Document jmLjR2dqe4BDz13DOVVej1p6R
INTERIM REPORT #1 - Analysis of Decatur Surface Water and Sediment Samples
STUDY TITLE Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and
Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, and Clams Using LC/MS/MS for the 3M Decatur Monitoring Program
DATA REQUIREMENTS EPA TSCA Good Laboratory Practice Standards 40 CFR 792
STUDY DIRECTOR Jaisimha Kesari P.E., DEE
Weston Solutions, Inc. 1400 Weston Way
West Chester, PA 19380 Phone: 610-701-3761
INTERIM REPORT COMPLETION DATE March 18, 2008
TESTING FACILITY MPI Research, Inc. 3058 Research Drive
State College, PA 16801 Phone: 814-272-1039
STUDY SPONSOR 3M Company
3M Building 42-02-E-27 St. Paul, MN 55144 Phone: 651-778-5200
PROJECT MPI Research Study Number: 0137.0219 ExyLIMS Protocol Number: P0003268
Total Pages: 107
Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
ExyLIMS Protocol Number P0003268, entitled "Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, and Clams Using LC/MS/MS for the 3M Decatur Monitoring Program," conducted for 3M Company, is being performed in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by MPI Research, Inc.
a Principal Investigator MPI Research, Inc.
/ Jaisimha Kesari P.E., DEE Study Director Weston Solutions, Inc.
tfi/w xvJj Q. Michael A. Sanjdr>iro Sponsor Representative 3M Company
MPI Research
Date Date
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MPI Study N o.: 0137.0219 ExyLIMS Protocol No.: P0003268
QUALITY ASSURANCE STATEMENT
MPI Research's Quality Assurance Unit reviewed ExyLIMS Protocol Number P0003268, entitled, "Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, and Clams Using LC/MS/MS for the 3M Decatur Monitoring Program". All reviewed phases1were inspected for conduct according to MPI Research's Standard Operating Procedures, the Study Protocol, the Study Method, and all applicable Good Laboratory Practice Standards. All findings were reported to the MPI Principal Investigator, Facility Management and to the Study Director.
Phase
3) Raw Data and Draft Report Review
4) Final Report Review
Date Inspected
08/13/07
Date Reported to Date Reported to
Principal
MPI Date Reported to
Investigator Management Studv Director
08/13/07
08/13/07
08/13/07
11/06/07
11/07/07
11/07/07
11/07/07
-i -o 8
Lynann Porter
Date
Quality Assurance Research Group Leader, Quality Assurance Unit
'Note: All in-lab inspections and the protocol review will be documented in the QA statement for the final analytical report at the conclusion of the study. This QA statement involves only the review of the interim report and associated raw data.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
CERTIFICATION OF AUTHENTICITY
This interim report, for ExyLIMS Protocol Number P0003268, is a true and complete representation of the raw data.
Submitted by: MPI Research, Inc. 3058 Research Drive State College, PA 16801 (814) 272-1039
Principal Investigator, MPI:
ren Risfya Manager Analytical MPI Research, Inc.
MPI Research Facility Management: i GL ionel
Kevin Lloyd General Manager, Analytical Sciences MPI Research, Inc.
Study Director, Weston Solutions, Inc.
Jaisimha Kesari P.E., DEE Weston Solutions, Inc.
Sponsor Representative, 3M Company:
03>|l8|o Date
sl
Date
m Date
Michael A. Sar; Director of Regulatory Affairs 3M Company
?k j m Date
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
STUDY IDENTIFICATION
Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, and Clams Using
LC/MS/MS for the 3M Decatur Monitoring Program
PROTOCOL NUMBER: MPI STUDY NUMBER: TYPE OF STUDY: SAMPLE MATRIX: TEST SUBSTANCES:
SPONSOR:
STUDY DIRECTOR:
STUDY MONITOR:
TESTING FACILITY:
ANALYTICAL PHASE TIMETABLE:
P0003268
0137.0219
Residue
Surface Water and Sediment
Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS)
3M Company 3M Building 42-02-E-27 St. Paul, MN 55144
Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380
Michael A. Santoro 3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144
MPI Research, Inc. 3058 Research Drive State College, PA 16801
Study Initiation Date:
06/14/07
Interim Analytical Start Date:
06/19/07
Interim Analytical Termination Date: 07/07/07
Interim Report Completion Date: 03/18/08
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PROJECT PERSONNEL
The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from MPI Research, Inc. were associated with various phases of this interim portion of the study:
Name Karen Risha Christine Edwards Krista Gallant Ellen Dashem Stacey Orso Nancy Saxton Mark Ammerman Eric Edwards
Title Manager Analytical Project Leader, Industrial Analysis Research Chemist Associate 1 Research Chemist Associate 1 Research Chemist Associate 1 Research Chemist Associate 1 Project Leader, Sample Control Sample Custodian 2
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TABLE OF CONTENTS
Page
TITLE PAGE....................................................................................................................... 1 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT..............................2 QUALITY ASSURANCE STATEMENT..........................................................................3 CERTIFICATION OF AUTHENTICITY...........................................................................4 STUDY IDENTIFICATION...............................................................................................5 PROJECT PERSONNEL....................................................................................................6 TABLE OF CONTENTS....................................................................................................7 LIST OF TABLES...............................................................................................................8 LIST OF FIGURES.............................................................................................................9 LIST OF APPENDICES.................................................................................................... 11 1.0 SUMMARY................................................................................................................ 12 2.0 OBJECTIVE............................................................................................................... 12 3.0 INTRODUCTION...................................................................................................... 13 4.0 ANALYTICAL TEST SAMPLES............................................................................. 13 5.0 REFERENCE MATERIAL........................................................................................ 14 6.0 DESCRIPTION OF ANALYTICAL METHOD........................................................ 15
6.1 Extraction Procedure for Surface W ater.................................................................. 15 6.2 Extraction Procedure for Sediment.......................................................................... 16 6.3 Preparation of Standards and Fortification Solutions.............................................. 16 6.4 Chromatography....................................................................................................... 17 6.5 Instrument Sensitivity............................................................................................... 17 6.6 Description of LC/MS/MS Instruments and Operating Conditions.........................17 6.7 Quantitation and Example Calculation.................................................................... 18 7.0 EXPERIMENTAL DESIGN......................................................................................20 8.0 RESULTS...................................................................................................................21 9.0 CONCLUSION...........................................................................................................21 10.0 RETENTION OF DATA AND SAMPLES.............................................................21
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Table I.
LIST OF TABLES Page
Summary of PFBS, PFHS, and PFOS in Surface Water Samples..................23
Table II. Summary of PFBS, PFHS, and PFOS in Sediment Samples.......................... 24
Table III. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Surface Water Samples...............................................................................................25
Table IV. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOSin Sediment Samples..........................................................................................................26
Table V. Total Percent Solids for Sediment Samples.................................................... 30
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Figure 1.
Pase
Typical Non-Extracted Calibration Curve for PFBS in 50:50 Acetonitrile:Water.................................................................................... ....32
Figure 2. Non-Extracted Standards of PFBS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively..................................................... ....33
Figure 3. PFBS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively....................... ....34
Figure 4. Chromatogram Representing a Surface Water Sample Analyzed for PFBS (ExyLIMS ID: C0227212, Data Set: 062007A)..................................35
Figure 5. PFBS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively..................................... ....36
Figure 6. Chromatogram Representing a Sediment Sample Analyzed for PFBS (ExyLIMS ID: C0226986, Data Set: 062507G)............................................37
Figure 7. Typical Non-Extracted Calibration Curve for PFHS in 50:50 Acetonitrile: W ater.................................................................................... .....38
Figure 8. Non-Extracted Standards of PFHS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively..........................................................39
Figure 9. PFHS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively............................40
Figure 10. Chromatogram Representing a Surface Water Sample Analyzed for PFHS (ExyLIMS ID: C0227208, Data Set: 061907A).................................41
Figure 11. PFHS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively..........................................42
Figure 12. Chromatogram Representing a Sediment Sample Analyzed for PFHS (ExyLIMS ID: C0226986, Data Set: 062507G)............................................43
Figure 13. Typical Non-Extracted Calibration Curve for PFOS in 50:50 Acetonitrile: W ater.................................................................................... .....44
Figure 14. Non-Extracted Standards of PFOS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively..........................................................45
Figure 15. PFOS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively............................46
Figure 16. Chromatogram Representing a Surface Water Sample Analyzed for PFOS (ExyLIMS ID: C0227212, Data Set: 062007A).................................47
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LIST OF FIGURES (continued) Page
Figure 17. PFOS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively..........................................48
Figure 18. Chromatogram Representing a Sediment Sample Analyzed for PFOS (ExyLIMS ID: C0226986, Data Set: 062507G)............................................ 49
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LIST OF APPENDICES
Page
Appendix A Study Protocol.......................................................................................... 50
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1.0 SUMMARY
MPI Research, Inc. successfully extracted and analyzed surface water and sediment samples for the determination of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) according to 3M Environmental Laboratory Method ETS-8-012 (V0003400) (Appendix A, pg. 65).
The limit of quantitation (LOQ) for the analytes in the surface water samples are listed in Table I. The nominal LOQ for the method for surface water samples was 0.025 ng/mL. The limit of quantitation (LOQ) for the analytes in the sediment samples are listed in Tables II. The target LOQ for the method for sediment samples was 0.20 ng/g. After evaluation of the reagent blanks (method blanks) used for the analysis, the LOQ was determined. In some cases, the LOQ was raised due to the evaluation. A discussion of the process used to evaluate the reagent blanks can be found in subsection 6.4.
Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS found in the surface water samples are summarized in Table I. Fortification recoveries for PFBS, PFHS, and PFOS in the surface water samples are detailed in Table III. The average percent recoveries standard deviations for PFBS, PFHS, and PFOS in the surface water samples were 118 9%, 102 12% and 95 21%, respectively. Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS found in the sediment samples are summarized in Table II. Fortification recoveries for PFBS, PFHS, and PFOS in the sediment samples are detailed in Table IV. The average percent recoveries standard deviations for PFBS, PFHS, and PFOS in the sediment samples were 102 10%, 102 13% and 101 21%, respectively.
The assessed accuracies for the majority of the samples reported are +/- 30%. The accuracies were assessed for each analyte for each sample by reviewing the matrix spike whose spiking level most closely matches the endogenous concentration found in the sample. Quantitative results were obtained for all samples and analytes except for PFBS in one surface water sample site, PFHS is one sediment sample, and PFOS in one sediment sample that are not reported (NR) due to quality control failures.
Total percent solid results for the sediment samples are detailed in Table V.
2.0 OBJECTIVE
The objective of the analytical part of this study was to determine levels of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) in surface water and sediment according to Protocol P0003268 (Appendix A).
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3.0 INTRODUCTION
This report details the results of the analysis for the determination of PFBS, PFHS and PFOS in surface water and sediment using the 3M Environmental Laboratory analytical method ETS-8-012.1 (V0003400) entitled, "Method of Analysis for the Determination of Perfluorobutanoic Acid (PFBA), Perfluoropentanoic Acid (PFPeA), Perfluorohexanoic Acid (PFHA), Perfluoroheptanoic Acid (PFFlpA), Perfluorooctanoic Acid (PFOA), Perfluorononanoic Acid (PFNA), Perfluorodecanoic Acid (PFDA), Perfluoroundecanoic Acid (PFUnA), Perfluorododecanoic Acid (PFDoA), Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil and Sediment by LC/MS/MS."
The study was initiated on June 14, 2007, when the study director signed protocol number P0003268. The analytical start date for this interim report was June 19, 2007, and the analytical termination date for this interim report was July 7, 2007.
4.0 ANALYTICAL TEST SAMPLES
A total of sixty-four samples (ExyLIMS ID C0226986 - C0227003, C0227200 C0227234, and C0227532 - C0227542, from login ID L00010412), forty surface waters and twenty-four sediments, were received on wet ice on December 20, 2006 from Charles Young at Weston Solutions, Inc. The forty surface water samples represented three rinse blanks, one trip blank, two associated trip blank field spikes, and eight surface water sites with their associated field spikes. All samples were logged in by MPI personnel and placed in refrigerated storage.
Sample identification (ID) codes for the surface water and sediment samples are of the form Dxx-Sx-LOCxxx-x(x)-06012xx and are composed of the strings described below:
The first string defines the sampling area where D indicates the Decatur, Alabama general study area and L3 = LOI Location 3 at Mallard Point Park, L2 = LOI Location 2 at Swan Creek mouth, LI = LOI Location 1 at Fox Creek mouth, MC = Mallard Creek mouth, BC = Bakers Creek mouth, OU = 3M outfall cove, and AA = Avenue A drainage.
The second string defines the sample matrix where SW = surface water and SD = sediment.
The third string indicates the specific sampling location.
The fourth string describes the sample aliquot where 0 = primary sample volume, DB = duplicate sample volume, LS = low spike, MS = mid spike, HS = high spike, and RB = equipment rinseate blank.
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The final string is the sample collection date in YYMMDD format.
Sample log-in and chain of custody information is located in the raw data package associated with this interim report. Storage records will be kept at MPI Research, Inc. (State College).
5.0 REFERENCE MATERIAL
The requisition information, lot, purity, and expiration date for the reference material used in this study is listed below. The reference material was stored refrigerated.
Compound
ExvLIMS Inventory No.
PFBS
SP0008058
PFBS
SP0008956
PFHS
SP0008057
PFHS PFOS
SP0008961 SP0002694
Supplier
3M Environmental
Laboratory 3M
Environmental Laboratory 3M
Environmental Laboratory 3M
Environmental Laboratory Fluka Corporation
Lot# 2
Puritv Expiration Received
(%) Date
Date
97.3 01/18/17 09/06/06
2 97.3 01/18/17 02/19/07
NB 120067-
69 NB 12006769 430180/
1
98.6 02/12/17 98.6 02/12/17 101.2 10/31/07
09/05/06 02/19/07 04/23/03
The molecular structure of the standard is given below:
PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt ^FgSCL'KT)
Transitions Monitored: 299 -->80, 99
Structure:
FF FF
F SO 3
FF FF
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PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (CeFnSCb'K*) Transitions Monitored: 399 - 80, 99 Structure:
FFF FFF
FFF
FFF
3
PFOS Chemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (CgFnSCL'K*)
Transitions Monitored: 499 -->80, 99
Structure:
FFFF FF
F SO3
FFFF FFFF
6.0 DESCRIPTION OF ANALYTICAL METHOD
The 3M Environmental Laboratory analytical method ETS-8-012.1 (V0003400) entitled, "Method of Analysis for the Determination of Perfluorobutanoic Acid (PFBA), Perfluoropentanoic Acid (PFPeA), Perfluorohexanoic Acid (PFHA), Perfluoroheptanoic Acid (PFHpA), Perfluorooctanoic Acid (PFOA), Perfluorononanoic Acid (PFNA), Perfluorodecanoic Acid (PFDA), Perfluoroundecanoic Acid (PFUnA), Perfluorododecanoic Acid (PFDoA), Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil and Sediment by LC/MS/MS" was used for the sample analysis in this study.
6.1 Extraction Procedure for Surface Water
A 10 mL aliquot of the water sample was used for the extraction procedure. The sample was measured into a 50 mL polypropylene centrifuge tube. The appropriate samples were fortified and 10 mL of acetonitrile was added. The samples were capped tightly and shaken. The samples were placed into an ultrasonic bath at room temperature for ~2 hours. The samples were then centrifuged at -3000 rpm for 10 minutes. A portion of the supemate was then transferred to an autosampler vial. Each sample was analyzed by LC/MS/MS electrospray.
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6.2 Extraction Procedure for Sediment
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
A 1 gram aliquot of the soil sample was used for the extraction procedure. The sample was weighed into a 15 mL polypropylene centrifuge tube. The appropriate samples were fortified and 8 mL of 80:20 acetonitrile:water was added. The samples were capped tightly and shaken. The samples were placed into an ultrasonic bath at room temperature for -2 hours. The samples were then centrifuged at -3000 rpm for 10 minutes. A portion of the supemate was then transferred to an autosampler vial. Each sample was analyzed by LC/MS/MS electrospray.
6.3 Preparation of Standards and Fortification Solutions
A stock standard solution was prepared for each analyte as specified in the method. The stock standard solutions were prepared at a concentration of 10,000 pg/mL by dissolving 1.0 g of the appropriate standard (corrected for purity and salt content, if necessary) in acetonitrile. From those solutions, a 1000 pg/mL mixed fortification standard solution was prepared by taking 10 mL of each stock and bringing the volume up to 100 mL with acetonitrile. By taking 10 mL of the 1000 pg/mL mixed fortification standard and bringing the volume up to 100 mL with acetonitrile, a 100 pg/mL mixed fortification standard was prepared. By taking 10 mL of the 100 pg/mL mixed fortification standard and bringing the volume up to 100 mL with acetonitrile, a 10 pg/mL mixed fortification standard was prepared. By taking 10 mL of the 10 pg/mL mixed fortification standard and bringing the volume up to 100 mL with acetonitrile, a 1.0 pg/mL mixed fortification standard was prepared. By taking 10 mL of the 1.0 pg/mL mixed fortification standard and bringing the volume up to 100 mL with acetonitrile, a 0.1 pg/mL mixed fortification standard was prepared. By taking 10 mL of the 0.1 pg/mL mixed fortification standard and bringing the volume up to 100 mL with acetonitrile, a 0.01 pg/mL mixed fortification standard was prepared.
A set of external calibration standards were prepared in 50:50 acetonitrile:water. The following concentrations were prepared:
Cone, of Fort. Solution (ng/mL) 100 100 100 5.0 2.5 1.0 0.5 0.25
Aliquot Volume
(mL) 5.0 2.5 1.0 10 10 10 10 10
Final Volume of
Solution (mL) 100 100 100 100 100 100 100 100
Final Cone, of Calibration Std.
(ng/mL) 5.0 2.5 1.0 0.50 0.25 0.10 0.05 0.025
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The stock standard solutions and the 1000 pg/mL mixed standard solution were stored in a freezer (-20 5C) when not in use. All other fortification and calibration standard solutions were stored in a refrigerator (4 2C) when not in use. Documentation of standard preparation is located in the raw data package associated with this interim report.
6.4 Chromatography
Quantification of the analyte was accomplished by LC/MS/MS electrospray. The retention times of PFBS, PFHS, and PFOS were 3.6 min, 3.8 min, and 4.0 min, respectively. Method blanks prepared for each data set were used to determine the LOQ. In instances where there were no peaks in the method blanks, the LOQ was determined by the concentration of the lowest standard injected in the analytical run that met the 70 130% recovery range of its known value. In instances where there were peaks detected in the method blanks, the blanks were evaluated. If the average of the responses of all the method blanks was less than 50 % of the response of the lowest standard meeting the recovery criteria, then the LOQ was determined by the lowest standard. If the average of the responses of all the method blanks was greater than 50 % of the response of the lowest standard meeting the recovery criteria, then the LOQ was raised to the standard that met the less than 50 % criteria.
6.5 Instrument Sensitivity
The smallest standard amount injected during the chromatographic run had a concentration of 0.0125 ng/mL for the surface water samples, and a concentration of 0.025 ng/mL for the sediment samples.
6.6 Description of LC/MS/MS Instruments and Operating Conditions
Instruments: API 5000 Biomolecular Mass Analyzer
Interface:
SCIEX Turbo Ion Spray Liquid Introduction Interface
Computer:
DELL Precision 360
DELL OptiPlex GX400
Software:
PE SCIEX Analyst 1.4.1
HPLC:
Hewlett Packard (HP) Series 1200
Hewlett Packard (HP) Series 1100
HP Quat Pump
HP Vacuum Degasser
HP Autosampler
HP Column Oven
HPLC Column: Phenomenex Luna C8 (2) Mercury, 2cm x 4 mm, 3pm
Column Temp.: -35 C Injection Voi.: 10 pL Mobile Phase (A): 2 mM Ammonium Acetate in water
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Mobile Phase (B): Methanol
Gradient:
Time (mini
%A
0.0 90
0.5 90
2.0 10
Time (min')
%A
5.0 10
5.1 0
6.0 0
6.1 90
10.0 90
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
%B 10 10 90 %B 90 100 100 10 10
Total run time: ~ 10 min
Flow Rate:
0.75 mL/min
Ions monitored:
Analvte
Mode
PFBS PFHS PFOS
negative negative negative
Transition Monitored 299 -> 80, 99 399 -+ 80, 99 499 -> 80, 99
Retention Time (min)
~3.6 min. ~3.8 min. ~4.0 min.
