Document OJMdya6akmMyqK5r1751qwp1j
INTERIM REPORT # 21 - Analysis of Soil Samples
STUDY TITLE Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum 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 November 6,2006
PERFORMING LABORATORY Exygen Research
3058 Research Drive State College, PA 16801
Phone: 814-272-1039
STUDY SPONSOR
3M Company
3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: 651-733-6374
PROJECT Protocol Number: P0001131 Exygen Study Number: P0001131
Total Pages: 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
Exygen Study Number P0001131, entitled "Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum 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 Exygen Research.
^Jo6hn Flaherty / Primncipal Investigator ' 1EHVxIWygen Research
Jaisimha Kesari P.E., DEE Study Director
Weston Solutions, Inc.
Sponsor Representative 3M Company
Exygen Research
Date
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QUALITY ASSURANCE STATEMENT
Exygen Research's Quality Assurance Unit reviewed Exygen Study Number P0001131, entitled, "Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program". All reviewed phases1 were inspected for conduct according to Exygen Research's Standard Operating Procedures, the Study Protocol, and all applicable Good Laboratory Practice Standards. All findings were reported to the Exygen Principal Investigator and Management and to the Study Director.
Phase 39. Raw Data and
Interim Report Review
Date Inspected
Date Reported to Date Reported to
Principal
Exygen
Date Reported to
Investigator Management Studv Director
10/18-24/06
10/31/06
11/02/06
11/06/06
41. Re-extract Raw Data and Interim Report Review
10/30-31/06
11/02/06
11/02/06
11/06/06
42. Final Interim Report Review
11/03/06
11/06/06
11/06/06
11/06/06
^ o te : All in-lab inspections 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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CERTIFICATION OF AUTHENTICITY
This interim report, for Exygen Study Number P0001131, is a true and complete representation of the raw data.
Submitted by:
Exygen Research 3058 Research Drive State College, PA 16801 (814) 272-1039
Principal Investigator, Exygen:
Date
Exygen Research Facility Management:
Date
Study Director, Weston Solutions, Inc.
Jaisimha Kesari P.E., DEE Weston Solutions, Inc.
Sponsor Representative, 3M Company:
i M c h J j & J jM W Michael A. Santoro/ Director of Regulatory Affairs
Exygen Research
Date 7
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STUDY IDENTIFICATION
Analysis of Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small Mammal Serum Using LC/MS/MS for the 3M Decatur
Monitoring Program
PROTOCOL NUMBER: EXYGEN STUDY NUMBER: TYPE OF STUDY: SAMPLE MATRIX: TEST SUBSTANCE:
P0001131
P0001131
Residue
Soil and Water
Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS)
SPONSOR: STUDY DIRECTOR:
3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144
Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380
STUDY MONITOR:
Michael A. Santoro 3M Company 3M Building 0236-01-B-10
St. Paul, MN 55144
PERFORMING LABORATORY: Exygen Research 3058 Research Drive State College, PA 16801
ANALYTICAL PHASE TIMETABLE:
Study Initiation Date:
11/05/04
Interim Analytical Start Date:
03/15/06
Interim Analytical Termination Date: 10/24/06
Interim Report Completion Date: 11/06/06
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PROJECT PERSONNEL
The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel from Exygen Research were associated with various phases of this interim portion of the study:
Name John Flaherty Karen Risha Eric Edwards Brian McAllister Christine Edwards Mark Ammerman Amy Sheehan Mindy Cressley Brittany Kravets Christa Gallant
Scott Crain Cameala Graybill
Ling Ling Liu Frances Crespi
Title Vice President
Scientist Sample Custodian Sample Custodian
Technician Sample Custodian Associate Scientist
Technician Technician Technician Technician Technician Technician Technician
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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....................................................................................................10 1.0 SUMMARY................................................................................................................11 2.0 OBJECTIVE.............................................................................................................. 12 3.0 INTRODUCTION.......................................................................................................12 4.0 ANALYTICAL TEST SAMPLES..............................................................................12 5.0 REFERENCE MATERIAL........................................................................................13 6.0 DESCRIPTION OF ANALYTICAL METHOD........................................................14
6.1. Extraction Procedure For Soil.................................................................................15 6.2 Percent Solids Procedure For Soil............................................................................15 6.3 Extraction Procedure for Water................................................................................15 6.4 Preparation of Standards and Fortification Solutions...............................................15 6.5 Chromatography.......................................................................................................16 6.6 Instrument Sensitivity...............................................................................................17 6.7 Description of LC/MS/MS Instrument and Operating Conditions...........................17 6.8 Quantitation and Example Calculation.....................................................................17 7.0 EXPERIMENTAL DESIGN......................................................................................19 8.0 RESULTS.................................................................................................................. 20 9.0 CONCLUSIONS........................................................................................................ 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 Soil Samples..................................23
Table H. Summary of PFBS, PFHS and PFOS in Re-extracted Soil Samples............ 29
Table in. Summary of PFBS, PFHS and PFOS in Trip Blank and Equipment
Rinseate Blank Samples ............................................................................33
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples........... 34
Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples.........................................................................................................45
Table VI. Matrix Spike Recovery of PFBS, PFHS and PFOS in Trip Blank Samples.......................................................................................................... 51
Table VII. Total Percent Solids in Soil Samples............................................................ 52
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Figure 1.
LIST OF FIGURES Page
Typical Calibration Curve for PFBS in Methanol......................................... 57
Figure 2. Extracted Standards of PFBS in Methanol, 25 ng/L and 50 ng/L, Respectively................................................................................................... 58
Figure 3. PFBS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively........................ 59
Figure 4. Chromatogram Representing a Soil Sample Analyzed for PFBS (Exygen ID: C0152904, Data Set: 031306C)................................................ 60
Figure 5. Typical Calibration Curve for PFHS in Methanol.........................................61
Figure 6. Extracted Standards of PFHS in Methanol, 25 ng/L and 50 ng/L, Respectively................................................................................................... 62
Figure 7. PFHS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively.................................... 63
Figure 8. Chromatogram Representing a Soil Sample Analyzed for PFHS (Exygen ID: C0152904, Data Set: 031306CR).............................................. 64
Figure 9. Typical Calibration Curve for PFOS in Methanol......................................... 65
Figure 10. Extracted Standards of PFOS in Methanol, 25 ng/L and 50 ng/L, Respectively................................................................................................... 66
Figure 11. PFOS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively.................................... 67
Figure 12. Chromatogram Representing a Soil Sample Analyzed for PFOS
(Exygen ID: C0152909, Data Set: 031406A R ).....................................................68
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LIST OF APPENDICES
Page
Appendix A Study Protocol P0001131 (Exygen Study No. P0001131) with Analytical Methods and Protocol Amendments....................................... 69
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1.0 SUMMARY
Exygen Research extracted and analyzed soil samples for the determination of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) according to Exygen Method V0001781 and water samples for the determination of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) according to Exygen Method V0001780 (Appendix A).
The limit of quantitation (LOQ) for PFBS, PFHS, and PFOS in soil was 0.2 ng/g. The LOQ provided is assuming wet weight values and the results for the actual samples are reported based on dry weight. The limit of quantitation for PFBS, PFHS, and PFOS in water was 25 ng/L.
Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS in soil samples based on dry weight are summarized in Table I. Matrix spikes that were in the appropriate concentration range (primary concentration must be less than or equal to three times the spiking level) were used to calculate the assessed accuracies. In instances of failed laboratory spike recoveries, the samples were not reported due to the quality control failure. Not all soil sample results met data quality objectives. A total of 102 samples had analytes designated as not reported (NR) due to quality control failures in initial analyses. Some of the samples failed because the analyte spike levels were too low relative to the endogenous level in the sample to allow assessment of matrix interference. Some samples failed for unknown reasons. A total of 98 samples were re-extracted and reanalyzed in an attempt to obtain quantitative results. Of these 98 samples, only 86 samples contained sufficient remaining sample mass to re-extract matrix spikes and an unspiked sample. Twelve samples contained only enough sample to allow for the re extraction of matrix spikes, and were therefore not reported. Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS in re-extracted soil
samples based on dry weight are summarized in Table II. Analytical results and assessed
accuracies for the analysis of PFBS, PFHS, and PFOS in water samples are summarized in Table III. Quantitative results were obtained for all samples and analytes except for PFBS in 19 samples, PFHS in 7 samples, and PFOS in 20 samples that are not reported due to quality control failures.
Fortification recoveries for the initial analysis of PFBS, PFHS, and PFOS in soil samples based on wet weight are summarized in Table IV. The overall average percent recovery standard deviation for PFBS, PFHS, and PFOS in the soil samples based on wet weight were 113 23%, 85 13%, and 95 16%, respectively. Fortification recoveries for the analysis of PFBS, PFHS, and PFOS in re-extracted soil samples based on wet weight are summarized in Table V. The overall average percent recovery + standard deviation for PFBS, PFHS, and PFOS in the re-extracted soil samples based on wet weight were 115 27%, 85 28%, and 97 23%, respectively. Fortification recoveries for the analysis of PFBS, PFHS, and PFOS in water samples are summarized in Table VI. The overall
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average percent recovery standard deviation for PFBS, PFHS, and PFOS in water samples were 107 9%, 97 6%, and 112 10%, respectively.
The total percent solids for soil samples are detailed in Table VII.
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 soil and water according to Protocol P0001131 (Appendix A).
3.0 INTRODUCTION
This report details the results of the analysis for the determination of PFBS, PFHS, and PFOS in soil using the analytical methods entitled, "V0001781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" and for the determination of PFBS, PFHS, and PFOS in water using the analytical methods entitled, "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS."
The study was initiated on November 05, 2004, when the study director signed protocol number P0001131. The analytical start date for this interim report was March 15, 2006 and the analytical termination date for this interim report was October 24,2006.
4.0 ANALYTICAL TEST SAMPLES
Five water sam ples, representing one trip blank and its associated spikes and two rinse
blanks (RB), and fifty-five soil samples (Exygen ED C0152885 - C0152944) were received on wet ice on February 17, 2006 from Tim Frinak at Weston Solutions, Inc. Three water samples representing one trip blank and its associated spikes and eleven soil samples (Exygen ID C0152946 - C0152959) were received at ambient temperature on February 21, 2006 from Tim Frinak at Weston Solutions, Inc. Six water samples, representing one trip blank and its associated spikes and three rinse blanks (RB), and forty-three soil samples (Exygen ID C0153313 - C0153361) were received on wet ice on February 22, 2006 from Tim Frinak at Weston Solutions, Inc. Please note that rinse blank (RB) samples do not have associated field spikes or laboratory spikes. The samples were logged in by Exygen personnel and placed in refrigerated storage.
The soil sample identification (ID) codes for the samples analyzed and reported are in the following format:
Dxxx-xx-xxxxxx-x(x)-xxxx
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For soil samples, the first character D represents Decatur, Alabama samples and the xxx defines the sampling location within the study area where:
DF05 = Field 5 DF11 = Field 11 DF13= Field 13
DF10 = Field 10 DF12 = Field 12
The second string, SB, defines the type of sample as a soil boring sample.
The third string defines the location within the sampling area as GPxx where GP refers to geoprobe.
The fourth string additionally defines the sample type where:
0 = Primary field sample
DB = Field duplicate sample
RB = Equipment rinseate blank sample
The fifth string defines the collection depth of the soil samples where:
0000 = 0 to 3 inches 0010 = 1 to 1.5 ft. 0050 = 5 to 6 ft.
0005 = 6 inches to 1 foot 0020 = 2 to 2.5 ft.
Sample login and chain of custody information is located in the raw data package associated with this interim report. Storage records will be kept at Exygen Research.
5.0 REFERENCE MATERIAL
The analytical standards, PFBS and PFHS, were supplied by 3M. PFBS was received from 3M at Exygen on May 13, 2005. PFHS was received from 3M at Exygen on January 20, 2003. PFOS was purchased from Fluka Corporation and was received at Exygen on April 23, 2003.
The available information for the reference materials is listed below. PFBS and PFHS were stored frozen and PFOS was stored refrigerated.
Compound PFBS PFHS PFOS
Exveen Inventorv No. SP0005726 SP0002401 SP0002694
Lot# 101 SE036 430180-1
Puritv (%) 96.7 98.6 101.2
Expiration Date 12/04/06 10/18/06 10/31/07
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The molecular structures of PFBS, PFHS and PFOS are given below:
PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (C4F9S03"K+) Transitions Monitored: 299 - 99 Structure:
FF FF
F SO 3
FF FF
PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (CFb SOs'K*) Transitions Monitored: 399 - 80 Structure:
FFF
PFOS Chemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (CgFnSOs'K'1') Transitions Monitored: 499 -> 80
Structure:
6.0 DESCRIPTION OF ANALYTICAL METHOD
The analytical methods "V0001781: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" and "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" were used for this study.
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6.1. Extraction Procedure For Soil
The soil samples were extracted using the procedure indicated in Exygen method V0001781. Before the samples were weighed for the extraction, they were placed into a new, clean Ziploc bag and mixed thoroughly. The samples were then transferred back to the sampling container. A 5-gram portion of soil was weighed into a fifty milliliter centrifuge tube for the extraction. After fortification of appropriate samples, 5 mL of methanol was added to the samples. The samples were allowed to shake on a wrist action shaker for -15 minutes and were then sonicated in an ultrasonic bath for ~15 minutes. The volume was taken to 40 mL with water and the samples were then centrifuged for ~10 minutes at -3000 rpm. The supernatant was then loaded onto a Cis SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray.
6.2 Percent Solids Procedure For Soil
Percent solids were determined using the procedure indicated in Exygen method V0000427. Approximately 20 grams of sample was weighed into a pan. The weight of the sample plus the pan was recorded. The samples was then dried in an oven overnight at 104 2 C. Then the samples were transferred to a dessicator and allowed to cool for -15 minutes. Each sample was then weighed again, including the weight of the pan. The percent solid for each sample was then calculated. See Table VII for percent solids results.
6.3 Extraction Procedure for Water
The water samples were extracted using the procedine indicated in Exygen method V0001780. A 40 mL aliquot of the water sample was used for the extraction procedure. After fortification of appropriate samples, the samples were loaded onto a Cis SPE cartridge conditioned with 10 mL of methanol and 5 mL of water. The eluate was discarded. Approximately five milliliters of methanol was added to the cartridge. Five milliliters of eluate was collected into a graduated 15 mL polypropylene centrifuge tube. Each sample was analyzed by LC/MS/MS electrospray.
6.4 Preparation of Standards and Fortification Solutions
Individual stock standard solutions of PFBS, PFHS and PFOS were prepared as specified in Exygen method V0001781. A mixed stock standard solution of PFBS, PFHS, and PFOS was prepared at a concentration of 1000 pg/mL by dissolving 100 mg of each of
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the standards (corrected for purity and salt content) in 100 mL of methanol. From this solution, the following fortificati on standards were prepared:
Cone, of
Fort
Stock or Fort. Volume
Solution
(mL)
(pg/mL)1
1000 10
100 10
10 10
1.0 10
0.1 10
1of PFBS, PFHS, and PFOS
Final Volume
(mL)
100 100 100 100 100
Final Cone, of Fortification Std.
(pg/mL)
100 10 1.0 0.1 0.01
A set of extracted calibration standards containing PFBS, PFHS, and PFOS were prepared. The following concentrations were prepared before extraction:
Cone, of Fort Fort
Volume of
Solution
Volume Fortified Sample
(ng/mL)'
(mL)
(mL)
- 40
10 0.1
40
10 0.2
40
10 0.4
40
100 0.1
40
100 0.2
40
100 0.4
40
1of PFBS, PFHS, and PFOS
Final Cone, of Calibration Std.
