1.3 M atrices: Rabbit, rat, bovine, monkey, and human serum, or other fluids as designated in the validation report.
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2.0 Summary of Method
2.1 This method describes the analysis o f fluorochemical surfactants extracted from serum or other fluids, using HPLC-electrospray/mass spectrometry, or similar system as appropriate. The analysis is performed by monitoring a single ion characteristic o f a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z= 499. Additionally, samples may be analyzed using a tandem mass spectrometer to further verify the identity o f a compound by detecting daughter ions o f the parent ion.
3.0 De fin itio n s_______________________________________________________________
3.1 Atmospheric Pressure Ionization (API): The Micromass Quattro II triple quadrupole systems allow for various methods o f ionization by utilizing various sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization process in these techniques occurs at atmospheric pressure (i.e., not under a vacuum).
3.2 Electrospray Ionization (ES, ESI): a method o f ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the application o f a strong electrical field.
3.3 Mass Spectrometry, Mass Spectrometer (MS), Tandem Mass Spectrometer (MS/MS): The API Quattro II triple quadrupole systems are equipped with quadrupole mass selective detectors. Ions are selectively discriminated by mass to charge ratio (m/z) and subsequently detected. A single MS may be employed for ion detection or a series (MS/MS) for more specific fragmentation information.
3.4 Conventional vs. Z-spray probe interface: The latest models o f Micromass Quattro II triple quadrupole systems (post 1998) utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonal to the cone aperture. In the conventional conformation it is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter electrode. Though the configuration is different, the methods o f operation, cleaning, and maintenance are the same. However, Z-spray components and conventional components are not compatible with one another, but only with similar systems (i.e., Z-spray components are compatible with some other Z-spray systems, etc.)
3.5 Mass Lynx Software: System software designed for the specific operation o f these Quattro II triple quadrupole systems. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (Micromass Quattro II triple quadrupole MassLynx or MassLynx NT U ser's Guide).
4.0 W arnings and Cautions______________________________________________
4.1 Health and Safety Warnings:
4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage o f approximately 5000 Volts.
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4.1.2 When handling samples or solvents wear appropriate protective gloves, eyewear, and clothing.
4.2 Cautions:
4.2.1 Do not operate solvent pumps above capacity of 400 bar (5800 psi) back pressure. If the back pressure exceeds 400 bar, the H P1100 will initiate automatic shutdown.
4.2.2 Do not run solvent pumps to dryness.
5.0 INTERFERENCES_________________________________________________________
5.1 To minimize interferences when analyzing samples, teflon should not be used for sample storage or any part o f instrumentation that comes in contact with the sample or extract.
6.0 Eq uipm ent_______ ;____________________
6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations.
6.1.1 6.1.2
Micromass Quattro II triple quadrupole Mass Spectrometer equipped with an electrospray ionization source
H P1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler
7.0 Supplies and M ater ials____________________________________________________
7.1 Supplies 7.1.1 High purity grade nitrogen gas regulated to approximately 100 psi (House air system)
7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data.
7.1.3 Capped autovials or capped 15 mL centrifuge tubes
8.0 Reagents and Standards__________________________________________________ 8.1 Reagents
8.1.1 Methanol, HPLC grade or equivalent
8.1.2 Milli-QTM water, all water used in this method should be Milli-QTM w ater or equivalent, and may be provided by a Milli-Q TOC Plus system or other vendor
8.1.3 Ammonium acetate, reagent grade or equivalent
8.2 Standards 8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. See ETS-8-4.1.
9.0 Sa m p le Ha n d lin g _________________________
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9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 mL centrifuge tubes until analysis.
9.2 If analysis will be delayed, extracted standards and samples can be refrigerated at approximately 4 C, or at room temperature, until analysis can be performed.
10.0 Qu a l it y Control___________________________________________________ _ 10.1 Solvent Blanks, Method Blanks and Matrix Blanks
10.1.1 Solvent blanks, method blanks and matrix blanks are prepared and analyzed with each batch to determine contamination or carryover.
10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve.
10.2 Matrix Spikes
10.2.1 Matrix spikes are prepared and analyzed to determine the matrix effect on the recovery efficiency.
10.2.2 Matrix spike duplicates are prepared and analyzed to measure the precision and the recovery for each analyte.
10.2.3 Analyze a matrix spike and matrix spike duplicate per forty samples, with a minimum o f 2 spikes per batch.
10.2.4 Matrix spike and matrix spike duplicate concentrations will fall in the mid-range o f the initial calibration curve. Additional spike concentrations may fall in the lowrange o f the initial calibration curve.
10.3 Continuing Calibration Verifications
10.3.1 Continuing calibration verifications are analyzed to verify the continued accuracy o f the calibration curve.
10.3.2 Analyze a mid-range calibration standard after every tenth sample, with a minimum o f one per batch.
11.0 Ca lib r a t io n and St a n d a r d iza t io n _________________________________________ 11.1 Analyze the extracted matrix standards prior to and following each set o f extracts. The
average o f two standard curves will be plotted by linear regression (y = my + b), weighted 1/x, not forced through zero, using MassLynx or other suitable software.
11.2 If the curve does not meet requirements, perform routine maintenance or reextract the standard curve (if necessary) and reanalyze.
11.3 For purposes o f accuracy when quantitating low levels o f analyte, it may be necessary to use the low end o f the calibration curve rather than the full range o f the standard curve. Example: when attempting to quantitate approximately 10 ppb o f analyte, generate a calibration curve consisting o f the standards from 5 ppb to 100 ppb rather than the full range o f the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting o f high concentration standards.
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12.0 Procedures 12.1 A cquisition Set up
12.1.1 Click on start button in the Acquisition Control Panel. Set up a sample list. Assign a filename using MO-DAY-last digit o f year-sample number, assign a method (MS) for acquiring, and type in sample descriptions.
12.1.2 To create a method click on scan button in the Acquisition control panel and select SIR (Single Ion Recording) or MRM. Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected along with the SIRs. Save acquisition method. If MS/MS instruments are employed, additional product ion fragmentation information may be collected. See Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM (Multiple Reaction Monitoring).
12.1.3 Typically the analytical batch run sequence begins with a set o f extracted matrix standards and ends with a set of extracted matrix standards.
12.1.4 Samples are analyzed with a continuing calibration check injected after every tenth i . sample. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered samples but may be included as such.
12.2 Using the A utosam pler
12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1,
12.2.2 Set-up the HP1100/autosampler at the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook:
12.2.2.1 Sample size = 10 pL injection
12.2.2.2 Inject/sample = 1
12.2.2.3 Cycle time =13.5 minutes
12.2.2.4 Solvent ramp =
Time
0.00 min. 8.50 min. 11.0 min. 12.0 min.
MeOH
40% 90% 90% 40%
2.0 mM Ammonium acetate
60% 10% 10% 60%
12.2.2.5 Press the "Start" button. 12.3 In stru m en t Set-up
12.3.1 Refer to ETS-9-24.0 for more details. 12.3.2 Check the solvent level in reservoirs and refill if necessary.
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12.3.3 Check the stainless steel capillary at th end o f the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. If the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary.
12.3.4 Set HPLC pump to "On" . Set the flow to 10 - 500 uL/min or as appropriate. Observe droplets coming out of the tip o f the probe. Allow to equilibrate for approximately 10 minutes.
12.3.5 Turn on the nitrogen. A fine mist should be expelled with no nitrogen leaking around the tip o f the probe. Readjust the tip o f the probe if no mist is observed.
12.3.6 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response:
12.3.6.1 Drying gas 250-400 liters/hour 12.3.6.2 ESI nebulizing gas 10-15 liters/hour 12.3.6.3 HPLC constant flow mode, flow rate 10 - 500 pL/min 12.3.6.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the
HPLC is operating correctly.)
12.3.7 Carefully guide the probe into the opening. Insert probe until it will not go any - . further. Connect the voltage cables to the probe.
12.3.8 Print the tune page, with its parameters, and store it in the study binder with a copy taped into the instrument log.
12.3.9 Using the cross-flow counter electrode in the ES/MS source is recommended for the analysis o f biological matrices.
12.3.10Click on start button in the Acquisition Control Panel (this may vary among MassLynx versions, see appropriate MassLynx USER'S GUIDE). Press the start button. Ensure start and end sample number includes all samples to be analyzed.
13.0 Da t a A n aly sis and Ca lc u la tio n s__________________________________________ 13.1 Calculations:
13.1.4 Calculate matrix spike percent recoveries using the following equation:
% Recovery =
Observed Result - Background Result x 100 Expected Result
13.1.5 Calculate percent difference using the following equation:
% Difference = Expected Cone. - Calculated Cone, x 100 Expected Cone.
13.1.6 Calculate actual concentration o f PFOS, or other fluorochemical, in matrix (pg/mL):
fng o f PFOS calc, from std. Curve x Dilution Factor) x 1 ue (Initial Volume o f matrix (mL) + mL o f Surrogate Standard! 1000 ng
Final Volume (mL)
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14.0 Method Performance 14.1 M ethod Detection Limit (MDL) and Limit o f Quantitation (LOQ) are method, analyte, and
matrix specific. Please see ETS-8-4.1, Attachment B, for a listing o f current validated MDL and LOQ values.
14.2 Solvent Blanks, Method Blanks, and Matrix Blanks
14.2.1 Solvent blanks, method blanks, and matrix blanks values are must be below the lowest standard in the calibration curve
14.3 Calibration Curves
14.3.1 The r2 value for the calibration curve must be 0.980 or better.
14.4 Matrix Spikes
14.4.1 Matrix spike percent recoveries are must be within 30% o f the spiked concentration.
14.5 Continuing Calibration Verifications
14.5.1 Continuing calibration verification percent recoveries must be 30% o f the spiked concentration.
14.6 If criteria listed in this method performance section isn't met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook.
14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text o f the report.
15.0 P o l l u t i o n P r e v e n t i o n a n d W a s t e M a n a g e m e n t _______________ __________ __ 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass
pipette waste is disposed in broken glass containers located in the laboratory.
16.0 R e c o r d s ___________________________________________________________________ 16.1 Each page generated for a study must have the following information included either in the
header or hand written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst.
16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument runlog.
16.3 Plot the calibration curve by linear regression, weighted 1/x, then print these graphs and store in the study folder.