6.7 Quantitation and Example Calculation
Ten microliters of sample or calibration standard was injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x fit weighted linear regression) by Analyst software using eight or nine concentrations of standards. The concentration was determined from the following equations.
Equation 1 calculated the amount of analyte found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program.
Equation 1:
Analyte found (ng/mL) = (Peak area - intercept) x EDF x PEDF slope
Where: EDF = Extraction Dilution Factor, factor by which the sample volume was diluted during the extraction (EDF =2 for water samples and EDF =1 for soil samples).
PEDF = Post Extraction Dilution Factor, factor by which the final volume was diluted, if necessary.
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For the sediment samples, equation 2 was used to convert the amount of analyte found in ng/mL to ng/g (ppb).
Equation 2:
Analyte found (ppb) = fanalvte found (ng/mL) x volume extracted (8 mL)l sample weight (1 g)
Equation 3 was then used to calculate the amount of analyte found in ppb based on dry weight.
Equation 3:
Analyte found (ppb) dry weight = analyte found (ppb) x [100% / total solids(%)]
NOTE: Total solids (%) = [dry weight (g) / wet weight (g)] x 100%
For samples fortified with known amounts of analyte prior to extraction, Equation 4 was used to calculate the percent recovery.
Equation 4: For water samples: Recovery (%) =
(total analyte found (ng/mL! - average analyte in sample (ng/mLV) xl00% analyte added (ng/mL)
For sediment samples (based on wet weight): Recovery (%) =
(total analyte found (ng/g! - average analyte in sample (ng/g')') xl00% analyte added (ng/g)
An example of a calculation using an actual sample follows:
Sediment sample Exygen ID: C0226989 Spike C (Set: 062507H), fortified at 2.0 ng/g
with PFHS where:
peak area
= 340745
intercept
= 16000
slope
1250000
extraction dilution factor
=1
post extraction dilution factor
=1
ng/g PFHS added (fort level)
= 2.0 ng/g
average amt in corresponding sample = ND (not detected)
total percent solid
= 48.16%
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From equation 1: Analyte found (ng/mL)
= r340745 - 160001 x 1 x 1 1250000
= 0.260 ng/mL
From equation 2: Analyte found, wet weight (ng/g)
= ('0.260 ng/mL x 8 mL)
1g = 2.08 ng/g
From equation 3: Analyte found (ng/g, ppb) dry weight = 2.08 ng/g x [100% / 48.16%]
From equation 4: % Recovery
= 4.32 ng/g
= (2.08 ng/g) x 100% 2.0 ng/g
= 104%
NOTE: Numbers may differ slightly from raw data due to rounding.
7.0 EXPERIMENTAL DESIGN
For water samples designated as field matrix spikes, PFBS, PFHS, and PFOS was added at a known concentration to the bottles in the laboratory before being shipped to the field. The samples were filled to a 200 mL volumetric fill line in the field. For the sediment samples designated as laboratory matrix spikes, PFBS, PFHS, and PFOS were added to the samples after they were aliquotted in the laboratory, before the extraction solvent was added to the samples.
The surface water samples were extracted in three sets. The sets included four reagent blanks (method blanks), three reagent blanks fortified at one lower level and three reagent blanks fortified at one higher level of known concentrations. The first two sets contained three surface water sample sites. The last set contained two sample sites, three equipment rinseate blanks, and one trip blank and associated trip blank field spikes. For each site, a sample, a field duplicate and a range of two to three matrix field spikes were collected and extracted.
The sediment samples were extracted in eight sets. Each set included four control blanks (method blanks), three control blanks fortified at one lower level and three control blanks fortified at one higher level of known concentrations. All eight sets contained three sample sites. For each sample site, a sample, a laboratory replicate, and three laboratory matrix spikes were prepared and extracted.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
8.0 RESULTS
The limit of quantitation (LOQ) for the analytes in the surface water samples are listed in Table I. The nominal LOQ for the method for surface water samples was 0.025 ng/mL. The limit of quantitation (LOQ) for the analytes in the sediment samples are listed in Tables II. The target LOQ for the method for sediment samples was 0.20 ng/g. After evaluation of the reagent blanks (method blanks) used for the analysis, the LOQ was determined. In some cases, the LOQ was raised due to the evaluation. A discussion of the process used to evaluate the reagent blanks can be found in section 6.4 of the report.
Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS found in the surface water samples are summarized in Table I. Fortification recoveries for PFBS, PFHS, and PFOS in the surface water samples are detailed in Table III. The average percent recoveries standard deviations for PFBS, PFHS, and PFOS in the surface water samples were 118 9%, 102 12% and 95 21%, respectively. Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS found in the sediment samples are summarized in Table II. Fortification recoveries for PFBS, PFHS, and PFOS in the sediment samples are detailed in Table IV. The average percent recoveries standard deviations for PFBS, PFHS, and PFOS in the sediment samples were 102 10%, 102 13% and 101 21%, respectively.
The assessed accuracies for the majority of the samples reported are +/- 30%. The accuracies were assessed for each analyte for each sample by reviewing the matrix spike whose spiking level most closely matches the endogenous concentration found in the sample. Quantitative results were obtained for all samples and analytes except for PFBS in one surface water sample site, PFHS is one sediment sample, and PFOS in one sediment sample that are not reported (NR) due to quality control failures.
Total percent solid results for the sediment samples are detailed in Table V.
9.0 CONCLUSION
The surface water and sediment samples were successfully extracted and analyzed for PFBS, PFHS, and PFOS according 3M Environmental Laboratory analytical method ETS-8-012.1.
10.0 RETENTION OF DATA AND SAMPLES
All original paper data generated by MPI Research, Inc. (State College) that pertains to this interim report will be shipped to the study director. This does not include facilityspecific raw data such as instrument or temperature logs. Exact copies of all raw data, as well as a signed copy of the final analytical report and all original facility-specific raw data, will be retained in the MPI Research, Inc. (State College) archives for the period of time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
TABLES
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
Table I. Summary of PFBS, PFHS, and PFOS in Surface Water Samples
ExvoenID
C0227200 C0227201
C0227204 C0227205
C0227208 C0227209
C0227212 C0227213
C0227216 C0227217
C0227220 C0227221
C0227224 C0227225
C0227229
C0227230
C0227231
C0227232
C0227538 C0227539
Client Sample ID
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutaoaaulfonate__________________ Perfluorohaxaneaulfonate___________________ Perfluorooctanaaulfonate
Acceptable Assessed
Acceptable Assessed
Acceptable Assessed
Analyte Found LOQ
Accuracy Analyte Found LOQ
Accuracy Analyte Found LOQ
Accuracy
(ppb, ng/mL) (no/mL)
(+/- %) (ppb, no/mL) (ng/mL)
(+/. %> (ppb. ng/mL) (ng/mL)
(+/- %)
DL3-SW-LOC001-0-061214 DL3-SW-LOC001-DB-061214
ND ND
0.025 0.025
30 30
ND
0.025
30
ND
0.025
30
ND
0.025
30
ND
0.025
30
DL2-SW-LOC001-0-061214 DL2-SW-LOC001-DB-061214
ND ND
0.025 0.025
30 30
ND
0.025
30
ND
0.025
30
ND
0.025
30
ND
0.025
30
DBC-SW-LOC001-0-061214 DBC-SW-LOC001-DB-061214
ND ND
0.025 0.025
30 30
ND
0.025
30
0.0506
0.025
40
ND
0.025
30
0.0554
0.025
40
DOU-SW-LOC001-0-061213 DOU-SW-LOC001-DB-061213
NR NR
1.42
0.025
30
1.41
0.025
30
5.63
0.025
30
6.08
0.025
30
DL1-SW-LOC001-0-061213 DU1-SW-LOC001-DB-061213
0.111A 0.0526A
0.025 0.025
30 30
ND
0.025
30
0.0615
0.025
30
ND
0.025
30
0.0660
0.025
30
DMC-SW-LOC001-0-061213 DMC-SW-LOC001-DB-061213
0.0292 0.0296
0.025 0.025
30 30
ND
0.025
30
0.0499
0.025
30
ND
0.025
30
0.0467
0.025
30
DAA-SW-LOC002-0-061215 DAA-SW- LOC002-DB-061215
0.320 0.307
0.025 0.025
30 30
0.284 0.293
0.025 0.025
30 30
1.59
0.026
30
1.64
0.025
30
DOU-F02-IPF004-RB-061212
ND
0.025
30
ND
0.025
30
ND
0.025
30
DL2-SW-LOC001-RB-061214
ND
0.025
30
ND
0.025
30
ND
0.025
30
DAA-SD-LOC006-R B-061215
ND
0.025
30
ND
0.025
30
ND
0.025
30
Trip Blank
ND
0.025
30
ND
0.025
30
ND
0.025
30
DAA-SW-LOC005-0-061215 DAA-SW- LOC005-DB-061215
13.9 14.4
0.025 0.025
30 30
46.4
0.025
30
46.5
0.025
30
105
0.025
30
110
0.025
30
ND = Not detected at or above the acceptable LOQ. NR = Not reported due to the endogenouse concentration of analyte in sample exceeds the range of applicability of the spike. ARelative Percent Difference >20%
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
Table II. Summary of PFBS, PFHS, and PFOS in Sediment Samples
Exvaen ID
C0226986 C0226986 Rep
C0226987 C0226987 Rep
00226988 C0226988 Rep
C0226989 C0226989 Rep
C0226990 C0226990 Rep
C0226991 C0226991 Rep
C0226992 C0226992 Rep
C0226993 C0226993 Rep
C0226994 C0226994 Rep
C0226995 C0226995 Rep
C0226996 C0226996 Rep
C0226997 C0226997 Rep
C0226998 C0226998 Rep
C0226999 C0226999 Rep
C0227000 C0227000 Rep
C0227001 C0227001 Rep
C0227002 C0227002 Rep
C0227003 C0227003 Rep
C0227532 C0227532 Rep
C0227533 C0227533 Rep
C0227534 C0227534 Rep
C0227535 C0227535 Rep
C0227536 C0227536 Rep
C0227537 C0227537 Rep
Client Sample ID
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutancsulfonatc__________________ PerfluorohexanesuKonate___________________Perfluorooctanesulfonate
Analyte Found Acceptable
(PPb. ng/g) Dry Weiaht
LOQ (ng/g)
Assessed Analyte Found Acceptable
Accuracy (ppb, ng/g)
LOQ
</-%> Dry Weiaht
(ng/g)
Assessed Analyte Found Acceptable
Accuracy (ppb. ng/g)
LOQ
(+/-%)
Dry Weiaht
(ng/g)
Assessed Accuracy
(*/-%>
DL3-SD-LOC001-0-061214 DL3-SD-LOC001 -0-061214*
ND ND
0.20 30 0.20 30
ND 0.20 30 ND 0.20 30
ND 0.20 30 ND 0.20 30
DL3-SD-LOC002-0-061214 DL3-SD-LOC002-0-061214*
ND ND
0.20 30 0.20 30
ND 0.20 30 0.477 0.20 30 ND 0.20 30 0.507 0.20 30
DL3-SD-L00003-0-061214
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL3-SD-LOC003-0-061214*
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC001-0-061214
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC001-0-061214*
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC002-0-061214
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC002-0-061214*
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC003-0-061214
ND
0.20 30
ND
0.20 30
ND
0.20 30
DL2-SD-LOC003-0-061214*
ND
0.20 30
ND
0.20 30
ND
0.20 30
DBC-SD-LOC001-0-061214 DBC-SD-LOC001-0-061214*
ND ND
0.20 30 0.556 0.20 30 0.20 30 0.455 0.20 30
61.2 0.40 30 55.2 0.40 30
DBC-SD-LOC002-0-061214 DBC-SD-LOC002-0-061214*
ND ND
0.20 30
ND1 0.20 30
0.20
30
0.4291
0.20
30
45.6 0.40 30 43.3 0.40 30
DBC-SD-LOC003-0-061214 DBC-SD-LOC003-0-061214*
ND ND
0.20 30 0.20 30
ND 0.20 30 26.9 0.40 50 ND 0.20 30 27.3 0.40 50
DOU-SD-LOC001 -0-061213
5.08
0.20 40
6.81A
0.20
30
488
0.20 30
DOU-SD-LOC001-0-061213*
5.20
0.20 40
3.62A
0.20
30
428
0.20 30
DOU-SD-LOC002-0-061213
0.853
0.20
30
3.51
0.20 50
DOU-SD-LOC002-0-061213*
0.753
0.20
30
4.15
0.20 50
NR NR
-
*
DOU-SD-LOC003-0-061213
0.658
0.20
30
DOU-SD-LOC003-0-061213*
0.536
0.20
30
1.17 0.20 30 1.07 0.20 30
69.7 0.20 30 87.3 0.20 30
DL1-SD-LOC001-0-061213 DL1-SD-LOC001-0-061213*
ND ND
0.20 30 0.20 30
ND 0.20 30 3.74 0.20 30 ND 0.20 30 3.05 0.20 30
DL1-SD-LOC002-0-061213 DL1-SD-LOC002-0-061213*
0.4871 ND'
0.20 0.20
30 30
ND
0.20
30
5.50A
0.20
30
ND
0.20
30
3.75A
0.20
30
DL1-SD-LOC003-0-061213
ND
0.20 30
ND
0.20 30 0.741 0.20 30
DL1-SD-LOC003-0-061213*
ND
0.20 30
ND
0.20 30 0.856 0.20 30
DMC-SD-LOC001 -0-061213 DMC-SD-LOC001-0-061213*
ND ND
0.20 30 0.20 30
ND
0.20
30
1.36A
0.20
30
ND
0.20
30
2.49A
0.20
30
DMC-SD-LOC002-0-061213 DMC-SD-LOC002-0-061213*
ND ND
0.20 30 0.20 30
ND
0.20
30
2.89A
0.20
30
ND
0.20
30
4.50A
0.20
30
DMC-SD-LOC003-0-061213 DMC-SD-LOC003-0-061213*
ND ND
0.20 30 0.20 30
ND
0.20
30
3.05A
0.20
30
ND
0.20
30
4.19A
0.20
30
DAA-SD-LOC006-0-061215
6.10
0.20
30
DAA-SD-LOC006-0-061215*
6.28
0.20
30
NR NR
- 554A 0.20 30 794A 0.20 30
DAA-SD-LOC005-0-061215
3.69
0.20
30
17.8 0.20 30
560
0.20 50
DAA-SD-LOC005-0-061215*
4.09
0.20
30
20.6 0.20 30
651
0.20 50
DAA-SD-LOC004-0-061215
85.6
0.20 30
674
0.20 30 2130 0.20 40
DAA-S D- LOC004-0-061215*
87.4
0.20 30
724
0.20 30
1910 0.20 40
DAA-SD-LOC003-0-061215
3.95
0.20
30
22.7
0.20 30
1290 0.20 30
DAA-SD-LOC003-0-061215*
4.77
0.20 30
26.1
0.20 30
1340 0.20 30
DAA-SD-LOC002-0-061215
0.372
0.20
30
0.540
0.20
30
31.7
0.20 30
DAA-SD-LOC002-0-061215*
0.388
0.20
30
0.511
0.20
30
33.3
0.20 30
DAA-SD-LOC001-0-061215
0.496
0.20
30
0.779
0.20
30
106 0.20 40
DAA-SD-LOC001-0-061215*
0.423
0.20
30
0.655
0.20
30
91.8
0.20 40
'Laboratory Duplicate ARelative Percent Difference > 30% 'Relative Percent Difference was not calculated due to the presence of a nondetect and resulting uncertainty.
ND = Not detected at or above the acceptable LOQ. NR = Not reported due to quality control failures.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
Table III. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Surface Water Samples
S am ple D escription
D L3-S W -LO C 001-LS -061214 (C0227202,0.25 ppb Field Spike)
DL3-SW -LO C 001-H S-061214 (C0227203, 5.0 ppb Field Spike)
Am ount S p ik e d (ng/m L)
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Peril u o r o b u ta n c tu ifo n a te ________ Perfluorohexaneiutfonate__________________________ Perfluorooctaneeuifonate
A m t Found in S am ple
(ng/m L)
Am ount R e cove re d
(ng/m L)
R e c o v e ry (%>
A m t Found in S am ple
(nfl/m L)
Am ount R e cove re d
(ng/m L)
R e cove ry <%)
A m t Found in Sam ple
(ng/m L)
Am ount R e c o v e re d
(ng/m L)
R e co ve ry (% )
0.25
ND
0.280
112
5.0 ND
NA NA
ND 0.226 90 ND NA NA
ND 0.192 77 ND NA NA
DL2-S W -LO C001 -LS -061214
(C0227206,0.25 ppb Field Spike)
0.25
ND
0.269
108
DL2-SW -LO C 001-H S-061214 (C0227207. 5.0 ppb Field Spike)
5.0
ND
NA NA
N D 0.233 93 ND NA NA
ND 0.176 70 ND NA NA
DBC -SW -LO C 001-LS-061214
(C0227210, 0.25 ppb Field Spike)
0.25
ND
0.295
118
D B C -SW -LO C 001-H S-061214 (C0227211. 5.0 ppb Field Spike)
5.0
ND
NA NA
ND
0.284
114
0.0530
0.381
131
ND NA NA 0.0530 NA NA
D O U -S W -LO C 001-LS -061213 (C0227214, 0.25 ppb Field Spike)
D O U -SW -LO C 001-H S-061213 (C0227215. 5.0 ppb Field Spike)
0.25 5.0
NR NR
N R NR 1.41 N A N A 5 .8 5 N A N A
NR
NR
1.41
5.85
89
5.85
11.0
103
D L1-S W -LO C 001-LS -061213 (C0227218,0.25 ppb Field Spike)
DL1-SW -LO C 001-H S-061213 (C0227219, 5.0 ppb Field Spike)
0.25 5.0
0.0819 0.0819
0.346 NA
106 NA
ND
0.306
122
0.0638
0.365
120
ND
NA
NA 0.0638 NA
NA
DM C -SW -LO C 001-LS-061213 (C0227222,0.25 ppb Field Spike)
D M C -SW -LO C 001-H S-061213 (C0227223. 5.0 ppb Field Spike)
0.25 5.0
0.0294 0.0294
0.339 NA
124 NA
ND
0.252
101
0.0483
0.275
91
ND
NA
NA 0.0483 NA
NA
DAA-SW -LO C 002-LS-061215
(C0227226, 0.25 ppb Field Spike)
0.25
0.313
0.634
128
0.288
0.514
90
D AA-S W -LO C 002-M S -061215 (C0227227, 5.0 ppb Field Spike)
5.0
0.313
NA
N A 0.288 N A
NA
DAA-S W -LO C 002-H S -061215 (C0227228,100 ppb Field Spike)
100
0.313
NA
NA 0.288 NA
NA
1.62 1.62 1.62
NA 6.51 NA
NA 98 NA
Trip Blank Low Spike (C0227233. 0.25 ppb Field Spike)
T rip B lank High S pike (C0227234, 5.0 ppb Field Spike)
0.25 5.0
ND ND
0.315 NA
126 NA
ND
0.275
110
ND NA NA
ND 0.182 73 ND NA NA
DAA-S W -LO C 005-LS -061215
(C0227540, 0.25 ppb Field Spike)
0.25
14.1
NA
NA 46.5
NA
NA
108
NA
NA
DAA-S W -LO C 005-M S -061215
(C0227541, 5.0 ppb Field Spike)
5.0
14.1
NA NA 46.5
NA
NA
108
NA
NA
D A A -S W -LO C 005-H S -061215
(C0227542,100 ppb Field Spike)
100
14.1
138 124 46.5 158 112
108
200
92
Average: Standard Deviation:
118 9
Average: Standard Deviation:
102 12
ND = Not detected at or above the acceptable LOQ reported on Table I. NA = Not applicable. This matrix spike concentration was not used to assess the accuracy for this analyte. NR = Not reported due to the endogenouse concentration of analyte in sample exceeds the range of applicability of the spike. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
Average: Standard Deviati
95 21
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
Table IV. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Sediment Samples
Sample Description
DL3-SD-LOC001 -0-061214 (C0226986 Spk C. 2.0 ppb Spike)
DL3-SD-LOC001 -0-061214 (C02269S6 sp k D, 40 ppb Spike)
DL3-S D- LOCOQ1-0-061214 (C0226986 Spk E, 800 ppb Spike)
Amount Spiked
(ng/0)
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutaneeuifonate____________________Perfluorohexaneeulfonate____________________ Perfluoroocteneeuifonate
Amt Found in Sample (ng/g) w et wt.