(ng/mL)
0 25 50 100 250 500 1000
The stock standard solution and all 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.
6.5 Chromatography
Quantification of PFBS, PFHS and PFOS was accomplished by LC/MS/MS electrospray. The retention times of PFBS, PFHS and PFOS were ~0.6 mins, -9.0 mins, and -11.6 mins, respectively. Peaks above the LOQ were not detected in any of the reagent blank samples corresponding to the analyte retention time.
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6.6 Instrument Sensitivity
The smallest standard amount injected during the chromatographic run had a concentration of 0.2 ng/mL of PFBS, PFHS and PFOS.
6.7 Description of LC/MS/MS Instrument and Operating Conditions
Instrument: API 4000 Biomolecular Mass Analyzer
Interface: Turbo Ion Spray Liquid Introduction Interface
Computer: DELL OptiPlex GX400
Software: Windows NT, Analyst 1.4.1
HPLC:
Hewlett Packard (HP) Series 1100
HP Quat Pump
HP Vacuum Degasser
HP Autosampler
HP Column Oven
HPLC Column: Thermo Fluophase RP, 50 mm x 2.1 mm
Column Temp.: ~30 C Injection Voi.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water
Mobile Phase (B): Methanol
Time /min') 0.0 1.0 8.0 10.0 11.0 18.0
Total run time: ~18 min
Flow Rate: 0.3 mL/min Ions monitored:
%A 65 65 25 25 65 65
Analvte
Mode
PFBS PFHS PFOS
negative negative negative
%B 35 35 75 75 35 35
Transition Monitored 299 99 399 -> 80 499 - 80
Approximate Retention Time
/min) -0.6 min. -9.0 min. -11.6 min.
6.8 Quantitation and Example Calculation
Fifteen microliters of sample or calibration standard were injected into the LC/MS/MS. The peak area was measured and the standard curve was generated (using 1/x fit weighted
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linear regression) by Analyst software using six concentrations of standards, except in a few cases of PFBS, where only five were used; as the 500 ng/L standard results were rejected as outliers. The concentration was determined from the following equations.
Equation 1 calculated the amount of analyte found (in ng/L, based on peak area) using the standard curve (linear regression parameters) generated by the Analyst software program.
Equation 1:
Analyte found (ng/L) = (Peak area - intercept! x DF Slope
Where: DF = Dilution Factor, factor by which the final volume was diluted, if necessary.
For samples fortified with known amounts of PFBS, PFHS and PFOS prior to extraction, Equation 2 was used to calculate the percent recovery.
Equation 2: Recovery (%) =
(analyte found (ng/Li - analyte in control (ng/L)) xl00% amount added (ng/L)
Note: For the analyte recovery calculation, the "control" is the unspiked aliquot of the primary field sample.
Equation 3 was used to convert the amount of PFBS, PFHS and PFOS found in ng/L to ng/g (ppb).
Equation 3:
Analyte found (ppb) = fAnalvte found (ng/L) x volume extracted (0.04L)] sample weight (5 g)
Equation 4 was then used to calculate the amount of PFBS, PFHS and PFOS found in ppb based on dry weight.
Equation 4: Analyte found (ppb) dry weight = Analyte found (ppb) x [100% / total solids(%)]
NOTE: Total solids (%) = [wet weight (g) / dry weight (g)] x 100%
An example of a calculation using an actual sample follows (calculation is for PFBS only):
Soil sample Exygen ID: C0153360 Spk G (Set: 032106F), fortified at 500 ng/L with where:
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peak area intercept slope dilution factor ng/L PFOA added (fort level) ng/L in corresponding sample volume extracted (L) sample weight (g) total solids (%)
= = = = = = = = =
290183 0.00192 471 1 500 132 0.04 5 84.00
From equation 1: Analyte found (ng/L) = f290183 - 0,00192] x 1 471
From equation 2: % Recovery
616 ng/L
(616 ng/L - 132 ng/L) x 100% 500 ng/L
From equation 3: Analyte found (ppb)
; 97%
= /616ng/Lx 0.04L 5g
4.93 ppb
From equation 4: Analyte found (ppb) dry weight = 4.93 ppb x (100% / 84.00%)
= 5.87 ppb
Note: Numbers may vary slightly 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 water sample bottles were filled to a 200 mL volumetric fill line in the field. For soil samples designated as laboratory matrix spikes, PFBS, PFHS, and PFOS were added at a known concentration to the samples in the laboratory after the samples were weighed for extraction.
The soil samples were initially extracted in twenty-two sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations. Each set
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contained five samples sites except for set 22, which contained four sample sites. One sample was collected for each sample site. For each site, two laboratory matrix spikes and a laboratory duplicate were extracted.
Ninety-eight samples were re-extracted in thirteen sets. Each set included one reagent blank and two reagent blanks fortified at known concentrations of PFBS, PFHS and PFOS. Three sets contained ten sample sites; one set contained nine sample sites; three sets contained eight sample sites; one set contained seven sample sites; three sets contained six sample sites; two sets contained five sample sites. One soil sample was extracted for each sample site, except for sixteen samples that were not re-extracted due to sample availability. For these sixteen samples, results from the initial analysis were used to calculate re-extracted matrix spike recovery. For each re-extracted site, variable matrix spikes of adequate concentration were extracted.
The water samples were extracted in three sets. Each set included one reagent blank, two reagent blanks fortified at known concentrations. Set one contained one trip blank and its associated spikes. Set two contained three rinse blanks and one trip blank with its associated spikes. Set three contained two rinse blanks and one trip blank with its associated spikes.
8.0 RESULTS
Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS in soil samples based on dry weight are summarized in Table I. Fortification recoveries for the initial analysis of PFBS, PFHS, and PFOS in soil samples based on wet weight are summarized in Table IV. The overall average percent recovery standard deviation for PFBS, PFHS, and PFOS in the soil samples based on wet weight were 113 23%, 85 13%, and 95 16%, respectively. Analytical results and assessed accuracies for the
analysis o f PFBS, PFHS, and PFOS in re-extracted soil samples based on dry weight are
summarized in Table II. Quantitative results were obtained for all samples and analytes except for PFBS in 19 samples, PFHS in 7 samples, and PFOS in 20 samples.
Fortification recoveries for the analysis of PFBS, PFHS, and PFOS in re-extracted soil samples based on wet weight are summarized in Table V. The overall average percent recovery standard deviation for PFBS, PFHS, and PFOS in the re-extracted soil samples based on wet weight were 115 27%, 85 28%, and 97 23%, respectively. Analytical results and assessed accuracies for the analysis of PFBS, PFHS, and PFOS in water samples are summarized in Table III. Fortification recoveries for the analysis of PFBS, PFHS, and PFOS in water samples are summarized in Table VI. The overall average percent recovery standard deviation for PFBS, PFHS, and PFOS in water samples were 107 9%, 97 6%, and 112 10%, respectively.
Accuracies were assessed for each sample by reviewing the individual QC results obtained for each sample site. For the soil samples, there were two laboratory matrix
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spike recovery results available for each sample. Matrix spikes that were in the appropriate concentration range (primary concentration has to be than or equal to three times the spiking level) were used to calculate the assessed accuracies. In instances of failed laboratory spike recoveries, the samples were not reported due to the quality control failure.
9.0 CONCLUSIONS
Except as noted above, the soil and water samples were successfully extracted and analyzed for PFBS, PFHS and PFOS according to analytical methods V0001781 and V0001780, respectively. There were no circumstances that would have affected the quality or integrity of the data.
10.0 RETENTION OF DATA AND SAMPLES
When the final analytical report is complete, all original paper data generated by Exygen Research will be shipped to the study director. This does not include facility-specific 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 Exygen Research archives for the period of time specified in EPA TSCA Good Laboratory Practice Standards 40 CFR 792.
Exygen Research
Page 21 o f 136
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Exygen Study No.: P0001131
TABLES
Exygen Research
Page 22 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table I. Summary of PFBS, PFHS and PFOS in Soil Samples
E x v a e n ID
C0152890 C0152890 Rep
C0152891 C0152891 Rep
C0152892 C0152892 Rep
C0152893 C0152893 Rep
C0152894 C0152894 Rep
C0152895 C0152895 Rep
CO152896 C0152896 Rep
CO152897 C0152897 Rep
CO152898 C0152898 Rep
C0152899 C0152899 Rep
C0152900 C0152900 Rep
C0152901 C0152901 Rep
C0152902 C0152902 Rep
CO152903 C0152903 Rep
C0152904 C0152904 Rep
C0152905 C0152905 Rep
C0152906 C0152906 Rep
C0152907 C0152907Rep
C0152908 C0152908 Rep
Client Sample ID
DF10-SB-GP01-0-0000 DF10-SB-GP01 -0-0000*
D F 10-S B -G P 0 1 -0 -00 0 5 DF10-SB-GP01-0-0005*
DF10-SB-GP01-0-0010 DF10-SB-GP01 -0-0010*
DF10-SB-GP01 -SB-0020 DF10-SB-GP01 -SB-0020*
DF10-SB-GP01-0-0020 DF10-SB-GP01 -0-0020*
DF10-SB-GP01 -0-0050 DF10-SB-GP01-0-0050*
DF10-SB-GP02-0-0000 DF10-SB-GP02-0-0000*
DF10-SB-GP02-0-0005 DF10-SB-GP02-0-0005*
DF10-SB-GP02-0-0010 DF10-SB-GP02-0-0010*
DF10-SB-GP02-0-0020 DF10-SB-GP02-0-0020*
DF10-SB-GP02-0-0050 DF10-SB-GP02-0-0050*
DF12-SB-GP01-0-0000 DF12-SB-GP01-0-0000*
DF12-SB-GP01-0-0005 DF12-SB-GP01-0-0005*
DF12-SB-GP01-0-0010 DF12-SB-GP01-0-0010*
DF12-SB-GP01-0-0020 DF12-SB-GP01-0-0020*
DF12-SB-GP01-0-0050 DF12-SB-GP01 -0-0050*
DF12-SB-GP01 -DB-0050 DF12-SB-GP01 -DB-0050*
DF12-SB-GP02-0-0000 DF12-SB-GP02-0-0000*
DF12-SB-GP02-0-0005 DF12-SB-GP02-0-0005*
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Psrffuorotxitsnesutfonst______ PerlluorohaxanasuHonata_______Psrfluorooctanssuttonst
Analyte Found (PPb, ng/g) Dry Weight
Assessed Accuracy
(+ /-% )
Analyte Found (ppb. ng/g) Orv Weight
Assessed Accuracy
(/-% )
Analyte Found (PPb, ng/g) Dry Weight
Assessed Accuracy
(/-% )
.
0.512
40
2.08
40
NR
0.314
40
2.03
40
NR
0.545
40
2.12
30
NR
-
0.499
40
2.00
30
NR
-
1.63 40 4.32
30
NR
-
1.50
40
5.00
30
NR
-
1.59 30 4.26 40 1.42 30 3.68 40
NR NR
-
-
1.10 1.03
30 30
3.91 3.67
30 30
NR NR
-
*
1.70 1.46
ND ND
30 30
40 40
17.1 14.8
1.68 1.75
30 30
30 30
NR NR
NR NR
-
-
ND 30 1.04 30 NR -
ND 30 1.09 30
NR
ND 30 1.09 30
NR
-
ND 30 1.09 30
NR
-
0.443
30
2.76
30
NR
-
0.464
30
2.93
30
NR
"
2.69
40
14.4
40
NR
-
3.19
40
18.5
40
NR
-
NR - NR - NR NR - NR - NR -
3.13
30
58.4
50
NR
-
3.02 30 51.5
50
NR
-
1.02
30
18.0
30
NR
-
1.18 30 23.6
30
NR
-
1.61 40 24.1
30
NR
-
1.68 40 22.8
30
NR
-
1.06 0.770
30 30
19.6 21.5
30 30
NR NR
-
0.527 0.553
40 40
20.4 19.0
30 30
NR NR
-
-
NR
_
36.3
40
NR
-
NR - 39.1 40 NR
3.79 30 31.0 3.67 30 29.9
40 40
NR NR
"
Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table II for re-extract data.
Exygen Research
Page 23 of 136
Interim Report #21 - Analysis of Soil Samples
Exygen Study No.: P0001131
Table I.
Summary of PFBS, PFHS and PFOS in Soil Samples Continued
ExvnenID
Client Sample ID
C0152909 CO152909 Rep
DF12-SB-GP02-0-0010 DF12-SB-GP02-0-0010*
C0152910 C0152910 Rep
DF12-SB-GP02-0-0020 DF12-SB-GP02-0-0020*
C0152911 C0152911 Rep
DF12-SB-GP02-0-0050 DF12-SB-GP02-0-0050*
C0152912 C0152912 Rep
DF12-SB-GP03-0-0000 DF12-SB-GP03-0-0000*
C0152913 C0152913 Rep
DF12-SB-GP03-0-0005 DF12-SB-GP03-0-0005*
C0152914 C0152914 Rep
DF12-SB-GP03-0-0010 D F 12-S B -G P 0 3 -0 -00 1 0*
C0152915 C0152915 Rep
DF12-SB-GP03-0-0020 DF12-SB-GP03-0-0020*
C0152916 C0152916 Rep
DF12-SB-GP03-0-0050 DF12-SB-GP03-0-0050*
C0152917 C0152917 Rep
DF12-SB-GP04-0-0000 DF12-SB-GP04-0-0000*
C0152918 C0152918 Rep
DF12-SB-GP04-0-0005 DF12-SB-GP04-0-0005*
C0152919 C0152919 Rep
D F 12-S B -G P 0 4 -0 -00 1 0 DF12-SB-GP04-0-0010*
C0152920 C0152920 Rep
DF12-SB-GP04-0-0020 DF12-SB-GP04-0-0020*
C0152921 C0152921 Rep
C0152922 C0152922 Rep
DF12-SB-GP04-DB-0020 DF12-SB-GP04-DB-0020*
DF12-SB-GP04-0-0050 DF12-SB-GP04-0-0050*
C0152923 C0152923 Rep
DF13-SB-GP01-0-0000 DF13-SB-GP01-0-0000*
C0152924 C0152924 Rep
DF13-SB-GP01-0-0005 DF13-SB-GP01-0-0005*
C0152925 C0152925 Rep
DF13-SB-GP01-0-0010 DF13-SB-GP01 -0-0010*
C0152926 C0152926 Rep
DF13-SB-GP01-0-0020 DF13-SB-GP01-0-0020*
C0152927
DF13-SB-GP01-0-0050
C0152927 Rep j DF13-SB-GP01-0-0050*
C4 Sulfonate PFBS Periluorobutanesulfonata
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
</-%)
Parfluorohaxenesulfonata
Analyte Found (ppb. ng/g) Dry Weight
Assessed Accuracy
(/-% )
NR NR
146 141
NR NR
0.632 0.609
ND ND
1.36 1.04
1.68 1.80
1.56 1.80
1.20 1.61
NR NR
4.19 3.71
2.15 1.61
1.65 2.55
4.00 3.76
10.4 11.1
NR NR
NR NR
NR NR
NR NR
-
50 50
-
50 50
40 40
30 30
30 30
30 30
40 40
_ -
30 30
30 30
40 40
30 30
30 30
-
. "
36.2 34.3
NR NR
NR NR
2.59 2.86
0.810 0.598
2.86 2.27
4.89 5.22
7.84 8.27
NR NR
NR NR
35.2 35.0
23.1 21.3
22.4 35.9
39.3 34.7
NR NR
58.4 56.7
48.0 35.9
NR NR
14.9 16.3
30 30
_ -
30 30
30 30
30 30
30 30
30 30 _ _ -
30 30
30 30
30 30
30 30
_ -
30 30
30 30 _ -
30 30
C8 Sulfonate PFOS Periluorooctaneiulfonats
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
<+/-%)
NR .