16.4 Print data integration summary, integration method, and chromatograms, from MassLynx, and store in the study folder.
16.5 Summarize data using suitable software (Excel 5.0) and store in the study folder, see Attachment A for an example o f a summary spreadsheet.
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16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location o f backup electronic data.
17.0 T ables. Diag r am s. Flo w charts, and V a l id a t io n Da t a _______________________ 17.1 Attachment A: ETS-8-5.1 Data summary spreadsheet.
18.0 References______________________________________________________________ 18.1 FACT-M -4.1, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical
compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry
18.2 ETS-9-24.0, "Operation and Maintenance o f the Micromass Atmospheric Pressure Ionization/Mass Spectrometer Quattro II triple quadrupole Systems"
18.3 The validation report-associated with this method is ETS-8-4.0 & 5.0-V -l.
19.0 A ffected D ocuments_______________________________________ ______________
19.1 ETS-8-4.1, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical Compounds from Serum for Analysis Using HPLC-Electrospray/Mass Spectrometry"
20.0 Revisions___________ _____
Revision Number.
1
Reason For Revision Section 6.1.2 Clarification o f H P1100 system components. Section 11.1 Average o f two curves, not standard values, are used for plotting linear regression and added the 1/x weighting o f the curve. Section 12.2.2.4 Clarification o f solvent ramp. Section 17.1 Changed from attachment B to A.
Revision Date
04/02/99
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Laboratory Study #
Study: Test Material: Matrix/Final Solvent: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y Intercept: Date of Extraction/Analyst: Date of Analysis/Analyst:
Group Sample# Concentration Dose ug/mL
V
Initial Vol. mL
Dilution Factor
Final Cone. ue/mL
Slope: Taken from linear regression equation. Group/Dose: Taken from the study folder. Sample#: Taken from the study folder. Concentration (ug/mL): Taken from the MassLynx integration summary. Initial Volume (mL): Taken from the study folder. Dilution Factor: Taken from the study folder. Final Cone. (ug/mL): Calculated by dividing the initial volume from the concentration
Attachment A: Summary Spreadsheet __
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3M Environm ental Laboratory
M ethod
A nalysis of P otassium Perfluorooctanesulfonate or Other Fluorochemicals in Liver Extracts U sing HPLC-Electrospray/M ass Spectrometry
M ethod Num ber: ETS-8-7.0
Author: Lisa Clemen, Glenn Langenburg (i/'
Approved By:
Adoption Date: Revision Date:
Laboratory Manager
Date
Group Leader
Date
Technical Reviewer
Date
1.0 Scope and A p p lic a t io n
1.1 Scope: This method is for the analysis o f liver extracts for fluorochemical surfactants using HPLC-electrospray/mass spectrometry.
1.2 A pplicable C om pounds: Fluorochemical surfactants or other fluorinated compounds, or other ionizable compounds.
1.3 M atrices: Rabbit, rat, bovine, monkey liver, or other tissues as designated in the validation report.
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2.0 Summary of Method
2.1 This method describes the analysis o f fluorochemical surfactants extracted from liver using HPLC-electrospray/mass spectrometry, or similar system as appropriate. The analysis is performed by monitoring a single ion characteristic o f a particular fluorochemical, such as the perfluorooctanesulfonate (PFOS) anion, m/z = 499. Additionally, samples may be analyzed using a tandem mass spectrometer to further verify the identity o f a compound by detecting daughter ions o f the selected parent ion.
3.0 D e f i n i t i o n s ___________________________________________________________
3.1 Atmospheric Pressure Ionization (API): The Micromass Quattro II triple quadrupole systems allow for various methods o f ionization by utilizing various sources, probes, and interfaces. These include but are not limited to: Electrospray Ionization (ESI), Atmospheric Pressure chemical Ionization (APcI), Thermospray, etc. The ionization process in these techniques occurs at atmospheric pressure (i.e. not under a vacuum).
3.2 Electrospray Ionization (ES, ESI): a method o f ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the application o f a strong electrical field.
3.3 Mass Spectrometry, Mass Spectrometer (MS), Tandem Mass Spectrometer (MS/MS): The API Quattro II triple quadrupole mass spectrometer is equipped with two quadrupole mass selective detectors and a collision cell. Ions are selectively discriminated by mass to charge ratio (m/z) and subsequently detected. A single MS may be employed for ion detection or an ion may be selected in the first quadrupole, fragmented in the collision cell, and these fragments may be analyzed in the second quadrupole.
3.4 Conventional vs. Z-spray probe interface: The latest models o f Micromass Quattro II triple quadrupole (post 1998) utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonal to the cone aperture. In the conventional conformation it is aimed directly at the cone aperture, after passing through a tortuous pathway in the counter electrode. Though the configuration is different, the methods o f operation, cleaning, and maintenance are the same. However, Z-spray components and conventional components are not compatible with one another, but only with similar systems (i.e. Z-spray components are compatible with other Z-spray systems, etc.)
3.5 Mass Lynx Software: System software designed for the specific operation o f these Quattro II triple quadrupole systems. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details refer to the manual specific to the instrument (Micromass Quattro II triple quadrupole MassLynx or MassLynx NT U ser's Guide).
4.0 W a r n i n g s a n d C a u t i o n s ______________________________________________ _
4.1 Health and Safety Warnings:
4.1.1 Use caution with the voltage cables for the probe. When engaged, the probe employs a voltage o f approximately 5000 Volts.
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4.1.2 When handling samples or solvents wear appropriate protective gloves, eyewear, and clothing.
4.2 C autions:
4.2.1 Operate the solvent pumps below a back pressure o f 400 bar (5800 psi). If the back pressure exceeds 400 bar, the HP 1100 will initiate automatic shutdown.
4.2.2 Do not run solvent pumps to dryness.
5.0 I n t e r f e r e n c e s _______________________________________________________________
5.1 To minimize interferences when analyzing samples, Teflon shall not be used for sample storage or any part o f instrumentation that comes in contact with the sample or extract.
6.0 E q u i p m e n t ___________________________________________________________________
6.1 Equipment listed below may be modified in order to optimize the system. Document any modifications in the raw data as method deviations.
6.1.1 6.1.2
Micromass Quattro II triple quadrupole Mass Spectrometer equipped with an electrospray ionization source.
H P 1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler
7.0 S u p p l i e s a n d M a t e r i a l s ______________________________________________________
7.1 Supplies 7.1.1 High purity grade air regulated to approximately 100 psi (house air system) 7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data 7.1.3 Capped autovials or capped 15 ml centrifuge tubes
8.0 Reagents and Standards__________________________________________________ 8.1 Reagents
8.1.1 Methanol, HPLC grade or equivalent
8.1.2 Milli-QTM water (ASTM type I), all water used in this method should be ATSM type I, or equivalent, and be provided by a Milli-Q TOC Plus system or other vendor
8.1.3 Ammonium acetate, reagent grade or equivalent
8.1.3.1 When preparing different amounts than those listed, adjust accordingly.
8.1.3.2
2.0 raM ammonium acetate solution: Weigh approximately 0.300 g ammonium acetate. Pour into a 2000 mL volumetric container containing 2000 mL Milli-QTM water, mix until all solids are dissolved. Store at room temperature.
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8.2 Standards
8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. Refer to ETS-8-6.0.
9.0 Sa m p le H a n d lin g _________________________________________________________ 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples
are stored in capped autovials or capped 15 ml centrifuge tubes until analysis.
9.2 If analysis will be delayed, extracted standards and samples may be stored at room temperature, or refrigerated at approximately 4 C, until analysis can be performed.
10.0 Qu a l it y Control________________________________________________________ 10.1 M ethod Blanks and.M atrix Blanks
10.1.1 Solvent blanks, method blanks, and matrix blanks are prepared and analyzed with each batch to determine contamination or carryover.
10.1.2 Analyze a method blank and a matrix blank prior to each calibration curve.
10.2 Matrix Spikes
10.2.1 Matrix spikes are prepared and analyzed to determine the matrix effect on the recovery efficiency.,
10.2.2 Matrix spike duplicates are prepared and analyzed to measure the precision and the recovery for each analyte.
10.2.3 Analyze a matrix spike and matrix spike duplicate per forty samples. With a minimum o f 2 spikes per batch.
10.2.4 Matrix spike and matrix spike duplicate concentrations will fall in the mid-range o f the initial calibration curve. Additional spike concentrations may fall in the lowrange o f the initial calibration curve.
10.3 C ontinuing Calibration Checks
10.3.1 Continuing calibration verifications are analyzed to verify the continued accuracy of the calibration curve.
10.3.2 Analyze a mid-range calibration standard every tenth sample, with a minimum o f one per batch.
11.0 Ca l ib r a t io n and Sta n d a r d iza tio n _________________________________________ 11.1 Analyze the extracted matrix standards prior to and following each set o f sample extracts.
The average o f two standard curves will be plotted by linear regression (y = mx + b), weighted 1/x, not forced through the origin, using MassLynx or other suitable software.
11.2 If the curve does not meet requirements perform routine maintenance or reextract the standard curve (if necessary) and reanalyze.
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11.3 For purposes o f accuracy when quantitating low levels o f analyte, it may be necessary to use the low end o f the calibration curve rather than the full range o f the standard curve. Example: when attempting to quantitate approximately 10 ppb o f analyte, generate a calibration curve consisting o f the standards from 5 ppb to 100 ppb rather than the full range o f the curve (5 ppb to 1000 ppb). This will reduce inaccuracy attributed to linear regression weighting o f high concentration standards.
12.0 P r o c e d u r e s ___________________________________________________________ _____
12.1 A cquisition Set up
12.1.1 Set up the sample list.
12.1.1.1 Assign a sample list filename using MO-DAY-last digit o f year-increasing letter o f the alphabet starting with a
12.1.1.2 Assign a method (MS file) for acquiring 12.1.1.3 Assign an HPLC program (Inlet file) 12.1.1.4 Type in sample descriptions and vial position numbers
12.1.2 To create a method click on method in the Acquisition control panel then mass spectrometer headings and select SIR (Single Ion Recording) or M RM (Multiple
. Reaction Monitoring). Set Ionization Mode as appropriate and mass to 499 or other appropriate masses. A full scan is usually collected along with the SIRs. Save acquisition method. If MS/MS instruments are employed, additional product ion fragmentation information may be collected. Refer to Micromass MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM.