Amount Recovered (ng/g) wet wt.
Recovery w
Amt Found in Sample (ng/a) w et wt.
Amount Recovered (ng/g) wet wt.
Recovery (%>
Amt Found in Sample (ng/g) w et wt.
Amount Recovered (ng/g) w et wt.
Recovery <%)
2.0 ND 40 ND 800 ND
2.10 105 ND NA NA ND NA NA ND
2.04 102 ND NA NA ND NA NA ND
2.35 118 NA NA NA NA
DL3-SD-LC002-0-061214 (C0226987 Spk F, 2.0 ppb Spike)
DL3-SD-LOC002-0-061214 (C0228987 Spk O, 40 ppb Spike)
DL3-SD-LOCQQ2-0-Q61214 (C0226987 Spk H, 800 ppb Spike)
2.0 40 800
ND ND ND
2,08 104 ND NA NA ND NA NA ND
1.93
97 0.214
2.12
95
NA NA 0.214 NA NA
NA NA 0.214 NA NA
DL3-SD-LOC003-0-061214 (C0226968 Spk 1, 2.0 ppb Spike)
DL3-SD-LOC003-0-061214 (C0226988 Spk J, 40 ppb Spike)
DL3-SD- LOC003-0-061214 (C0226988 Spk K, 800 ppb Spike)
2.0 40 800
ND ND ND
1.88 94 ND NA NA ND NA NA ND
1.96 98 ND NA NA ND NA NA ND
2.01 101 NA NA NA NA
DL2-S D-LOC001-0-061214 (C0226989 Spk C, 2.0 ppb Spike)
DL2-SD-LOC001-0-061214 (C0226969 Spk D, 40 ppb Spike)
DL2-SD-LOC001-0-061214 (C0226969 Spk E. 800 ppb Spike)
2.0 40 600
ND ND ND
2.01 101 ND NA NA ND NA NA ND
2.08 104 ND NA NA ND NA NA ND
2.25 113 NA NA NA NA
DL2-SD-LOC002-0-061214 (C0226990 Spk F, 2.0 ppb Spike)
DL2-SD-LC002-0-061214 (C0226990 Spk G, 40 ppb Spike)
DL2-SD-LOC002-0-061214 (C0226990 Spk H, 800 ppb Spike)
2.0 40 800
ND ND ND
2.09 105 ND NA NA ND NA NA ND
2.08 104 ND NA NA ND NA NA ND
2.14 107 NA NA NA NA
DL2-SD-LOC003-0-061214 (C0226991 Spk I, 2.0 ppb Spike)
DL2-SD-LOC003-0-061214 (C0226991 Spk J, 40 ppb Spike)
DL2-S D- LOC00 3-0-061214 (C0226991 Spk K, 800 ppb Spike)
2.0 40 800
ND ND ND
1.99 100 ND NA NA ND NA NA ND
2.03 102 ND NA NA ND NA NA ND
2.05 103 NA NA NA NA
DBC-S D-LOC001-0-061214 (C0226992 Spk C, 2.0 ppb Spike)
DBC-S D-LOC001-0-061214 (C0226992 Spk D, 40 ppb Spike)
DBC-S D-LOC001-0-061214 (C0226992 Spk E, 800 ppb Spike)
2.0 40 800
ND ND ND
2.04 102 0,232 2.30 103 26.7 NA NA 0.232 NA NA 26.7 NA NA 0.232 NA NA 26.7
NA NA 72.5 115 NA NA
ND = Not detected at or above the acceptable LOQ reported on Table II. NA = Not applicable. This matrix spike concentratrion was not used to assess the accuracy for this analyte. NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
Table IV. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Sediment Samples (continued)
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
__________________________________ Periluorobutanesulfonate__________________________________ Perfluorohexaneiulfonete____________________ Perfluorooctanoeulfonate
Sample Description
Amount Spiked (ng/o)
Amt Found in Sample (ng/g) w et wt.
Amount Recovered (na/a) w et wt.
Recovery <%)
Amt Found in Sample (ng/g) wet wt.
Amount Recovered (ng/g) wet wt.
Recovery (%>
Amt Found in Sample (ng/g) w et wt.
Amount Recovered (ng/g) w et wt.
Recovery <%)
DBC-S D-LOC002-0-061214 (C0226993 Spk F, 2.0 ppb Spike)
DBC-S D-LOC002-0-061214 (C0226993 Spk O, 40 ppb Spike)
DBC-S D-LOC00 2-0-061214 (C0226093 Spk H, 800 ppb Spike)
2.0 40 800
ND ND ND
2.04 102 0.201 NA NA 0.201 NA NA 0.201
2.24 102 21.0 NA NA 21.0 NA NA 21.0
NA NA 64.2 108 NA NA
DBC-S D-LOC003-0-061214 (C0224994 Spk I. 2.0 ppb Spike)
DBC-S D-LOC003-0-061214 (C0226994 Spk J, 40 ppb Spike)
DBC-S D-LOC003-0-061214 (C0226994 Spk K, 800 ppb Spike)
2.0 40 800
ND ND ND
2.03 102 ND NA NA ND NA NA ND
2.14 107 11.7 NA NA 11.7 NA NA 11.7
NA NA 68.8 143 NA NA
DOU-S D-LOC001 -0-061213 (C0226995 Spk C, 2.0 ppb Spike)
DOU-SD-LOCOQ1-0-061213 (C0229995 Spk D, 40 ppb Spike)
DOU-SD-LOC001-0-061213 (C0226995 Spk E, 800 ppb Spike)
2.0 40 800
3.47 3.47 3.47
4.75 NA NA
64 3.52 NA 3.52 NA 3.52
5.05 NA NA
77 309 NA 309 NA 309
NA NA NA NA 948 80
DOU-S D-LOC002-0-061213 (C0226996 Spk F, 2.0 ppb Spike)
DOU-S D-LOC002-0-061213 (CO220996 Spk G, 40 ppb Spike)
DOU-SD-LOC002-0-061213 (C0226996 Spk H, 800 ppb Spike)
2.0 40 800
0.459 0.459 0.459
2.61 NA NA
108 2.19 NA 2.19 NA 2.19
5.10 146 NR NA NA NR NA NA NR
NR NR NR NR NR NR
DOU-S D-LOC003-0-061213 (C0226997 Spk 1, 2.0 ppb Spike)
DOU-S D-LOC003-0-061213 (C0226997 Spk J. 40 ppb Spike)
DOU-S D-LOC003-0-061213 (C0226997 Spk K, 800 ppb Spike)
2.0 40 800
0.468 0.468 0.468
2.60 NA NA
107 0.875 NA 0.875 NA 0.875
3.01 NA NA
107 61.5 NA 61.5 NA 61.5
NA NA 95.6 85 NA NA
D L1-S D-LOC001-0-061213 (C0226998 Spk C, 2.0 ppb Spike)
DL1-SD-LOC001-0-061213 (CO220998 Spk D, 40 ppb Spike)
DL1-SD-LOC001-0-061213 (C0226998 Spk E, 800 ppb Spike)
2.0 40 800
ND ND ND
2.05 103 ND NA NA ND NA NA ND
2.09 105 1.81 NA NA 1.81 NA NA 1.81
3.45 82 NA NA NA NA
DL1-SD-LOC002-0-061213
(C0220999 Spk F, 2.0 ppb Spike)
2.0
0.210
2.26
103
ND
DL1-S D-LOC002-0-061213
(C0229999 Spk G, 40 ppb Spike)
40
0.210
NA
NA ND
DL1-SD-LOC002-0-061213
(C0226999 Spk H, 600 ppb Spike)
800
0.210
NA
NA ND
2.02 101 2.08 NA NA 2.08 NA NA 2.08
4.41 117 NA NA NA NA
ND = Not detected at or above the acceptable LOQ reported on Table II. NA = Not applicable. This matrix spike concentratrion was not used to assess the accuracy for this analyte. NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
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Table IV. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Sediment Samples (continued)
Sample Description
DL1-SD-LOC003-0-061213 (C0227000 Spk 1, 2.0 ppb Spike)
DL1-SD-LOC003-0-061213 (C0227000 Spk J, 40 ppb Spike)
DL1-SD-LOC003-0-061213 (C0227000 Spk K, 600 ppb Spike)
Amount Spiked (ng/g)
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Pertluorobutanesulfonate____________________ Perfluorohexartetulfonate____________________ Perfluorooctaneeulfonatc
Amt Found
Amount
in Sample Recovered
(na/o1) wet wt. (hqIq) wet wt.
Recovery i%)
Amt Found in Sample (ng/gl wet wt.
Amount Recovered (ng/g) wet wt.
Amt Found
Amount
Recovery in Sample Recovered
(%> (no/p) wet wt. (ng/tf) wet wt.
Recovery (%)
2.0 ND 40 ND 800 ND
2.07 104 ND NA NA ND NA NA ND
2.07 104 0.631 3.05 121 NA NA 0.631 NA NA NA NA 0.631 NA NA
DMC-S D-LOC001-0-061213 (CQ227Q01 Spk C, 2.0 ppb Spike)
DMC-SD-LOC001-0-061213 (C0227001 Spk D, 40 ppb Spike)
DMC-SD-LOC001-0-061213 (00227001 Spk E. 800 ppb Spike)
2.0 40 800
ND ND ND
2.08 104 ND NA NA ND NA NA ND
2.10 105 1.26 3.24 99
NA NA 1.26
NA NA
NA NA 1.26
NA NA
DMC-S D-LOC002-0-061213 (C0227002 Spk F, 2.0 ppb Spike)
DMC-S D-LOC002-0-061213 (C0227002 Spk G, 40 ppb Spike)
DMC-S D-LOC002-0-061213 (C0227002 Spk H, 600 ppb Spike)
2.0 40 800
ND ND ND
2.15 108 ND NA NA ND NA NA ND
2.00 100 1.67 NA NA 1.67 NA NA 1.67
3.40 87 NA NA NA NA
DMC-SD-LOC003-0-061213 (C0227003 Spk I, 2.0 ppb Spike)
DMC-S D-LOC003-0-061213 (C0227003 Spk J, 40 ppb Spike)
DMC-S D-LOC00 3-0-061213 (C0227003 Spk K, 800 ppb Spike)
2.0 40 800
ND ND ND
2.01 101 ND NA NA ND NA NA ND
1.94 97 1.60 NA NA 1.60 NA NA 1.60
3.63 102 NA NA NA NA
DAA-S D- LOC006-0-061215 (C0227532 Spk C, 2.0 ppb Spike)
DAA-SD-LOC006-0-061215 (C0227532 Spk D, 40 ppb Spike)
DAA-SD-LOC006-0-061215 (C0227532 Spk E, 800 ppb Spike)
2.0 40 800
4.51 4.51 4.51
NA NA NR 40.8 91 NR NA NA NR
NR NR 491
NA NA
NR NR 491
NA NA
NR
NR 491
1140
81
DAA-SD-LOC005-0-061215 (C0227S33 Spk F, 2.0 ppb Spike)
DAA-SD-LOC0Q5-0-061215 (C0227533 Spk G, 40 ppb Spike)
DAA-S D-LOC005-0-061215 (C0227S33 Spk H, 600 ppb Spike)
2.0 40 800
2.79 2.79 2.79
4.79 100 13.8 NA NA 13.8 NA NA 13.8
NA 49.1 NA
NA 434 88 434 NA 434
NA NA NA NA 893 57
DAA-S D-LOC004-0-061 2 1 5 (C0227534 Spk i. 2.0 ppb Spike)
DAA-S D-LOC004-0-061 2 1 5 (C0227S34 Spk J, 40 ppb Spike)
DAA-S D-LGC004-0-061215 (C0227S34 Spk K, 600 ppb Spike)
2.0 40 800
26.4 26.4 26.4
NA NA 213 63.6 93 213 NA NA 213
NA NA 616
NA NA
NA NA 616
NA NA
805
74 616
1150
67
ND = Not detected at or above the acceptable LOQ reported on Table II. NA = Not applicable. This matrix spike concentratrion was not used to assess the accuracy for this analyte.
NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
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Table IV. Matrix Spike Recovery Summary of PFBS, PFHS, and PFOS in Sediment Samples (continued)
Sample Description
DAA-SD-LOC003-0-061215 (C022753S Spk C. 2.0 ppb Spike)
DAA-S D- LOC003-0-061 2 1 5 (C0227535 Spk D, 40 ppb Spike)
DAA-SD-LOC003-0-061215 (C0227S35 Spk E.800 ppb Spike)
Amount Spiked (ng/g)
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutaneulfonate____________________ Parfluorohexaneaulfonate___________________ Periluorooctaneaulfonate
Am t Found in Sample (ng/g) wet wt.
Amount Recovered (na/a) w et wt.
Recovery (%>
Amt Found in Sample (ng/g) w et wt.
Amount Recovered (ng/g) w et wt.
Recovery (%)
Am t Found in Sample (ng/g) wet wt.
Amount Recovered (ng/g) wet wt.
Recovery (%)
2.0 1.94 40 1.94 800 1.94
4.29 118 10.8 NA NA 10.8 NA NA 10.8
NA NA 585 52.3 104 585 NA NA 585
NA NA 1380
NA NA 99
DAA-S D-LOC002-0-061215 (C0227S36 Spk F, 2.0 ppb Spike)
DAA-SQ-LOCOQ2-Q-Q61215 (C0227536 Spk G. 40 ppb Spike)
DAA-SD-LOC002-0-061215 (C0227536 Spk H. 800 ppb Spike)
2.0 40 800
0.273 0.273 0.273
2.52 NA NA
112 0.378 NA 0.378 NA 0.378
2.65 NA NA
114 23.4 NA 23.4 NA 23.4
NA NA 63.7 101 NA NA
DAA-SD-LOC001 -0-061215 (C0227537 Spk 1, 2.0 ppb Spike)
DAA-S D-LOC001-0-061215 (C0227537 Spk J, 40 ppb Spike)
DAA-SD-LOC001 -0-061215 (C0227537 Spk K, 800 ppb Spike)
2.0 40 800
0.257 2.58 116
0.257 NA NA
0.257
NA
Average: Standard Deviation:
NA
102 10
0.402
2.60
0.402
NA
0.402
NA
Average: Standard Deviation:
NO = Not detected at or above the acceptable LOQ reported on Table II. NA = Not applicable. This matrix spike concentratrion was not used to assess the accuracy for this analyte. NR = Not reported due to quality control failures. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
110
NA
NA 102 13
55.3 NA NA
55.3 111 139
55.3 NA
Average: Standard Deviation:
NA
101 21
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Table V. Total Percent Solids for Sediment Samples
Exygen ID C0226986 C0226987 C0226988 C0226989 C0226990 C0226991 C0226992 C0226993 C0226994 C0226995 C0226996 C0226997 C0226998 C0226999 C0227000 C0227001 C0227002 C0227003 C0227532 C0227533 C0227534 C0227535 C0227536 C0227537
Client Sam ple ID D L3 -S D -L O C 0 0 1 -0 -0 6 1 2 1 4 D L 3 -S D -L O C 0 0 2 -0 -0 6 1 214 D L3 -S D -L O C 0 0 3 -0 -0 6 1 2 1 4 D L2 -S D -L O C 0 0 1 -0 -0 6 1 2 1 4 D L 2 -S D -L O C 0 0 2 -0 -0 6 1 214 D L2-S D -LO C 003-0-061214 D B C -S D -L O C 0 0 1 -0 -0 6 1 2 1 4 D B C -S D -L O C 0 0 2 -0 -0 6 1 214 D B C -S D -L O C 0 0 3 -0 -0 6 1 2 1 4 DOU-SD-LOC001 -0-061213 D O U -S D -L O C 002-0-061213 D O U -S D -L O C 003-0-061213 D L1 -S D -L O C 0 0 1 -0 -0 6 1 2 1 3 D L 1-S D -L O C 0 0 2 -0 -0 6 1213 D L 1 -S D -L O C 0 0 3 -0 -0 6 1 213 DMC-SD-LOC001 -0-061213 D M C -S D -L O C 002-0-061213 D M C -S D -L O C 0 0 3 -0 -0 6 1 213 D A A -S D -L O C 006-0-061215 D A A -S D -L O C 0 0 5 -0 -0 6 1 215 D A A -S D -L O C 004-0-061215 D A A -S D -L O C 0 0 3 -0 -0 6 1 215 D A A -S D -L O C 0 0 2 -0 -0 6 1 215 DAA-S D -LO C 001-0-061215
Total Percent Solids (%) 65.16 43.41 46.81 48.16 46.49 47.72 45.84 47.17 43.10 67.54 57.12 78.34 53.47 45.02 78.98 65.71 45.17 44.17 72.89 71.69 30.49 44.39 71.91 56.02
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FIGURES
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Figure 1. Typical Non-Extracted Calibration Curve for PFBS in 50:50 Acetonitrile:Water
062007A.rdb (PFBS): "Linear*1Regression C'1 fyi' weighting): y 8.03e+005 x + 0.18e+OO3 (r = 0.9982)
Area, counts
Concentration, ng/mL
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Figure 2.
Non-Extracted Standards of PFBS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively
SS0022274-PFBS (Standard) 299.0/80.0 am u, 299.0/99.0 s iwy sam p/9 1 o f 40 from 0S2M7A.wtff Area: 1222 co u n ts Height; 2.70+003 cp RT: 3.57 mitt 3.57
Time, min
SS0022273 - PFBS (Standard) 299.0/80.0 amu.299.0/99.0 amu sample 2 of 40 from 062007A.vuiff Area: 20717 counts Height: 4.71e+003 cps RT:3.57 min
3.57
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Figure 3.
PFBS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively
R a a g oatB lank 1 -P FB S (Ukkaown) 299.W90.0amuf299.0/99,0amu sam pte 11 o f 4Q from 062C07A.wiff A n a : 2797 counts Height: 5.29e+O02cp* RT: 3.57 m;'
Time, min
Reagent Spike A1 PFBS (00)299.0/80.0 amu.299.0/99.0 amu - sample 12 of 40 from 062007A.uuiff Area: 92440 counts Height: 2 .14e+004 cps RT: 3.57 min 3.57
Time, min
Reagent Sp ike B1 -P F B S (QC) 299.0/80.Kamil,299.0/99.0 am n sam ple 15o f 4 from 062087A.wiff Area: 848233 counts Height: f.96e+005 cps RT: 3.57 m in 3.67
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Figure 4.
Chromatogram Representing a Surface Water Sample Analyzed for PFBS (ExyLIMS ID: C0227212, Data Set: 062007A)
I C0227272 PFBS (Unknown) 299.0V80.0 a m a ,299.0/99.0 amir -sa m p fe 21 o f 40 from 062007A.w/ff Area: 6683256 counts Height: 7.52e+006cp* RT: 3L57 m in
Intensity, cps
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Figure 5.
PFBS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively
C0221520 B lank 1 PFBS (Unknown) 299.0/90.0 a m u ,299.0/99.0 am u -sam ple 10 o f 42 from 02507G.wiff A n a : 1502 co u n ts Height: 3.319+002 cps RT: 3.57 m in
Time, min C0221520 Spike A1 PFBS (QC)299.0/80.0 amu.2Q9.0/99.0 am u- sample 11 of 42 from 062507O.wiff Area: 441409 counts Height: 9.9Ce+004 cps R T :3.58m in
3.58
8.0 e4 CQ<_O). 6.Qe4-
55 cn 4.0 e4_c
2.0 e4 -
0.0
8 9 10
Time, min
I C0221520 Sp ike 8 1 - PFBS (QC) 299.0/90,0 a m u ,299.0/99.0 am u -sa m p le 1 4 o f 42 from 062507G.wiff A n a : 2232640counts Height: 5b02e+005cp* RT: 3.57 m in
3.57
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Figure 6. Chromatogram Representing a Sediment Sample Analyzed for PFBS (ExyLIMS ID: C0226986, Data Set: 062507G)
C 0 22S 9& 6 P F B S (Unknown) 29&frVA0aivrir,299L0/99L0ffjfr<f - sa m p le 20 o f 4 2 from 062507G.wv7f Area: 7379 co u n ts Height: 3.3ie+O02 cps RT: 3.57 m in 0.26
intensity, cps
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Figure 7. Typical Non-Extracted Calibration Curve for PFHS in 50:50 Acetonitrile:Water
061907A.rdb (PFHS): "Lineai" Regression C'1 / x " weighting): y 1.59e+006 x + 1.18e+004(r 0.9968)
Area, counts
Concentration, ng/mL
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Figure 8.