NR -
NR . NR -
NR _ NR
NR . NR -
NR _ NR -
87.3 92.6
NR NR
79.0 71.2
30 30
-
30 30
NR NR -
NR NR -
NR NR
NR NR
NR . NR -
NR NR -
NR . NR -
NR NR -
NR NR -
NR NR *
NR NR
` Laboratory Duplicate ND = Not detected lit or above the Limit of Quantitation (LOQ) o f 0.2 nfl/g (wet weight). NR = Not reported due to quality control failures, see Table II for re-extract data.
Exygen Research
Page 24 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study N o.: P0001131
Table I.
Summary of PFBS, PFHS and PFOS in Soil Samples Continued
ExvaenID
CO152928 C0152928 Rep
C0152929 C0152929 Rep
C0152930 C0152930 Rep
C0152931 C0152931 Rep
C0152932 C0152932 Rep
C0152933 C0152933 Rep
C0152934 C0152934 Rep
C0152935 C0152935 Rep
C0152936 C0152936 Rep
C0152937 C0152937 Rep
C0152938 C0152938 Rep
C0152939 C0152939 Rep
C0152940 C0152940 Rep
C0152941 C0152941 Rep
C0152942 C 0152942 Rep
C0152943 C0152943 Rep
C0152944 C0152944 Rep
C0152949 C0152949 Rep
C0152950 C0152950 Rep
C 01 52951 C0152951 Rep
Client Sample ID
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Porfluorobutan--ulfooate______ Pffluorohaxanesulfonata_______ Perfluorooctanesulfonate
Analyte Found (ppb, ng/g) Div Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb. ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found
(ppb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
DF13-SB-GP02-0-0000 DF13-SB-GP02-0-0000*
3.30 2.03
30 30
67.6 47.9
40 40
NR NR
-
DF13-SB-GP02-0-0005
NR
_
10.2
50
NR
_
DF13-SB-GP02-0-0005*
NR
-
9.26
50
NR
-
DF13-SB-GP02-0-0010 DF13-SB-GP02-0-0010*
0.838 0.922
40 40
20.7 19.5
30 30
NR NR
_
-
DF13-SB-GP02-DB-0010 DF13-SB-GP02-DB-0010*
1.42 1.45
30 30
30.7 31.9
30 30
NR NR
-
-
DF13-SB-GP02-0-0020 DF13-SB-GP02-0-0020*
2.76 2.51
30 52.3 30 51.0
40 40
NR NR
_
DF13-SB-GP02-0-0050
1.91
30
40.1
40
NR
.
DF13-SB-GP02-0-0050*
2.06
30
41.8
40
NR
-
DF13-SB-GP03-0-0000
ND
30 2.80
30
NR
-
DF13-SB-GP03-0-0000*
ND
30 1.74
30
NR
-
DF13-SB-GP03-0-0005
ND
30 0.994
30
NR
.
DF13-SB-GP03-0-0005*
ND
30
1.02
30
NR
-
DF13-SB-GP03-0-0010
ND
30 1.13
30
NR
_
D F 1 3 -S B -G P 0 3 -0 -0 0 10*
ND
30 1.01
30
NR
-
DF13-SB-GP03-0-0020 DF13-SB-GP03-0-0020*
0.857 0.841
30 30
4.79 5.14
30 30
NR NR
-
-
DF13-SB-GP03-0-0050 DF13-SB-GP03-0-0050*
0.520 0.609
30 30
4.25 5.56
30 30
NR NR
-
-
DF13-SB-GP04-0-0000
3.73
30
59.4
30
NR
.
DF13-SB-GP04-0-0000*
3.78
30
59.4
30
NR
*
DF13-SB-GP04-0-0005 DF13-SB-GP04-0-0005*
0.380 0.369
40 40
5.08 5.21
30 30
NR NR
-
DF13-SB-GP04-0-0010
0.730
30
7.58
30
NR
_
DF13-SB-GP04-0-0010*
0.678
30
7.42
30
NR
-
DF13-SB-GP04-0-0020 DF13-SB-GP04-0-0020*
0.891 0.932
40 40
5.31 5.26
30 30
NR NR
_
'
DF13-SB-GP04-0-0050
1.34
40
7.99
30
NR
-
DF13-SB-GP04-0-0050*
1.25
40
7.04
30
NR
-
DF13-SB-GP04-DB-0050 DF13-SB-GP04-DB-0050*
1.70 1.69
40 40
8.16 8.25
30 30
NR NR
-
-
DF05-SB-GP03-0-0000 DF05-SB-GP03-0-0000*
ND ND
30 0.393 30 0.333
30 30
NR NR
-
-
DF05-SB-GP03-0-0005
0.384
30
2.35
30
NR
-
DF05-SB-GP03-0-0005*
0.334
30
2.08
30
NR
DF05-SB-GP03-0-0010 D F 0 5 -S B -G P 0 3 -0 -00 1 0*
0.541 0.397
30 30
2.46 2.13
30 30
NR NR
-
"
"Laboratory Duplicate ND h Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table II for re-extract data.
Exygen Research
Page 25 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study N o.: P0001131
Table I.
Summary of PFBS, PFHS and PFOS in Soil Samples Continued
Exvaen ID
C0152952 C0152952 Rep
CO152953 C0152953 Rep
C0152954 C0152954 Rep
CO152955 C0152955 Rep
C0152956 C0152956 Rep
C0152957 C0152957 Rep
C0152958 C0152958 Rep
C 0 152959 C0 152959 Rep
C0153316 C0153316 Rep
C0153317 C0153317 Rep
C0153318 C0153318 Rep
C0153319 C0153319 Rep
C0153320 C0153320 Rep
C0153321 C0153321 Rep
C 0 153322 C0153322 Rep
C0153323 C0153323 Rep
C0153324 C0153324 Rep
CO153325 C0153325 Rep
CO153326 C0153326 Rep
CO153327 C0153327 Rep
CO153328 C0153328 Rep
Client Sample ID
DF05-SB-GP03-DB-0010 D F 0 5 -S B -G P 0 3 -D B -0 0 10*
DF05-SB-GP03-0-0020 DF05-SB-GP03-0-0020*
DF05-SB-GP03-0-0050 DF05-SB-GP03-0-0050*
DF05-SB-GP04-0-0000 DF05-SB-GP04-0-0000*
DF05-SB-GP04-0-0005 DF05-SB-GP04-0-0005*
DF05-SB-GP04-0-0010 DF05-SB-GP04-0-0010*
DF05-SB-GP04-0-0020 DF05-SB-GP04-0-0020*
DF05-SB-GP04-0-0050 DF05-SB-GP04-0-0050*
DF10-SB-GP03-0-0000 DF10-SB-GP03-0-0000*
DF10-SB-GP03-0-0005 DF10-SB-GP03-0-0005*
DF10-SB-GP03-0-0010 DF10-SB-GP03-0-0010*
DF10-SB-GP03-0-0020 DF10-SB-GP03-0-0020*
DF10-SB-GP03-0-0050 DF10-SB-GP03-0-0050*
DF10-SB-GP04-0-0000 D F1O-SB-G P04-0-0000*
DF10-SB-GP04-0-0005 DF10-SB-GP04-0-0005*
D F 10-SB-G P04-0-0010 DF10-SB-GP04-0-0010*
DF10-SB-G P04-0-0020 DF10-SB-GP04-0-0020*
DF10-SB-GP04-0-0050 DF10-SB-GP04-0-0050*
DF10-SB-GP04-DB-0050 DF10-SB-GP04-DB-0050*
DF11-SB-GP01-0-0000 D F11-SB-GP01-0-0000*
DF11-SB-GP01 -0-0005 DF11-SB-GP01 -0-0005*
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
PerfluorobutanesuHonate_______ Perfluorohexanesulfonita________Psrfluorooctanesulfonata
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy _W-%>
Analyte Found (ppb, ng/g) Drv Weight
Assessed Accuracy
<+/-%)
Analyte Found (ppb. ng/g) Drv Weight
Assessed Accuracy
(+/- %)
.
0.440
30
2.49
30
NR
0.440
30
2.81
30
NR
-
1.13 0.453
30 30
6.56 3.44
40 40
137 30 143 30
NR
7.01
30
66.6
30
NR
7.78
30
95.6
30
ND 40 ND
30
78.1
30
ND 40 ND
30 67.6 30
ND 30 ND 30 NR ND 30 ND 30 NR
-
ND
30 0.367 30
63.9
30
ND
30 0.322
30
61.9
30
0.254 30 0.867 ND 30 0.744
30 30
NR NR
-
0.279 40 0.368
30
0.310
30
0.317 40 0.428 30 0.425 30
NR - 121 30 NR - 138 30
NR
-
74.5
30
NR
-
70.9
30
15.4 16.2
30 30
NR NR
-
NR
-
21.6
30
NR
-
19.7
30
NR NR
NR NR
NR NR
NR NR
19.3
30
139
30
21.7
30
156
30
NR NR
2.48 40 42.8 3.00 40 32.8
50 50
NR NR
1.78 40 22.8 1.59 40 20.0
NR . NR NR - NR
NR . 33.1 NR - 34.0
39.1 36.4
30 30
NR NR
41.2 38.9
30 30
NR NR
50 50
_ -
40 40
-
-
NR NR
NR NR
NR NR
NR NR
NR NR
38.3
30
NR
-
NR -
46.2
30
NR
-
NR -
31.3
30
NR
-
NR -
29.5
30
NR
"
NR
'
'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table II for re-extract data.
Exygen Research
Page 26 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table I.
Summary of PFBS, PFHS and PFOS in Soil Samples Continued
Exygen ID
CO153329 C0153329 Rep
CO153330 C 0153330 Rep
CO153332 C0153332 Rep
C0153333 C0153333 Rep
C0153334 C0153334 Rep
C0153335 C0153335 Rep
CO153336 C0153336 Rep
C0153337 C0153337 Rep
C 0 153338 C0153338 Rep
CO153339 C0153339 Rep
C0153340 C0153340 Rep
C0153341 C0153341 Rep
CO153342 C0153342 Rep
CO153344 C0153344 Rep
C 0 153345 C0153345 Rep
C0153346 C0153346 Rep
C0153347 C0153347 Rep
C0153348 C0153348 Rep
CO153349 C0153349 Rep
Client Sample ID
DF11-SB-GP01-0-0010 DF11-SB-GP01-0-0010*
DF11-SB-GP01-0-0020 DF11-SB-GP01-0-0020*
D F11-SB-GP01 -0-0050 DF11-SB-GP01-0-0050*
DF11-SB-GP02-0-0000 DF11-SB-GP02-0-0000*
DF11-SB-GP02-0-0005 DF11-SB-GP02-0-0005*
DF11-SB-GP02-0-0010 DF11-SB-GP02-0-0010*
DF11-SB-GP02-0-0020 DF11-SB-GP02-0-0020*
DF11-SB-GP02-0-0050 DF11-SB-GP02-0-0050*
DF11-SB-GP03-0-0000 D F11-SB-GP03-0-0000*
D F 1 1-SB-GP03-0-0005 DF11-SB-GP03-0-0005*
DF11-SB-GP03-0-0010 D F 1 1-SB -G P03-0-0010*
DF11-SB-GP03-DB-0020 D F11-SB-GP03-DB-0020*
D F11-SB-GP03-0-0020 DF11-SB-GP03-0-0020*
DF11-SB-GP03-0-0050 DF11-SB-GP03-0-0050*
DF11-SB-GP04-0-0000 DF11-SB-GP04-0-0000*
DF11-SB-GP04-0-0005 DF11-SB-GP04-0-0005*
DF11-SB-GP04-0-0010 DF11-SB-GP04-0-0010*
DF11-SB-GP04-0-0020 D F 1 1-SB-GP04-0-0020*
DF11-SB-GP04-0-0050 D F11-SB-GP04-0-0050*
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutanasulfonata_______Partiuorohexanaiulfonata_______ Parfluorooctanaaulfonata
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
(+ /.% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
< /.% )
.4 0 . 4
30
NR
NR
33.2
30
NR
-
NR
27.1 50 121
30
31.3
50
126
30
NR NR
NR NR
0.489 0.440
.
-
40 40
85.5 78.0
2.49 2.23
30 30
30 30
NR NR
NR NR
2.70 2.89
40 40
NR NR
NR NR
4.93 5.34
30 30
NR NR
NR NR
55.1 55.3
30 30
NR NR
NR NR
NR NR NR NR
NR NR
NR
2.87
30
NR
NR
2.35
30
NR
NR
8.76
30
NR
NR
9.20
30
NR
NR
21.1
30
NR
NR
23.6
30
NR
122 40 109 40
NR NR
-
NR NR
62.7
40
NR
-
NR
69.1
40
NR
-
NR
34.4 30 83.8
30
25.4
30
64.2
30
ND 30 1.22 30 ND 30 1.32 30
NR NR
NR NR
-
ND
30 0.427
30
75.6
30
ND
30 0.642
30
89.4
30
ND
30 0.994
30
NR
-
0.265 30 0.953
30
NR
1.10 30 4.37 1.19 30 4.56
30 30
NR NR
-
-
1.72 30 4.69 1.67 30 4.59
30 30
NR NR
'
` Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR - Not reported due to quality control failures, see Table II for re-extract data.
Exygen Research
Page 27 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table I.
Summary of PFBS, PFHS and PFOS in Soil Samples Continued
Exvqen ID
CO153350 C0153350 Rep
C0153351 C0153351 Rep
C0153352 C015 3 3 5 2 Rep
CO153353 C0153353 Rep
C0153354 CO153354 Rep
CO153356 C0153356 Rep
C0153357 C015 3 3 5 7 Rep
CO153358 C0153358 Rep
C0153359 C0153359 Rep
C0153360 C0153360 Rep
C0153361 C0153361 Rep
Client Sample ID
DF05-SB-GP01-0-0000 DF05-SB-GP01 -0-0000*
DF05-SB-GP01 -0-0005 DF05-SB-GP01-0-0005*
DF05-SB-GP01-0-0010 DF05-SB-GP01-0-0010*
DF05-SB-GP01-0-0020 DF05-SB-GP01 -0-0020*
DF05-SB-GP01-0-0050 DF05-SB-GP01-0-0050*
DF05-SB-GP02-0-0000 DF05-SB-GP02-0-0000*
DF05-SB-GP02-0-0005 DF05-SB-GP02-0-0005*
DF05-SB-GP02-0-0010 DF05-SB-GP02-0-0010*
DF05-SB-GP02-0-0020 DF05-SB-GP02-0-0020*
DF05-SB-GP02-0-0050 DF05-SB-GP02-0-0050*
DF05-SB-GP02-DB-0050 DF05-SB-GP02-DB-0050*
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Prt1uorobutnuifonte______ Psrtluorohexanesulfbnate_______PsrtluorooctanasuHbnata
Analyte Found (ppb. ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (PPb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
</-%)
4.42 5.00
30 30
NR NR
-
ND 50 0.393 30 ND 50 0.465 30
ND
30 0.657
30
ND
30 0.571
30
0.335 0.286
40 40
1.53 1.55
30 30
NR
.
85.2
50
NR
-
90.9
50
ND 30 ND ND 30 ND
30 30
ND
30 0.379
30
ND
30 0.364
30
NR
.