12.1.3 Typically the analytical batch run sequence begins and ends with a set o f extracted matrix standards.
12.1.4 Samples are analyzed with a continuing calibration verification injected standard after every tenth sample. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered samples but may be included as such.
12.2 Using the A utosam pler
12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1,
12.2.2 Set-up the HP1100/autosampler at the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditions in the instrument logbook:
12.2.2.1 Sample size = 10 jiL injection
12.2.2.2 Inject/sample = 1
12.2.2.3 Cycle time = 9 minutes
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12.2.2.4 Solvent ramp conditions
Time
MeOH
0.00 min. 1.0 min. 4.5 min. 6.5 min. 7.0 min. 9.0 mi.
40% 40% 95% 95% 40% 40%
2.0 mM Ammonium acetate
60% 60% 5% 5% 60% 60%
12.2.2.5 Press the "Start" button.
12.3 Instrument Set-up
12.3.1 Refer to ETS-9-24.0, "Operation and Maintenance o f the Micromass Quattro II Triple Quadrupole Mass Spectrometer Fitted with an Atmospheric Pressure Ionization Source," for more details.
12.3.2 Check the solvent level in reservoirs and refill if necessary.
12.3.3 Check the stainless steel capillary at the end o f the probe. Use an eyepiece to check the tip. The tip should be flat with no jagged edges. I f the tip is found to be unsatisfactory, disassemble the probe and replace the stainless steel capillary.
12.3.4 Turn on the nitrogen.
12.3.5 Open the tune page. Clicks on operate to initiate source block and desolvation heaters.
12.3.6 Open the Inlet Editor.
12.3.6.1 Set HPLC pump to "On" 12.3.6.2 Set the flow to 10 - 500 uL/min or as appropriate 12.3.6.3 Observe droplets coming out o f the tip o f the probe. A fine mist should be
expelled with no nitrogen leaking around the tip o f the probe. Readjust the tip o f the probe if no mist is observed 12.3.6.4 Allow to equilibrate for approximately 10 minutes.
12.3.7 The instrument uses these parameters at the following settings. These settings may change in order to optimize the response:
12.3.7.1 Drying gas 250-400 liters/hour 12.3.7.2 ESI nebulizing gas 10-15 liters/hour 12.3.7.3 HPLC constant flow mode flow rate 10 - 500 pL/min 12.3.7.4 Pressure <400 bar (This parameter is not set, it is a guide to ensure the
HPLC is operating correctly.)
12.3.7.5 Source block temperature 150
12.3.7.6 Desolvation temperature 250
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12.3.8 Print the tune page, with its parameters, and store it in the study binder with a copy taped into the instrument log.
12.3.9 Click on start button in the Acquisition Control Panel (this may vary among MassLynx versions, refer to appropriate MassLynx U ser's Guide). Ensure start and end sample number includes all samples to be analyzed.
13.0 Data Analysis and Calculations________________________________________ __ 13.1 Calculations:
13.1.4 Calculate matrix spike percent recoveries using the following equation:
% Recovery =
Observed Result - Background Result x 100 Expected Result
13.1.5 Calculate percent difference using the following equation:
% Difference = Expected Cone. - Calculated Cone, x 100 Expected Cone.
13.1.6 Calculate actual concentrations in matrix (pg/g):
(ng o f PFOS calc, from std. Curve x Dilution Factor") (Initial Weight o f Liver ('g') Final Volume (mL)
x 1 ug 1000 ng
14.0 Method Performance_____________________________________________________ 14.1 M ethod Detection Limit (MDL) and Limit o f Quantitation (LOQ) are method, analyte, and
matrix specific. Refer to ETS-8-6.0, Attachment B for a listing o f current validated MDL and LOQ values.
14.2 Solvent Blanks, Method Blanks and Maitrix Blanks
14.2.1 Solvent blanks, method blanks, and matrix blanks must be below the lowest standard in the calibration curve.
14.3 Calibration Curves
14.3.1 The r2 value for the calibration must be 0.980 or better.
14.4 Matrix Spikes
14.4.1 Matrix spike percent recoveries must be within 30% o f the spiked concentration.
14.5 Continuing Calibration Verification
14.5.1 Continuing calibration verification percent recoveries must be within 30% o f the spiked concentration.
14.6 If criteria listed in the method performance section are not met, maintenance may be performed on the system and samples reanalyzed or other actions as determined by the analyst. Document all actions in the appropriate logbook.
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14.7 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text o f the report.
15.0 Po llu t io n Pr even tio n and W aste M an a g em en t_________________________ _ 15.1 Sample extract waste and flammable solvent is disposed in high BTU containers, and glass
pipette waste is disposed in broken glass containers located in the laboratory.
16.0 Records_________________________________________________________________ 16.1 Each page generated for a study must have the following information included either in the
header or hand written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. 16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument runlog.
16.3 Plot the calibration curve by linear regression, weighted 1/x, then print these graphs and store in the study folder.
16.4 Print data integration summary, integration method, and chromatograms from MassLynx and store in the study folder.
16.5 Summarize data using suitable software (Excel 5.0+) and store in the study folder, refer to Attachment A for an example o f a summary spreadsheet.
16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location o f backup electronic data.
17.0 T ables. Diag r a m s . Flo w c h ar ts, and V a l id a t io n Da t a ______________________ 17.1 Attachment A: ETS-8-7.0 D ata summary spreadsheet
18.0 References______________________________________________________________ 18.1 FACT-M -2.1, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical
Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry"
18.2 ETS-9-24.0, "Operation and Maintenance o f the Micromass Atmospheric Pressure Ionization/Mass Spectrometer Quattro II triple quadrupole Systems"
18.3 The validation report associated with this method is ETS-8-6.0 & 7.0-V -l
19.0 A ffected Documents____________________________________________________ 19.1 ETS-8-6.0, "Extraction o f Potassium Perfluorooctanesulfonate or Other Fluorochemical
Compounds from Liver or Fluid for Analysis Using HPLC-Electrospray/Mass Spectrometry"
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20.0 Revisio n s
Revision Number
Reason For Revision
Revision Date
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Laboratory Study #
Study: Test Material: Matrix/Final Solvent: Method/Revision: Analytical Equipment System Number: Instrument Software/Version: Filename: R-Squared Value: Slope: Y Intercept: Date of Extraction/Analyst: Date o f Analysis/Analyst:
Group Sample# Concentration Dose ng/g
Initial Wt. g
Dilution Factor
-'t
Final Cone, ug/g
Slope: Taken from linear regression equation. Group/Dose: Taken from the study folder.
Sample#: Taken from the study folder. Concentration (ng/g): Taken from the MassLynx integration summary. Initial Wt. (g): Taken from the study folder. Dilution Factor: Taken from the study folder. Final Cone, (ug/g): Calculated by dividing the initial volume from the concentration
Attachment A: Summary Spreadsheet
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oooo9e
3M E nvironm ental Laboratory
M ethod
Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane SULFONYLAMIDE (PFOSA), ANDPERFLUOROOCTANOATE (POAA) INWATER BYLlQUIDSolid Extraction and High-Performance Liquid Chromatography/Tandem Mass
Spectrometry (HPLC/MS/MS)
Method Number: ETS-8-154.0
Adoption Date:
Author: Kristen J. Hansen/Harold O. Johnson
Revision Date:
Approved By: William K. Reagen, Kent R. Lindstrom
William K Reagen, Laboratory Management
Date
Kristen J. Hansen, Ph.D., Group Leader
Date
Kent R. Lindstrom, Technical Reviewer
Date
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1.0 Scope and Application
1.1 This method provides collection, extraction, and analytical procedures for the determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane Sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in groundwater, surface water, and drinking water samples.
1.2 This method was prepared according to the EPA document, "Guidelines and Format for Methods to be Proposed at 40 CFR Part 136 or Part 141" (see Reference 18.1), and is based in part on the report "Method of Analysis for the Determination of Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in W ater" (see Reference 18.2).
2.0 Summary of Method_____________________________________________________
2.1 W ater samples are collected from a site of interest and shipped cold to an analytical facility. PFOS, PFOSA, and POAA are extracted from 40mL water samples using C lg solid phase extraction (SPE) cartridges. The compounds are eluted from the C 18cartridge, using methanol. Separation, identification, and measurement are accomplished by highperformance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis using multiple response monitoring (MRM).
The concentration o f each identified component is measured by comparing the MS response o f the quantitation ion produced by that compound to the MS response o f the quantitation ion produced by the same compound in an extracted calibration standard (external standard).
3.0 Definitions_________________________________________________________ _
3.1 Analytical Sample-- A portion o f an extracted Laboratory sample prepared for analysis.
3.2 Calibration Standard-- A solution prepared from the Working Standard (WS) and extracted according to this method. The calibration standard solutions are used to calibrate the instrument response with respect to analyte concentration.
3.3 Duplicate Sample (DS)-- A separate aliquot o f a sample, taken in the analytical laboratory and analyzed separately with identical procedures. Analysis o f DSs compared to that of the first aliquot give a measure of the precision associated with laboratory procedures, but not with sample collection, preservation, or storage procedures.
3.4 Field Blank Control Sample (FB)-- Type I water placed in a sample container in the laboratory and treated as a sample in all respects, including exposure to sampling site conditions, storage, preservation and all analytical procedures. The purpose o f the FB is to determine if test substances or other interferences are present in the field environment.
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3.5 Field Duplicate (FD)-- A sample collected in duplicate at the same time as the sample and placed under identical circumstances and treated exactly the same throughout field and laboratory procedures. Analysis o f FD compared to that o f the first sample gives a measure o f the precision associated with sample collection, preservation and storage, as well as with laboratory procedures.
3.6 Field Matrix Spike (FMS)-- A sample collected in duplicate to which known quantities of the target analytes are added in the field at the time of sample collection. The FMS should be spiked at approximately 50-150% of the expected analyte concentration in the sample. The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issues may complicate the interpretation of the sample analysis.
3.7 Field Spike Control Sample (FSCS)-- An aliquot o f type I water to which known quantities o f the target analytes are added in the field at the time o f sample collection (at an appropriate concentration to be determined by the project lead). The FSCS is extracted and analyzed exactly like a sample to determine whether a loss o f analyte could be attributed to sample storage and/or shipment.