Non-Extracted Standards of PFHS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively
S S 9022274 - PFHS (Standard) 399L/80.0 a*itf,399L0/99.0 am u -ta m p fe 1 o f 40 front 061907A.wiff Area: 25924 co u n ts Height: 5.88+003 cp RT; X 83 m in
7.83
Time, min
I SS0022273 PFHS (Standard) 309.0/80.0 amu,390.0/99,0 am u- sample 2 of 40 from 061Q07A.iaiM Area: 46860 counts Height: 1.0Se+004cps RT:3.83 min
3.83
7.83
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Figure 9.
PFHS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively
I Ragant BlanA 1 -PFHS (Unknown) 399.0/90.0 am u,399.0/99.0 antu -sam ple 11 o f 40 from 061907A.wiff p ea k not found)
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Figure 10. Chromatogram Representing a Surface Water Sample Analyzed for PFHS (ExyLIMS ID: C0227208, Data Set: 061907A)
60227208 PFHS (Unknown) 399.0*80.0 a * ,3 9 f t 0/9ft 0am u sam ple 35 o f 40 fro m 067907A w iff Area: 3034counts H eight: 7.74+002 cp* RT: 3.03mia 7.84
Intensity, cps
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Figure 11. PFHS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively
C0221520Blank 1 -PFHS (U nknow n)39910/80L0 am u,399.(V9%0 am u sam ple 10o f 42 from 062507G.wiff Area: 12418 co u n ts Height: f.99e+003cp* RT: 3.90 m in
7.99
Tim, min
I C0221520 Spike A1 PFHS (QC) 399.0/30.0 amu.399 0/99.0 amu - sample 11 of 42 from 062507G.wiff Area: 402514 counts Height: 8.66e+004 cps RT:3.81 min
3.81
I C0221520Spike B 1-P F H S (QC)399.(VS0.0amu,3S9.<V9ft.0 am u -sa m p le U o f 42 from 0S25TG.mff Area: W95041 counts Height: 4.13e *005 cps RT: 3.80 m/ 3.80
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Figure 12. Chromatogram Representing a Sediment Sample Analyzed for PFHS (ExyLIMS ID: C0226986, Data Set: 062507G)
I C 0 2 2 6 9 B 6 - P F H S ( U n k n o w n ) 39910/9910 a m a , 3 9 9 .0/99. d a m n - s a m p l e 20 o f 4 2 f r o m 0 6 2 S 7 G .w if f A r e a : 1 4 S 9 9 c o a n ts H e ig k t: 2 .4 *+ 0 0 3 c p s A T : 3.80 m in
7.90
Intensity, cps
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Figure 13. Typical Non-Extracted Calibration Curve for PFOS in 50:50 AcetonitrilerWater
062007A.rdb (PFOS): "Linear" Regression ('1 / * ' weighting): y 1.48e+000 x + 1.04e+004(r = 0.9073)
Area, counts
Concentration, ng/mL
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Figure 14. Non-Extracted Standards of PFOS in 50:50 Acetonitrile:Water, 0.0125 ng/mL and 0.025 ng/mL, Respectively
I SS0022274 P F O S ( S t a n d a r d ) 4 9 9 . M 0 . 0 a m a , 4 9 9 . 0 m . 0 a m a - s a m p l e 1 o f 4 0 f r o m 02007A -w H T A r e a : 23382 c o u n ts H e ig h t: 4 .9 2 e +003 c p s R T : 3 9 8 m in 3.88
Time, min
I SS0022273 - PFOS (Standard) 4Q9.G#0.Q amu,40Q,0/99.Q amu sample 2 of 40 from 062007A.imff Area: 46630 counts Height: 0.70e+003 cps RT:3.98 min
3.98
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Figure 15. PFOS in a Reagent Blank, a 0.25 ng/mL Fortified Reagent Spike A, and a 2.5 ng/mL Fortified Reagent Spike B, Respectively
R eagent B ia n t 1 - PFOS (Unknown) 499.0/90.0 am u,499.0/99.0 am u -*am p/e 11 o f 40 from 062007A.w iff A n a : 1294count* Heigkt: Z61e+002cp* RT: 3.99 mi/t
Time, min
Reagent Spike A1 - PFOS (00)400.0/80.0 amu.409.0/90.0 am u- sample 12 of 40 from 062007A.wiff Area: 224171 counts Height: 4.75e+004 ops RT: 3.08 min
3.98
Reagent S p ite B1 - PFOS (QC) 499.0/90.0 am a,499.0/99.0 am n -sam pfe 15 o f 40 from 062007A.wiff Area: 2045992 count* Heigkt: 4.29e*005 cp* RT: 3.99 m in 3.98
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Figure 16. Chromatogram Representing a Surface Water Sample Analyzed for PFOS (ExyLIMS ID: C0227212, Data Set: 062007A)
C 0 2 2 7 2 1 2 - P F O S ( U n k n o w n ) 499.0/80.0 a m u .4 9 9 . 0 /9 9 .0 a m u - s a m p l e 2 1 o f 4 0 f r o m 0 6 2 0 0 7 A .w if f A r e a : 4J72976 c o u n t s H e ig h t: 8.57s+005 cp# R T : 3.98 m in 3 98
Intensity, cps
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Figure 17. PFOS in a Control Blank, a 2.0 ng/g Fortified Control Spike A, and a 20 ng/g Fortified Control Spike B, Respectively
10C 022T520B l a n k 1 P F O S ( U n k n o w n ) 4 9 9 .0 /9 0 .0 a m u , 4 9 9 .0 /9 9 .0 a m u - s u m p f e o f 4 2 f r o m 062507 G .w if f A r e a : 0936 c o u n t s H e ig h t: 1 .3 9 e + 0 0 3 c p s R T : 3. 97 m in 3.97
C 0 2 2 1 S 2 0 S p ik e B 1 - P F O S (Q C ) 4 9 9 .0 /8 0 .0 a m u , 4 9 9 . 0 /9 9 .0 a m u - s a m p l e 1 4 o f 4 2 f r o m 062507G.w;7f A rea: 1909004co u n ts H eigh t: X 9 1 e+ 0 0 5 cp s RT: X 9 6 m in 3.96
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Figure 18. Chromatogram Representing a Sediment Sample Analyzed for PFOS (ExyLIMS ID: C0226986, Data Set: 062507G)
C02269B6 - PFOS (Unknown) 499.0/80.# antu,499.0/99.0 am n -sam ple 20 o f 42 from 062507G.wiff A n a : 51921 coant* Height: 1.07e*004cps RT: 3.96 min 3.96
Intensity, cps
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APPENDIX A
Study Protocol
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STUDY PROTOCOL
Study Title:
Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and
Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, and Clams Using LC/MS/MS for the 3M Decatur Monitoring Program
MPI Research Study Number.: 0137.0219 ExyLIMS Protocol Number: P0003268
P erfo rm in g L aboratory: M P I R esearch, Inc. S tate C ollege 3058 R esearch D rive S tate C o lleg e, PA 16801 Phone: (814) 272-1039
S po n so r R epresentative: M ichael A . S antoro D irector o f R eg u lato ry A ffairs 3M B uilding 0236-01-B -10 St. P au l, M N 5 5 1 4 4 Phone: (651)733-6374
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DISTRIBUTION:
ExyLIMS Protocol Number: POOQ3268
1) Jaisim ha K esari, S tudy D irector, W eston Solutions, Inc. 2) K aren R isha, P rincipal Investigator, M PI R esearch, Inc. 3) M ichael A . Santoro, Sponsor R epresentative, 3M C om pany 4) M PI R esearch Q uality A ssurance U nit
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PRO TO CO L APPROVAL
S tudy Title: Analysis o f Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate
(PF H S ), and P erfluorooctanesulfonate (PF O S ) in W ater. Soil, S edim ent,
Fish, and Clam:. U sing LC/M S-'M S for the .i.Vi D e c ^ u r M o n ito rin g
Program
i
M PI Research Study Num ber: 0137.0219 ExyLIM S Protocol Num ber: P0003268
A PPR O V A LS
Jaislm ha Kesari, Study Director W eston Solutions
D ate
Oll&lo7 D ate
MPI Research, Inc.
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TABLE OF CONTENTS
TITLE PA G E................................................................................................................................................................... 1 D IS T R IB U T IO N ..............................................................................................................................................................2 PROTOCOL APPROVAL.............................................................................................................................................3 TABLE OF CONTENTS............................................................................................................................................... 4 INTRODUCTION........................................................................................................................................................... 5 TEST M ATERIALS....................................................................................................................................................... 5 OBJECTIV E.................................................................................................................................................................... 6 TESTING FACILITY.....................................................................................................................................................7 STUDY DIRECTOR.......................................................................................................................................................7 SPONSOR REPRESENTATIVE.................................................................................................................................. 7 PRINCIPAL INVESTIGATOR.................................................................................................................................... 7 PROPOSED EXPERIMENTAL START AND TERMINATION D A TE S.......................................................... 8 IDENTIFICATION AND JUSTIFICATION OF THE TEST SY STEM ...............................................................8 SAMPLE PROCUREMENT, RECEIPT AND RETENTION.................................................................................8 SAMPLE IDENTIFICATION...................................................................................................................................... 9 ANALYTICAL PROCEDURE SUMMARY............................................................................................................. 9 VERIFICATION OF ANALYTICAL PROCEDURE............................................................................................... 10 METHOD FOR CONTROL OF B IA S........................................................................................................................ 11 STATISTICAL M ETHODS.......................................................................................................................................... 11 GLP STATEM ENT........................................................................................................................................................ 11 R EPO R T........................................................................................................................................................................... 12 SAFETY AND HEALTH.............................................................................................................................................. 13 AMENDMENTS TO PROTOCOL..............................................................................................................................13 DATA RECORD K EE PIN G .........................................................................................................................................13 QUALITY A SSURANCE............................................................................................................................................. 14 RETENTION OF DATA AND ARCHIVING........................................................................................................... 14 APPENDIX I, ANALYTICAL METHODS................................................................................................................15
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INTRODUCTION
T he purpose o f this study is to perform analysis for perfluorobutanesulfonate (PFB S), perfluorohexanesulfonate (PFH S) and perfluorooctanesulfonate (PFO S) in w ater, soil, sedim ent, fish, and clam s using L C /M S/M S for the 3M D ecatur M onitoring Program .
The study w ill be audited for com pliance w ith E PA T SC A G ood Laboratory Practice Standards 40 C FR 792 by the Q uality A ssurance U nit o f M PI R esearch .
TEST MATERIALS
T he test m aterials are perfluorobutanesulfonate (PFB S), perfluorohexanesulfonate (PFH S) and perfluorooctanesulfonate (PFOS).
PFBS C hem ical N am e: Perfluorobutanesulfonate M o le c u la r W e ig h t: 3 3 8 s u p p lie d a s th e p o ta s s iu m s a lt ^ F S O sT C ) T ransitions M onitored: 299 -> 8 0 ,9 9 S tru c tu re :
F FF
S03
FF FF
PFHS C hem ical N am e: Perfluorohexanesulfonate M o le c u la r W e ig h t: 4 3 8 su p p lie d a s th e p o ta s s iu m s a lt (C F ^ S O T C 1) T ransitions M onitored: 399 -> 80, 99 S tru c tu re :
FF
F F
F F
F SO3
FF FF
F F
PFOS C hem ical N am e: Perfluorooctanesulfonate M olecular W eight: 538 supplied as the potassium salt (C sF n S O sT O
T ransitions M onitored: 499 -> 80, 99
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S tru c tu re :
F
SOF F F F
F3
FFFF FFFF
OBJECTIVE
T he purpose o f this study is to perform analysis for perfluorobutanesulfonate (PFB S), perfluorohexanesulfonate (PFH S) and perfluorooctanesulfonate (PFO S) in w ater, soil, sedim ent, fish, and clam s for the 3M D ecatur M onitoring Program using the current versions o f the follow ing 3M C om pany and M P I R esearch analytical m ethods:
ETS-8-012: "M ethod o f A nalysis for the D eterm ination o f Perfluorobutanoic
A cid
(PFB A ),
P e rflu o ro p e n ta n o ic
A cid
(PF P eA ),
Perfluorohexanoic A cid (PFH A ), Perfluoroheptanoic A cid
(PFH pA ), Perfluorooctanoic A cid (PFO A ), Perfluorononanoic
A cid
(PFN A ),
P e rflu o ro d e c a n o ic
A cid
(PFD A ),
Perfluoroundecanoic A cid (PFU nA ), Perfluorododecanoic A cid
(PFD oA ),
P e rflu o ro b u ta n e su lfo n a te
(P F B S ),
Perfluorohexanesulfonate (PFH S), and Perfluorooctanesulfonate
(PFO S) in W ater, Soil and Sedim ent by L C /M S/M S"
V 0001780: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in W ater b y L C /M S/M S"
V 0001781: "M ethod o f A nalysis for the D eterm ination o f P erfluorooctanoic
A cid (PFO A ) in Soil by L C /M S/M S"
V 0001782: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Sedim ent by L C /M S/M S"
V 0001783: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Fish and C lam s b y L C /M S/M S"
The prim ary analytical m ethod for the analysis o f the w ater, soil, and sedim ent sam ples is the ETS-8-012 m ethod. In instances w here quantitative analytical results are not obtained by this m ethod, the Study D irector m ay direct the perform ing laboratory to analyze the sam ples by alternate m ethods (V 0001780 for w ater sam ples, V 0001781 for soil sam ples, and V 0001782 for sedim ent sam ples).
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TESTING FACILITY
M PI Research, Inc. (form erly E xygen R esearch, Inc.) State College 3058 R esearch D rive State C ollege, PA 16801 P hone:(814)272-1039 Fax: (814)231-1580
STUDY DIRECTOR
Jaisim ha K esari P.E ., D EE W eston S olutions, Inc. 1400 W eston W ay W est C hester, PA 19380 Phone: (610) 701-3761 Fax: (610) 701-7401 j .k e s a ri@ w e sto n so lu tio n s.c o m
SPONSOR REPRESENTATIVE
M ichael A . Santoro 3M Com pany D irector o f R egulatory A ffairs 3M B uilding 0236-01-B -10 St. P aul, M N 55144 P h o n e :(6 5 1 )7 3 3 -6 3 7 4
PRINCIPAL INVESTIGATOR
K aren R isha M P I R esearch, Inc. State College 3058 R esearch D rive State College, P A 16801 Phone: (814) 272-1039 karen.risha@ m piresearch.com
ExyLIMS Protocol Number: P0003268
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PROPOSED EXPERIMENTAL START AND TERMINATION DATES
It is proposed that the analytical portion o f this study b e conducted from June 14, 2007 to D ecem ber 31, 2007. T he actual experim ental start and term ination dates w ill be included in the interim reports and the final report.
IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM
T he follow ing are the test system s for this study: W ater (groundw ater and surface w ater) Soil Sedim ent Fish C lam s
T he sam ples w ill be collected by W eston Solutions. T he control sam ples w ill be purchased and prepared by the testing facility. Purchase and processing details for the control sam ples w ill be included in the appropriate interim report and/or final report associated w ith this study.
T he analysis o f PFBS, PFH S, and PFO S are included in a voluntary effort by 3M to gain additional characterization o f sam ples collected to com ply w ith the 3M L etter o f Intent (LO I) w ith the U .S. E PA to collect P F O A data. A dditional sam ples not part o f the LO I m ay be analyzed at the discretion o f the S tudy D irector.
SAMPLE PROCUREMENT, RECEIPT AND RETENTION
W ater, soil, sedim ent, fish, and clam sam ples are being received at M PI R esearch directly from W eston Solutions. The details o f sam ple procurem ent procedures for this study are outlined in the 3M w ork plan entitled "Phase 2 W ork Plan for Sam pling Environm ental M edia for PFO A at the 3M D ecatur, A L P lant." T he num ber and types o f sam ples received and analyzed w ill vary depending on data needs for additional characterization o f PFB S, PFH S, and PFO S concentrations in various environm ental m edia and availability in the field. T he total num ber o f sam ples received and analyzed for each m atrix w ill be docum ented in the appropriate interim report associated w ith this study.
W ater, soil, and sedim ent sam ples w ill be used as received at M P I R esearch. T hese sam ples w ill b e hom ogenized before the extraction process by vigorously shaking the sam ple bottles. T hese sam ples w ill be stored refrigerated at 2C -8C . F ish and clam sam ples w ill b e processed according
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<,to th e a p p ro p ria te a n a ly tic a l m e th o d (se e A p p e n d ix I)- T h e s e s a m p le s w ill b e
s to r e d fro z e n a t -1 0 "C . T h e receip t and processing o f the sam ples w ill b e docum ented in the appropriate interim report and raw data associated w ith the study.
SAMPLE IDENTIFICATION
P rior to analysis, each sam ple w ill be assigned a laboratory sam ple reference num ber. T he reference num ber w ill be unique and w ill distinguish each laboratory sam ple that is processed throughout the analytical procedure. C hrom atographic data w ill b e identified by the laboratory sam ple reference num ber.
Sam ple storage conditions and locations w ill be docum ented throughout the study.
ANALYTICAL PROCEDURE SUMMARY
R eferences:
ETS-8-012: "M ethod o f A nalysis for the D eterm ination o f Perfluorobutanoic
A cid
(PFB A ),
P e rflu o ro p e n ta n o ic
A cid
(PFPeA ),
Perfluorohexanoic A cid (PFH A ), Perfluoroheptanoic A cid
(PFH pA ), Perfluorooctanoic A cid (PFO A ), Perfluorononanoic
A cid
(PFN A ),
P e rflu o ro d e c a n o ic
A cid
(PFD A ),
Perfluoroundecanoic A cid (PFU nA ), Perfluorododecanoic A cid
(PFD oA ),
P e rflu o ro b u tan esu lfo n ate
(P F B S ),
Perfluorohexanesulfonate (PFH S), and Perfluorooctanesulfonate
(PFO S) in W ater, Soil and Sedim ent b y L C /M S /M S "
V 0001780 "M ethod o f A nalysis for the D eterm ination o f P erfluorooctanoic
A cid (PFO A ) in W ater by L C /M S/M S"
V0001781 "M ethod o f A nalysis for the D eterm ination o f P erfluorooctanoic
A cid (PFO A ) in Soil by L C /M S/M S"
V 0001782 "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Sedim ent by L C /M S/M S" V 0001783 "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Fish and C lam s b y L C /M S/M S"
The prim ary analytical m ethod for the analysis o f the w ater, soil, and sedim ent sam ples is the ETS-8-012 m ethod. In instances w here quantitative analytical results are not obtained by this m ethod, the Study D irector m ay direct the perform ing laboratory to analyze the sam ples by alternate m ethods (V 0001780 for w ater sam ples, V 0001781 for soil sam ples, and V 0001782 for sedim ent sam ples).
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T he above referenced m ethods are to be follow ed for the analysis o f the sam ples w ith the follow ing exceptions:
I f sludge sam ples are received by M P I R esearch to be included in the study, the sludge sam ples w ill be extracted and analyzed follow ing the m ethod for soil sam ples.
I f necessary for the preparation o f appropriate standards levels, alternate w eights o f test m aterial and alternate volum es o f suitable solvent (acetonitrile, m ethanol, etc.) m ay be used,
If necessary, the duplicate bottle o f a w ater sam ple m ay b e used in an extraction if the prim ary bottle does n ot h ave enough volum e available.
VERIFICATION OF ANALYTICAL PROCEDURE
A laboratory control sam ple w ill b e used for th e preparation o f fortified control sam ples. T he test substance w ill be m ade into solutions as per the m ethod, and added to the m atrices via a m icropipette.