1.05
30
NR * 0.790 30
NR
-
0.698
30
NR - 0.671 30
NR
.
1.26
30
NR * 0.833 30
NR - 4.83 30 NR " 4.66 30
NR NR
NR NR
NR NR
NR NR
NR NR
NR NR
NR NR
NR NR
NR NR
NR NR
7.55 6.22
. . -
-
-
-
-
-
-
-
30 30
'Laboratory Duplicate ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table II for re-extract data.
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table II. Summary of PFBS, PFHS and PFOS in Re-extracted Soil Samples
Exvaen ID C0152890 C 0 1 52891 C0152892 C0152893 C0152894 C0152895 C0152896 C0152897 C0152898 C0152899 CO152900 C0152901 C0152902 C0152903 C0152904 C0152905 C0152906 C0152907 C0152908 C0152909 C0152910 C 0 1 52911 C0152912 C0152913 C0152915 C0152919 C0152921
Client Sample ID D F 10-SB-GP01 -0-0000 D F 10-SB-GP01 -0-0005 DF10-SB-GP01 -04)010 DF10-SB-GP01-DB-0020 DF10-SB-GP01 -04)020 DF10-SB-GP01-0-0050 DF10-SB-GP02-0-0000 DF10-SB-GP02-0-0005 DF10-SB-GP02-0-0010 DF10-SB-GP02-0-0020 DF10-SB-GP02-0-0050 DF12-SB-GP01-0-0000 DF12-SB-GP01-0-0005 DF12-SB-GP01-0-0010 DF12-SB-GP01-0-0020 DF12-SB-GP01-0-0050 DF12-SB-GP01-DB-0050 DF12-SB-GP02-0-0000 DF12-SB-GP02-0-0005 DF12-SB-GP02-0-0010 DF12-SB-GP02-0-0020 DF12-SB-GP02-0-0050 DF12-SB-GP03-0-0000 DF12-SB-GP03-0-0005 DF12-SB-GP03-0-0020 DF12-SB-GP04-0-0010 DF12-SB-GP04-DB-0020
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Poffluorobutareutfonato______ Pwfluorohaxanwutfonata_______Pertluorooctan-- ultbnate
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
</.%)
Analyte Found
(ppb, ng/g) Dry Weight
Assessed Accuracy
---
-
1870
30
---
---
---
---
---
---
---
---
---
--14.1 30 227
---
---
---
---
8.93
30
-
---
NR NR
-
- - 414
262 40 686
-
1440
30
- 980 30
-
1050
30
- 988 40
-
6340
30
- 231 30
- 495 30
- 357 30
- 364 30
- 271 40
30
254000
30
-
33000
30
-
8020
30
-
22000
30
-
12900
40
-
10700
40
-
16800
30
-
6910
30
-
6300
30
50
1100
30
40 871 30
-----------
- 426 30
- 445 30
-
88.8
50
- NR NR
-
11700
30
N R = Not reported due to quality control failures.
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table II. Summary of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Exvaen ID C0152922 C0152923 C0152924 C0152925 C0152926 C0152927 C0152928 C0152929 C0152930 C 01 52931 C0152932 C0152933 C0152936 C0152937 C0152938 C0152940 C 0 152941 C 0152942 C0152943 C0152944 C0152949 C0152950 C 0 1 52951 C0152952
Client Sample ID DF12-SB-GP04-0-0050 DF13-SB-GP01 -0-0000 DF13-SB-GP01-0-0005 DF13-SB-GP01-0-0010 DF13-SB-GP01-0-0020 DF13-SB-GP01-0-0050 DF13-SB-GP02-0-0000 DF13-SB-GP02-0-0005 DF13-SB-GP02-0-0010 DF13-SB-GP02-DB-0010 DF13-SB-GP02-0-0020 DF13-SB-GP02-0-0050 DF13-SB-GP03-0-0010 DF13-SB-GP03-0-0020 DF13-SB-GP03-0-0050 DF13-SB-GP04-0-0005 DF13-SB-GP04-0-0010 DF13-SB-GP04-0-0020 DF13-SB-GP04-0-0050 DF13-SB-GP04-DB-0050 DF05-SB-GP03-0-0000 DF05-SB-GP03-0-0005 DF05-SB-GP03-0-0010 DF05-SB-GP03-DB-0010
C4 Sulfonate PFBS
C0 Sulfonate PFHS
C8 Sulfonate PFOS
PertluofotHitannilfoiurte_______Periluorohsxanesulfbnats_______ PsrfluorooctsnssurfonaU
Analyte Found
(ppb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
</-*>
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
(+/-% )
---
-
26800
30
-
-
284
30
285000
30
NR NR
-
-
76900
40
2.68
40
-
-
18600
30
5.83
30
287
50
22000
30
0.754
40
-
-
7560
30
---
-
102000
30
0.790
30
-
-
38300
40
----
-
8550
30
-
7160
30
--
-
8870
30
--
-
8340
30
--
- 223 30
--
- 735 30
--
- 543 50
--
-
5940
30
---
-
3470
50
-
2160
30
--
-
1840
30
--
-
1760
30
--
-
-
83.1
30
- - - - 194 30
---
- 151 30
--
- 121 30
N R = Not reported due to quality control failures.
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Page 30 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table II. Summary of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Exvqen ID C0152954 C0152956 C0152958 C0153319 C0153320 C0153321 CO153322 C0153325 CO153326 C0153329 C0153330 C0153332 C0153333 C0153334 C0153335 C0153336 C0153337 C0153338 C0153340 C 0 1 53341
Client Sample ID DF05-SB-GP03-0-0050 DF05-SB-GP04-0-0005 DF05-SB-GP04-0-0020 DF10-SB-GP03-0-0020 DF10-SB-GP03-0-0050 DF10-SB-GP04-0-0000 DF10-SB-GP04-0-0005 DF10-SB-GP04-0-0050 DF10-SB-GP04-DB-0050 DF11-SB-GP01-0-0010 DF11-SB-GP01-0-0020 DF11-SB-GP01-0-0050 D F11-SB -GP02-0-0000 D F 1 1-SB -GP02-0-0005 DF11-SB-GP02-0-0010 DF11-SB-GP02-0-0020 D F 1 1-SB-GP02-0-0050 D F11-SB-GP03-0-0000 DF11-SB-GP03-0-0010 D F 1 1-SB-GP03-DB-0020
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Parfluorobutanesulfonate_______ Parfluorohexangsulfonata_______ Perfluorooctan-- ulfonata
Analyte Found Assessed Analyte Found Assessed Analyte Found Assessed
(ppb, ng/g) Dry Weight
Accuracy (+ '-% )
(ppb, ng/g) Dry Weight
Accuracy (+/-% )
(ppb, ng/g) Dry Weight
Accuracy (+/-% )
NR NR
-
--
---
113 30
---
7.36
50
NR NR
-
4470
30
---
5260
40
---
84800
30
---
13800
30
-
-
274
50
5890
30
-
-
268
30
1610
50
-
-
140
30
10700
40
---
-
10600
50
NR NR
-
- NR NR
---
-
10800
30
-
-
603
40
1960
30
-
-
332
30
14500
30
-
-
524
30
3500
40
NR NR 463
40
2110
50
NR NR
-
- 817 30
NR NR
-
-
19800
30
-
-
393
30
1410
30
NR = Not reported due to quality control failures.
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table II. Summary of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Exvaen ID C0153342 C0153344 C0153345 C0153347 C0153348 C0153349 CO153350 C0153351 C0153352 C0153353 C0153354 C0153356 C0153357 C0153360 C 01 53361
Client Sample ID DF11-SB-GPO3-0-0020 DF11-SB-GP03-0-0050 DF11-SB-GP04-0-0000 DF11-SB-GP04-0-0010 DF11-SB-GP04-0-0020 D F 1 1-SB -G P04-0-0050 DF05-SB-GP01 -0-0000 DF05-SB-GP01-0-0005 DF05-SB-GP01-0-0010 D F 05-S B -G P 0 1 -0-0020 DF05-SB-GP01-0-0050 DF05-SB-GP02-0-0000 DF05-SB-GP02-0-0005 DF05-SB-GP02-0-0050 DF05-SB-GP02-DB-0050
C4 Sulfonate PFBS
C6 Sulfonate PFHS
C8 Sulfonate PFOS
Perfluorobutan--ulfbnato______Pffluorohaxan--utfonata_______PerfluorooctanesuHbnate
Analyte Found (ppb. ng/g) Dry Weight
Assessed Accuracy
(+/-% )
Analyte Found (ppb, ng/g) Dry Weight
Assessed Accuracy
</-%>
Analyte Found
(ppb. ng/g) Dry Weight
Assessed Accuracy
(/-% )
- - 408 30 NR NR
---
- 420 30
---
- 183 30
---
- 192 30
---
- NR NR
---
- NR NR
- - NR NR 16000 40
-----
- 619 30
-
1010
30
---
-
1980
30
NR NR
-
-
1430
30
---
- 156 30
-NR NR
-
- 333 30 - 126 30
NR NR
-
-
--
N R = Not reported due to quality control failures.
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table III. Summary of PFBS, PFHS and PFOS in Trip Blank and Equipment Rinseate Blank Samples
Exvaen ID C0152885 C0152888 C0152889 C0152946 C0153313 C0153331 C0153343 C0153355
Client Sample ID DF13-SB-TRIP1-0-060209 DF12-SB-GP03-RB-0000 DF12-SB-GP01-0-RB-0000 DF05-SB-TRIP3-0-060216 DF10-SB-TRIP2-0-060216 DF11-SB-GP01-RB-0040 DF11-SB-GP03-RB-0040 DF05-SB-GP02-RB-000
C 4 S u lfo n ate PFBS
C 6 S u lfo n a te PFHS
C 8 S u lfo n a te PFOS
PeffluofofauUnasulfoiwt______ Perfluorohexanesulfonat______ Paffluorooctan-- utfonate
Analyte Found (ng/L)
Assessed Accuracy
(*/%)
Analyte Found (ng/L)
Assessed Accuracy
(+/-%)
Analyte Found (ng/L)
Assessed Accuracy
(+/-% )
ND 30 ND
30 ND 30
ND - ND - ND -
ND - ND
- ND -
ND 30 ND
30 ND 30
ND 30 ND
30 ND 30
ND - ND
- ND -
ND - ND - ND -
ND - ND
- ND -
ND = Not detected at or above the Limit of Quantitation (LCJQ) of 25 ng/L.
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Page 33 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples
Sample Description
O l^^n at^FB S W rtW o jgh^^^B S uj^n a^^FH S W etW e lgh^^^C S S ijton at^FO S JW etW o lg^
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(ng/g) (ng/a)
(ng/g)
(%>
(ng/g)
(ng/g)
(%)
(ng/g)
(ng/g)
(%)
D F 10-SB-GP01-0-0000
(C0132S90Spk E, 4 ppb Spike)
4
0.437
5.63
130
1.78
DF 10-SB-GP01-0-0000
(C0132890Spk F, 40 ppb Spike)
40
0.437
54.2
134
1.78
4.54 69 42.2 101
NR NR
NR NR NR NR
DF 10-SB-GP01-0-0005
(C0152S91 Spk 6 ,4 ppb Spike)
4
0.457
5.72
132
1.78
5.16
85
DF10-S8-GP01 -0-0005
(C01S2991 Spk H, 40 ppb Spike)
40
0.457
52.6
130
1.78
34.8
83
NR NR
NR NR NR NR
DF10-SB-GP01 -0-0010
(C01S2892Spk 1,4 ppb Spike)
4
1.35
6.54 130 3.58
7.35
94
DF10-SB-GP01-0-0010
(00192092 Spk J, 40ppb Spike)
40
1.35
54.3 132 3.58
35.8
61
NR NR
NR NR NR NR
DF10-SB-GP01-DB-0020
(00132803Spk K, 4ppb Spike)
4
1.33
6.44 128 3.55
6.17
66
DF10-SB-GP01-DB-0020
(00132893Spk L, 40 ppb Spike)
40
1.33
53.0 129 3.55
34.9
78
NR NR
NR NR NR NR
DF10-SB-GP01-0-0020
(C01S2884Spk 0 ,4 ppb Spike)
4
0.912
4.80
97
3.23
6.93
93
DF10-SB-GP01-0-0020
(00132884Spk D, 40 ppb Spike)
40
0.912
44.2
108
3.23
37.8
86
NR NR
NR NR NR NR
DF10-SB-GP01-0-0050
(C0152889Spk E, 4 ppb Spike)
4
1.41
5.82 110 14,2
27.5
*
NR
NR NR
DF 10-SB-GP01-0-0050
(00132883Spk F, 40ppb Spike)
40
1.41
29.3
70
14.2
45.6
79
NR
NR NR
DF 10-SB-GP02-0-0000 (COI32888Spk 6 .4 ppb Spike)
DF 10-SB-GP02-0-0000 (C01S2888 Spk H, 40ppb Spite)
4 40
ND ND
2.66 67 1.40 4.69 87 30.4 76 1.40 35.1 84
NR NR
NR NR NR NR
DF 10-SB-GP02-0-0005 (CM32897Spk 1,4 ppb Spike)
DF 10-SB-GP02-0-0005 (C0192897Spk J, 40 ppb Spite)
DF10-SB-GP02-0-0010 (C0152888Spk K, 4 ppb Spike)
DF 10-SB-GP02-0-0010 (C0132888Spk L, 40 ppb Spike)
4 40 4 40
ND ND ND ND
4.10 103 0.872 4.46 37.1 93 0.872 35.2 3.77 94 0.888 4.16 40.8 102 0.886 35.4
90 86 82 86
NR NR NR NR
NR NR NR NR NR NR NR NR
DF10-SB-GP02-0-0020
(00132888Spk C. 4 ppb Spike)
4
0.356
4.94
115
2.22
DF 10-SB-GP02-0-0020
<00152899 Spk 0,40 ppb Spite)
40
0.358
44.6
111
2.22
5.70 36.8
87 86
NR NR
NR NR NR NR
'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated NO = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values mayvary slightly from the values In the raw date.
Exygen Research
Page 34 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C4 Sulfonate PFBS Wet W eight C6 Sulfonate PFHS Wet W eight C8 Sulfonate PFOS Wet Weight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery (ng/g) (ng/g) wet wt.'ng/g) wet wt. <%> fno/o) wet wU'ng/g) wet wt. (%) (ng/g) wet wt/ng/g) wet wt. (%)
D F 10-SB-GP02-0-0050
(00152900Spk E, 4ppb Spike)
4
2.12
4.88 69
11.4
14.3
73
DF 10-SB-GP02-0-0050
(C01S2900Spk F, 40ppb Spike) 40
2.12
32.7
76
11.4
36.8
64
NR NR
NR NR NR NR
DF 12-SB-G P01-0-0000 (C01S2901 Spk6,4 ppb Spike)
DF12-SB-GP01 -0-0000 (Cd929d SpkH, 40ppb Spike)
4 40
NR NR
NR NR NR NR NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 12-SB-GP01-0-0005 (Cd52902Spk 1,4ppb Spike)
4
2.70
6.90 105 50.4
43.6
*
DF 12-SB-G P01-0-0005
(C0152902SpkJ. 40ppbSpike) 40
2.70
52.8 125 50.4
71.0
52
NR NR
NR NR NR NR
DF12-SB-GP01-0-0010
(C d52903Spk K, 4 ppb Spike)
4
0.656
5.88
126
15.0
24.4
*
DF12-SB-GP01-0-0010
(Cd52903Spk L, 40ppb Spike) 40
0.856
51.1
126
15.0
51.3
91
NR NR
NR NR NR NR
DF 12-SB-GP01-0-0020 (C0152904SpkC, 4ppb Spike)
DF12-SB-GP01 -0-0020 (C d52904Spk 0,40ppb Spike)
4 40
1.34 1.34
6.82 137 20.1 55.3 135 20.1
17.8 49.3 73
NR NR
NR NR NR NR
DF 12-SB-GP01 -0-0050
(C0152905SpkE, 4ppb Spike)
4
0.886
5.98
127
16.3
25.1
.