3.8 Laboratory Control Sample (LCS)-- An aliquot o f type I water to which known quantities of the target analytes are added in the laboratory. Two levels are included, one at the LOQ (approx. 25Pg/mL), the other at a concentration of approx. 100-250Pg/mL or another concentration to be determined by the project lead. The LCS is extracted and analyzed exactly like a laboratory sample to determine whether the methodology is in control, and whether the laboratory is capable o f making accurate measurements at the required method detection limit and higher.
3.9 Laboratory Sample-- A portion o f a sample received from the field for testing.
3.10 Limit of Detection (LOD)-- The lowest concentration o f an analyte that can be measured and reported with 99% confidence that the analyte concentration is greater than zero. The LOD can be determined in several ways, including signal-to-noise ratio and statistical calculations.
3.11
Limit of Quantitation (LOQ)-- The lowest concentration (LLOQ) or highest concentration (ULOQ) that can be reliably achieved within the specified limits of precision and accuracy during routine operating conditions.
Note: The L L O Q is generally 5 -1 0 times the LO D . For many analytes, the L L O Q analyte
concentration is selected as the lowest non-zero standard in the calibration curve. H owever, it m ay be nominally chosen within these stated guidelines to sim plify data reporting. Sample LLO Q s are matrix-dependent.
3.12 Matrix Spike (MS)-- An aliquot o f a sample, to which known quantities o f target analytes are added in the laboratory. The MS is extracted and analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The background concentrations of the analytes in the sample matrix must be determined in a separate aliquot and the measured values in the MS corrected for background concentrations.
3.13 Method Blank-- An aliquot o f type I water that is treated exactly like a laboratory sample including exposure to all glassware, equipment, solvents, reagents, internal standards, and surrogates that are used with other laboratory samples. The method blank
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is used to determine if test substances or other interferences are present in the laboratory environment, the reagents, or the apparatus.
3.14 Method Detection Limit (MDL) Determination-- One o f several processes that may be used to establish a LOD value. The statistically calculated minimum amount o f an analyte that can be measured with 99% confidence that the reported value is greater than zero. This term is usually associated with the EPA definition in 40 CFR Part 136 Appendix B.
3.15 Sample-- A sample is a small portion collected from a larger quantity o f material intended to represent the original source material.
3.16 Spiking Stock Standard (SSS)-- A solution prepared from stock standards used to prepare the working standard.
3.17 Stock Standard (SS)-- A concentrated solution o f a single analyte prepared in the laboratory with an assayed reference compound.
3.18 Working Standard (WS)-- A solution o f several analytes prepared in the laboratory from SSs and diluted as needed to prepare calibration standards and other required analyte solutions.
4.0 Warnings and Cautions__________________________________________________
4.1 Health and Safety Warnings 4.1.1 The acute and chronic toxicity o f the standards for this method have not been precisely determined; however, each should be treated as a potential health hazard. 4.1.2 Unknown samples may contain high concentrations o f volatile toxic compounds. Sample containers should be opened in a hood and handled with gloves to prevent exposure. 4.1.3 The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulations regarding the handling o f the chemicals used in this method. A reference file of material safety data sheets (MSDS) should be available to all personnel involved in these analyses.
5.0 Interferences_______________________________________________________ _
5.1 During extraction and analysis, major potential contaminant sources are reagents and liquid-solid extraction devices.
5.2 All materials used in the analyses shall be demonstrated to be free from interferences under conditions of analysis by running method blanks.
5.3 Teflon containing materials (e.g. caps, wash bottles) contain fluorocompounds which may cause interferences and should not be used during collection, storage, extraction, or analysis of the samples.
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6.0 Equipment, Supplies, and Materials
Note: Brand names, suppliers, and part numbers are for illustrative purposes only. Equivalent
performance m ay be achieved using apparatus and materials other than those specified here, but demonstration o f equivalent performance that meets the requirements o f this method is the responsibility o f the laboratory.
6.1 Sampling Equipment 6.1.1 Sample collection bottles-- LDPE (e.g., NalgeneTM) narrow-mouth bottles with screw cap. Note: Do not use Teflon bottles or Teflon lined caps. 6.1.2 Coolers for sample shipment. 6.1.3 Ice for sample shipment. 6.1.4 Bottles must be lot-certified to be free of artifacts by running Method blanks according to this method.
6.2 Laboratory Equipment (Extraction and Analytical) 6.2.1 Balance, analytical (display at least 0.000lg), Mettler. 6.2.2 Vacuum pump, Bchi. 6.2.3 Visiprep vacuum manifold, Supelco. 6.2.4 Sep Pak Vac 6cc (lg ) tC 18cartridges (part # WAT 036795),Waters. 6.2.5 50mL disposable polypropylene centrifuge tubes, VWR. 6.2.6 15mL disposable polypropylene centrifuge tubes, VWR. 6.2.7 Disposable micropipettes (50-100pL, 100-200qL), Drummond. 6.2.8 Class A pipettes and volumetric flasks, various. 6.2.9 Hypercarb drop-in guard column (4mm) (part # 844017-^100), Keystone. 6.2.10 Stand-alone drop-in guard cartridge holder, Keystone. 6.2.11 125mL LDPE narrow-mouth bottles, Nalgene. 6.2.12 HPLC pump (LC10AD), Shimadzu. 6.2.13 2mL clear HPLC vial kit (cat #5181-3400), Hewlett Packard. 6.2.14 Standard lab equipment (graduated cylinders, disposable tubes, etc.), various. 6.2.15 LC/MS/MS and HPLC systems, as described in section 10.1.
6.3 Equipment Notes 6.3.1 In order to avoid contamination, the use o f disposable labware is highly recommended (tubes, pipettes, etc.). 6.3.2 Teflon or Teflon-lined containers or equipment, including Teflon-lined HPLC vials or caps for the HPLC auto sampler must not be used. 6.3.3 Type I water used during the sample and standard extraction should be filtered through a Hypercarb guard column using a HPLC pump. This water is referred to as "filtered type I water", hereafter in this report. 6.3.4 It is necessary to check the solvents (methanol) for the presence o f contaminants (especially POAA) by LC/MS/MS prior to use. Certain lot numbers have been found to be unsuitable for use. 6.3.5 Use disposable micropipettes or pipettes to aliquot standard solutions to make calibration standards and matrix spikes.
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7.0 Reagents and Standards
Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent performance
may be achieved using chemicals obtained from other suppliers. D o not use a lesser grade o f chemical than those listed.
7.1 Chemicals 7.1.1 Methanol (MeOH), HPLC grade, JT Baker, Catalog No. JT9093-2. 7.1.2 Ammonium Acetate, Reagent grade, Sigma-Aldrich, Catalog No. A-7330. 7.1.3 Water, type I, prepared in-house. 7.1.4 Sodium Thiosulfate, Reagent grade, JT Baker.
7.2 Standards 7.2.1 Potassium perfluorooctane sulfonate (see Attachment A, Figure 1). 7.2.2 Perfluorooctane sulfonylamide (see Attachment A, Figure 2). 7.2.3 Ammonium perfluorooctanoate (see Attachment A, Figure 3).
7.3 Reagent Preparation 7.3.1 250mg/mL sodium thiosulfate solution (Extraction)-- Dissolve 25g o f sodium thiosulfate in lOOmL reagent water. 7.3.2 40% methanol (Extraction)-- Measure 400mL methanol and adjust the volume to 1,0L with reagent water. 7.3.3 lOOmM ammonium acetate solution (Analysis)-- Weigh 7.7 lg o f ammonium acetate and dissolve in 1.0L o f reagent water. Dilute the lOOmM solution by a factor of 50 to make the 2mM ammonium acetate solution used for mobile phase A.
Note: Alternative volum es m ay be prepared as long as the ratios o f the solvent to solute ratios are
maintained.
7.4 Spiking Stock Standard (SSS) Preparation 7.4.1 lOOpg/mL each PFOS, PFOSA, and POAA SSSs-- Weigh out lOmg of analytical standard (corrected for percent salt and purity-- i.e., 10 mg C8F )7S03K purity 90% = 8.35mg C8F,7S03- ) and dilute to lOOmL with methanol in a lOOmL volumetric flask. Transfer to a 125mL LDPE bottle. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 42C for a maximum period o f 6 months from the date o f preparation. 7.4.2 lpg/mL mixed SSS-- Add l.OmL each of the 100pg/mL SSSs (from 7.4.1) to a lOOmL volumetric flask and bring up to volume with methanol. 7.4.3 O.lpg/mL mixed SSS-- Add lO.OmL o f the l.Opg/mL-mixed solution (from 7.4.2) to a lOOmL volumetric flask and bring up to volume with methanol. 7.4.4 0.01pg/mL mixed SSS-- Add lO.OmL o f the 0.1 pg/mL-mixed solution (from 7.4.3) to a lOOmL volumetric flask and bring up to volume with methanol. 7.4.5 Storage Conditions-- Store all SSSs in a refrigerator in 125mL LDPE bottles at 42C for a maximum period of 3 months from the date o f preparation.
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7.5 Calibration Standards 7.5.1 lOOpg/mL each PFOS, PFOSA, and POAA stock standard solutions-- Weigh out lOmg o f analytical standard (corrected for percent salt and purity) and dilute to lOOmL with methanol in a lOOmL volumetric flask. Transfer to a 125mL LDPE bottle. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 42C for a maximum period o f 6 months from the date of preparation. 7.5.2 ljxg/mL Working Standard-- Add l.OmL each o f the lOOpg/mL SS solutions (from 7.5.1) to a lOOmL volumetric flask and bring up to volume with methanol. 7.5.3 O.lpg/mL Working Standard -- Add lO.OmL o f the 1.Opg/mL mixed solution (from 7.5.2) to a lOOmL volumetric flask and bring up to volume with methanol. 7.5.4 O.Olpg/mL Working Standard -- Add lO.OmL o f the 0. lpg/m L mixed solution (from 7.5.3) to a lOOmL volumetric flask and bring up to volume with methanol. 7.5.5 Storage Conditions-- Store all WSs in a refrigerator (in 125mL LDPE bottles) at 42C for a maximum period of 3 months from the date o f preparation. 7.5.6 Calibration Standard-- Prepare a minimum o f five calibration solutions in filtered type I water according to the following table:
Concentration Volume of Final Calibration Standard
of WS, pg/mL WS, pL
Volume, mL
0.0 0
40
0.010
100
40
0.010
200
40
0.010
400
40
0.10 100
40
0.10 200
40
0.10 300
40
0.10 400
40
1 May be prepared to extend the range beyond 500Pg/mL. 2 May be prepared to extend the range beyond 750Pg/mL.
Final Concentration of Calibration Standard, Pg/mL
0 25 50 100 250 500 7501 10002
Note: The absolute volum es o f the standards m ay be varied b y the analyst as long as the correct
proportions o f solute to solvent are maintained.