For w ater sam pling, M P I R esearch w ill supply one bottle per sam ple collected. T he bottles w ill be 500 m L precleaned Sci/Spec P rem ier w ide m outh H D PE bottles. T hese bottles have been routinely used for fluorochem ical sam ple collection at the testing facility and have been show n to b e free o f P F B S , PFH S and PFO S. S am ples w ill b e added to each co n tainer to a volum etric fill line at 200 m L. A field duplicate, a low field spike, a m id field spike (optional, at the discretion o f the S tudy D irector), and a high field spike o f each sam ple w ill be collected. T he low , m id (optional), and high field spike bottles w ill contain PFB S, PFH S and PFO S as w ell as p e r flu o ro o c ta n o ic a c id (P F O A ) a n d 1 ,2 -13C p e r flu o r o o c ta n o ic a c id ( 13C P F O A , o p tio n a l). P F O A a n d 13C P F O A (o p tio n a l) a r e in c lu d e d in th e so lu tio n s u se d to sp ik e th e sa m p le s. T h e re s u lts fo r P F O A a n d ,3C P F O A (optional) w ill not be reported in this study. M P I R esearch w ill supply one field blank (control w ater) and tw o o r three field blank spikes (control w ater fortified w ith PFB S, PFH S and PFO S at a low , m id (optional) and high level) for every tw enty aqueous sam ples collected. A t the testing facility, each w ater sam ple (excluding field duplicates and field spikes) m ay b e extracted in duplicate and m ay also be fortified at a low , m id (optional), and high concentration and processed through the described procedure. The use o f the in lab duplicate and the in lab fortified spikes is not a requirem ent b ut m ay be requested to determ ine m ethod accuracy and to check for b ias at the discretion o f the Study D irector.
F or soil, sedim ent, fish, and clam s, M PI R esearch w ill supply one 500 m L precleaned Sci/Spec Prem ier w ide m outh H D PE bottle p er sam ple collected or a zip-seal bag. A ll containers/bags used for sam ple collection w ill be shipped to th e sam ple location. Sam ples w ill be added to each container or b ag in the
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field. A t the testing facility, each sam ple w ill b e extracted in duplicate and w ill also be fortified at a know n concentration w ith PFB S, PFH S and PFO S at a low , m id (optional, at the discretion o f the S tudy D irector), and high level and processed through the described procedure to determ ine m ethod accuracy and to check for bias.
L ow , m id, and high spiking levels for each m atrix are d efined below :
M atrix
Low PFOA
M id P F O A
H igh PFO A
S p ik in g Levels S p ik in g L evels S p ik in g Level
W ater
0.25 ng/m L
5.0 ng/m L
100 ng/m L
Soil
2 n g /g
40 ng/g
800 ng/g
Sedim ent
2 ng/g
40 ng/g
800 ng/g
Fish
2 ng/g
40 ng/g
-
C lam s
2 ng/g
40 ng/g
-
L ow , m id and high spiking levels o f the analytes for each m atrix m ay be altered depending on sam ple size, sam ple availability, and/or to cover analyte concentrations expected in the sam ples. In instances w here the expected analyte concentrations exceed the relevant range o f the low er level spikes, the low er level spikes m ay be deferred.
R ecoveries are anticipated to be betw een 70% and 130% o f the fortified levels; how ever, the exact precision and accuracy w ill b e determ ined b y the analysis o f the quality control sam ples described above. A statem ent o f accuracy w ill be included in the appropriate interim report.
METHOD FOR CONTROL OF BUS
C ontrol o f bias w ill be addressed by taking representative sub-sam ples from a hom ogeneous m ixture o f each m atrix from untreated control sam ples, and by analyzing at least tw o levels o f fortifications.
STATISTICAL METHODS
Statistics w ill be lim ited to those specified in the subject m ethods and to the calculation o f average recoveries, as applicable.
GLP STATEMENT
A ll aspects o f this study shall be perform ed and reported in com pliance w ith E P A T SC A G ood L aboratory Practice S tandards 4 0 C F R 792. T he final report or data package (supplied to the Sponsor) shall contain a statem ent that
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the study w as conducted in com pliance w ith current and applicable GLP standards and w ill outline any deviations in the study from those standards, T his statem ent w ill be signed by the Study D irector and Sponsor R epresentative.
REPORT
Interim reports w ill be prepared by the principal investigator or their designee for specific sam pling sites and sam ple m atrices due to the size o f the data sets and the phased nature o f sam ple collection activities. A final report w ill be prepared by the principal investigator o r their designee at the conclusion o f the study. A ll reports w ill include, but w ill not be lim ited to, the follow ing:
The nam e and address o f the Study D irector, Sponsor R epresentative, and o f the testing facility.
A statem ent o f G LP com pliance (any related docum entation, such as chain-of-custody records, m ust be in the study records).
T he signed and dated statem ent by the M P I R esearch Q uality A ssurance U nit regarding dates o f study inspections and dates findings w ere reported to the Study D irector and M anagem ent.
A description o f the exact analytical conditions em ployed in the study. If the subject m ethod w as follow ed exactly, it is n ecessary to include o nly a copy o f the analytical m ethod. A ny m odifications to this m ethod w ill be incorporated into the report. I f the m ethod is photo-reduced, the project num ber and page num ber m ust be included on each page.
D escription o f the instrum entation used and operating conditions.
A ll results from all sets analyzed. C ontrol and fortified sam ples w ill be identified and the data table w ill include sam ple num ber and fortification level.
R epresentative chrom atogram s for each analyte in each m atrix, including chrom atogram s o f a standard and a control sam ple, and a chrom atogram at a fortification level. T he location o f the analyte peaks w ill be clearly identified in all chrom atogram s.
A ll circum stances that m ay have affected the quality or integrity o f the data w ill be docum ented in the report.
L ocations w here raw data, interim reports, and the final report are to be archived.
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A dditions or corrections to the final report shall b e in the form o f an am endm ent signed b y the Study D irector. T he am endm ent shall clearly identify that part o f the report that is being altered and the reasons for the alterations. The am endm ent w ill be signed and dated by the Study D irector and the Sponsor Representative.
A ll applicable requirem ents for reporting o f study results as per 40 C FR 792.185.
SAFETY AND HEALTH
L aboratory personnel w ill practice good sanitation and health habits.
E very reasonable precaution shall be taken to prevent inadvertent exposure o f personnel and the environm ent to the test or reference substance(s).
AMENDMENTS TO PROTOCOL
A ll significant changes to the analytical protocol outlined here w ill be expressed in w riting, signed and dated by the S tudy D irector and Sponsor R epresentative. A m endm ents usually w ill b e issued prior to initiation o f study plan change. H ow ever, w hen a change is required w ithout sufficient tim e for the issue o f a w ritten am endm ent, that change m ay be effected verbally w ith supporting docum entation signed and dated by the Study D irector and follow ed w ith a w ritten am endm ent as soon as possible. In this case, the effective date o f the w ritten am endm ent w ill b e the date o f the docum ented change. C opies o f the signed am endm ents w ill b e appended to all distributed study plan copies. T he original am endm ent w ill be m aintained w ith the original study plan. A ny deviations from the study plan or from the analytical m ethod as provided w ill be docum ented and reported prom ptly to the Sponsor R epresentative.
DATA RECORD KEEPING
R ecords to be m aintained include the follow ing (as appropriate): Sam ple tracking sheet(s) Sam ple receipt records, storage history, and chains o f custody H istory and preparation o f standards (stock, fortification, calibration) D escription o f any m odifications to the m ethod Instrum ent run sheets, bench-sheets or logs A nalytical data tables A ll chrom atographic and instrum ental conditions
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Sam ple extraction and analysis dates A com plete listing o f study personnel, signatures and initials C hronological presentation o f all study correspondence A ny other docum entation necessary for the reconstruction o f the study
Chromatograms- A ll c h ro m a to g ra m s w ill c o n ta in th e fo llo w in g :
Sam ple identification, injection date, arrow or o ther indication o f the area o f interest, and injection num ber corresponding to the run.
A dditionally, fortifications w ill include the am ount o f analyte added and the sam ple num ber o f the sam ple that w as fortified.
A nalytical standard chrom atogram s w ill additionally include the concentration (e.g., pg/m L).
A s part o f the docum entation the follow ing sheets w ill be included in each analytical set: a run sheet listing the sam ples to b e run in the set, and an instrum ent conditions sheet describing the in stalm ent type and operating conditions.
QUALITY ASSURANCE
T he Q A U nit o f M PI R esearch w ill inspect the study at intervals adequate to a s s u re c o m p lia n c e w ith G L P 's, a n d w ill r e p o rt th e fin d in g s o f a u d its to th e Study D irector, M PI R esearch M anagem ent, and the Sponsor R epresentative.
RETENTION OF DATA AND ARCHIVING
A ll hard copy raw data, including, but not lim ited to, the original chrom atogram s, w orksheets, correspondence, and results shall be included w ith the data package subm itted to the S tudy D irector. T hese w ill b e archived w ith the original study plan, am endm ents, final report, and all pertinent inform ation from the Sponsor.
T he testing facility shall keep all electronic raw data and any instrum ent, equipm ent, and storage logs for the period o f tim e specified in 40 C FR 792.195. A n exact copy o f the m aterials subm itted to th e study director will also be kept at M PI R esearch, State College.
M PI R esearch w ill obtain perm ission from the study director before discarding o r retam in g sam ples.
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APPENDIX I ANALYTICAL METHODS
ETS-8-012: "M ethod o f A nalysis for the D eterm ination o f Perfluorobutanoic
A cid
(PFB A ),
P e rflu o ro p e n ta n o ic
A cid
(PFPeA ),
Perfluorohexanoic A cid (PFH A ), Perfluoroheptanoic A cid
(PFH pA ), Perfluorooctanoic A cid (PFO A ), Perfluorononanoic
A cid
(PFN A ),
P e rflu o ro d e c a n o ic
A cid
(PFD A ),
Perfluoroundecanoic A cid (PFU nA ), Perfluorododecanoic A cid
(PFD oA ),
P e rflu o ro b u tan esu lfo n ate
(P F B S ),
Perfluorohexanesulfonate (PFH S), and Perfluorooctanesulfonate
(PFO S) in W ater, Soil and Sedim ent by L C /M S/M S"
V 0001780: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in W ater by LC /M S/M S"
V 0001781: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Soil by LC /M S/M S"
V 0001782: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Sedim ent by L C /M S/M S"
V 0001783: "M ethod o f A nalysis for the D eterm ination o f Perfluorooctanoic
A cid (PFO A ) in Fish and C lam s by L C /M S/M S"
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3M Environmental Laboratoiy
. Method
.
Method o f Analysis for the Determination o f Perfluorobutanolc Acid (PFBA), Pertluoropentanoic Acid (PFPeA), Perfluorohexanoic Acid (PFHA), Perfiuoroheptanoic Acid (PFHpA), Perfiuorooctanoic A cid (PFOA), Perfluorononanoic Acid (PFNA), Perfluorodecanolc A cid (PFOA),
Perfluoroundecanoic Acid (PFUnA), Ptrfluorododecanolc Acid(PFDoA), Perfiuorobutanesuffonate (PFBS), Perfluorohexanesulfonate (PFHS), end Perfluorooctanesutfonate (PFOS) In Water, Soil and Sediment by LC/MS/MS
Method Number: BTS-8-012.1
Adoption Date: Upon Signing
Effective Date:
Approved By:
VVfHiamK. Reagan,TechnicalManager
Date
ETS-S-C12.1
Page I ofJ2
MMhod ofAnafyaia torIho Odfomdnuion ofPardwrebutandc ACM(PFBAJ, Pffluoropantanoic A d i (PFPaA). Paritooroftaxandc Add (PFHA), Pofduorohopunoto Add (PFHpA). Pvtuorooctande Add (PFOA). PorftooranenanoleAdd (PFNA).Padtoorodocmoic
Add (PFOA).PartuorouidacaraleAdd (PFUnA).ftrawrododacandc Add (PFQqA), PtdtoorabuttnoM tondto (PFBS). Pariuoroheianewltoruto (PFHS). and Paduonoctanoaultonsle (PFOS)In Wafer, Sod and Sediment by LC/MS/MS
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ExyLIMS Protocol Number: P0003268
1 Scope and Application
This method to be used to quantity Perfluorobutanofc Add (PFBA), Perfluoropentanoic Add (PFPeA), Perfluorobexanoic Add (PFHA), Perfboroheptonofc Add (PFHpA), Perfluorooctanofc Add (PFOA), Perfluofononanoic Add (PFNA), Perfluorodecsnoic Add (PFDA), Perfluoroundecanofc Add (PFUnA), Perfluorododecenoic Add (PFDoA), Pertuorobutanesutfbnate (PFBS), PerfluorohexanssuHbnate (PFHS), and Perfluorooctanesulfonato (PFOS) by High Performance Liquid Chromatography coupled to a tandem Mass
Spectromebic Detedor (LC/MS/MS) inwater, soil and sediment Themethod Isdesignedto target a Inver Emit
of quantitation (LOQ)of0.025 ng/mL(water)and0.20ng/g(soil and sediment).
2 Method Summary
Aqueous samples aremixedwith equalvolumesofacetonitrile, thoroughly mixed, centrifuged if necessary, and afiquoted for analysis by LC/MS/MS. Simiarty, one-gram aliquots ofsoil and sediments are mixed with 80:20 acetonftriieiwatermixture, thoroughlymixed,centrifuged, and aliquotedforanalysis by LC/MS/MS. -
This is a performance4ased method. Method accuracy is determined for each sample set using multiple laboratory control spikes at multipie concentrations. This methodalso requires that the precision and accuracy for each sample be determined using field matrix spikes (aqueous samples) or laboratory matrix spices (sol and sediment)to verifythattoe methodIs applicableto eachsamplematrix. Sample results forspikes outside
of70%to 130%,will notbe reported dueto non-compliantqualitycontrolsamples.
Fortification levels for field matrix spikes and for laboratory matrix spikes should be at least 50% of the
endogenous level and less than 10 times the endogenous level to be used to determine the statement of
accuracyforanalytical results.
'
3 Definitions
3,1 Calibration Standard
A solution prepared by spSdnga knownvolumeofthe Working Standard (WS) into a predeterminedamount of ASTM type I or HPLC grade water, diluted with acetonitrile, and analyzed accordingto this method. Calibration standardsareusedto cafibrate theinstrument responsewith respect to analyte concentration.
3.2 Laboratory Duplicate Sample (LOS, or Lab Dup)
A laboratory dupficate sample is a separate aliquot of a sample taken in the analytical laboratory that is
extracted and analyzed separately with identical procedures. Analysis of LDSs compared to that of tha first
afiquot give a measure of the precision associated with laboratory procedures, but not with sample collection,
preservation, or.stprage procedures.
.
3.3 Field Blank (FB)/Trip Blank
ASTMTypeI orHPLCgradewaterplaced ina samplecontainerin the laboratory and treated asa samplein sB respects, Including exposure to sampling site conditions, storage, preservation and a l analytical procedures. The purpose of the FB is to determine If tost substances or other interferences are present in the field environment This sample is also referred to as e Trip Blank. Trip blanks are not e requirement for sod or sediment samples.
3.4 Field Duplicate Sample (FDS, Field Dup)
A sample collected In dupficete at the same time from the same location as the sample. The FDS b placed under identical circumstances and treated exactly the same throughout field and laboratory procedures.
ETS-8-012.1
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Malhod o fAnalysis fw ito OetermlnoUono f PerSuorabutanoic Add (PFBA). Pedluoropenunoie Add (PFPeA). PerSuwoheunoic Add (PFHA).PtrikM cdtoptanoie Add (PFHpA). Pettuoroodanoic Add (PFOA),Perttarononanoic Add (PFNA).Perthjorodecanole
Add (PFOA). Periboroundeeonofc Add (PFUnA). Pwfluorododocenote Add (PFOoA), PertuorcO U inew lfanele (PFBS). PeriluorotoxetosuRenste (PFHS). end Perfluoraocaneudonsie (PFOS) in W star. Sod end Sodimeni by LC/MS/MS
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Analysis of the FDS compared to that of the first sample gives a measure of the precision associated with samplecolection,preservation and storage, aswell aswih laboratory procedures.
3.5 Field Matrix Spike (FMS)
A sample to which known quantities of the target analytes are added to the sample bottle In the laboratory beforethe bottles aresent to the field foroollection ofaqueous samples. A known, specific volume of sample must be added to the sample container without rinsing. This may be accompfished by making e "fill to this leveHneon the outside ofthe samplecontainer. The FMSshould be spiked betweenapproximately 50%and 10 limes the expected analyte concentration in the sample. If the greeted rangeofanalyte concentrations is unknown, multiple spikes at varying levels may be prepared to increase the likefihood that a spike at an appropriate'leveils made. The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issuesmaycon*>Bcatethe interpretationofthe sampleanalysis-
3.6 Trip Blank Spike (Field Spike Control Sample, FSCS)
An adquof ofASTMType f or HPLC gradewaterto which knownquantities ofthe targetanalytes are added in
the laboratory prior to the shipmentofthe coledion bottles. The FSCSis extracted and analyzed exactly like a
sampleto help determine if the method is in control andwhethera loss ofanalyte could be attributed to holding
time, sample storage end/Of shipment issues. A low end high FSCS ere appropriate when expected sample
concentrations are not known or may vary. At least one separate, un-spiked sample must be taken at the
sametimeandplaceas eachFMS.
.
3.7 Laboratory Control Sample (LCS)
An aliquotof control matrix to which knovn quantities ofthe target analytes are added in the laboratory atthe
time of sampleextraction. At least two levelsareincluded, onegeneraRyatthe low endof the csBbretion curve
and one near the mid to upper range of the cures. The LCSs are attracted and analyzed exactly like a
laboratorysampleto determinewhetherthe method is in control LCSs should be prepared eachday samples
are attracted-
-
3.8 Laboratory Matrix Spike (LMS)
A laboratorymatrixspike is an aliquotofasampleto which knownquantities oftargetanalytes areadded Inthe laboratory. The IMS is attracted and analyzed exactly fike a laboratory sample lo determine whether the samplematrixconUfoutes bias to the analyticalresults. The endogenousconcentrations cfthe analytes in the sample matrix must be determined in a separate aliquot and the measured values in the LMS corrected for these concentrations. LMSs are required for soils and sediments and are optional for analysis of aqueous sample.
3.9 Laboratory Sample
.
A portion oraiquof ofasample receivedfromthe field for testing.
3.10 Limit of Quantitation (LOQ)
Thelower limit of quantitation (LLOQ) for a dataset is the lowest concentration that can be reliably quantitated within the specified Smite of precision and accuracy during routine operating conditions. To simplify data reporting, the LLOQ is generally selected as the lowest non-zero standard in the calibration curve that meets methodcriteria. Sample LLOQs arematrix-dependent
TheupperBmfcofquantitation (ULOQ)lor s datasetis the highestconcentrationthat canbereiably quantitated within the specified Smiteof precision and accuracy during routine operating conditions. The highest standard in the calibrationcurve that meetsmethodcriteria isdefinedas the ULOQ.
ETS-8-012.1
Page 3o ft2
Method o fAnalysis for the Detannination o fPerSuorodutanolc Add (PFBA), Pertuoropentanote Add (PFPaAJ. Perfuorotannofc Add (PFHA). Portluorohaptande Add (PFHpA), Portuoroodanoie Add (PFOA). PariborononanoJc Add (PFNA). Parfuprodacandc
Add (PFOA). PaflU nroundteatoic Add(PFU \A ), Pw 4uorafcx)tcandeAdd(PfD aA), PerflucroOutanesutfsnata (PFBS),
PeftuoroM ianeauJfonat* (PFMS), and PartuoroodanewlfonaM (PFOS) In W attr, Sod mt i Sedfananl by LC/MS/MS
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3.11 Method Blank
.
An aliquotofcontrol matrixthat it treated exactly Ike a taboretoiy sampleIncludingexposure to a l glassware, equipment, solvents, end reagents that are used with other laboratory samples. The method blank is used to determine If test substances or other interferences are present in the laboratory environment, the reagents, or the apparatus.