DF12-SB-GP01-0-0050
(Cd52905Spk F, 40ppb Spike)
40
0.888
51.4
126
16.3
46.5
76
NR NR
NR NR NR NR
DF12-SB-GP01-DB-0050
(Cd52906Spk G, 4 ppb Spike)
4
0.455
5.71
131
17.6
23.5
*
DF12-SB-GP01-DB-0050
(C d52906SpkH, 40ppb Spike) 40
0.455
49.8
123
17.6
48.6
78
NR NR
NR NR NR NR
DF 12-SB-GP02-0-0000 (Cd52907 Spk 1,4ppb Spike)
DF12-SB-GP02-0-0000 (Cd52907Spk J, 40ppb Spike)
4 40
NR NR
NR NR 31.2 33.6
NR NR 31.2 57.1 65
NR NR
NR NR NR NR
DF 12-SB-GP02-0-0005 (Cd52906Spk K, 4ppb Spike)
4
3.18
7.37 105 26.0
29.3
DF12-SB-GP02-0-0005
(Cd52908 Spk L, 40ppb Spike)
40
3.18
49.2 115 26.0
53.0
68
NR NR
NR NR NR NR
DF 12-SB-GP02-0-0010 (C d52909Spk C, 4ppb Spike)
DF 12-SB-GP02-0-0010 (Cd52909Spk 0.40 ppb Spike)
4 40
NR NR
NR NR 30.4 33.5
*
NR NR 30.4 65.4 88
NR NR
NR NR NR NR
' Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND * Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table show s rounded results, recovery values may vary slightly from die values in die raw data.
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Page 35 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C A S u lfo n a teP FB S W etW elg h t^C eS u lfo n a teP FJ^^
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(no/o) (no/o) wet wt.'ng/g) wet wt. (%) fno/a) wet wt/na/a) wet wt. (%) (ng/g) wet wt.[ng/g) wet wt. (%)
D F 12-SB-GP02-0-0020 (C0152910Spk E, 4ppb Spike)
DF 12-SB-GP02-0-0020 (C0152910Spk F, 40 ppb Spike)
4 40
113 113
113 171 145
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 12-SB-GP02-0-0050 (C0152911 Spk 0 ,4 ppb Spike)
DF12-SB-GP02-0-0050 (00152911 Spk H, 40 ppb Spike)
4 40
NR NR
NR NR NR NR NR NR
NR NR NR NR NR NR
NR NR NR NR
DF12-SB-GP03-0-0000
(00152912Spk 1,4 ppb Spike)
4
0.515
6.47
149
2.11
DF 12-SB-GP03-0-0000
(00192912Spk J, 40 ppb Spike)
40
0.515
56.5
145
2.11
5.66 89 35.0 82
NR NR
NR NR NR NR
DF 12-S8-GP03-0-0005 (C019291S Spk K, 4 ppb Spike)
DF 12-SB-GP03-0-0005 (COt52913Spk L, 40 ppb Spike)
4 40
ND ND
2.48 62 0.707 3.87 79 28.0 70 0.707 32.9 80
NR NR
NR NR NR NR
DF 12-SB-GP03-0-0010
(00152914Spk C, 4 ppb Spike)
4
1.14
4.24
78
2.38
5.50 78
72.9 72.2
*
DF 12-SB-GP03-0-0010
(00152914Spk D, 40ppb Spike)
40
1.14
33.4 81
2.38
36.0
84
72.9
110
93
DF 12-SB-GP03-0-0020
(C0152915Spk E, 4 ppb Spike)
4
1.43
4.28
71
4.16
7.39
81
DF 12-SB-GP03-0-0020
(CM52915Spk F, 40 ppb Spike)
40
1.43
20.2
47
4.16
30.1
65
NR NR
NR NR NR NR
DF 12-SB-GP03-0-0050
(CM52919 Spk 6 ,4 ppb Spike)
4
1.30
4.14
71
6.50 9.68 80
65.5 71.8
*
DF 12-SB-GP03-0-005Q
(CM52916Spk H, 40ppb Spike)
40
1.30
29.2
70
6.50
40.3 85
65.5
110 111
DF12-SB-GP04-0-0000 (CM52917 Spk 1,4 ppb Spike)
DF12-SB-GP04-0-0000 (CM52917Spk J, 40ppb Spike)
DF 12-SB-GP04-Q-0005 (CM 52916Spk K, 4 ppb Spike)
DF12-SB-GP04-0-0005 (CMS2918 Spk L, 40 ppb Spike)
4 40 4 40
0.984 0.984
NR NR
6.50 38.5 NR NR
138 94 NR NR
NR NR NR NR
NR NR NR NR NR NR NR NR NR NR NR NR
NR NR NR NR NR NR NR NR
DF 12-SB-GP04-0-0010 (CM52919Spk C, 4 ppb Spike)
DF 12-SB-GP04-0-0010 (CM 52919 Spk D, 40ppb Spike)
4 40
3.51 3.51
8.16 116 29.5 54.8 128 29.5
36.8 * 77.0 119
NR NR
NR NR NR NR
`Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summarytable shows rounded results, recovery values mayvary slightlyfrom the values in the raw data.
Exygen Research
Page 36 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C4ShilfonatoBPFBSW etW el2htii^ iC e S u l ^ ^
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery In Sample Recovered Recovery
(ng/g) ing/o) wet wt.!no/g) wet wt. <%) (ng/g) wet wt/ng/g) wet wt. <%> (ng/g) wet wt/ng/g) wet wt. (%>
DF12-SB-GP04-0-0020
(C0152920Spk E, 4 ppb Spike)
4
1.82
6.56 119
19.6
22.3
D F 12-SB-GP04-0-0020
(C0152920Spk F, 40 ppb Spike)
40
1.82
50.6 122 19.6
56.0
91
NR NR
NR NR NR NR
DF12-SB-GP04-DB-0020 (C0152921 Spk 6 ,4 ppb Spike)
DF12-SB-GP04-DB-0020 (C0152921 Spk H, 40 ppb Spike)
4 40
1.45 1.45
6.98 138 19.7 51.2 124 19.7
29.4 62.4 107
NR NR
NR NR NR NR
DF12-SB-GP04-0-0050
(C0152922Spk 1,4 ppb Spike)
4
3.18
6.01
71
31.2
34.6
DF 12-SB-GP04-0-0050
(C0152922Spk J, 40ppb Spike)
40
3.18
44.2 103 31.2
60.9
74
NR NR
NR NR NR NR
DF13-SB-GP01-0-0000 (C0152923Spk K, 4ppb Spfce)
DF13-SB-GP01-0-0000 (C0152923 Spk L, 40 ppb Spike)
4 40
8.40 8.40
11.8 85 59.4 128
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF13-SB-GP01-0-0005 (C01S2924Spk C, 4 ppb Spike)
DF 13-SB-GP01-0-0005 (C01S2924Spk D, 40 ppb Spike)
4 40
NR NR
NR NR 50.0 54.1 NR NR NR 50.0 101 128 NR
NR NR NR NR
DF 13-SB-GP01 -0-0010 (C0152925Spk E, 4 ppb Spike)
DF13-SB-GP01 -0-0010 (C01S2925Spk F, 40 ppb Spike)
4 40
NR NR
NR NR 41.1 43.4
*
NR NR 41.1 75.3 86
NR NR
NR NR NR NR
DF13-SB-GP01-0-0020 (C0152929Spk 0 ,4 ppb Spike)
DF13-SB-GP01-0-0020 (C0152929 Spk H, 40ppb Spike)
4 40
NR NR
NR NR NR NR NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 13-SB-GP01-0-0050 (C01S2927Spk 1,4 ppb Spike)
DF13-SB-GP01-0-0050 (C01S2927Spk J, 40ppb Spike)
DF 13-SB-GP02-0-0000 (C01S292S Spk K, 4 ppb Spike)
DF13-SB-GP02-0-0000 (00152929Spk L. 40 ppb Spike)
4 40 4 40
NR NR 2.50 2.50
NR NR 12.6 NR NR 12.6 6.78 107 51.1 56.8 136 51.1
20.0 50.6 33.8 77.8
. 95 . 67
NR NR NR NR
NR NR NR NR NR NR NR NR
DF 13-SB-GP02-0-0005 (C0192929Spk C, 4 ppb Spike)
DF 13-SB-GP02-0-0005 (C0152929 Spk 0,40 ppb Spike)
4 40
NR NR
NR NR 8.24 10.5 57 NR NR 8.24 43.0 87
NR NR
NR NR NR NR
Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND 8 Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR 8 Not reported due to qualitycontrol failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightlyfrom the values In the raw data.
Exygen Research
Page 37 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C4 Sulfonate PFBS Wet W eight C6 Sulfonate PFHS Wet W eight C8 Sulfonate PFO S W et W eight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
inaia) (no/p) wet wt.'np/p) wet wt. (%) (no/p) wet wt-'np/p) wet w t <%> (no/p) wet wt.np/p) wet w t (%)
DF13-SB-GP02-0-0010
(C01S2930Spfc E, 4ppb Spil)
4
0.699
5.38
130
17.3
20.6
*
DF13-SB-GP02-0-0010
(C0152930Spk F, 40ppb Splke)
40
0.699
54.4
134
17.3
50.6
83
NR NR
NR NR NR NR
DF13-SB-GP02-DB-0010
(C01S2931 SpkG, 4ppb Splke)
4
1.18
5.88 118 25.4
28.5
*
DF 13-SB-GP02-DB-0010
(COI52931 Spk H, 40ppb Splke) 40
1.18
52.3 128 25.4
58.5
83
NR NR
NR NR NR NR
DF 13-SB-GP02-0-0020
(COI52932Spk 1,4ppb Splke)
4
2.31
6.90 115 43.8
48.4
*
DF 13-SB-GP02-0-0020
(COI52932spk J , 40ppb Splke)
40
2.31
50.6 121 43.8
69.9
65
NR NR
NR NR NR NR
DF 13-SB-GP02-0-0050
(C0152933Spk K, 4ppb Splke)
4
1.57
6.06 112 32.9
35.6
*
DF13-SB-GP02-0-0050
(C0152933Spk L, 40ppb Splke) 40
1.57
53.0 129 32.9
60.3
69
NR NR
NR NR NR NR
DF13-SB-GP03-0-0000 (COI52034Spk C, 4ppb Splke)
DF13-SB-GP03-0-0000 (C0152934Spk D, 40ppb Splke)
4 40
ND ND
5.06 127 2.26 5.73 87 45.4 114 2.28 36.3 85
NR NR
NR NR NR NR
DF 13-SB-GP03-0-0005 (COI52935Spk E, 4ppb Splke)
DF 13-SB-GP03-0-0005 (C0152935Spk F, 40ppb Splke)
4 40
ND ND
4.67 117 0.824 4.26 50.2 126 0.824 35.6
86 87
NR NR
NR NR NR NR
DF 13-SB-GP03-0-0010 (COI52939Spk G, 4ppb Splke)
DF13-SB-GP03-0-0010 (COI52939Spk H, 40ppb Splke)
4 40
ND ND
4.87 122 0.920 4.02 47.5 119 0.920 33.8
78 82
NR NR
NR NR NR NR
DF13-SB-GP03-0-0020
(C01529S7Spk 1,4ppb Splke)
4
0.639
4.98
109
3.58
DF 13-SB-GP03-0-0020
(C O I52937 Spk J , 40 ppb Splke)
40
0.639
44.3
109
3.58
6.63 37.6
76 85
NR NR
NR NR NR NR
DF 13-SB-GP03-0-0050
(C0152939Spk K, 4ppb Splke)
4
0.414
3.59
79
3.39
6.42
76
DF 13-SB-GP03-0-0050
(COt52930Spk L, 40ppb Splke) 40
0.414
32.6
80
3.39
34.8
79
NR NR
NR NR NR NR
DF 13-SB-GP04-0-0000 (COI52938Spk C, 4ppb Splke)
4
3.08
7.60 113 49.1
42.5
*
DF13-SB-GP04-0-0000
(COI52939Spk D, 40ppb Splke) 40
3.08 .
51.4
121
49.1
78.3 73
NR NR
NR NR NR NR
`Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated
ND * Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
Exygen Research
Page 38 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No. : P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C ^ulfonrtoJP FB S W etW dght^^^^ujfO flg^JFH SW etW elgh^^^C ^ijjfona^^FO S JW etW giglrt
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked In Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery (no/o) (no/a) wet wt.'na/g) wet w t (%) (na/a) wet wt/no/g) wet wt. <%) (no/o) wet wt.:na/g) wet w t (%)
DF 13-SB-GP04-0-0005
(C0152*40 Spk E, 4 ppb Spike)
4
0.310
5.63
133
4.14
772
90
DF13-SB-GP04-0-0005
(CM 52*40 Spk F. 40 ppb Spike)
40
0.310
52.6
131
4.14
37.8
84
NR NR
NR NR NR NR
DF13-SB-GP04-0-0010
(CM 52041 Spk 6 ,4 ppb Spike)
4
0.602
5.38
119
6.26
D F 13-SB-G P04-0-0010
(CM 52*41 Spk H, 40 ppb Spike)
40
0.602
46.2
114
6.26
10.1 41.0
96 87
NR NR
NR NR NR NR
DF13-SB-GP04-0-0020
(CM52942 Spk 1,4 ppb Spike)
4
0.714
6.04
133
4.26
7.70
86
DF13-SB-GP04-0-0020
(CM 52942 Spk J, 40ppb Spike)
40
0.714
46.6
115
4.26
35.6
78
NR NR
NR NR NR NR
DF 13-SB-GP04--0050 (CM 52949Spk K, 4 ppb Spike)
DF 13-SB-GP04-0-0050 (CM 52943 Spk L, 40 ppb Spike)
4 40
1.11 1.11
6.46 134 6.62 53.2 130 6.62
10.8 105 39.0 81
NR NR
NR NR NR NR
DF 13-SB-GP04-DB-0050 (CMS2944 Spk C, 4 ppb Spike)
DF 13-SB-GP04-DB-0050 (CM 52944Spk D, 40 ppb Spike)
4 40
1.46 1.46
6.68 131 6.99 50.0 121 6.99
10.4 39.6
65 82
NR NR
NR NR NR NR
DF05-SB-GP03-0-0000 (C0152949 Spk C, 4 ppb Spike)
DF05-SB-GP03-0-0000 (CM 52949 Spk D, 40 ppb Spike)
DF05-SB-GP03-0-0005 (CM52*50Spk E, 4 ppb Spike)
DF05-SB-GF03-0-0005 (C01529S0Spk F, 40 ppb Spike)
4 40
4 40
ND ND
0.310 0.310
4.70 118 0.330 38.1 95 0.330
5.16 121 1.90 52.2 130 1.90
3.85 33.8
4.98 34.3
86 84
77 61
NR NR
NR NR
NR NR NR NR
NR NR NR NR
DF05-SB-GP03-0-0010
(CM 52951 Spk Q, 4 ppb Spike)
4
0.425
4.94
113
1.94
DF05-SB-GP03-0-0010
(C01S2951 Spk H, 40 ppb Spike)
40
0.425
53.5
133
1.94
5.09 36.3
79 86
NR NR
NR NR NR NR
DF05-SB-GP03-DB-0010
(C0152952 Spk 1,4 ppb Spike)
4
0.338
5.35
125
1.91
DF05-SB-GP03-DB-0010
(C0152952Spk J, 40ppb Spike)
40
0.338
52.4
130
1.91
5.22 83 35.9 85
NR NR
NR NR NR NR
DF05-SB-GPQ3-0-0020
(CM 52953Spk K, 4 ppb Spike)
4
0.664
4.22
84
5.00
7.43
61
105
152
*
DF05-SB-GP03-0-0020
(C0152953 Spk L, 40 ppb Spike)
40
0.864
46.6
114
5.00
37.4
81
105
138 83
`Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated
ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values In the raw data.