7.5.7 The standards are processed through the extraction procedure (Section 9.0), identical to the laboratory samples. The extracted concentration o f the calibration standard is equal to 8X the initial concentration, due to the concentration o f the standard during the extraction process.
7.5.8 Storage Conditions-- Store all extracted calibration standards in 15mL polypropylene tubes at 42C, for a maximum period o f two weeks from the date of preparation.
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8.0 Sam ple Co llec tio n , Preservation and Storage___________________________
Note: Sampling equipment, including automatic samplers, must be free o f Teflon tubing, gaskets,
and other parts that m ay leach interfering analytes into the water sample. Autom atic samplers that composite samples over time should use refrigerated polypropylene sample containers i f possible. Sample bottles should not be rinsed before sample collection.
8.1 Tap Water-- Open the tap and allow the system to flush until the water temperature (1510C) has stabilized (usually about two minutes). Adjust the flow to about 500mL/min and collect samples from the flowing stream.
8.2 Ground Water-- Purge the well o f standing water using a pump or a bailer. Collect the sample directly from the pump or from the bailer.
8.3 Surface Water-- When sampling from an open body o f water, fill the sample container with water from a representative area.
8.4 Sample Dechlorination--All samples should be iced or refrigerated at 42C and kept in the dark from the time o f collection until extraction. Residual chlorine should be reduced by adding 200pL of a 250mg/mL sodium thiosulfate solution to each water sample, FB, and FSCS (which may be placed in each bottle before leaving for the sampling site.).
8.5 Holding Time (HT)-- Results o f the time/storage study of all target analytes showed that the three compounds are stable for 14 days in water samples when the samples are dechlorinated and stored as described in section 8.4 (see also reference 18.3). Therefore, laboratory samples must be extracted within 14 days and the extracts analyzed within 30 days o f sample collection. If the HT exceeds 14 days, great care is used when evaluating field spikes to avoid misrepresentation o f the sample concentration.
8.6 Field Blanks 8.6.1 Process a Field Blank Control Sample (FB) along with each sample set (samples collected from the same general sample site at approximately the same time). At the laboratory, prior to sample collection, fill a sample container with filtered type I water, seal, and ship the FB to the sampling site along with the empty sample containers. Return the FB to the laboratory with the filled sample bottles. 8.6.2 When sodium thiosulfate is added to samples, use the same procedure to preserve the FB.
8.7 Field Duplicates 8.7.1 Collect a Field Duplicate (FD) for every ten (10) samples collected or per each sampling set, if less than 10 samples are collected. 8.7.2 Separate FDs must be collected for each type of water sample (ground, tap, etc.) collected. 8.7.3 Collect the FD immediately after the sample. 8.7.4 Preserve, store and ship FD using the same procedures as used for the samples.
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8.8 Field Spike Control Sample (FSCS) 8.8.1 A Field Spike Control Sample (FSCS) must be prepared for each sample shipment. If multiple coolers are used to ship a set o f samples, each cooler must contain a FSCS. 8.8.2 At the laboratory, fill a sample container with 1OOmL o f type I water. Seal and ship to the sampling site along with the empty sample containers and FBs. 8.8.3 When sodium thiosulfate is added to samples, use the same procedure to add the same amounts to the FSCS. 8.8.4 Seal and gently invert the FSCS to mix. Store and ship the FSCS using the same procedures as used for the samples.
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9.0 Extraction Procedure
9.1 E xtraction Scheme 9.1.1 Allow samples to equilibrate to room temperature. Thoroughly mix samples by gently inverting the sample bottle. 9.1.2 Measure 40mL o f sample into 50mL polypropylene centrifuge tubes (Spike the QC and Matrix spikes as required*, replace lid and mix well).
Note: * Samples m ay need to be prescreened to determine an appropriate matrix spike level (typically 50 -150 % o f sample concentration).
9.1.3 Condition the C 18 SPE cartridges (lg , 6mL) by passing lOmL methanol followed by 5mL filtered type I water (~2drop/sec). Do not let column run dry.
Note: For the follow ing steps, maintain a ~ldrop/sec flow rate. D o not allow the column to run dry at any time.
9.1.4 Load the analytical sample onto the C,8 SPE cartridge. Discard eluate. 9.1.5 Wash with ~5mL 40% methanol in water. Discard eluate. 9.1.6 Elute with ~5mL 100% methanol. Collect 5mL o f eluate into graduated 15mL
polypropylene centrifuge tubes. This is the target elution fraction (final volume = 5mL). 9.1.7 Analyze a portion o f the target elution fraction eluent using negative electrospray HPLC/MS/MS (Section 10.2).
N ote: Samples are concentrated by a factor o f eight during the extraction; Initial V o l = 40m L -> Final Vol. = 5mL.
9.1.8 Samples are stable at room temperature for at least 24 hours. Analytical samples may be stored in a refrigerator at 42C until analysis.
9.1.9 Standardization of Ci8SPE columns-- If poor recoveries are observed, it may be necessary to standardize the C 18SPE columns in the following manner before analyzing samples.
9.1.9.1 Use a standard with an analyte concentration between 1000 and 4000 Pg/mL. Follow the extraction scheme as outlined from steps 9.1.1 to 9.1.6, except, collect the eluate fraction separately (approx. 5mL), as well as the target elution fraction.
9.1.9.2 After step 9.1.6, collect a post-elution fraction by, eluting with an additional 5mL of 100% methanol.
9.1.9.3 Analyze all three fractions by HPLC/MS/MS. If the target fraction contains a minimum of 85% of the respective analytes, it may be considered acceptable.
9.1.9A If the wash contains significant standard (>15%), either the wash volume or percentage of MeOH should be decreased.
9.1.9.5 If the post-elution fraction contains significant standard (>15%), the target elution volume should be increased.
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10.0 10.1
Calibration and Standardization (Analytical Setup)_____________________
Note: Other instruments m ay be used and the equipment and conditions m ay be very different as
long as the method criteria are met. The operator must optimize and document the equipment and settings used.
Establish the LC/MS/MS system and operating conditions equivalent to the following: Mass Spec: Micromass Quattro Ultima (Micromass) Interface: Electrospray (Micromass) Mode: Electrospray Negative, Multiple Response Monitoring (MRM) Harvard infusion pump (Harvard Instruments), for tuning Computer: COMPAQ Professional Workstation AP200 Software: Windows NT, MassLynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven
Note: A 4 x 10mm Hypercarb drop-in guard cartridge (Keystone, part # 844017-400) is attached
on-line after the purge valve and before the sample injector port to trap any residue contaminants that m ay be in the m obile phase and/or H PL C system.
HPLC Column: Genesis Cg(Jones Chromatography), 2.1mm x 50mm, 4pm Column Temperature: 35C Injection Volume: 15pL Mobile Phase (A): 2mM Ammonium Acetate in filtered type I water (See 7.3.1) Mobile Phase (B): Methanol
HPLC Gradient Program:
Time, Percent Mobile Percent Mobile
min Phase A
Phase B
0.0 60
40
0.4 60
40
1.0 10
90
7.0 10
90
7.5 0
100
9.0 0
100
9.5 60
40
13.5 60
40
14.0 60
40
Flow Rate, mL/min
0.3 0.3 0.3 0.3 0.3 0.4 0.4 0.4 0.3
Note: Other H PLC gradients m ay be used as long as the method criteria are met.
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It may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g. 2.1mm x 30mm) and columns from different manufacturers (Keystone Betasil C18etc.) may be used.
Ions Monitored: Analyte Primary Ion Product Ion
POAA PFOS PFOSA
413.0 499.0 498.0
169.0 99.0 78.0
Approximate Retention Time
5.0 5.2 5.8
Other product ions may be chosen at the discretion o f the analyst, although m/z 99 is suggested for PFOS. Use of the suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch o f mobile phase etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run.
10.2 Tune File Parameters 10.2.1 The following values are provided as an example. Actual values may vary from instrument to instrument. Also, these values may be changed from time to time in order to optimize for greatest sensitivity.
Analyte POAA PFOS PFOSA
Dwell, sec 0.2-0.4 0.2-0.4 0.2-0.4
Collision Energy, eV
10-25 30-60 20-50
Cone, V 20-30 50-80 30-60
Source Capillary Hexapole 1 Aperture 1 Hexapole 2 Source Block Temp. Desolvation Temp.
Set 2.56-3.5kV
0.5V 0.2V 0.8V 100-150C 250--400C
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Analyzer LM Res 1 HM Res 1 Energy 1 Entrance
Exit LM Res 2 HM Res 2 IEnergy 2 Multiplier
Set
12.0-15.0V 12.0-15.0V
0.7V -2V IV
11.ov 11.ov
1.0V 650V
Gas Flows Cone Gas Desolvation
Set 150L/hr 700L/hr
Pressures Gas Cell
Set 3.0e-3m bar
11.0 Analytical Quality Control_________________________________________
11.1 Analytical results o f the FB, FMS, FD, and FSCS should be evaluated at the conclusion of the study to help interpret the data quality o f samples data. Analytical results for these control/duplicate samples must be reported with the sample data.
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D eterm ination o f PFO S, PFO SA , PO A A in Water by Liquid-Solid Extraction and LC /M S/M S
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12.0 A n a l y t i c a l P r o c e d u r e
12.1 Sample Analysis 12.1.1 Set up analysis sample queue. 12.1.2 Inject the same aliquot (between 5-25pL) o f each standard, analytical sample, recovery, control etc. into the LC/MS/MS system. 12.1.3 All samples showing a response for one or more analytes above the response of the highest, active calibration curve level must be diluted and reanalyzed.