3.12 Sample
A sample is an stiquot removed from a largerquantity of material Intended to represent the original source material.
3.13 Stock Standard Solution (SSS) A concentrated solution ofasingle-analytepreparedinthe laboratorywithan essayed referencecompound.
3.14 Surrogate
A compound sintier to the targetanalytefs) in chemicalcompositionand behaviorthat Is not normally found in the samples). A surrogate compound is typicaly a target analyte with at least one atom containing an feotopicatly labeled substitution, tf used, surrogates) are added to ati samples and quality control samples (except solvent blanks and hatf of the prepared method blanks). Surrogates) are added to quantitatively evaluate the entire analytical procedure Inducting sample collection, extraction, and analysts. Inclusion of a surrogateanalyte is anoptionalqualitycontrolmeasureand NOTrequired.
.
3.15 Working Standard (WS)
A solution of several analytes prepared in the laboratory from SSSs and diluted as needed to prepare calibration standardsandotherrequiredanalytesolutions.
4 Warnings and Cautions___________
_________________
4.1 Health and Safety
Theacute and chronic toxicity ofthe standardsfor this method have not been precisely determined; however, each should be treated as a potential health hazard. The analyst should wear gloves, a lab coat, and safety glassestopreventexposureto chemicalsthatmightbepresent.
The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulationsregarding the handing ofthe chemicalsused in this method. A referencefile ofmaterial safetydata sheets (MSOS)should be availableto al personnelinvolvedin.theseanalyses.
4.2 Cautions
.
Theanalyst must be familiar with the laboratory equipmentand potentialhazards Including, but not limited to. the use of solvents, pressurized gas and solvent lines, high voltage, and vacuum systems. Refer to the appropriateequipmentprocedureoroperatormanualtor additionalinformation andcautions.
5 interferences
During extraction and analysis, majorpotentialcontaminant sources are reagents are}glassware. A l materiab
used In the analyses shelf be demonstrated to be free from interferences under conditions of analysis by running method blanks.
ETS-8-012.1
Page 4 of 12
Method otA w iy ii* to r& DeWmJrM&an oTPn1tuorH*o)cAdd (PfB A), Partuwopenisnote Add (PFPsA), PsrtuorbhbM nolc . Add (PfH A), Psriluorghdptanaic Add (PFNpA). Psrfluorooctanofc Add (PfOAJ, Psrfluoranonanofc Add (PFNA). PwSuorwtoesnolc
Add <PFOA), Perftuoraunddcanoic Add (PFUnA). Pertuorododecandc Add (PPOoAJ. PerduorrtU snm dfencM (P fB S ), PeiS uorotacw S uitonalt (PFHS). in d P trS uoroodinasiito n a li (PFOS)M VUibir, Sod and S o d v w l by IC/MS/MS
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ExyLIMS Protocol Number: POOQ3268
Part and supples that contain Teflon should be avoided or minimteod due to the possfeiQtyof interference and/or contamination. These may include, but are not limited to: wash bottles, Teflon* lined caps, autovial caps, HPLCparts, etc. The use cf disposable micropipettes or pipettes to aliquot standard solutions is recommended to make calfcration standardsandmatrix spfcea.
6 Instrumentation, Supplies, and Equipment
6.1 Instrumentation and Equipment
A high performance Squidchromatograph capable ot pumping up to 2 solvent and equipped with a variable
volume vectorcapableofinjecting 5-200p i collected toa tandemMassSpectrometer(LC/MS/MS). Applied
BiosystemsSciexAPI 5000instrumentation is requiredto meetthe LOGSof 0.025 ng/ml (water)and0.20 ngfg
(sol and sediment). If analyte concentrations require dilutions tor one or more aneiytas that predude the
targeted LOQs from being reached, Applied Biosystems SciexAPI 4000 instrumentation may be utilized since
the LOQswilatoady beraised.
'
Analytical balancecapableofreadingto 0.0001 gram.
A device to collectrawdatator peakintegration andquantitation.
15-mL end 50-mLdisposable polypropylene centrifuge tubes.
Deposable micropipettes(1040y l. 2W 0 pL, 50-100pL. 100500pi).
l2frmL LDPEnarow^nouth bottles.
2-mL dearHPLCvia)kit
Disposablepipettes, polypropyleneorglass as appropriate.
Ultrasonic bath.
Centrifugecapableof spinning 15-mLand50-mLpolypropylenetubes at3000rpm
6.2 Chromatographic System
AnalyticalColumn; Luna3pmC8(2) Mercury(Phenomenon).2rrwnx 4mm, 3 pm (P/tf. 00M-4248-D0-CE)
Temperature: 35*C
'
McMe Phase(A): 2mMAmmoniumAcetatein Water MobilePhase(B): Methanol Gradient Program:
'
Tme ftnlnl
2l
%B
0.0 90 10
0.5 90 10
2.0 10 90
5.0 10 90
5.1 0 100
6.0 0 100
6.1 90 10
10.0 90 10
Ir^ectionVolume: 5p i (canbe increased toasmuchas 50pL),
Flow Rate ' fmL/minl
' 0.75 0.75 0.75
0.75 0.75 0.75 0.75 0.75
Quantitation: PeakAres- externalstandard calibrationcurve, 1/xweighted. 6TS-6-012.1
Page 5of 12
Method or Andyali for V tt DetemlnaOoA ofP trikm obuU nolc Add (P ifiA ). PwfluoroponUnaic Add (PFPeAJ, PorSuorohuanoie
Add (PFHA), Portuorehaptanoic Add (PFHpA), PwSuorooctanolc Add (PFOA). Perfluorononande Add (PfN A), PvSuorodacdW le Add (PFOAJ. PdrSuoroundoeandc Add (PfU nA), Portuorododocanoie Add (PFOoA), Parauorobutarvstefenati (PfB S), Pemuoroheionoodranati (PFHS). end PerSuorooetanoMifonalo (PfO S) In W Uaf. S o land Sadimantby LCfMSMS
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RunTime; -1 0 minutes.
The previous information is Intended as a fluide; alternate conditions and equipment may be used
provided that data quality objectives aremet
6.3 MS/MS System
6.3.1 Mode- ElectrosprayNegath*MRMmode,monfcvinglhefotawinfltransi6m
A/iaMe
Transition Monitored
PFBA
213-169
PFPeA
263 -219
PFHA
313 -*269 and 313 -119
PFHpA .
363-319. 363-169 and 363-119
PFOA
413 - 369.413 - 219 and 413 -169
PFNA
463 - 419.463 -* 169 and463 - 219
PFDA
$13 - 469. 513 -* 219 and 513 - 269
PFUnA
563 -519. 563 -269 and 563 -> 219
PFOoA
613 -569.613 - 169and 613 -319
PFBS
299-80 and 299-99
PFHS
399-80 and 399-99
PFS
499 -80. 499-99 and 499-130
Multiple transitions for monitoring the analytes is an option beeaust summing multiple transitions
may provide quantitation of Somers that more closely matches NMRdata and may have the
added benefit of increased analyte signal. The use of.one daughter ion is acceptable if method
sensitivity is achieved, provided that retention time criteria are met to assure adequate specificity.
The previous information is intended as a guide, alternate instruments and equipment may be used.
7 Reagents and Standards
____ _______________
Water- HPLC grade Methanol- HPLC grade Ammonium Acetate - AC S. Reagent Grade Acetonitrile - HPLC grade Pertuorobutanoic Acid (PFBA) - Oakwood Products, Inc Pertuoropentanot Add (PFPeA) - StynaAJdrich Pertuorohexancic Add (PFHA) - Oakwood Products, Inc Perituoroheptanoic Acid (PFHpA)-Oakwood Products, Inc PerfluofooctanoicAct'd (PFOA) - OakwoodProducts, (nc Pertuoiononandc Acid (PFNA)- OakwoodProducts,ine Pertuorodecanoic Add (PFOA)-Oakwood Products, ine Pertuoraundecanoic Add (PFUnA)- Oakwood Products, ine Pertuorododecanoic Add (PFOoA)- Oakwood Products, Inc
ETS-8-012.1
PagedoftZ
M atted ofAnalysis for tha Oatarmlnatton ofPartuorobUanolc Add (PFBA). ParSuoropanlanoic Add (PFPaA). Parfluorohexanoic
Add (PFHA). Parfluorohaplanqlc Add (PFHpA),Pariluaroodanoic Add (PFOA). PerSuoranonanoleAdd (PFHA),PtrSuorodecanok Add (PFOA). Partuoroundacanoic Add (PFUnA). PartworododaeandcAdd (PFOo*), PartuerobutanasuKenala (PFBS). PartuorohasanasuKanata (PFHS).and P artaarooctanatallonale (PFOS) In W alar, 80Sand Sadlmem te IC /M SM S
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ExyLIMS Protocol Number: P0003268
Perliuorebutanesulfonate(PF0S)- 3M
PerfiuorohexanesuVonate(PFHS)- 3M
Perfluoraoctanesulfonato (PFOS)- Fluka
The previous information is Intended as a guide. Reagents and standards from alternate sources may be used.
8 Sample Handling
________________________ ______________________
6.1 Water Sample Extraction
8.1.1 Measure 10mLofsampleintoa50mLpolypropylenecentrifugeMm. Fortificationsareto bedoneatMs point,?necessary. Otherflumes andcontainersmaybeusedasappropriate.
8.12 Add1QmL<rfacetonilrietothesrpte\thecentrifogetebe. Captightlyandshake.
8.1.3 Sonicatesamptefor-2 hoursatroomtemperature. Thisstepisoptional, but b recommendedfparticutaies
appearto bepresent
.
8.1.4 CertrifogeforMOminutasat-3000rpm. Thisstepisoptional, butis recommended?samplesare sonicated.
8.1.5 Transferaportionofthesupernatantto anautosamplervialforanalysis.
6.1.6 Mute sample, ?necessary,wfth$050acetontritavrater.
8.2 Soil and Sediment Extraction 82.1 Weigh1gofsampleintoa 15-mLpo/ypropytenecentrifugetube. Fortificationsarefobedoneatthispoint?
622 Add0m l of80:20acetoniWe.vraterto thesamplein(heoentrifogetube. CaptighOyandshake 82.3 Sonicatesamplefor-2 hoursatroomtemperature. 82.4 Centrifugefor-IOn^utes at-3000rpm 82JS TransforaportfoncfthasupematanttoanautosarTpiervialforanalysis. 82.6 Mute sample,?necessary,with50:50acetonlrle.'waler. 82.7 Analyzesamples usingelectrosprayICAAS/MS
Other welgMs end volumes can be used as long as the QC elements specified fo this method era satisfied and all samplepreparationproceduresmaybe reconstructed.
9 Sample Analysis LC/MSfflIS___________________________ '____________
ETS-8-012.1
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M atto* o fAnalysis fo r tha OatarmtnaUono f PaifluPO M anoic Add (PF6A). Partluoropentanato Add (PFPaA), PaiSuorotoxanote
Add (PFHA). Partuorchaptawte Add (PFHpA), Pertuorooctatoic Add <PfOA). ParSuorononanoM Add (PFNA). PartuorodacanoM Add (PFDA). Parfluorouneacandc Add {PFUnA).Parftuonidodacanofc Add (P fOCA). Partuwobutanasultanala (PFBS), PartuorotoxanatutConata (PFKS1.M PatSuoroodanesulfonala (PFOS)In W alar.Son and S adrrant by LCTMSMS
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9.1.1 A mintonumofskcalferaliontarelsmustbepresenttoithefinalcaferationcurve.
9.122 Anentiresalofcafcrationstandardsbtotjedadatthe boginningofasamplesetpriortosampleanalysis. As an atematfve. an entire setofeafibralionstandards may beinjectedat the beginningofa samplesat Wowed bycafeaGcnstandardsinterspersedvaryMO samples{toaccounttorasecondsatofstandards), tot eithercase,cafitretion standardsora continuing calbration verification standardmust bracketthe first sampleandthelastsample. AnalyticalstudesmayonlyrequirethatanwtWcaptationcur beanalyzed priortosamplesandta t thecontinuedaccuracyoftheinitialcafemfoncurvebecorttomadbythe analysisof continuingcaferalionverificaficnstartdaids. Thesarmapproachshouldbeusedtirou^xuttheentirestudy unlessreasonsaredocumertedandtechnicel^otfcsdon tochangeIsapprovedpriortosampleanalysis.
9.1.3 Thestandardcun* isplottedbyquadraticH(y ax*+bx*cX weightedV* orunwanted, orusingaInear fiLweighted1/Ausingsutafalesoftware. Thecafibretioncurvesmayincludebutshouldnotbeforced throughzero. Themathematicalmethodusedtocalculatethectfbrafloncurveshouldbeapplied consistentlythroughoutastudy. Anychangeshouldbedocumentedintherawdata.
9.1.4 Samplescontaininganalytesmatarequantitatedabovetheconcentrationofthehigheststandardin the curveshouldbe Mherdiluted and reanalyzedor reinjected usinga smallervolume.
10 Quality Control__________
10.1 Data Quality Objectives.
This method and required quafity control samples Is designed to generate data accurate to +/-3Q% with a targeted LOO of 0.025 ngrtmL (water) or 0.20 ngrg, Wet weight (soil end sediment). Any deviations from the quality controlmeasuresspeled outbelowwiBbedocumentedin therawdata andfootnoted Inthe finalreport
10.2 Blanks
MethodBlank
Method blanks mustbepreparedwith eachextraction batch. A rangeofthree to seven methodblanks must be prepared. These method blanks must be hterspersed throughout the extraction batch and analyzed interspersedthroughouttheanalyticalsequence.
The meanareacountfor each analyte in the method blanksmust be less than 50%of the area count of the LOQ standard. The standard deviation of the area counts of these method blanks should be calculated end reported. If the mean area counts of the method blanks exceed 50% of the LOQ standard, then the LOQ must be raised to the.first standard level in the curve that meets criteria, or alternatively, the method blanks must be evaluated statistically to determine outliers or technical justification to eliminateoneormoreresults should bemade.
1022 SolventBlank
Solver blanks mustbeanalyzedthroughout the analyticalsequence. Solventblanks that show ansa counts greateir than 50% of the area counts of the LOO standard must be evaluated to determine if analytical results are significantly impactedby samplecarry-overor an unacceptable bufctup ofanalyte In the instrumental background.
10.3 Sample Replicates
Samples should be prepared tot dupficate tot the lab (soil and sediment) or collected In duplicate tot the field (water). The relative percent difference, RPD, should be reported. RPD results greater than 20% (water) or
ETS-e-012.1
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Method or AralyM for <w OManwrettaw o t Puaiabul>ftolc Add (PFBA),Prtiuorapnttiw tc Add (PFPA).P re *yo h m re ic Add (PFHA), PerfluorcftepMnoic Add (PFHpA), PwSuorooctanoieAdd (PFOA). Pertuorononanoic Add (PFNA), Pwfluorodacsnoto
Add (PFOA). PerSueroundeeiPdcAdd (PFUnA). PwDuorododecanolc Add (PFDeA), PtrttuerabuunMuSonaW (PFB8). PwtkwronexanetulfDnate (PFHS), end PefluorooctenesulfonMe (PFOS) In WHar, S ol and Sedimentby LC/MS/MS
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30% (sol and sediment}will beflagged in the report, but wil not be excluded from reporting. The requirement forrepBcates excludesfieldWanksand rinseblanks.
10.4 Surrogate Spikes Surrogatespikes arenot requiredbutmay be usedon project specificrequirements.
10.5 Lab Control Sample
Triplicate tab control spites at a minimum of two different concentrations are to be prepared with each extraction batch. Low lab control spikes should be prepared at concentrations in the range offive to ten times higher than the targeted LOQand high lab control spites should be prepared at concentrations near the mid* point of the curve. The relative standard deviation of the control spices for these six pooled control samples must be less than or eqiat to 20% andthe avenge recovery must be 80*120%. Hthe above criteria arenot met, the entire setofsamplesshould bere-injected orre-extractedasappropriate.
10.6 Field Matrix Spike / Lab Matrix Spikes
Recoveries of field matrix spikes (aqueous samples) end laboratory matrix spices (soft and sediments) are anticipated to be between70% and 130%of the fortified levels. Kthe recoveries of the spikes fait outsidethe acceptable range, the sampleresuRwDbereported as'NR?(not reporteddueto non-compKantQCresuRs).
The targeted fortification levels should be at least 50% of the endogenous level and less than 10 times the endogenousleveltobe usedwithoutjustification todeterminethe statementofaccuracyfor analytical resuts.
The average ofthe sampleand the labduplicate (sols and sediment)orfield duplicate (water) should be used
to calculate therecovery.
.
10.7 Standard Preparation
10.7.1 StandardStockSolutions
Prepare Individual stock solutions of each analyte at 1000 pmL by weighing 100 mg of analytical standard
(corrected for purity and salt content, if necessary) and dKute to 100 m l with methanol or aoetonrtrfie In
separate 100-ml volumetric flasks. For purity correction, take the amount of analyte that is needed for
weighing (i.a.100 mg) and divide by the purity Indecimalform(Le. 99.6%* 0.996). Theresultis theweightthat
is needed to makethe solution corrected for pirtty (Le. weigh 100.4 mg). For salt correction, calculatethe safl
contentby taking the molecularweight ofthe target compound (le. PROS* 499)anddividing bythe molecular
weight of the entire compound (Le. PFOS potassium salt (C*FitS0j-K* 536). The result is the sat content in
dcima}form (le. 0.9275). Take the amountof analytethat is needed for weighing (le. 100 mg)and dkrideby
th salt content in decimalform. Theresult Is the weight that is needed to make the solution corrected for salt
content (i.e. weigh 107.6 mg). Store at stock solutions In appropriate containers and at appropriate storage
conditions for upto 6monthsfromthedateofpreparation.
'
Atemate stocksolutionconcentrationscanbe made,if necessary,usingalternative massesandvolumes.
10.7.2 StandardFortificationSolutions
A 10 pgAnLmixedfortification solutioncontainingsti oftheanalytes is prepared bybringing 1mLof each ofthe 1000 pg/mL stock solutions to a final volume of 100 mLwith acetonitrle in a 10CknL volumetric flask. A 1.0 pgfrnL mixed fortification solution containing at of the analytes is prepared by bringing 10 mL of the 10pgfrnL mixed solution to a final volume of 100 with acetonitrile in a 100-ml volumetric flask. A 0.1 uotnL mixed
fortification solution containing at of theanalytes is prepared by bringing 10m l of the 1.0 pgftnLmixed solution to a final volumeof 100 with acetonitrile in a 100-mlvolumetric flask. A 0.01 pgfrnLmixedfortification solution containing at of the analytes is prepared by bringing 10 mL of the 0.1 pgftnL solution to a final volume of 100
with acetonitrile in a 100-rri volumetric flask. Store at fortification standards up to 6 months from the date of preparation .
ETS-8-012.1
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Method oTAnalyeit lor the DeiermlnotiQne l PerfluorobuUnoto Add (PFBA), PorSuorepenlenoic Add (PFPaA). Partuorohexanoic Add (PFHA), PeifluoreheptaneicAdd (PFHpA). Perfluoroodonote Add (PfOAJ. PerSuorononanofc Add (PFNA), PorSuorodoeanoic
Add (PfDAJ. Perfcm undecandc Add tPFVJo^.Pvfworododoeondc Add (W DoA), PertluorotaianwuHonate (PFBS). PwSuorohvwiMUdbnoto (PFHS). and Porfluoroocunesufonata (PFOS) In WMer. S o l and Sodmonlby LCIMSlMS
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Alternatefortification solution concentrations canbemade,if necessary. likewise, alternativevolume* maybe used.
10.8 Instrument Calibration
10.8.1 LG/MSMScaBrttionstBndaTis arepreparedeidamallyin 50:50aratonitribvraterandamtoad toconstruct tie caferationcurvesforthewater,soi, andsedimentprocedures.
10.8.2 Thefotowingisatypicalexampleofacaftration set Alternateoradditionalconcentrationsmaybeprepared asneeded.
Concentration Initial Solution
Final Concentration of
of Initial Solution Aliquoled Volume Final Solution
(ng/mL)
{mL> Volume (mL)
100 5
100
Calibration Standard (ng/mL) 5.0
100
2.5
100
2.5
10 10 100
1.0
5 10 100
0.5
2.5 10 1 10
100 100
0.25 0.1
0.5 1
10 2.5
100 100
0.05 0.025
Store all caftrration standards for upto threemonths. ABematevolumes andconcentrations ofstandardsmay
bepreparedas needed.