Exygen Research
Page 39 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked In Sample Recovered Recovery In Sample Recovered Recovery In Sample Recovered Recovery
(nfl/g) (no/o) wet wt no/o) wet w t (%> (no/n) wet wt.'no/a) wet wt. (%) (na/a) wet wt.'no/a) wet wt. (%)
DF05-SB-GP03-0-0050 (00132054 Spk C, 4 ppb Spike)
DF05-SB-GP03-0-0050 (C0132934Spk D, 40ppb Spike)
4 40
NR NR
NR NR 5.43 9.92 112 51.6 74.6
*
NR NR 5.43 39.3 85 51.6 86.4 87
DF05-SB-GP04-0-0000
(C01529SS Spk E, 4 ppb Spike)
4
ND
2.75 69
ND
3.42 86 66.5 80.8
*
DF05-SB-GP04-0-0000
(00132953Spk F, 40 ppb Spike)
40
ND
32.1 80
ND
36.2 91 66.5
107 101
DF05-SB-GP04-0-0005 (C01S2936Spk 6 ,4 ppb Spike)
DF05-SB-GP04-0-0005 (0013295 Spk H, 40 ppb Spike)
4 40
ND ND
3.90 98 44.8 112
ND ND
3.74 94 38.9 97
NR NR
NR NR NR NR
DF05-SB-GP04-0-0010 (00152937 Spk 1,4 ppb Spike)
DF05-SB-GP04-0-0010 (00152937Spk J, 40ppb Spike)
4 40
ND ND
4.87 122 0.312 4.00 92 54.3 60.2
.
54.1
135 0.312
37.3
92
54.3
100 114
DF05-SB-GP04-0-0020 (0015295 Spk K, 4 ppb Spike)
DF05-SB-GP04-0-0020 (00132958 Spk L, 40 ppb Spike)
4 40
0.218 0.218
5.15 52.0
123 0.745 129 0.745
4.30 37.7
89 92
NR NR
NR NR NR NR
DF05-SB-GP04-0-0050
(C01S2959 Spk 0 ,4 ppb Spike)
4
0.238
5.72
137 0.314
3.90
90 0.264
3.59
83
DF05-SB-GP04-0-0050
(COIS29S9 Spk D, 40 ppb Spike)
40
0.238
52.0
129 0.314
33.7
83 0.264
30.4
75
DF10-SB-GP03-0-0000 (C0153316Spk E, 4 ppb Spike)
D F 10-SB-GP03-0-0000 (COI33313 Spk F, 40 ppb Spike)
4 40
NR NR
NR NR 93.6 NR NR 93.6
154 . 123 74
NR NR
NR NR NR NR
DF10-SB-GP03-0-0005 (00133317 Spk G, 4 ppb Spike)
DF 10-SB-GP03-0-0005 (00163317SpkM, 40ppb Spike)
4 40
NR NR
NR NR 57.2 50.4
.
NR
NR NR 67.2 88.0 77 NR
NR NR NR NR
DF10-SB-GP03-0-0010 (C0153318 Spk 1,4 ppb Spike)
DF10-SB-GP03-0-0010 (00133318 Spk J, 40 ppb Spike)
4 40
12.6 12.6
21.3 62.5 125
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 10-SB-GP03-0-0020 (00133319 Spk K, 4 ppb Spike)
DF10-SB-GP03-0-0020 (00153319Spk L. 40ppb Spike)
4 40
NR NR
NR NR 17.4 24.4
NR NR 17.4 52.8 89
NR NR
NR NR NR NR
Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated NO * Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ngfg (wet weight).
NR = Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the valuee In the raw data.
Exygen Research
Page 40 o f 136
Interim Report #21 - Analysis of Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked In Sample Recovered Recovery In Sample Recovered Recovery In Sample Recovered Recovery
(nfl/g) (ng/g) wet wt;ng/g) wet wt. (%) (ng/g) wet wt.'ng/g) wet wt. (%) (ng/g) wet wt/no/g) wet wt. (%)
DF 10-SB-GP03-0-0050 (C0153320 Spk C, 4 ppb Spike)
DF10-SB-GP03-0-0050 (C01S3320 Spk D, 40 ppb Spike)
4 40
15.1 15.1
17.6 60.6 114
109 109
100 NR 150 103 NR
NR NR NR NR
DF10-SB-GP04-0-0000 (C0153321 Spk E, 4 ppb Spike)
DF10-SB-GP04-0-0000 (C0153321 Spk F, 40 ppb Spike)
4 40
1.97 1.97
7.36 135 34.0 51.4 124 34.0
27.0 54.2
. 51
NR NR
NR NR NR NR
DF10-SB-GP04-0-0005
(00153322 Spk 0 ,4 ppb Spike)
4
1.49
6.96 137 19.1
21.5
.
D F10-SB-GP04-0-0005
(00153322 Spk H, 40ppb Spike)
40
1.49
54.2 132 19.1
39.4
51
NR NR
NR NR NR NR
DF10-SB-GP04-0-0010 (C0153323 Spk 1,4 ppb Spike)
DF10-SB-GP04-0-0010 (C0153323Spk J, 40 ppb Spike)
4 40
NR NR
NR NR NR NR NR NR
NR NR NR NR NR NR
NR NR NR NR
DF10*SB-GP04-0-0020 (00153324Spk K, 4 ppb Spike)
DF10-SB-GP04-0-0020 (00153324 Spk L, 40 ppb Spike)
4 40
NR NR
NR NR 27.8 30.9 NR
NR NR 27.8 54.4 67
NR
NR NR NR NR
DF10-SB-GP04-0-0050 (00153325Spk C, 4 ppb Spike)
DF 10-SB-GP04-0-0050 (00153325Spk D, 40 ppb Spike)
4 40
32.9 32.9
40.2 66.1
83
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 10-SB-GP04-DB-0050 (00153326Spk E, 4 ppb Spike)
4
35.0
37.7
*
DF10-SB-GP04-DB-0050
(00153326Spk F, 40ppb Spike)
40
35.0
67.8
82
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP01-0-0000 (C0153327 Spk 6 ,4 ppb Spike)
DF11-SB-GP01-0-0000
(C01S3327 Spk H, 40 p p b Splka)
4 40
30.4 30.4
39.0 71.9
104
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 11-SB-GP01-0-0005 (C0153320 Spk 1,4 ppb Spike)
DF11-SB-GP01-0-0005 (00153328 Spk J, 40 ppb Spike)
4 40
25.8 25.8
36.7 * 67.7 105
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP01 -0-0010 <00153320Spk K. 4 ppb Spike)
DF11-SB-GP01-0-0010 (00153328 Spk L, 40 ppb Spike)
4 40
32.6 32.6
33.4 . 75.0 106
NR NR
NR NR NR NR NR NR
NR NR NR NR
'Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated
ND * Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recoveryvalues mayvary slightly from the values in the raw data.
Exygen Research
Page 41 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C 4 S u lfo n a t PFBS Wet Weight C6 Sulfonate PFHS W et W eight C8 Sulfonate P FO S Wet Weight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(nfl/g) (ng/g) wet wt'no/a) wet wt (%) (ng/g) wet wt.'ng/g) wet wL (%) (ng/g) wet wt.'ng/g) wet wt <%)
DF11-SB-GP01-0-0020 (C0153330Spk C, 4 ppb Spike)
D F 11-SB-GP01-0-0020 (C0153330 Spk D, 40 ppb Spike)
4 40
22.6 22.6
33.1 79.8 143
101 101
106 138 93
NR NR
NR NR NR NR
DF 11-SB-GP01-0-0050 (C0153332 Spk E, 4 ppb Spike)
D F11-SB-GP01-0-0050 (C0153332Spk F, 40ppb Spike)
4 40
NR NR
NR NR 67.8 74.5
NR NR 67.8
104 91
NR NR
NR NR NR NR
DF11-SB-GP02-0-0000
(C0153333Spk G, 4 ppb Spike)
4
0.401
5.98
139
2.04
5.52
87
DF11-SB-GP02-0-0000
(C0153333 Spk H, 40 ppb Spike)
40
0.401
56.6
140
2.04
35.2
83
NR NR
NR NR NR NR
DF11-SB-GP02-0-0005 (C0153334Spk 1, 4ppb Spike)
DF 11-SB-GP02-0-0005 (C01S3334Spk J, 40ppb Spike)
4 40
2.23 2.23
7.62 135 59.8 144
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP02-0-0010 (C0153335 Spk K, 4 ppb Spike)
DF 11-SB-GP02-0-0010 (C0153335 Spk L, 40 ppb Spike)
4 40
4.01 4.01
8.08 102 56.6 131
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF 11-SB-GP02-0-0020 (C0153336 Spk C. 4 ppb Spike)
DF 11-SB-GP02-0-0020 (C015333 Spk 0,40 ppb Spike)
4 40
44.3 44.3
46.8 73.1
. 72
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP02-0-0050 (C01S3337Spk E, 4 ppb Spike)
DF11-SB-GP02-0-0050 (C0153337 Spk F, 40 ppb Spike)
4 40
NR NR
NR NR NR NR NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP03-0-0000 (C01S3330Spk G, 4 ppb Spike)
DF 11-SB-GP03-0-0000 (COI5333 Spk H, 40ppb Spike)
4 40
NR NR
NR NR 2.42 6.69 107 NR
NR NR 2.42 42.2 99
NR
NR NR NR NR
DF11-SB-GP03-0-0005 (C01S3330 Spk 1,4 ppb Spike)
DF 11-SB-GP03-0-0005 (COI53339 Spk J, 40 ppb Spike)
4 40
NR NR
NR NR 7.36 11.8 111 NR
NR NR 7.36 44.0 92
NR
NR NR NR NR
DF11-SB-GP03-0-Q010 (COi53340Spk K. 4 ppb Spike)
D F 11-SB-GP03-0-0010 (COI53340Spk L. 40ppb Spike)
4 40
NR NR
NR NR 17.6 17.1
.
NR NR 17.6 54.6 93
NR NR
NR NR NR NR
`Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND = Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to quality control failures, see Table V for re-extract data. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
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Page 42 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
C4 Sulfonate PFBS Wet W eight C6 Sulfonate PFHS Wet W eight C8 Sulfonate PFO S Wet W eight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(n0/9) ino/a) wet wt.'ng/a) wet wt. (%) (ng/g) wet wt.'ng/g) wet wt. <%) (ng/g) wet wt. na/o) wet w t (%)
DF11-SB-GP03-DB-0020 (C0193341 Spk C, 4 ppb Spike)
DF11-SB-GP03-DB-0020 (00155341 Spk D, 40 ppb Spike)
4 40
100 100
95.2 155 138
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF11-SB-GP03-0-0020 <00153342 Spk E, 4 ppb Spike)
DF11-SB-GP03-0-0020 (C0153342 Spk F, 40ppb Spike)
4 40
51.9 51.9
47.6 * 108 140
NR NR
NR NR NR NR NR NR
NR NR NR NR
D F11-SB-GP03-0-0050
(C0153344 Spk G, 4 ppb Spike)
4
27.6
33.4
67.3 65.6
.
DF 11-SB-GP03-0-0050
(C0153344Spk H, 40 ppb Spike)
40
27.6
66.9
98
67.3
105
94
NR NR
NR NR NR NR
DF11-SB-GP04-0-0000 (C01S3345 Spk 1,4 ppb Spike)
DF11-SB-GP04-0-Q000 <00153345Spk J, 40 ppb Spike)
4 40
ND ND
3.46 87 1.05 3.95 73 37.8 95 1.05 34.9 85
NR NR
NR NR NR NR
DF 11-SB-GP04-0-0005 (00153344 Spk K. 4 ppb Spike)
DF11-SB-GP04-0-0005 (C0153346 Spk L, 40 ppb Spike)
4 40
ND ND
5.09 127 0.357 3.80 86 63.2 70.6
*
45.8 115 0.357 33.8 84 63.2
114 127
DF 11-SB-GP04-0-0010 (C0153347 Spk C, 4 ppb Spike)
DF11-SB-GP04-0-Q010 (C0153347 Spk D, 40 ppb Spike)
4 40
ND ND
3.24 81 0.816 5.03 105 NR
22.6 57 0.816 29.5 72
NR
NR NR NR NR
DF 11-SB-GP04-0-0020
(00153340Spk E, 4 ppb Spike)
4
0.880
5.16
107
3.50
DF 11-SB-GP04-0-0020
(C01S3346Spk F, 40ppb Spike)
40
0.880
35.9
88
3.50
7.25 94 32.7 73
NR NR
NR NR NR NR
DF11-SB-GP04-0-0050 (00153340Spk 6 ,4 ppb Spike)
DF11-SB-GP04-0-0050 <C01S334eSpk H, 40 ppb Split*)
4 40
1.41 1.41
6.23 121 3.85 48.3 117 3.85
7.06 33.4
80 74
NR NR
NR NR NR NR
DF05-SB-GP01-0-0000 <00153350Spk 1,4 ppb Spike)
DF05-SB-GP01-0-0000 (C0153350 Spk J, 40 ppb Spike)
4 40
3.80 3.80
7.12 83 68.1 161
NR NR
NR NR NR NR NR NR
NR NR NR NR
DF05-SB-GP01-0-0005 (C0153351 Spk K, 4 ppb Spike)
DF05-SB-GP01-0-0005 (C0153351 Spk L, 40ppb Spike)
4 40
ND ND
5.66 142 0.337 3.92 53.0 133 0.337 32.3
90 80
NR NR
NR NR NR NR
Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated NO * Not detected at or atx)ve the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR = Not reported due to quality control failures, see Table V for re-extract data. Note: S ince th is sum m ary ta b le show s rounded re su lts, recovery values m ay vary s lig h tly fro m the values In the raw data.