12.2 Calibration Curve 12.2.1 Starting with the standard o f lowest concentration, inject the same size aliquot (between 10-25pL) of each extracted calibration standard according to Section 12.1 and tabulate the response (peak height or area) versus the concentration in the standard. Use linear standard curves for quantitation generated for each analyte by linear regression with 1/x weighting o f peak area versus calibration standard concentration. The correlation coefficient (r) for the calibration curves must be >0.990 (^>0.980). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation and the standards reanalyzed. 12.2.2 Curve-- The measured value for each curve point must be within 30% of theoretical values when curve is evaluated over a range appropriate to the data. High or low points may be deactivated to achieve these criteria, but an acceptable curve must contain at least five active curve points. 12.2.3 Continuing Curve Verification (CCV)-- Mid- and low-level calibration checks should be analyzed every 5-10 injections. The analyte level measured in the CCVs should be within 30% o f theoretical values. If CCVs fall outside of this range, data collected subsequent to the last passing CCV should not be used. Only data collected between acceptable CCVs or the initial curve can be used.
13.0 Data Analysis and Calculations
13.1 Calculate the analytical sample (extract) concentration from the standard curve using the following equation: Extract Concentration, pg/mL= (Peak area - intercept) (slope)
13.2 Calculate the percent recovery o f the FSCS using the following equation:
(FSCS cone., Pg/mL ) 1QQ FSCS % ree.
(Cone, added, Pg/mL)
13.3 Calculate the percent recovery o f the MSs using the following equation:
MS%rec. (M Sconc.,Pg/mL-SampleConc,, Pg/mL) ; ^ (Cone, added, Pg/mL)
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D eterm ination o f PFO S, PFO SA , P O A A in W ater by Liquid-Solid Extraction and LC /M S/M S
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14.0 Method Performance Parameters
N ote: A n y method performance parameters that are not achieved must be considered in the evaluation o f the data. Nonconformance to any specified parameters must be described and discussed in any reporting o f the data.
14.1 Linearity-- Linear standard curves for quantitation generated for each analyte by linear regression with 1/x weighting o f peak area versus calibration standard concentration. The correlation coefficient (r) for the calibration curves must be >0.990 (^>0.980).
14.2 Calibration Curve Standards-- The measured value for each curve point must be within 30% o f theoretical values when curve is evaluated over a range appropriate to the data. High or low points may be deactivated to achieve these criteria, but an acceptable curve must contain at least five active curve points.
14.3 CCV Performance-- Mid and low level calibration checks to be analyzed every 5-10 injections. The analyte level measured in the CCVs should be within 30% of theoretical values. If CCVs fall outside o f this range, data collected subsequent to the last passing CCV should not be used. Only data collected between acceptable CCVs can be used.
14.4 Limit of Detection (LOD)-- The lowest calibration standard with a peak area at least 2X the peak area of the extraction blank that can be measured at a concentration greater than zero.
14.5 Limits of Quantitation (LOQ)-- The lower LOQ (LLOQ) is the lowest non-zero active standard in the calibration curve; the peak area o f the LLOQ must be at least 2X that o f the extraction blank. By definition, the measured value of the LLOQ must be within 30% o f the theoretical value.
14.6 Matrix Spikes-- Matrix spike percent recoveries must be within 30% o f the spiked concentration.
14.7 Solvent Blanks, Method Blanks, and Matrix Blanks-- Values must be below the lowest non-zero active standard in the calibration curve. Matrix blanks are considered compliant if no test substance is detected above the LOD for that analyte.
14.8 Reproducibility-- Reproducibility o f the method is defined by the results o f the matrix spikes and matrix spike duplicates. The MS/MSD should be reproducible to within 20%.
14.9 Use of Confirmatory Methods--None
14.10 Demonstration of Specificity-- Specificity is demonstrated by chromatographic retention time (within 3% of standard) and the mass spectral response o f unique product ions generated from a characteristic primary ion.
14.11 Documentation 14.11.1 If criteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as determined by the analyst. Document all actions in the appropriate logbook. 14.11.2 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text o f the report.
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Determination o f PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LC/MS/MS
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15.0 Pollution Prevention and Waste Management
15.1 Sample extract waste and flammable solvent is discarded in high BTU containers, and glass pipette waste is discarded in broken glass containers located in the laboratory.
16.0 Records________________________________________________________________
16.1 Each page generated for a study must have the following information included, either in the header or hand-written on the page: study or project number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst.
16.2 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument run log.
16.3 Plot the calibration curves as described in this method, then print these graphs and store in the study folder.
16.4 Print data integration summary, integration method, and chromatograms, from MassLynx, and store in the study folder.
16.5 Summarize data using suitable software (MS Excel 97) and store in the study folder.
16.6 Back up electronic data to appropriate medium. Record in study notebook the file name and location o f backup electronic data.
17.0 Attachments_________________________________________________________________ 17.1 Attachment A: Figures-- Fluorochemical Compounds
18.0 References_____________________________________________________________
18.1 "Guidelines and Format for Methods to be Proposed at 40 CFR Part 136 or Part 141", U.S. Environmental Protection Agency, Office o f Science and Technology Office o f Water, Washington, D.C. Draft 1996.
18.2 "Method o f Analysis for the Determination o f Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc., State College, Pennsylvania, January 2000.
18.3 Validation report for the "Method o f Analysis for the Determination o f Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in W ater", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc., State College, Pennsylvania, (Approval pending)
19.0 Revisions___________________________________________________________
Revision Number.
Reason For Revision
Revision Date
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Figure 1: PFOS
Chemical Name Molecular ion
= =
0
Perfluorooctane sulfonate 499 (CgF |7S 0 3~)
CgFi7S-0-
0
PFOS
N ote: Standards are made from the salt, potassium perfluorooctane sulfonate [C 8F 17S 0 3K ], m/w 538.
Figure 2: PFOSA
Chemical Name Molecular ion
= =
Perfluorooctane sulfonylamide 498 (C8F 17S 0 2NH2)
o
C8F17S----- nh2
o
PFOSA
Figure 3: POAA
Chemical Name Molecular ion
= =
o
Perfluorooctanoate 413 (C7F 15COO~)
C7F15CO POAA
Note: Standards are made from the salt, ammonium perfluorooctanoate [C 7F i5C O O N H 4], m/w 431
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0 0 0 1 1 3Determination o f PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LC/MS/MS
Compound-Specific, Quantitative Characterization of Organic Fluorochemicais in Biological Matrices
K risten J. H ansen*; Lisa A. Clem en; M ark E. Ellefson; H arold O. Johnson ^C o rresp o n d in g author em ail: kihansen@ m m m .com : phone 651-778-6018; fax 651-778-6176) 3M Environm ental Laboratory Building 2-3E-09 P.O . B ox 33331 St. Paul, M N 55133-3331
Abstract
Since the early 1980s, there has been a steady increase in the use o f nonvolatile fluorinated organic compounds for a variety o f industrial applications. The industrial use of these relatively stable compounds has initiated debate over the fate of fluorochemicais in the environment and, ultimately, the bioavailability o f these compounds (1,2). Until recently, levels o f organic fluorochemicais in biological matrices have been determined by non-chemical specific analytical methods such as total fluoride analysis (2-6). In this manuscript, we present a compound-specific method for the extraction o f extremely low levels of several commercial organic fluorochemicais from sera and liver with quantitative detection by negative ion electrospray tandem mass spectrometry. This technique represents a robust, previously undescribed approach to quantifying specific organic fluorochemicais in biological matrices. This method should prove useful in future studies designed to determine the levels of organic fluorochemicais in humans and the environment. Results from a study o f 65 human sera samples purchased from biological supply companies and the details of the analytical method for the quantitative analysis of specific organic fluorine containing compounds are described.
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Introduction The unique chemical/physical properties of fluorine make fluorinated organic compounds useful for many commercial applications, and industrial production o f these compounds has increased significantly since the early 1980s. Fluorinated organics are used as refrigerants, surfactants, and polymers and as components of pharmaceuticals, fire retardants, lubricants, and insecticides (1). Fluorochemical compounds that are not perfluorinated may be susceptible to partial chemical breakdown at functional group bonds (7). However, given the energy o f the carbon-fluorine bond, it is expected that many organic fluorochemical compounds will be resistant to hydrolysis, photolysis, biodegradation, or metabolism (8). For example, even in the high-energy environment o f the stratosphere, the carbon-fluorine bonds in chlorofluorocarbons are exceptionally stable (9).
In 1974, Guy et al. reported results for the determination o f organic fluorine levels in plasma from 106 individuals from five cities in the United States (2). These researchers demonstrated that although levels of inorganic fluorine in human plasma could be correlated to fluoride levels in drinking water, organic fluorine levels showed no such correlation. Guy et al. showed that the organic fluorine levels measured from samples collected within a particular city were, basically, log normally distributed with few outliers. The average organic fluorine level in human plasma samples included in the study was reported to be 1.35 0.85 micromolar R-F (approximately 26 ppb organic fluorine). By concentrating the organic fluorine from 20 liters of plasma and performing nuclear magnetic resonance (NMR) analysis, Guy et al. postulated that the
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perfluorooctanoate anion (PFOA) or a structurally related compound may be the source of the organic fluorine. Further, they suggested that there may be three or more different components of organic fluorine in plasma samples collected from the general population. Although the source of the organic fluorine in general population blood has been debated and never definitively determined, some have postulated that contamination of the environment with industrial fluorochemicals is the source o f the organic fluorine compounds (4). Others suggest that the organic fluorine is likely to have a natural source (10). Despite the route of exposure, researchers agree that such low levels of organic fluorochemicals are unlikely to cause toxic effects (2,11,12).
A large number of studies in both humans and animals have been conducted to study the toxicity associated with PFOA. In these studies, when determinations of the PFOA levels in tissues were necessary, a total organic fluorine method was employed or a study using radiolabeled material was designed, because easy, sensitive, compound-specific methods have not been available (11-17). Historically, low-level detection o f fluorochemicals such as PFOA and perfluorooctanesulfonate (PFOS) has been limited to relatively insensitive or non-mass-specific detection methods, such as gas chromatography-flame ionization detection, gas chromatography-electron capture detection and high performance liquid chromatography (HPLC)-ultraviolet detection (18-20).