.
11 Data Analysis and Calculations
Use the following equationto calculate the amountof analyte found(in nghnl, basedon peak area) using th standardcurve(linear regression parameters) generatedbyan appropriatesoftwareprogram:
Analyte found(ng/ml)
; OF
DFfactorbywhichthe finalvolumewasdiluted, if necessary.
For samplesfortified with knownamounts ofanalyte prior to extraction, use the following equation to calculate the percentrecovery.
,,R e c o. v eTortaylanalyte fo" und (ng*/Am1nl)a-.lAy.tveeardadgeed.a.(nnga/.lryrtre.t..)foundin sample(na/mL) xi.u.o.
Usethe followingequationto convertthe amountofanalyte (bundinngftnLto ngfg(ppb).
Analytefound(nsymL)* Volume extracted|mL)
' lr r '
Karnnla uiainhlm\
Usethe foltowingequation to calculatethe amountofanalyte found in ppbbased ondryweight. Analyte found(ppb) dry weightAnalyte found(ppb) x(100%/ totalsolids(%)]
12 Method Performance
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Method o fA n e iy iti 1erthe Determination o i Perfluorobutanote Add (PFBA), Perfknropenlanole Add (PFPeA), PertJooroheMndc
Add (PFHAJ. PerSuorelteptanoie Add (PFHpA). Pertuorooctande Add (PFOA).PerfluorononMigfc Add (PFNAJ, PerSuorodeonofc Add (PFOA). Perfuoroundecandc Add (PFUnA), PerSuerododecenoie Add (PFDoA).PerflgorobUaneautfonala (PFBS), Perfluorohcunaadronale (PFHS). end PerAioroodanaaultonate (PFOS) in W ater. S o lend Sedbrwrt by IC/USTMS
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Any method performance parameter* that are not achieved must be considered in the evaluation ofthe data. Nonconformanceto anyspecifiedparametersmustbe describedanddiscussed ifthe Technical Manager(nonGLP study) or StudyDirector(GLPstudy)choosesto reportthedata.
Hcriterialisted in this methodperformancesection arenot met, maintenancemay beperformedon the system andsamplesreanalyzed, orotheractionstakenesappropriate. Documental!actions inthe rawdata. .
Ifdata areto be reported when performance criteria havenot been met, the data must be footnoted ontables enddiscussed in thetext ofthe report
12.1 System Suitability
.
.System Suitability standards are not a requiredcomponent of this method. If required by protocol or by the technical manager, a minimum of three system sustabBty samples are injected at the beginning of each analytical run priorto the calibration cuve. Typicallythesesamplesare ata concentrationnearthe midlevel of the calibration curve and are repeated injections from one autosampler vial. The system suitabiity injections must haveareacounts with an RSDof5%andaretentiontime RSDofS2%tobe compBant
12.2 Quantitation
Calibration Curve: The coefficient of determination (r*) value forthe castration curve mustbe greaterthan or equal to 0.985. The concentration of each point in the curve must back calculate to be within 125% of the theoretical concentration with the swaption ofthe UOQ, which may be within 30%. Calibration curve points thatarenotat the highor lowend ofthecurvemaynotbedeactivated.
CCV Performance: The continued accuracy of (he catibcafon curve must be demonstrated b/analyzing continuing caibration verification (CCV) standards. Each CCV may be a caibration curve point that is reflected or a separately prepared standard, and is typically near the middle of the caibration curve. Alternative concentrations or multiple concentrations may be chosen depending on project requirements. Not more than ten samples or spires may be analyzed between the Initial calibration and a CCV or bracketing CCV*. Theaccuracy c4eachanalytemustbewithin25%ofthe theoreticalvalue. Samplesthat are bracketed byCCVi not meetingthese criteria mustbereanalyzed.
Demonstration of Specificity: Specificity is demonstrated by chromatographic retention time (within 4% of standard)andthe massspectral responseofuniqueions.
12.3 Sensitivity
Thetargeted limit ofquantitationforal analytes is 0.025 ngArt (water) and0.20 ng/g (soils andsediments, wet weight). TheLOQforany specificanalyte mayvarydependingonthe evaluation ofappropriate blanks and the accuracyofthe Iwr-teveicalibrationcurve points. Referto Section 10for additionaldetails.
12.4 Accuracy
This methodand requiredquality control samples are designed to generate data that are accurate to +A30%. Section 10 contains additional information regarding the required accuracy of laboratory contra! spikes, field matrixspikes (watersamples)and laboratorymatrixspikes (soilsandsediments).
12.5 Precision
Samples should be prepared in duplicate in the fob (soi and sedvnent) or colacted in duplicate in the field (aqueous). The relative percentdifference, RPD, shouldbereported. RPD results greaterthan20% (water) or 30% (soil and sediment) will be flagged in the report, butw i not be excluded from reporting. Therequirement forreplicates excludesfield blanks or rinse blanks.
Section10contains additional informationregarding the requiredprecision oflaboratory control spires.
13 Pollution Prevention and Waste Management ' T&WU.1 "
Page lie f
Method ofAnalyse tor Via Oelermfnstton o f Perfluorobuienolc Add (P fBA). Porfluoropentanoto Add (PFPoA). Parfluorohasenolc Acid (PFHA). Perituorohoptsnoic Add (PFHpA). Perfluoroodanoic Add (PFOA1, Periluoraron*noie Add (P fNA1. Parituorodecaneic
Add (PFOA). PertuoroundecanoicAdd (PFUnA), P e rito rododecendic Add (PFOoA), PerttorobvUnesuHonate (PfB S).
Perftorohexenetulfooete (PFHS). end Perituoroteanuulfonete(PFO S) In WWer, Sc* and Sedtaent by IC/MS/MS
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Waste generatedwhen performing this methodwil be deposed ofappropriately- The original samples will be archived atthe 3MEnvironmentalLaboratoryin accordancewith internal procedures.
14 Records
Each datapackage generatedtora study mustinclude al supporting informationfor reconstruction ofthe data. Information forthe datapackagemuetInclude,butis not Srrdtedtothefoltowlng Items: studyor projectnumber, sample and standard prepsheets/records,instrumentamlog (mawment batchrecords, instrumentacquisition
method, summarypages), instrumentresults files, chromatograms, catibrationcurves, enddeta calculations.
15 References
Exygen ResearchAnalytical Method V0003305-1 details method development and verification of recovery for multipleanalytes Ingroundwater, surfacewater, soil, andsediment Thiswill be archived ss partofEQ6-0549.
16 Affected Documents
None.
17 Revisions____________ .
Revision Number
RevisionDescription .
Revision Q&
Thesonicalion andcentrifugation stepsofthewater samplepreparationwere
1 1/10/06
made optional. Theaccuracyrequirementsofthe analyticalbalancewere updated.
Themethodwas updatedto allowthe useofa linear 1/xweightedcalibration curve.
Thetable ofrequireddaughterIonsforthe analyteswas updated. Themethod
was updatedto alowdifferentsamplevolumesto be extractedendto allow
differentstandard bottles andstoragecond&ionsto beused. Calibrationrequirements
wereupdated to showthatanalyzingcontinuing calibration verification standardsatone
concentration wasan acceptableaftametivato reloadingall curvepoints usedto construct
thecalibrationcurve. Minorwordingchangesweremadethroughout
ETS4-012.1
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MatPode fAnalysis fo r the DeterminaDono l PtrSoorcfeUanoic Add (PFBA). ParSuoropantanolcAdd (PFPeA), Parfluoroftexarefc Add (PFHA). Perfloorohpianole Add (PPHpA). Partuoroocunoie Add (PFOA). Pwlluorenenaneie Add (PFNA). Perfluorodecanoie
Add (PFDAJ. Pertuoroundacande Add (PFUnA), Parftuorododecanoic Acid (PFDoA), Periluorobutanasuironata (PFBS). Pertluorohaunasuttonata (PFHS), and PsrSuoroodanesdfonata (PFOS) n w ater. Sod and Sadmerit by LC/MS/MS
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MPI Research
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ANALYTICAL METHOD Method Number: V0001780
Method of Analysis for the Determination of Perfluorooctanoic Acid (PPOA) in Water by LC/MS/MS
Analytical Testing Facility:
Exygen Research 3058 Research Drive State College. PA 16801
Approved By:
TAX
_________
Paul Connolly
'
Technical Leader, LC-MS, Exygen Research
2
John Flaherrtty / Vice President, Operations, Exygen Research
>oiz.w/otf
Date
Date
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Exygen Research
Method NumberV00OI780
ANALYTICAL m e t h o d
]
Method ofAnalysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Water by LC/MS/MS
1.0 Scope
This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in water.
2.0 Safety
2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At least 40 mL of test sample for extraction. 3.2 No sample processing is needed for water samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room
temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Any samples containing particles should be centrifuged at -3000 rpm for -5
minutes and the supernatant used for the extraction. 3.6 Sample collection procedures will be specified in the sampling plan for this
project.
Reagents and Standards
4.1 Water-HPLC grade 4.2 Methanol - HPLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Perfluorooctanoic Acid - Sigma-Aldrich
5.0 Instrument and Equipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5*200 pL connected to a tandem Mass Spectrometer (LC/MS/MS).
5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable of reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-100uL, 100*200uL). 5.7 125-mL LDPE narrow-mouth bottles. 5.8 2 m L clear HPLC via) kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10*100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (1g) tC1S SPE cartridges.
Pge 2 o f 7
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Exygen Research
Method Number V0001780
ANALYTICAL METHOD
Method of Analysis for the Determination of Periluorooctanoic Acid (PFOA) in Water by LC/MS/MS
5.12 SPE vacuum manifold. 5.13 Centrifuge capable ofspinning 50 mL polypropylene tubes at 3000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130)
6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program:
Time (mini
0.0 1.0 8.0 20.0 22.5
'A h 65 65
25
25 65
Flow Rate % B ImL/miri) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3
6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes.
The above conditions are intended as a guide and may be changed in order to optimize the HPLC system.
7.0 MS/MS System 7.1 Mode: Electrospray Negative MRM mode, monitoring 413 --369 m/z.
The above conditions are intended as a guide and may be changed in order to optimize the MSMS system.
8.0 Preparation of Solutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL of water.
Alternate volumes may be prepared.
Pkge 3ot ?
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Exygen Research
Method Number V000I780
ANALYTICAL METHOD
Method ofAnalysis for the Determination of Periluorooctanoic Acid (PFOA) in Water by LC/MS/MS
9.0 Standard Preparation
9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution o f-100 pg/mL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 10 pg/mL fortification solution ofPFOA is prepared by bringing 10 mL of the 100 pg/mL solution to a final volume of 100 with methanol in a 125 raL LDPE bottle. 9.1.3 A 1.0 jig/mL fortification solution of PFOA is prepared by bringing 10 mLofthe 10 pg/mL solution to a final volume o ft 00 with methanol in a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 1.0 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date ofpreparation.
9.2 Standard Calibration Solutions
9.2.1 LC/MS/MS calibration standards are prepared in HPLC water The calibration standards are processed through the extraction procedure, identical to samples.
9.2.2 The following is a typical example*, additional concentrations may be prepared as needed.
Final
Concentration Fortification Volume of Concentration of Calibration
o f Fortification Volume Fortified Control Calibration
Standard ID
Solution (t>pb)
(PL)
Sample (mL) Standard (ppt)*
(example)
0 0 40
0 XCmmddyy-Q
10 100 40
25 XCmmddyy-1
10 200 40
50 XCmmddyy-2
10 400 40
100 XCmmddyy*3
100 100 40
250 XCmmddyy-4
100 200
40
500 XCmmddyy-5
100 400
40
1000 XCmmddYY-6
* The extracted concentration of the calibration standard is equal to 8x its initial
concentration, due to the concentration of the standard during the extraction (SPE).
XC = extracted calibration standard.
Ptgc4 o f 7
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Exygen Research
Method Number VOOOI783
a x a l y tic .vl m e t h o d
Method ofAnalysis for the Detennination ofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
14.3 For samples fortified with known amounts of PFOA prior to extraction, use the following equation to calculate the percent recovery.
Recovery (%)
[ total analyte found (ng/g) - analyte found in control (ng/g)] .. ^ analyteadded (ng/g)
MPI Research
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Exygen Research
Method Number VO00I780
__________________________ ANALYTICAL M ETHOD___________________________
Method ofAnalysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS
versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system.
12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed.
13.0 Acceptance Criteria
13.1 Chromatogram must show a peak of a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide.
13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 50 ng/L, then a new blank sample must be obtained and the entire set must be re-extracted.
13.3 Recoveries of control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set of samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted.
13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation of the calibration curve. However, the total number of extracted calibration standards that could be excluded must not exceed 20% of the total number of extracted standards injected. The correlation coefficient (R) for calibration curves generated must be 0.992 (R2 0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set of samples should be reanalyzed.
13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed.
14.0 Calculations
14.1 Use the following equation to calculate the amount of PFOA found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program:
PFOA found (ng/L) (Peak area - intercept) x DF slope
DF *factor by which the final volume was diluted, if necessary.
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Exygeo Research
Method NumberV0001780
ANALYTICAL METHOD
Method of Analysis for the Determination of Periluorooctanoic Acid (PFOA) in Water by LC/MS/MS
14.2 For samples fortified with known amounts of PFOA prior to extraction, use the following equation to calculate the percent recovery.
Recovery (%) -
[ total analyte found (ng/L) - analyte foundin control (ng/L)] analyte added (ng/L)
MPI Research
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ANALYTICAL METHOD Method Number: V0001781
Method of Analysis for the Determination of Perfluorooctaooic Acid (PFOA) in Soil by LC/MS/MS
Analytical Testing Facility:
Exygen Research 3058 Research Drive State College, PA 16801
Approved By:
"PA co L
Paul Connolly
'
Technical Leader, LC-MS, Exygen Research
loln/p/
Date
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Exygea Research
MethodNumber V00Q17I
ANALYTICAL METHOD
Method of Analysts for the Determination ofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
1.0 Scope
This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in soil.
2.0 Safety
2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 Atleast 15 goftest sample for extraction. 3.2 No sample processing is needed for soil samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room
temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this
project.
4.0 Reagents and Standards
4.1 Water -KPLC grade 4.2 Methanol - HPLC grade 4.3 Ammonium Acetate - A.C.S. Reagent Grade 4.4 Perfluorooctanoic Acid - Sigma-Aldrich
5.0 Instrument and Equipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5*200 pL connected to a tandem Mass Spectrometer (LC/MS/MS).
5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable of reading to 0.00001 g. 5.4 50 mL disposable polypropylene centrifuge tubes. 5.5 15 mL disposable polypropylene centrifuge tubes. 5.6 Disposable micropipets (50-100uL, 100-200uL). 5.7 125-mL LDPE narrow-mouth bottles. 5.8 2 mL clear HPLC via] kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (lg) tC18 SPE cartridges. 5.12 SPE vacuum manifold. 5.13 Ultrasonic bath.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygeo Research
Method NumberV000178I
| ANALYTICAL METHOD
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
5.14 Wrist-action shako1. 5.15 Centrifuge capable of spinning 50 mL polypropylene tubes at 5000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: FtuophaseRP (Keystone Scientific), 2.1 mm x 50 mm. 5p (P/N: 82505-052130)
6.2 Temperature: 30C 6.3 Mobile Phase (A): 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B): Methanol 6.5 Gradient Program:
0.0Time (min) 1.0 8.0 20.0 22.5
%A 65 65 25 25 65
Flow Rate % B (mL/min) 35 0.3 3S 0.3 75 0.3 75 0.3 35 0.3
6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 RunTime: ~ 23minutes.
The above conditions are intended as a guide and may be changed in order to optimize the HPLC system.
7.0 MS/MS System
7.1 Mode: Electrospray Negative MRM mode, monitoring 413 --369 m/z for PFOA.
The above conditions are intended as a guide and may be changed in order to optimize the MSMS system.
8.0 Preparation of Solutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL ofwater.
Alternate volumes may be prepared.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygea Research
Method Number V0001781
_____________ .________ANALYTICAL METHOD
Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
9.0 Standard Preparation
9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution of ~100 pg/mL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPEbottle. 9.1.2 A 10 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 100 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 10 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of fhe 1.0 pg/mL solution to a final volume of 100 with methanol in a 125 mLLDPE bottle. 9.1.5 A 0.01 pg/raL fortification solution of PFOA is prepared by bringing 10 mL of the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months fromthe date of preparation.
Standard Calibration Solutions
9.2.1 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through the extraction procedure, identical to samples.
9.2.2 The following is a typical example: additional concentrations may be prepared as needed.
Final
Concentration Fortification Volume of Concentration of Calibration
o f Fortification Volume Fortified Control Calibration
Standard ID
Solution (nob) (HL> Sample (mL) Standard fppt)* (example)
0 0 40
0 XCmmddyy*0
10 100 40
25 XCmmddyy-1
10 200 40
50 XCmmddyy-2
10 400 40
100 XCmmddyy-3
100 100 40
250 XCmmddyy-4
100 200
40
500 XCmmddyy-5
100 400
40
1000 XCnunddyv-6
* The extracted concentration of the calibration standard is equal to 8x its initial
concentration, due to the concentration of the standard during the extraction (SPE).
XC = extracted calibration standard.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number VOOOI781
ANALYTICAL METHOD
Method ofAnalysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Soil by ' IC/MS/MS
9.2.3 A zero standard solution (reagent blank) must be prepared with each set of standards extracted.
9.2.4 Store all extracted calibration standards in 15-mL polypropylene tubes at 2C to 6C, up to two weeks.
9.2.5 Alternate volumes and concentrations of standards may be prepared as needed.
10.0 Batch Set Up
10.1 Each batch of samples extracted (typically 20 or less) must include at least one reagent control (method blank using 5 mL of methanol) and two reagent controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project.
11.0 Sample Extraction
11.1 Weigh 5 g of sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well).
11.2 Add 5 mL of methanol and shake on a wrist action shaker for -15 minutes. 11.3 Transfer the tubes to an ultrasonic bath and sonicate for -15 minutes. 11.4 Bring the volume up to 40 mL with water in the 50 mL polypropylene
centrifuge tube. 11.5 Centrifuge for -10 minutes at -3000 rpm. 11.6 Condition the Cis SPE cartridges (1 g, 6 mL) by passing 10 mL methanol
followed by 5 mL of HPLC water 2 drop/sec). Do not let column run dry 11.7 Load (decant) the sample on the conditioned Ci SPE cartridge. Discard
eluate. 11.8 Elute with -5 mL 100% methanol. Collect 5 mL of eluate into graduated
15 mL polypropylene centrifuge tubes (final volume - 5 mL). 11.9 Analyze samples using electrospray LC/MS/MS.
12.0 Chromatography
12.1 Inject the same amount of each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples.
12.2 Standards of PFOA corresponding to at least five or more concentration levels must be included in an analytical set.
12.3 An entire set of extracted calibration standards must be included at the beginning and at the end of a sample set. Extracted standards must be interspersed between every 5-10 samples. As an alternative, an entire set of
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MPI Study N o.: 013 7.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V000178)
ANALYTICAL METHOD
1
Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
extracted calibration standards may be injected at the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to account for a second get of extracted standards). In either case, extracted calibration standards must be the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent)
software system. V2.5 'Sample response should not exceed standard responses. Any samples that
exceed standard responses should be further diluted and reanalyzed.
13.0 Acceptance Criteria
13.1 Chromatogram must show a peak of a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide.
13.2 Method blanks must not contain PFOA at levels greater than die LOQ- If a blank contains PFOA at levels greater than 50 ng/L, then a new blank sample must be obtained and the entire set must be re-extracted.
13.3 Recoveries of control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set of samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation of foe calibration curve. However, foe total number of extracted calibration standards that could be excluded must not exceed 20% of foe total number of extracted standards injected.
13.5 The correlation coefficient (R) for calibration curves generated must be 0.992 (R2 0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set of samples should be reanalyzed.
13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. irretention time drift exceeds this limit within an analytical run then the set must be reanalyzed.