Exygen Research
Page 43 of 136
Interim Report #21 - Analysis of Soil Samples
Exygen Study No.: P0001131
Table IV. Matrix Spike Recovery of PFBS, PFHS and PFOS in Soil Samples Continued
Sample Description
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked In Sample Recovered Recovery In Sample Recovered Recovery in Sample Recovered Recovery
(ng/fl) fno/a) wet wL'ng/g) wet wt. (%) Inala) wet wt.'no/o) wet w t <%) inala) wet wt. na/a) wet w t (%)
DF05-SB-GP01-0-0010 (C0153352 Spk C, 4 ppb Spiles)
DF05-SB-GP01 -0-0010 (C0153352 Spk D, 40 ppb Splks)
4 40
ND ND
5.01 125 0.546 4.06 51.8 130 0.546 35.0
88 86
NR NR
NR NR NR NR
DF05-SB-GP01-0-0020
(C01S3353 Spk E, 4 ppb Splks)
4
0.271
5.65 134 1.24
4.74
88
DF05-SB-GP01-0-0020
(CO153353Spk F, 40 ppb Splks)
40
0.271
58.5
146
1.24
36.6
89
NR NR
NR NR NR NR
DF05-SB-GP01-0-0050 (C01S3394Spk G. 4 ppb Splks)
DF05-SB-GP01-0-0050 (C01S33S4 Spk H, 40 ppb Splks)
4 40
NR NR
NR NR 68.8 81.6 NR
NR NR 68.8
126 143 NR
NR NR NR NR
DF05-SB-GP02-0-0000
(C01533SSSpk 1.4 ppb Splks)
4
ND
3.80 95
ND
3.68 92
NR
DF05-SB-GP02-0-0000
(C015335SSpk J, 40ppb Splks)
40
ND
32.6 82
ND
33.5 84
NR
NR NR NR NR
DF05-SB-GP02-0-0005 (C0153357 Spk K, 4 ppb Splks)
DF05-SB-GP02-0-00Q5 (00133357Spk L, 40 ppb Splks)
4 40
ND ND
3.98 100 0.318 3.83 88 35.8 90 0.318 34.0 84
NR NR
NR NR NR NR
DF05-SB-GP02-0-0010 (00153359Spk C. 4 ppb Splks)
DF05-SB-GP02-0-0010 (00153354 Spk D, 40 ppb Splks)
4 40
NR NR
NR NR 0.880 4.48 90 NR NR 0.880 35.4 86
NR NR
NR NR NR NR
DF05-SB-GP02-0-0020 (C d 53359 Spk E, 4 ppb Splks)
DF05-SB-GP02-0-0020 (C0153359 Spk F. 40ppb Splks)
4 40
NR NR
NR NR 0.622 4.78 104 NR
NR NR 0.622 36.2 69
NR
NR NR NR NR
DF05-SB-GP02-0-0050 (00153300 spk G, 4 ppb Splks)
DF05-SB-GP02-0-0050
(C o is a s s o Spk H, 40 pp b Spika)
4 40
NR NR
NR NR 1.06 4.93 97
NR
NR NR 1.06 38.2 93 NR
NR NR NR NR
DF05-SB-GP02-DB-0050 (00153341 Spk 1,4 ppb Splks)
DF05-SB-GP02-DB-0050 (00153301 Spk J. 40 ppb Splks)
4 40
NR NR
NR NR 4.02 7.66 91 6.29 9.76 87 NR NR 4.02 38.5 86 6.29 41.8 89
Average: 113 Standard Deviation: 23
A v e ra g e : Standard Deviation:
85 13
`Sample residue exceeds the spiking level significantly (3x spiking level); therefore, an accurate recovery value cannot be calculated ND * Not detected at or above the Limit of Quantitation (LOQ) of 0.2 ng/g (wet weight). NR * Not reported due to qualitycontrol failures, see Table V for re-extract data. Note: Since this summary table show s rounded results, recovery values may vary slig h tly from die values In the raw data.
Average: Standard Deviation:
95 16
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Page 44 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples
Sample Description
C4 Sulfonate PFBSWst Weight
C6 Sulfonate PFHS Wot Weight
C8 Sulfonate PFQ3 Wet Weight
Amount Amt Found Amount
Amt Found Amount
Amt Found
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample
(ng/fl) (ng/g)wetwt. (na/g)wetwt. i% ) (Ha'll) wet wt. (ng/g)wetwt. 1% ) (no/a)wet wt.
Amount Recovered
Recovery
(% )
DF10-SB-GP01-0-0000 (00152600Spk C, 2000ppb Spike) 2000
..
.
-
1590
3490
95
D F10-S B -G P 01-0-0005 (COI52601Spk D, 2000ppb Spike)
2000
-
--
-
-
1210
2910
85
D F10-S B -G P 01-0-0010 (COI52602Spk E. 2000ppb Spike)
2000
--
-
816
2580
88
D F10-SB-GP01-DB-0020 (COI52603Spk F, 2000ppbSpike)
2000
--
-
-
872
2400
76
DF10-SB-GP01-0-0020 (COI52664Spk Q, 2000ppb Spike) 2000
-
--
-
-
816
2600
89
D F10-S B -G P 01-0-0050 (C0152895Spk H. 8000ppb Spike) 8000
--
-
-
5240
10400
65
DF 10-SB-GP02-0-0000 (COI52696Spk 1,600ppb Spike)
D F 10-SB-GP02-0-0005 (COI52997Spk J, 600ppb Spike)
DF10-SB-GP02-0-0010 (C0152896Spk K. 400ppb Spike)
DF10-SB-GP02-0-0020 (COI52996Spk L, 400ppbSpike)
D F 10-SB-GP02-0-0050 (COI52900Spk C. 400ppb Spike)
800 800 400 400 400
- -- -----
- - 192 960 96
-
416
1440
128
-
- 291
683 98
-
- 294
747 113
- 214
486 68
DF12-SB-GP01-0-0000 (COI52901Spk 0,4 ppb Spike)
D F12-S B -G P 01-0-0000 (COI52901Spk E, 200ppb Spike)
D F 12 -S B -G P 0 1-0-0000 (COI52901Spk F. 60000ppb8pike)
4 200 80000
11.9 -
15.8 98 - - 102 - --
- --
374 91
--
-
-
214000
272000
73
DF12-5B-GP01-0-0005 (COI52602Spk 0 , 60000ppb Spike)
D F12-S B -G P 01-0-0010 (COI52903Spk H. 8000ppbSpike)
80000 8000
DF12-SB-GP01-0-0020 (COI52904spk 1,20000ppbSpike) 20000
-
-- -- --
-
-
28500
92000
79
-
-
6700
13000
79
-
-
18300
37000
94
DF 12-SB-GP01-0-0050 (COI52905Spk J. 9000ppb Spike)
8000
--
-
-
10800
21800
138
DF 12-SB-GP01-DB-0050 (00152909Spk K. 20000ppb Spike) 20000
- --
-
-
9280
35800
133
DF 12-SB-GP02-0-0000 (COI52907Spk L, 4ppb Spike)
DF12-SB-GP02-0-0000 (COI52907spk M.20000ppb Spike)
4 20000
7 .6 6
10.0 81 -
-
- --
-
14400
31900
88
Note: Since th is summary table shows rounded results, recovery values may vary sliQ htly from the values In the raw data.
Exygen Research
Page 45 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No. : P0001131
Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Sample Descrtotion
C4 Sulfonate PFBS Wat W ight
C6 Sulfonate PFHS Wet Weight
CB Sulfonate PFOS Wat Weight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery (ng/g) <nnta>wet wt. ina/a) wet wt. (%) (nota) wet wt. fnofa) wet wt. (*>> (no/aiwet wt. (na/a)wet wt. (%>
DF12-SB-GP02-0-0005
(00192908 Spk C. 8000 ppb Spite)
8000
.
._
5790
14800
113
DF12-SB-GP02-0-0010 (C01S2S08 Spk D. 4 ppb Spite)
DF12-SB-GP02-0-0010
(00192900 Spk E, 40 ppb Spite)
DF12-SB-GP02-0-0010 (COI92909 Spk F. 8000 ppb Spite)
4 40 6000
NR NR -
NR NR NR NR --
.
. -
. ..
._
. --
--
-
-
5290
13700
105
DF12-SB-GP02-0-0020
(COI52910 spk 0,400 ppb Spite)
DF12-SB-GP02-0-0020
( C d 52910 spk H, 800 ppb Spite)
400 800
_
-
_ . 319 542 56
..
- --
- - 846 1590 93
DF12-SB-GP02-0-0050
(C01S2911 Spk 1.200 ppb Spite)
DF 12-SB-GP02-0-0050
(COI82911 Spk J , 600 ppb Spite)
200 800
212 -
492 140 - - 556
.. 1670 139 706
1370
. 83
DF12-SB-GPO3 -0 -0 0 0 0
(C0152912 Spk K, 800 ppb Spite)
800
-
--
-
-
347
1280
117
DF12-SB-GPO3-0-0005
(COI32913 Spk L. 800 ppb Spite)
800
--
-
-
369
1120
91
DF12-SB-GP03-0-0020
(COI92915 Spk M. 40 ppb Spite)
40
--
- - 75.5 99.2 59
DF12-SB-GPO4-0-0010 (COI92919 Spk F, 20000 ppb Spite) 20000
--
-
- NR
NR NR
DF12-SB-GP04-DB-0020
(C0192921 spk H, 8000 ppb Spite)
8000
--
-
-
10200
19400
115
DF12-SB-GP04-0-0050
(COI92922 Spk 1.20000 ppb Spite)
20000
--
-
-
21100
41900
104
DF13-SB-GP01-0-0000 (C0182023 Spk J, 200 ppb Spite)
DF13-SB-GP01-0-0000
(COI82923 S p k K. 90000 p p b S p ite)
200 80000
. 229 428 100
-
--
-
230000
290000
75
DF13-SB-GP01-0-0005
(C0192924 Spk L. 4 ppb Spite)
DF13-SB-GP01-0-0005
(COI92924 Spk M. 80000 ppb Spite)
4 80000
NR
NR NR --
-
..
-
-
-
65800
175000
137
DF13-SB-GP01-0-0010
(COI92929 Spk N, 4 ppb Spite)
DF13-SB-GP01-0-0010
(C0152929 Spk 0.20000 ppb Spite)
4 20000
2.30 -
7.89 135 --
-
..
-
-
15900
41300
127
NR Not reported due to qualitycontrol failures. Note: Since M s summarytable shows rounded results, recovery values may vary slightly from the values In the raw data.
Exygen Research
Page 46 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Sample Description
C4 Sulfonate PFBS Wet Weight
C6 Sulfonate PFH8 W t Witaht
Amount Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery
(ntfo) ino/a) wet wt. ina/a) wet wt. (% ) (nafa)wetwt. (ng/g)wetwt. (% )
C8 Sulfonate PFOS Wat Wataht
Amt Found Amount in Sample Recovered Recovery
(na/al wet wt. (% )
DF13-SB-GP01-0-0020 (C0192926Spk C. 4ppb Spike)
D F13-S B -G P 01-0-0020 (C01S292SSpk D,200ppb 8pike)
D F13 -S B -G P O 1-0-0020 (C0192S2SSpk E, 40000ppbSpike)
4 200 40000
4.85 . -
7.79 74 . . 238 - --
._
346 54
-
--
-
-
18300
54600
91
D F13-S B -G P 01-0-0050 (C01S2S27Spk F. 4ppb Spike)
DF13-SB-GP01-0-0050 (C01S2027Spk 0 .40ppbSpike)
DF13-SB-GP01-0-0050 (C0152927Spk H, 8000ppb 8pike)
4 40 8000
0.641 0.641
-
6.26 140 51.4 127
--
. . -
. ..
--
- --
--
-
-
6420
13800
62
D F 13-SB-GP02-0-0000 (C0152926Spk 1,80000ppb Spike) 80000
- --
-
-
77500
166000
111
D F 13-SB-GP02-0-0005 (C01S2928spk J, 4ppbSpike)
D F 13-SB-GP02-0-0005 (00192929Spk K. 40000ppbSpike)
4 40000
0.637
5.78 129 --
. -
_..
.-
-
-
30900
64800
135
D F 13-SB-GP02-0-0010 (C0192930Spk L, 8000ppbSpike)
8000
- --
-
-
7130
13900
85
DF13-SB-GP02-DB-0010 (C0152931Spk M, 8000ppb Spike) 8000
- --
-
-
5910
14600
109
DF 13-SB-GP02-0-0020 (C0192932Spk N. 8000ppb Spike)
8000
- --
-
-
7440
15400
100
DF13-SB-GP02-0-0050 (C0192933Spk 0 ,8000ppb Spike)
8000
- --
-
-
6830
13900
88
DF13-SB-GP03-0-0010 (00152938Spk E, 400ppb Spike)
400
- --
- - 182 654 118
DF 13-SB-GP03-0-0020 (C0192937Spk F, 800ppb Spike)
800
- --
-
-
548
1120
72
DF13-SB-GP03-0-0050 (CCHS29 Spk 0.400 ppb Spike)
D F 13-SB-GP04-0-0005 (COI92940Spk i, 20000ppb Spike)
400 20000
-- --
-
-
433
1030
149
-
-
4850
21600
84
DF 13-SB-GP04-0-0010 (00192941spk J, 4000ppb Spike) 4000
- --
-
-
2860
8800
149
DF 13-SB-GP04-0-0020 (COI52942Spk K, 4000ppb Spike) 4000
- --
-
-
1730
5420
92
DF 13-SB-GPO4-0-0050 (COI92943Spk L, 4000ppbSpike) 4000
- --
-
-
1530
5160
91
DF13-SB-GP04-DB-0050 (COI52944Spk C. 4000ppb Spike)
4000
- --
-
-
1500
4910
85
DF05-SB-GPO3-0-0000 (COI52949Spk D,40ppb Spike)
40
- --
-
- 69.8
121 128
Note: Sine* th is summary tabIs shows roundsd rssults, recovery values may vary slightly from the values in the raw data.
Exygen Research
Page 47 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Sample Description
C4 Sulfonate PFBS Wat Weight
C6 Sulfonate PFHS Wot Weight
C6 Sulfonate PFOS Wet Weight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(ng/a) ina/a)wetwt. (ng/g)wet wt. f%) fnatal wet wt. (natal wet wt. (%i (ng/g)wetwt. (na/fl)wet wt. (%)
DF13-SB-GP04-DB-0050
(C01S2S44 Spk C . 4000 ppb Splka)
4000
. ..
.
1500
4910
85
DF05-SB-GP03-0-0000
(C01S2948 Spk D. 40 ppb Splka)
40
- --
-
- 69.8
121 128
DF05-SB-GP03-0-0005 ( C d 52950 Spk E, 200 ppb Splka)
200
- --
- - 156 305 75
DF05-SB-GP03-0-0010
(00152991 Spk F. 200 ppb Splka)
200
-
- 118 298 90
DF05-SB-GP03-DB-0010
(C0152S52 Spk 0,200 ppb Splka)
200
-
- 92.8 290 99
DF05-SB-GP03-0-0050
(00152954 Spk H, 4 ppb Splka)
4
NR
NR NR
- --
DF05-SB-GP04-0-0005
(00152950 Spk 1.200 ppb Splka)
200
-
- 100 313 107
DF05-SB-GP04-0-0020
(COI52958 Spk J. 4 ppb Splka)
4
-
- 6.32 8.56 56
DF10-SB-GP03-0-0020
(C015S319 Spk 0 .4 ppb Splka)
DF10-SB-GPO3-0-0020
(COI53319 Spk H, 4000 ppb Splka)
4 4000
NR -
NR NR --
-
.-
-
-
3610
8160
114
DF10-SB-GP03-0-0050
(COI53320 Spk 1,8000 ppb Splka)
8000
-
-
4120
9600
69
DF10-SB-GP04-0-0000 (C0153321 Spk J , 80000 ppb Spike) 80000
-
-
-
67300
138000
68
DF10-SB-GP04-0-0005 (COI53322 Spk K, 20000 ppb Splka) 20000
-
-
11600
31600
100
DF10-SB-GP04-0-0050
(C0153325Spk E, 200 ppb Splka)
DF10-SB-GPO4-0-0050
(COI53325 Spk F. 2000 ppb Splka)
D F 1 0 -S B -G P 0 4 -D B -0 0 5 0
(COI53328 Spk0.200 ppb Spike) DF10-SB-GP04-DB-005Q
( C d 53329 Spk H. 2000 ppb Splka)
200 2000 200 2000
-
_ 230 -
330 50
.