In the work presented here, a new method for the analysis o f several low-level fluorinated organic compounds in sera and liver tissue is described. After initial extraction o f the tissue with an ion-pairing reagent, extracts are analyzed with HPLC-negative ion electrospray tandem mass spectrometry (HPLC-ESMSMS). The ability to select a unique
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product ion upon fragmentation of the molecular ion provides a very selective analysis that is not as likely to be affected by biological interferences.
Detection limits and the linear range of the method were determined for four fluorinated organic compounds [PFOA, PFOS, perfluorooctanesulfonylamide (PFOSA), and perfluorhexanesulfonate (PFFIS)] in both liver and sera by spiking each matrix with standard material and quantitatively recovering the compounds. Although detection limits can be improved by concentrating sample extracts, extraction o f non-concentrated sera produced detection limits for all target analytes o f 1-3 ppb.
The method presented here has been used to quantitatively analyze four organic fluorochemicals in 65 human sera samples collected from several biological supply companies in the United States. High-resolution time-of-flight mass spectrometry was used to confirm the identity of PFOS, PFOA, PFHS, and PFOSA extracted from a single representative sera sample.
Experimental Materials and Methods Rabbit and rat sera were purchased from Sigma (St. Louis, MO). HPLC-grade methyltert-butyl-ether (MTBE) and methanol were purchased from E.M. Science (Gibbstown, NJ); the tetra-butyl ammonium (TBA) hydrogen sulfate was purchased from Kodak (Rochester, NY); the pH o f the TBA solution was adjusted with sodium hydroxide (J.T. Baker; Phillipsburg, NJ). Before use, water was purified with a Milli-Q system (Millipore; Bedford, MA). Human sera samples were purchased from the following biological supply companies: Sigma (St. Louis, MO); Golden West Biologicals
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(Temecila, CA); Biological Specialty Corporation (Colmar, PA); and Lampire Biological Laboratories (Pipersville, PA).
New Zealand White [Hra:(NZW)SPF] rabbit liver was obtained from Covance Laboratories, Inc., in Madison, WI.; Sprague Dawley rats were purchased from Harlan (Indianapolis, IN), and rat liver samples were harvested by 3M Toxicology personnel (St. Paul, MN).
The PFOS and PFOA used as standards and as matrix spikes were purchased from Fluka (Milwaukee, WI); standards of PFHS and PFOSA were made available from 3M Company (St. Paul, MN). The internal standard, 1H,1H,2H,2H perfluorooctane sulfonate (THPFOS), was purchased from ICN (Costa Mesa, CA). Extraction Procedure: One half mL o f sera, 5 pL o f internal standard, 1 mL o f 0.5 M TBA solution (adjusted to pH 10), and 2 mL o f 0.25 M sodium carbonate buffer were added to a 15-mL polypropylene tube for extraction. After thorough mixing, 5 mL of MTBE was added to the solution, and the mixture was shaken for 20 minutes. The organic and aqueous layers were separated by centrifugation, and an exact volume of MTBE (4.0 mL) was removed from the solution. The aqueous mixture was rinsed with MTBE and separated twice more; all rinses were combined in a second polypropylene tube. The solvent was allowed to evaporate under nitrogen before being reconstituted in 0.5 mL o f methanol. The sample was vortex mixed for 30 seconds and passed through a 0.2 pm nylon mesh filter into an autovial. Depending upon the species o f test animal, the serum extract was typically either colorless or light yellow.
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For the extraction of liver samples, a liver homogenate of 1 gram of liver to 5 mL of Milli-Q water was prepared. One mL o f the homogenate was added to a polypropylene tube, and the sample was extracted according to the procedure for sera (described above).
Teflon or glass containers were avoided in this procedure; the former may cause analytical interferences, and the latter may bind the surfactants in an aqueous solution. Disposable polypropylene or plastic lab wear was used to minimize the possibility of sample contamination that can occur when glassware is reused. Any glassware used in the preparation of the reagents was thoroughly rinsed with methanol prior to use.
To ensure that target analytes were not introduced to the matrix prior to extraction, blood collection supplies were extracted and analyzed. Blood bags were purchased from Baxter (Deerfield, IL), and five different types of Vacutainers (two labeled "gel and clot activator," two labeled "K3EDTA," and one labeled "no activator") were purchased from Becton Dickinson (Franklin Lakes, NJ). Additional blood collection materials tested consisted o f 3-cc and 10-cc syringes, 19G1 1/2 Precision Guide sterile needles, multiple sample Vacutainer sterile needles, and Terumo winged infusion sets, all o f which were obtained from Becton Dickinson.
The inside surfaces of all blood collection supplies were exposed to methanol (from 0.5 mL to 80 mL, depending on the particular supply) for 1 hour. The extraction solvent was dried and reconstituted to exactly 1 mL of methanol. A second set of samples was
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spiked with analyte and extracted in exactly the same way as the first set to ensure that analyte could be recovered.
Extraction blanks were prepared using Milli-Q water, and matrix blanks were prepared from rabbit or rat tissue spiked with THPFOS.
Analyte separation was performed using a Hewlett-Packard H P1100 liquid chromatograph modified with low dead-volume internal tubing. Prior to the autosampler, a 1 cm Hypercarb cartridge from Keystone (Bellefonte, PA) was added. Ten pLs of extract were injected onto a 50 x 2mm (5 pm) Keystone Betasil Cjg column with a 2 mM ammonium acetate/methanol mobile phase starting at 45% methanol. At a flow rate of 300 pL/minute, the gradient increased to 90% methanol before reverting to original conditions at 9 minutes. Column temperature was maintained at 25 C.
For quantitative determination, the HPLC system was interfaced to a Micromass (Beverly, MA) Quattro II atmospheric pressure ionization tandem mass spectrometer operated in the electrospray negative mode. Instrumental parameters were optimized to transmit the [M-H] ion for all analytes. When possible, multiple daughter ions were monitored, but quantitation was based on a single product ion. Refer to Table 2 for a summary o f transitions monitored.
In all cases, the capillary was held between 1.6- 3.2 kV. For PFOA determination, the quantitation ion (m/z=169) corresponds to C5F9'; the product ion m/z=99 corresponds to
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FSO3' for quantitative determination o f PFOS. Quantitation o f PFOSA occurs at m/z=78, corresponding to SO2N'; quantitation o f PFHS occurs at m/z=80 (SO3").
In the ESMSMS system, the 499 Da.-> 80 Da. transition can provide a stronger signal than the 499 Da.-> 99 Da. transition of the PFOS analysis. However, in the analysis of tissue samples collected from some species of animals, an unidentified interferent was present in the 499 Da.-> 80 Da. transition. Although this interferent was rarely observed, to ensure complete selectivity, quantitation was based on the 499 Da. -> 99 Da. transition.
Exact mass determination was achieved by interfacing the chromatographic system to either a Micromass LCT; product ion spectra were collected with a Micromass Q-TOF. Both the LCT and Q-TOF are high-resolution time-of-flight mass spectrometers. An 800 ng/mL solution o f raffinose in 50/50 ACN/water was infused into the source at 20 pL/hr as a lock mass (503.1612 Da). PFOS, PFOSA, and PFHS were measured at a cone voltage o f 70 V; PFOA was measured at a 10-V cone voltage. For analysis o f all analytes, the capillary was maintained at 3200 V.
Results and Discussion Characterization of the M ethod A series of experiments, described in more detail below, was designed to characterize the analytical method. In general, all curves, extracted or unextracted, were plotted using linear regression, weighted 1/X. Tables 3 and 4 show the extraction efficiency, limit o f detection, and linear range for the target analytes.
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With the exception of PFOA, the extraction efficiency was determined by extracting and analyzing six replicate rat or rabbit sera samples spiked at approximately the following levels: 10 ng/mL, 50 ng/mL, 100 ng/mL, and 500 ng/mL. For PFOA, only the three higher levels were used for extraction efficiency calculations. Extraction efficiency in liver was determined by extracting samples spiked at 50 ng/g, 100 ng/g, and 500 ng/g. For both sera and liver, the extracted samples were evaluated versus the average curve produced by two unextracted solvent curves analyzed before and after the extracts. The extraction efficiency for PFHS and PFOSA from liver was determined to be significantly lower than those determined for the PFOS and POAA. However, because sample analysis is conducted using extracted curves, the relatively low recoveries should not affect the results. Table 3 shows the compiled average for all spike levels along with the standard deviation.
For both sera and liver analyses, the internal standard was used for quantitative determination o f PFOS and PFOA, only.
The limit o f detection was determined as per EPA Regulation 40 CFR part 136, Appendix B. For each analyte, seven low-level spikes were prepared and analyzed. Based on the standard deviation associated with the replicate analysis, a limit o f detection was calculated. This calculated limit of detection was verified by analyzing a sample that was spiked at that level and extracted.
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The linear range was determined by analyzing duplicate curves extracted from each matrix over a wide range. Starting with the highest standard, points were removed from the curve until the correlation coefficient for the 1/x weighted fit was greater than 0.99. For the sera curves, all points except for the lowest standard level were evaluated to be within 20% of the expected value. For the standard curves extracted from liver, all points except the lowest point were within 30.
Characterization of Blanks Method blanks were prepared from Milli-Q water. Because analyte-free (less than 1 ng/mL) human sera matrix could not be located, surrogate matrix blanks were prepared from rabbit sera. None o f the analytes were detected in either set of blanks. Instrument blanks, consisting of HPLC-grade methanol, were analyzed after high-level-standardcurve points and after periodic calibration checks, to monitor potential carry-over. No carry-over was observed. Methanol extracts of blood collection supplies were analyzed; none of the target analytes were detected in these extracts. In addition, the Teflon cap liners o f glass jars used for reagent storage were extracted with methanol. Low-levels of PFOS and PFOA were detected in some o f the extracts of the Teflon liners. These materials were removed from the extraction procedure.
Figure 1 compares the results of the multiple response monitoring (MRM) analysis for PFOS in an extraction blank, in unspiked rabbit sera, and in unspiked, commercially available human sera.