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Interim Report #1 - Analysis of Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method NumberV0001781
L ANALYTICAL METHOD Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
14.0 Calculations 14.1 Use the following equation to calculate the amount of PFOA found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program:
PFOA found (ng/L) = (Peak area - intercept! x DF slope
DF = factor by which the final volume was diluted, if necessary.
14.2 For samples fortified with known amounts of PFOA prior to extraction, use the following equation to calculate the percent recovery.
Recovery (%)
[ total analyte found(ng/L) analyte found in control (ng/L)] ^ analyte added (ng/L)
14.3 Use the following equation to convert the amount of PFOA found in ng/L to ng/g (ppb).
PFOA found (ppb) - [PFOA found (ng/L) x volume extracted (0.04D1 sample weight (5 g)
14.4 Use the following equation to calculate the amount of PFOA found in ppb based on dry weight.
PFOA found (ppb) dry weight **PFOA found (ppb) x [100% / total solids(%>)
Pgc7of7
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
ANALYTICAL METHOD Method Number: V0001782
Method of Analysis for the Determination of Perfluorooctanoie Acid (PFOA) in Sediment by LC/MS/MS
Analytical Testing Facility:
Exygen Research
3058 Research Drive State College, PA 16801
Approved By:
V -S- c u L
Paul Connolly
I
Technical Leader, LC-MS, Exygen Research
'Jmtk l/
ohn Flaherty Vice President, Operations, Exygen Research
__lolzbM
Date Date
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V0001782
f ANALYTICAL METHOD
Method of Analysis foTthe Determination ofPerfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS
1.0 Scope
This method is to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectromtrie Detector (LC/MS/MS) in sediment.
2.0 Safety
2.1 Always observe safe laboratory practices. 2-2 Consult the appropriate MSDS before handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At least 30 g of test sample for extraction. 3.2 No sample processing is needed for sediment samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room
temperature. 3.4 All samples must be thoroughly mixed before being sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this
project.
Reagents and Standards
4.1 Water-HPLC grade 4.2 Methanol - HPLC grade 4.3 Acetic Acid- Reagent grade 4.4 Ammonium Acetate - A.C.S. Reagent Grade 4.5 Perfluorooctanoic Acid - Sigma-Aldrich
5.0 Instrument and Equipment
5.1 A high performance liquid chromatograph capable of pumping up to 2
solvents equipped with a variable volume injector capable of injecting 5-200
pL connected to a tandem Mass Spectrometer (LC/MS/MS).
5.2 A device to collect raw data for peak integration and quantitation.
5.3 Analytical balance capable of reading to 0.00001 g.
5.4 50 mL disposable polypropylene centrifuge tubes.
5.5 15 mL disposable polypropylene centrifuge tubes.
5.6 Disposable micropipets (50-10GuL, 100-200uL).
5.7 125-mL LDPE narrow-mouth bottles.
5.8 2 mL clear HPLC vial kit.
.
5.9 Disposable pipettes.
5.10 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips.
5.11 Waters Sep Pale Vac 6 cc (Ig) tC18 SPE cartridges.
5.12 SPE vacuum manifold.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0Q03268
Exygen Research
Method Number V0001782
[___________________________ ANALYTICAL M ETH OD
1
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS
5.13 Vortexer. 5.14 Wrist-action shako'. 5.15 Centrifuge capable of spinning 50 mL polypropylene tubes at 3000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, 5p (P/N: 82505-052130)
6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program:
0.0Time (mini 1.0 8.0 20.0 22.5
%A
65 65 25 25 65
Flow Rate % B ImL/min) 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3
6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: ~ 23 minutes.
The above conditions are intended as a guide and may be changed in order to optimize the HPLC system.
7.0 MS/MS System
7.1 Mode: Electrospray Negative MRM mode, monitoring 413 -+ 369 m/z for PFOA.
The above conditions are intended as a guide and may be changed in order to optimize the MSMS system.
8.0 Preparation of Solutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL of water.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V00017S2
I ANALYTICAL METHOD
|
Method ofAnalysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by
LC/MS/MS
8.2 Extraction Solutions
8.2.1 ]% acetic acid in water is prepared by adding 10 mL of acetic acid to 1000 mL ofwater.
Alternate volumes may be prepared.
9.0 Standard Preparation
9.1 Standard Stock/Fortification Solution 9.1.1 Prepare a stock solution of ~100 pg/mL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle. 9.1.2 A 10 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 100 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.3 A 1.0 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 10 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.4 A 0.1 pg/mL fortification solution ofPFOA is prepared by bringing 10 mL of the 1.0 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.5 A 0.01 pg/mL fortification solution ofPFOA is prepared by bringing 10 mL of the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle. 9.1.6 The stock and fortification solutions are to be stored in a refrigerator at approximately 4C and are stable for a maximum period of 6 months from the date of preparation.
9.2 Standard Calibration Solutions
9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution ofthe 0.1 pg/mL fortification solution.
9.2.2 The following is a typical example: additional concentrations may be
Concentration
o f Fortification Solution na/mU
100 100 100 10 5 2
Volume fmL)
1 5 2 10 10 10
Diluted to (mL)
100 100 100 100 100 100
Final
Concentration
(nfi/m L)
10.0 5.0 2.0 1.0 0.5 0.2
Ptgt4 o f7
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V0001782
JI ANALYTICAL METHOD
Method ofAnalysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS
9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles at 2*C to 6*C, up to six months.
9.2.4 Alternate volumes and concentrations of standards may be prepared as needed.
10.0 Batch Set Up
10.1 Each batch of samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project.
11.0 Sample Extraction
11.1 Weigh 5 g of sample into SOmL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well).
11.2 Add 3S mL of 1% acetic acid, cap, vortex and shake on a wrist action shaker for -60 minutes.
11.3 Centrifuge the tubes at -3000 rpm for -20 minutes. 11.4 Condition the Cu SPE cartridges (1 g, 6 mL) by passing 10 mL methanol
followed by 20 mL ofHPLC water (- 2 drop/sec). Do not let column run dry 11.5 Load (decant) the sample on the conditioned Cu SPE cartridge. Discard
eluate. 11.6 Add 20 mL of methanol to the sediment left in the bottom of the 50 mL
centrifuge tube. Cap, vortex and shake on a wrist action shaker for -30 minutes. 11.7 Centrifuge the tubes at -3000 rpm for -20 minutes. 11.8 Decant the methanol onto the same SPE cartridge. Collect the eluate. 11.9 Wash the column with 4 mL of methanol. Collect the eluate and add it to the eluate collected in step 11.8. 11.10 Condition a second C| SPE cartridge (1 g, 6 mL) by passing 10 mL methanol followed by 20 mL ofHPLC water (- 2 drop/sec). Do not let column run dry 11.11 Add the methanol to -200 mL of water and load on the second conditioned SPE cartridge. 11.12 Elute with -5 mL 100% methanol. Collect 5 mL of eluate into graduated IS mL polypropylene centrifuge tubes (final volume " 5 mL). 11.13 Analyze samples using electrospray LC/MS/MS.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygcn Research
Method Number V000I782
| ______________ANALYTICAL M ETH O D
|
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS
12.0 Chromatography
12.1 Inject the same amount of each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples.
12.2 Standards ofPFOA corresponding to at least live or more concentration levels must be included in an analytical set.
12.3 An cnlire set of extracted calibration standards must be included at the beginning and at the end of a sample set. Standards must be interspersed between every 5-10 samples. As an alternative, an entire set of calibration standards may be injected at the beginning of a set followed by calibration standards interspersed every 5-10 samples (to account for a second set of standards). In either case, calibration standards must be the first and last injection in a sample set.
12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system.
12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed.
13.0 Acceptance Criteria
Chromatogram must show a peak of a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide. Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 0.2 ng/mL, then a new blank sample must be obtained and the entire set must be re-extracted. Recoveries of control spikes and matrix spikes must be between 70-130% of their known values, if a control spike falls outside the acceptable limits, the entire set of samples should be re-extracted. Any matrix spike outside 70 130% should be evaluated by the analyst to determine if re-extraction is warranted. 13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation of the calibration curve. However, the total number of extracted calibration standards that could be excluded must not exceed 20% of the total number of extracted standards injected. 13.5 The correlation coefficient (R) for calibration curves generated must be 0.992 (R2 0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set of samples should be reanalyzed.
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Reieuch
MethodHunter V000H12
I ANALYTICAL METHOD
|
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by
LC/MS/MS
13.6 Retention times between standards and samples must not drift more than
4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed.
14.0 Calculations
14.1 Use the following equation to calculate the amount of PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program:
PFOA found (ng/mL) (Peak area - intercept! x DF slope
DF factor by which the final volume was diluted, if necessary.
14.2 For samples fortified with known amounts of PFOA prior to extraction, use the following equation to calculate the percent recovery.
Recovery (%) =
[ total analyte found (ng/mL) - analyte foundin control (ng/mL)] xlOO
analyte added(ng/mL)
14.3 Use the following equation to convert the amount of PFOA found in ng/mL to ng/g (ppb).
PFOA found (ppb) [PFOA found fae/mL) x final volume f5 mL)) sample weight (5 g)
14.4 Use the following equation (if necessary) to calculate the amount of PFOA found in ppb based on dry weight.
PFOA found (ppb) dry weight- PFOA found (ppb) x [100%/ total solids(%)]
Pag 7 o f 7
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
ANALYTICAL METHOD Method Number V0001783
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
Analytical Testing Facility:
Exygen Research 3058 Research Drive State College, PA 16801
Approved By:
Paul Connolly Technical Leader, LC-MS, Exygen Research
2l1/n fk /y
John Flaherty ' Vice President, Operations, Exygen Research
Date Date
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Method Number VOOOI783
[ ANALYTICAL m eth o d Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
1.0 Scope
This method is to be employed for the isolation and quantitation of perfiuorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass Spectrometric Detector (LC/MS/MS) in fish and clams.
2-0 Safety
2.1 Always observe safe laboratory practices. 2.2 Consult the appropriate MSDS before handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At least 20 g of test sample for extraction. 3.2 Samples should be processed before extraction. Place the frozen sample in a
food processor and homogenize with dry ice. Place the samples in containers arid leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time of analysis. 3.3 Sample collection procedures will be specified in the sampling plan for this project
4.0 Reagents and Standards
Water - HPLC grade Acetonitrile - HPLC grade Carbon (120-400 mesh) - Reagent grade Methanol - HPLC grade Silica gel (60-200 mesh) - Reagent grade 4.6 FlorisU. (60-IQQmesh) - Reagent grade 4.7 Superclean LC-NHj - Reagent grade 4.8 1-Octanol - HPLC grade 4.9 L-Ascorbie acid - Reagent grade 4.10 Dtmethyldichlorosilane - Reagent grade 4.11 Toluene - Reagent grade 4.12 Ammonium Acetate - A.C.S. Reagent Grade 4.13 Perfiuorooctanoic Acid - Sigma-Aldrich
S.O Instrument and Equipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume injector capable of injecting 5-200 pL connected to a tandem Mass Spectrometer (LC/MS/MS).
5.2 A device to collect raw data for peak integration and quantitation. 5.3 Analytical balance capable of reading to 0.00001 g.
Page 2 of 8
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MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V0001783
ANALYTICAL METHOD
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
$.4 Rotary evaporator. 5.5 Tissumizer. 5.6 12S mL pear-shaped flasks. 5.7 50 mL disposable polypropylene centrifuge tubes.
5.8 15 mL disposable polypropylene centrifuge tubes. 5.9 Disposable micropipets (50-IOOuL, 100-200uL). 5.10 125-mL LDPE narrow-mouth bottles. 5.11 2 mL clear HPLC vial kit. 5.12 Disposable pipettes. 5.13 Autopipettes (100-1000 pL and 10-100 pL), with disposable tips. 5.14 SPE tubes (20mL) (Supelco cat. no. N057I77). 5.15 Wrist action shaker.
5.16 Centrifuge capable of spinning 50 mL polypropylene tubes at 2000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: Fluophase RP (Keystone Scientific), 2.1 mm x 50 mm, Sp (P/N: 82505-052130)
6.2 Temperature: 30C 6.3 Mobile Phase (A) : 2 mM Ammonium Acetate in Water 6.4 Mobile Phase (B) : Methanol 6.5 Gradient Program:
Time (mini 0.0 1.0 8.0 20.0 22.5
%A 65 65 25 25 65
Flow Rate % B mL/min) 35 0.3 35 0.3 75 0.3
75 0.3 35 0.3
6.6 Injection Volume: 15 pL (can be increased to as much as 50 pL). 6.7 Quantitation: Peak Area - external standard calibration curve. 6.8 Run Time: -2 3 minutes.
The above conditions are intended as a guide and may be changed in order to optimize the HPLC system.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V0001783
ANALYTICAL METHOD
Method ofAnalysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
7.0 MS/MS System
7.1 Mode: Electrospray Negative MRM mode, monitoring 413- 369 m/z for PFOA.
The above conditions are intended as a guide and may be changed in order to optimize the MSMS system.
8.0 Preparation ofSolutions
8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammonium acetate to 1000 mL ofwater.
8.2 Extraction Solutions
8.2.1 2% ascorbic acid in methanol is prepared by dissolving 2 g of ascorbic acid in 100mL of methanol.
8.2.2 30% Dimethyldichlorosilane in toluene is prepared by bringing 3 mL of dimethyldichlorosilane to a linai volume of 10 mL with toluene.
Alternate volumes may be prepared.
9.0 Standard Preparation
9.1 Standard Stock/Fortification Solution
9.1.1 Prepare stock solution o f-100 pg/mL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mL LDPE bottle.
9.1.2 A 1.0 pg/mL fortification solution of PFOA is prepared by bringing 1 mL of the 100 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle.
9.1.3 A 0.1 pg/mL fortification solution of PFOA is prepared by bringing 10 mL ofthe 1.0 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle.
9.1.4 A 0.01 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 0.1 pg/mL solution to a final volume of 100 with methanol in a 125 mL LDPE bottle.
9.1.5 The stock and fortification solutions are to be stored in a refrigerator ai approximately 4C and are stable for a maximum period of 6 months from the date ofpreparation.
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Interim Report #1 - Analysis of Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number VOOO1783
1 ANALYTICAL METHOD
Method ofAnalysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
9.2 Standard Calibration Solutions
9.2.1 LC/MS/MS calibration standards are prepared in methanol via dilution of the 1.0 pg/mL fortification solution.
9.2.2 The following is a typical example: additional concentrations may be prepared as needed.
Concentration
Final
of Fortification Volume Dilutedto Concentration
Solution(uft/mL) (mL)
(mL)
(lig/mL)
1.0 5.0 100
0.05
1.0 2.5 100 1.0 1.0 too 0.05 10 too
0.025
0.01 0.005
0.025 10
100
0.0025
0.1 10 100
0.001
0.005 10
100
0.0005
9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles
at 2C to 6C, up to six months.
9.2.4 Alternate volumes and concentrations of standards may be prepared as
10.0 Batch Set Up
10.1 Each batch of samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations (lab control spike) to verify procedural recovery for the batch. Requirements for field and laboratory duplicates and spikes will be specified in the quality assurance plan for this project.
11.0 Sample Extraction
11.1 Weigh 5 g of frozen sample into 50 mL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well).
11.2 Add 30 mL of acetonitrile and shake on a wrist action shaker for ~15 minutes. 11.3 Place the tubes in a freezer for-1 hour. 11.4 Pack and condition the SPE tubes and silaiuze the pear-shaped flasks. 11.5 Pack the 20 mL SPE tubes in sequence with 2 g florisil, 2 g silica gel, 2 g
carbon, and 1 g LC-NHj. Condition the columns with 20 mL of methanol, then 20 mL of acetonitrile. Discard all washes, Do not allow the column to dry. 11.6 Silanize the 125 mL pear-shaped flasks by rinsing with the 30% dimethyldichlorosilane in toluene solution. Rinse the flask with toluene once, followed by methanol (three times). Dry the flasks completely before use, either by air-drying or with a stream of nitrogen.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V0001783
ANALYTICAL METHOD
Method ofAnalysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
11.7 Centrifuge the SO mL polypropylene tubes containing sample at -2000 rpm for -10 minutes.
11.8 Decant the extract on to a conditioned SPE column fitted inside the mouth of the pear-shaped flask. Collect the eluate in the 125 mL silanized pear-shape flask.
11.9 Add 10 mL of acetonitrile to the sample in the 50 mL centrifuge tube. Homogenize the frozen fat phase using a tissumizer for -30 seconds and rinse the tissumizer with -10 mL of acetonitrile into the tube.
11.10 Shake the sample again for-10 minutes on a wrist-action shaker. 11.11 Place the tubes in a freezer for - 1hour more. 11.12 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm
for -10 minutes. 11.13 Decant the extract onto the same SPE column. Collect the eluate into the
same pear-shaped flask and combine with the eluent from the initial extraction. 11.14 Pass 20 mL of acetonitrile through the SPE column and combine the eluate in the same pear-shaped flask. 11.15 Add 3-4 drops of 1-octanol to the extract in the pear-shaped flask and evaporate at reduced pressure using a rotary evaporator (at < 40C). 11.16 Make the final volume, by adding 2 mL of 2% ascorbic acid in methanol to the pear-shaped flask and swirl to mix/dissolve. 11.17 Transfer the extracts to HPLC vials using disposable pipets. 11.18 Analyze samples using electrospray LC/MS/MS.
12.0 Chromatography
12.1 Inject the same amount of each standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples.
12.2 Standards of PFOA corresponding to at least five or more concentration levels must be included in an analytical set.
12.3 An entire set of calibration standards must be included at the beginning and at the end of a sample set. Standards must be interspersed between every 5*10 samples. As an alternative, an entire set of calibration standards may be injected at the beginning of a set followed by calibration standards interspersed every 5-10 samples (to account for a second set of standards), in either case, calibration standards must be the first and last injection in a sample set.
12.4 Use linear standard curves for quantitation. Linear standard curves are generated for the analyte by linear regression using 1/x weighting of peak area versus calibration standard concentration using MassLynx 3.3 (or equivalent) software system.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number V000H83
| ANALYTICAL METHOD
Method of Analysis for the Determination ofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
12.5 Sample response should not exceed standard responses. Any samples that exceed standard responses should be further diluted and reanalyzed.
13.0 Acceptance Criteria
13.1 Chromatogram must show a peak of a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion (369 amu) represents the loss of carbon dioxide.
13.2 Method blanks must not contain PFOA at levels greater than the LOQ. If a blank contains PFOA at levels greater than 0.5 ppb, then a new blank sample must be obtained and the entire set must be re-extracted.
13.3 Recoveries of control spikes and matrix spikes must be between 70-130% of their known values. If a control spike falls outside the acceptable limits, the entire set of samples should be re-extracted.
13.4 Any calibration standard found to be a statistical outlier by using the Huge Error Test, may be excluded from the calculation of the calibration curve. However, the total number of calibration standards that could be excluded must not exceed 20% ofthe total number of standards injected.
13.5 The correlation coefficient (R) for calibration curves generated must be 0.992 (RJ 0.985). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or the relevant set ofsamples should be reanalyzed.
13.6 Retention times between standards and samples must not drift more than 4 % within an analytical run. If retention time drift exceeds this limit within an analytical run then the set must be reanalyzed.
14.0 Calculations
14.1 Use the following equation to calculate the amount of PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generated by the Mass Lynx software program:
PFOA found (ng/mL) (Peak area intercept! slope
14.2 Use the following equation to converi the amount of PFOA found in ng/mL to ng/g (ppb).
PFOA found (ppb) - fPFOA found fne/mLl x final volume (mLI x DF1 sample weight (g)
DF factor by which the final volume was diluted, if necessary.
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Interim Report #1 - Analysis o f Decatur Surface Water and Sediment Samples
MPI Study No.: 0137.0219 ExyLIMS Protocol No.: P0003268
ExyLIMS Protocol Number: P0003268
Exygen Research
Method Number VOOOI78J
AMAi.VYIC.VL M ETHOD
Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
14.3 For samples fortified with known amounts of PFOA prior to extraction, use the following equation to calculate the percent recovery.
Recovery (%) =
[ total analyte found (ng/g) - analyte found in control (ng/g)] ^ ^ analyteadded (ng/g)
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