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Identification of Target Analytes The retention times of the analytes extracted from human sera were matched to within 2% o f the retention time of standard material spiked into and extracted from rabbit sera. MRM analysis was used for verification of analyte identity. For each analyte except PFOSA, at least two characteristic product ions were monitored, although quantitation was based on the response o f a single product ion. PFOSA was detected at such low levels, only a single product ion could be monitored, even for qualitative purposes. In the human sera samples, for all analytes except PFOSA, the relative abundances o f two or more product ions collected by MRM were confirmed to within 20% of standards as criteria for analyte verification (21).
To further verify the identity o f the detected analytes, a 30-fold concentrated extraction of one sera sample was prepared. This concentrated extract was used for exact mass determination o f all four analytes using high-resolution time-of-flight mass spectrometry. The concentration o f the detected analytes were confirmed to within 5 ppm for all target analytes. Figure 2 shows the results of the PFOS and PFOSA high-resolution analysis. Additionally, using high-resolution time-of-flight mass spectrometry, full product ion spectra were collected for each analyte in the concentrated extract. The product ion spectra for the PFOS identified in human sera is shown in Figure 3.
Quantitation of Target Analytes in H um an Sera Quantitation o f the analytes was based on comparison o f a single product ion peak area to the response o f two standard curves, weighted 1/X, bracketing each sample set. Mid-level calibration checks were analyzed every five to ten samples. Based on the precision
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determined from repeat injections o f the standard curves, results were considered quantitative to 30%. Quantitative results, presented as compound-specific average concentrations in sera are presented in Table 5. In addition to the average analyte concentration, the concentration o f organic fluorine represented by each compound is presented. For example, by weight, PFOS is 65% fluorine; for samples reported here, the average PFOS concentration was determined to be 33 ng/mL o f PFOS. This corresponds to about 22 ng/mL o f organic fluorine.
Added together, the four specific fluorochemicals measured in this small set o f samples account for approximately 31 ng o f organic fluorine per milliliter o f sera. Within experimental error associated with each technique, this value compares closely to the value obtained by Guy et al. (approx. 26 ng/mL) more than 20 years ago (2).
Also in accordance with Guy et al., PFOA has specifically been identified in the sera samples, although not necessarily as the major component. For the 65 samples reported here, PFOS was present at the highest concentration. Each analyte measured was detected in every sample, with the following significant exceptions: PFOSA was not measured above the detection limit in 60 of the 65 samples; PFHS was not detected in one sample.
A combination of extraction and analytical methods that do not require chemical derivitization, use small volumes of samples, and are highly sensitive and mass specific were developed for the low-level analysis o f several fluorinated organic compounds in sera and liver. Using these methods, samples o f human sera collected from biological supply companies were analyzed for four separate fluorochemicals, PFOA, PFOS, PFHS,
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and PFOSA. Taken together, these fluorochemicals account for 31 ng/mL o f organic fluorine in an examination o f 65 human sera samples from biological supply companies, consistent with historical reports of total organic fluorine studies.
Although this study comprises a relatively small sample set, it does suggest the possibility of a more complete characterization of the organic fluorine compounds present in human sera. Additionally, these compound-specific analyses should be paired with a total organic fluorine analysis to determine what fraction of the total organic fluorine present is due to the four fluorochemicals quantified in this study.
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A cknowledgements The authors are grateful to Dr. Andrew Seacat and Deanna Luebker o f 3M Toxicology for supplying rat liver for method development, and to Dr. George Moore for providing standard materials. Dr. Robert Voyksner is acknowledged for his thorough and timely review o f this work.
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References
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2 Guy, W.S., Taves, D.R., and Brey, W.S., Jr. Organic Fluorocompounds in Human Plasma: Prevalence and Characterization. Biochemistry Involving Carbon-Fluorine Bonds, Edition num ber; Publisher; Place of Publication; Y ear; Volume num ber; pp. 117-XXX.
3 Taves, D. Evidence that there are Two Forms o f Fluoride in Human Serum. Nature 1968, 217, 1051.
4 Taves, D.R. Comparison o f `Organic' Fluoride in Human and Nonhuman Serum. Nature 1971, 50, 783.
5 Belisle, J., and Hagen, D.F. Method for the Determination o f the Total Fluorine Content o f Whole Blood, Serum/Plasma, and Other Biological Samples Anal. Biochem. 1978, 87, 545.
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7 7 Hagen, D.F.; Belisle, J.; Johnson, J.D.; Venkateswarlu, V. Characterization o f Fluorinated Metabolites by a Gas Chromatgraphic Helium Microwave Plasma Detector - The Biotransformation o f lH,lH,2H,2H-Perfluorodecanol to Perfluorooctanoate. Anal. Biochem. 1 9 8 1 ,118, 336.
8 Organofluorine Chemistry Principles and Commercial Applications', Banks, R.E.; Smart, B.E.; Tatlow, J.C., Eds.; Pelnum Press: New York, 1994.
9 Atmospheric Chemistry', Finlayson-Pitts, B.J., Pitts, J.N. Jr., Eds; John Wiley & Sons: New York, 1986.
10 Belisle, J. Organic Fluorine in Human Serum: Natural Versus Industrial Sources. Science 1981, 212, 1509.
11 Gilliland, F.D., and Mandel, J.S. Serum Perfluorooctanoic Acid and Hepatic Enzymes, Lipoproteins, and Cholesterol: A Study of Occupationally Exposed Men Am. J. Ind. Med. 1996, 29, 560.
12 Griffith, F.D., and Long, J. E. Animal Toxicity Studies with Ammonium Perfluorooctanoate. Am. Ind. Hyg. Assoc. J. 1980, 41, 576.
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13 Kennedy, J.R., and Gerald, L.; Dermal Toxicity o f Ammonium Perfluorooctanoate. Toxicol. Appl. Pharmacol. 1985, 81, 348.
14 Kennedy, G.L., Jr., Hall, G.T., Brittelli, M.R., and Chen, H.C. Inhalation Toxicity o f Ammonium Perfluorooctanoate. Food Chem. Toxicol. 1986, 24, 1325.
15 Hanhijarvi, H., Ophaug, R.H., and Singer, L. The Sex-related Difference in Perfluorooctanoate Excretion in the Rat. Proceedings o f the Societyfor Experimental Biology and Medicine 1 982,171, 50.
16 Venden Heuvel, J.P., Kuslikis, B.I., Van Rafelghem, M.J., and Peterson, R.E. Tissue Distribution, Metabolism, and Elimination o f Perfluorooctanoic Acid in Male and Female Rats. J. Biochem. Toxicol. 1991, 6, 83.
17 Venden Heuvel, J.P., Davis, J.W., II, Sommers, R., and Peterson, R.E.; Renal Excretion o f Perfluorooctanoic Acid in Male Rats: Inhibitory Effect of Testosterone./. Biochem. Toxicol. 1992, 7, 31.
18 Ohya, T., Kudo, N., Suzuki, E., and Kawashima, Y., Determination o f Perfluorinated Carboxylic Acids in Biological Samples by High-performance Liquid Chromatography. J. Chromatogr. 1998, 720, 1.
19 Belisle, J., and Hagen, D.F. A Method for the Determination o f Perfluorooctanoic Acid in Blood and Other Biological Samples. Anal. Biochem. 1 9 8 0 ,101, 369.
20 Ylinen, M., Hanhijarvi, H., Peura, P., Ramo, O. Quantitative Gas Chromatographic Determination o f PFOA as the Benzyl Ester in Plasma and Urine. Arch. Environ. Contam. Toxicol. 1 9 8 5 ,14, 713.
21 Lily Y.T., Campbell, D.A., Bennett, P.K., and Henion, J. Acceptance Criteria for Ultratrace HPLC-Tandem Mass Spectrometry: Quantitative and Qualitative Determination o f Sulfonylurea Herbicides in Soil, Anal. Chem. 1996, 68, 3397.
00012^
16
C om pound
P r im a r y ion (D a )
P r o d u c t ions (D a )
O ptim a l Com ;
V o l t a g e (V )
O ptim a l C o llisio n
E n e r g y (eV)
PFOA
413 119,169*,219 25
20
PFOS
499 80, 99*, 130 60
45
PFHS
399 80, 99*, 130 60
45
PFOSA
498
78*
60
45
THPFOS
427
80*
60
40
*Product ions were used for quantitation.
Table 2. Summary of Primary Ions, Product Ions, and ESMSMS Conditions
000130
17
A nalyte
PFOA PFOS PFOSA PFHS
E x t r a c t io n E ffic ien c y
Standard D e v ia t io n
101 9 % 93 9 % 95 6 % 85 7 %
L im it o f D etec tio n
1.0 ppb 1.7 ppb 1.5 ppb 2.0 ppb
L in ea r R anch, EXTRACTED
5-1000 ppb 5-1000 ppb 5-1000 ppb 5-1000 ppb
C o r r e l a t io n C o effic ien t
0.998 0.995 0.998 0.998
Table 3. Method Characteristics for the Analysis o f Specific Organic Fluorochemicals in Sera. (All concentrations are expressed as ng/g.)
000131
18
A naivii:
PFOA PFOS PFOSA PFHS
KX'I'RA C TIO N L ffic ifn c y
L im it o r D ftfc tio n
87 12%
5.0 ppb
100 13 %
8.5 ppb
56 11 %
3.5 ppb
71 2 3 %
2.0 ppb
i_____________________________________________
Lim a r R anci f , LXTRACTFI)
10-1000 5-1000 5-1000 ppb 5-1000 ppb
C O R R F I.A I ION C O F I FIC IF N I
0.989 0.991 0.995 0.994
Table 4. Method Characteristics for the Analysis o f Specific Organic Fluorochemicals in Liver. (All concentrations are expressed as ng/g.)
GG013Z
19
A nalyte
PFOS PFOA PFHS PFOSA
A verage C O N C E N T R A T I O N IN
S ira
33 15 6.6 3* 6.4 5* | 1.8 0.3*
Range of C O N C E N T R A T IO N IN
S era
5-85 1-13 1-13 <1-2
C o n c e n t r a t io n of O r g a n ic F lu o r in e
R epresented
22 4.6 3.7 1.1
* Several samples were determined to contain the target analyte below the limit of quantitation, therefore, average concentration is estimated.
Table 5. Concentrations (ng/mL) o f Various Organic Fluorochemicals in Human Sera
000133
20
Figure 2. High Resolution Analysis o f PFOS and PFOSA
000134
Figure 3. Product Ion Spectra for PFOS Endogenous in Human Sera
G G CI35
22