-
4960
7170
111
. 228
455 114
-
- --
-
-
1370
2390
51
DF11-SB-GP01-0-0010
( C d 53329 Spk M. 200 ppb Splka)
DF11-SB-GP01-0-0010
( C d 53329 Spk N. 20000 ppb Splka)
200 20000
-
. 113 268 78
-
- --
-
-
8640
22500
69
DF11-SB-GP01-0-0020
( C d 53330 Spk 0 , 8000 ppb Splka)
8000
-
-
-
8860
13200
54
DF11-SB-GP01-0-0050
( C d 53332 Spk C . 40 ppb Spike)
DF11-SB-GP01-0-0050 (C0153332 Spk D. 4000 ppb Spike)
40 4000
NR -
NR NR --
-
--
-
- NR
NR NR
DF11-SB-GP02-0-0000 ( C d 53333 Spk E. 20000 ppb Splka) 20000
-
- --
-
-
8880
32200
117
NR = Not reported due to qualitycontrol failures. Not: Sine this summarytab! shows rounded results, recoveryvalues mayvaryslightlyfromthe values Inthe rawdata.
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Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Sample Description
DF11-S8-GP02-0-0005 (COI93334 Spk H, 300 ppb Spite)
(C4^ulfonat^FB3JW tW rtght_^___C6^ujfOM tiPnHSW2nWlht_B^ _ D C88u|fonat9PFOS>W tWjghti
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(nata) (noto) wet wt ino/a) wetwt. (%) (na/a) wet wt. (oq/q)wet wt. (%) Inala) wet wt. (na/a) wet wt. <*>
800 .
.
.
498
1570 134 1620
2500
110
DF11-SB-GP02-0-0010 (COI53335 Spk 1,400 ppb Spite)
DF11-SB-GP02-0-0010
(COI53335 Spk J , 20000 ppb Spite)
400 20000
_ -
. . 270 611 85 .
..
- -
-
-
11800
27700
80
DF11-SB-GP02-0-0020
(C0153333 Spk K, 400 ppb Spite)
DF11-SB-GF02-0-0020
(COI53333 Spk L, 4000 ppb Spite)
400 4000
. -
. 422 715 73
..
- -
-
-
2820
8160
134
DF11-SB-GP02-0-0050
(C0153337 Spk M, SO ppb Spite)
DF11-SB-GP02-0-0050 (COI53337 Spk N. 400 ppb Spite)
DF11-SB-GP02-0-0050
(COI93337 Spk 0 , 4000 ppb Spite)
80 400 4000
NR . -
NR NR . . 376 - --
..
-
617 60
--
-
-
1710
4070
59
DF11-SB-GP03-0-0000
(COI53336 Spk C , 4 ppb Spite)
DF11-SB-GPO3-0-0000 (COI93339 spk D. 40 ppb Spite)
DF11-SB-GP03-0-0000
(COI93339 Spk E. 2000 ppb Spite)
4 40 2000
NR NR -
NR NR NR NR --
-
..
..
--
-
-
-
690
2980
115
DF11-SB-GP03-0-0010
(COI53340 Spk 1,4 ppb Spite)
DF11-SB-GP03-0-0010
(C0153340 Spk J , 20000 ppb Spite)
4 20000
NR -
NR NR --
-
- -
.-
-
-
16600
35400
94
DF11-SB-GP03-DB-0020
(COf53341 Spk K, 400 ppb Spite)
DF11-SB-GP03-DB-0020
(COI93341 Spk L, 900 ppb Spite)
400 800
. -
. 322 620 75 -
--
- --
-
-
1150
1770
78
DF11-SB-GP03-0-0020
(C0153342 Spk M, 400 ppb Spite)
DF 11-S B -G P O 3-0-0020 (COI53342 Spk N. 4000 ppb Spite)
400
4000
_ -
. 338 658 80
--
- --
-
- NR
NR NR
DF11-SB-GP03-0-0050 (COI53344 Spk 0,400 ppb Spite)
400
-
- --
- - 337 615 70
DF11-SB-GP04-0-0000 (COI53345 Spk C, 200 ppb Spite)
200
-
- -
- - 157 329 66
DF11-SB-GP04-0-0010
(C0193347 Spk D, 200 ppb Spite)
200
-
- --
- - 158 323 83
DF11-SB-GP04-0-0020
( C d 93349 Spk E, 200 ppb Spite)
200
-
- --
-
- NR
NR NR
DF11-SB-GP04-0-0050
(C0193349 Spk F, 90 ppb Spite)
80
-
- --
-
- NR
NR NR
DF05-SB-GP01-0-0000
(COI93350 Spk 0,40 ppb Spite)
DF05-SB-GP01-0-0000 (COI93350Spk H, 20000ppb Spite)
40 20000
.
-
.
. NR
NR NR
-
- --
-
-
13800
40200
132
NR 3 Not reported due to qualitycontrol failures. Not*: Sine* M s summarytable shows rounded results, recovery values mayvary slightly from the values In the raw data.
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Table V. Matrix Spike Recovery of PFBS, PFHS and PFOS in Re-extracted Soil Samples Continued
Sample Description
C4 Sulfonate PFBSW1 Wight
C8 Sulfonate PFHS Wat Weight
C8 Sulfonate PFOS Wet Wight
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered Recovery in Sample Recovered Recovery in Sample Recovered Recovery
(natal (natal wet wt. (natal wet wt. (%l (natal wet wt. (natal wet wt. (%> (natal wet wt. (natal wet wt. (%)
DF05-SB-GP01-0-0005
(C0153391 Spk 1.400 ppb Spike)
400
.
. ..
.
530 1040 128
DF05-SB-GP01-0-0010
(C0153332 Spk J , S00 ppb Spike)
800
-
- --
- 840 1700 108
DF05-SB-GP01-0-0020
(C01S33S3 Spk K, 2000 ppb Spike)
2000
-
- --
-
-
1600
3310
86
DF05-SB-GP01-0-0005
(C01S3331 Spk 1,400 ppb Spike)
400
- --
-
-
530
1040
128
DF05-SB-GP01-0-0010
(C0153352 Spk J . 000 ppb Spike)
800
-
- --
- - 840 1700 108
DF05-SB-GP01-0-0020
(C01S33S3 Spk K, 2000 ppb Spike)
2000
-
---
-
-
1600
3310
86
DF05-SB-GP01-0-0050
(C01333S4 Spk L, 40 ppb Spiks)
DF05-SB-GP01-0-0050
(C0153394 Spk M, 2000 ppb Spike)
40 2000
NR -
NR NR --
.
-
..
--
-
-
1150
2740
60
DF05-SB-GP02-0-0000
(C01533SS Spk N, SO ppb Spike)
80
- --
- - 134 221 109
DF05-SB-GP02-0-0005
(C0153337 Spk C. 400 ppb Spike)
400
- --
- - 279 642 91
DF05-SB-GP02-0-0050
(00153340 Spk J , 4 ppb Spike)
DF05-SB-GP02-0-0050
(C0153300 Spk K, 40 ppb Spike)
DF05-SB-GP02-0-0050
(C01S3300 Spk L, SO ppb Spike)
DF05-SB-GP02-DB-0050
(C01533S1 Spk M, 4 ppb Spike)
DF05-SB-GP02-DB-0050
(C0153301 Spk N, 40 ppb Spike)
4 40 80
4 40
NR NR
-
NR NR
NR NR NR NR
--
NR NR NR NR
.
. -
-.
--
..
-
- - 106 172 83
..
--
- -
--
A vm g i: 115 Standard Deviation: 27
Avwaga: 85 Standard Deviation: 28
NR * Not reported due to quality control failures. Note: Since thle summarytable shows rounded results, recovery values mayvary slightly from the values in the raw data.
Average: 97 Standard Deviation: 23
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Table VI. Matrix Spike Recovery of PFBS, PFHS and PFOS in Trip Blank Samples
S am ple D e sc rip tio n
C4 Sulfonate P FB S _______________ C6 Sulfonate P FH S _______________ C8 S ulfonate P F O S
Amount Amt Found Amount
Amt Found Amount
Amt Found Amount
Spiked in Sample Recovered R ecovery in Sample Recovered Recovery in Sample Recovered Recovery
(no/U
(nn/L)
(ng/L)
(%)
(ng/L)
(ng/L)
<%)
(ng/L)
<ng/L>
D F 13-SB-TRIP1 -LS-060209 (00152806,1.0 ppb Field Spike)
1000
ND
1190
119
ND
957 96
ND
1150
115
DF13-SB-TRIP1-HS-060209 (00152887,10 ppb Field Spike)
10000
NO
9920
99
ND
10200
102
ND
12200
122
D F 0 5 -S B -T R IP 3 -L S -0 6 0 2 16 (C01S2M7,1.0 ppb Field Spik.)
1000
ND
1010
101
ND
961 96
DF05-SB-TRIPW1S^>60216
(C0152848,10ppb Field Spike) 10000
ND
11700
117
ND
8710
87
ND
1170
117
ND
9650
97
D F 1 < L S B -T R IP 2 -LS -0 6 0 2 1 6 <00153314,1.0 ppb Field Spik.)
D F 1CFSB-TR1P2-HS-060216 (C0153315,10ppb Field Spike)
1000 10000
ND ND
1060 10100
106 101
ND 974 97
ND
10300
103
ND
1010
101
ND
11800
118
Average: Standard Deviation:
107 9
Averaga: Standard Deviation:
97 6
NO * Not detectedat or above the Limit of Quantitation (LOQ) of25 ng/L. Note: Since this summary table shows rounded results, recovery values may vary slightly from the values in the raw data.
Averaga: Standard Deviation:
112 10
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Table VII. Total Percent Solids in Soil Samples
Exygen
C0152890 C0152891 C0152892 C0152893 C0152894 C0152895 C0152896 C0152897 C0152898 C0152899 C0152900 C0152901 C0152902 C0152903 C0152904 C0152905 C0152906 C0152907 C0152908 C0152909 C0152910 C0152911 C0152912 C0152913 C0152914 C0152915 C0152916 C0152917 C0152918
S a m p le _ _ =J J e s c rlg J k > n _ _ _ _
DF10-SB-GP01-0-0000 DF10-SB-GP01-0-0005 DF10-SB-GP01-0-0010 DF10-SB-GP01 -DB-0020 DF10-SB-GP01-0-0020 DF10-SB-GP01-0-0050 DF1 0-SB-GP02-0-0000 DF10-SB-GP02-0-0005 D F 10-S B-G P02-0-0010 DF10-SB-GP02-0-0020 DF10-SB-GP02-0-0050 DF12-SB-GP01-0-0000 DF 12-SB-GP01 -04)005 DF12-SB-GP01-0-0010 DF12-SB-GP01-04)020 DF12-SB-GP01-0-0050 DF12-SB-GP01 -DB-0050 DF 12-SB-G P02-0-0000 DF12-SB-GP02-0-0005 DF12-SB-GP02-0-0010 DF12-SB-GP02-0-0020 DF12-SB-GP02-0-0050 DF12-SB-GP03-0-0000 DF12-SB-GP03-0-0005 DF12-SB-GP03-0-0010 DF12-SB-GP03-Q-0020 DF12-SB-GP03-0-0050 DF12-SB-GP04-0-0000 DF12-SB-GP04-0-0005
Total_SoJW 8j%^ 85.25 83.75 83.04 83.40 82.61 82.69 83.23 84.05 81.66 80.70 78.71 84.44 86.29 83.62 83.30 83.44 86.37 85.85 83.83 83.95 77.10 81.01 81.46 87.31 83.48 85.00 82.94 81.74 82.92
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Table VII. Total Percent Solids in Soil Samples Continued
Exygen ID
C0152919 CO152920 C 0 152921 C 0 152922 C 0 152923 C0152924 C0152925 C0152926 CO152927 CO152928 C 0152929 C 0152930 C0152931 C0152932 C0152933 C0152934 CO152935 C0152936 C0152937 C0152938 C0152939 CO152940 C 0 1 52941 C0152942 CO152943 C 0152944 C0152949 C0152950 C0152951
S a m p le D e s c rip tio n DF12-SB-GP04-0-0010 DF12-SB-GP04-0-0020 DF12-SB-GP04-DB-0020 DF12-SB-GP04-0-0050 DF13-SB-GP01-0-0000 DF13-SB-GP01-0-0005 D F 13-S B -G P 0 1 -0 -00 1 0 D F 13-SB-GP01 -0-0020 D F 13-S B -G P 0 1 -0 -00 5 0 D F 13-S B -G P 0 2 -0 -00 0 0 DF13-SB-GP02-0-0005 DF13-SB-GP02-0-0010 DF13-SB-GP02-DB-0010 DF13-SB-GP02-0-0020 DF13-SB-GP02-0-0050 DF13-SB-GP03-0-0000 D F 1 3 -S B -G P 0 3 -0 -00 0 5 DF13-SB-GP03-0-0010 D F 1 3 -S B -G P 0 3 -0 -00 2 0 DF13-SB-GP03-0-0050 DF13-SB-GP04-0-0000 D F 13-S B -G P 0 4 -0 -00 0 5 D F 13-S B -G P 0 4 -0 -00 1 0 D F 13-S B -G P 0 4 -0 -00 2 0 D F 13-S B -G P 0 4 -0 -00 5 0 DF13-SB-GP04-DB-0050 D F 05-S B -G P 0 3 -0 -00 0 0 D F 05-S B -G P 0 3 -0 -00 0 5 DF05-SB-GP03-0-0010
Total Solids (%) 83.85 84.73 87.71 79.37 80.68 85.57 85.70 83.10 85.00 75.63 80.61 83.40 82.56 83.87 81.95 80.93 82.87 81.48 74.58 79.74 82.68 81.56 82.53 80.19 82.85 85.68 84.06 80.60 78.56
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Table VII. Total Percent Solids in Soil Samples Continued
Exygen ID
CO152952 CO152953 CO152954 C0152955 CO152956 CO152957 C0152958 CO152959 C0153316 C0153317 C0153318 C0153319 CO153320 C 01 53321 C0153322 CO153323 C0153324 CO153325 CO153326 C 0153327 CO153328 C0153329 C 0153330 C0153332 CO153333 C0153334 C0153335 CO153336 C0153337
S a m p le D es c rip tio n DF05-SB-GP03-DB-0010 DF05-SB-GP03-0-0020 D F 05-S B -G P 0 3 -0 -00 5 0 DF05-SB-GP04-0-0000 D F 05-S B -G P 0 4 -0 -00 0 5 DF05-SB-GP04-0-0010 DF05-SB-GP04-0-0020 DF05-SB-GP04-0-0050 DF10-SB-GP03-0-0000 DF10-SB-GP03-0-0005 DF10-SB-GP03-0-0010 DF10-SB-GP03-0-0020 DF10-SB-GP03-0-0050 DF10-SB-GP04-0-0000 DF10-SB-GP04-0-0005 DF10-SB-GP04-0-0010 DF10-SB-GP04-0-0020 D F 10-S B -G P 0 4 -0 -00 5 0 DF10-SB-GP04-DB-0050 DF11-SB-GP01-0-0000 D F11-SB-GP01 -043005 DF11-SB-GP01-0-0010 DF11-SB-GP01-0-0020 DF11-SB-GP01-0-0050 D F 1 1-S B -G P02-0-0000 D F 11-S B -G P 0 2 -0 -00 0 5 DF11-SB-GP02-0-0010 DF11-SB-GP02-0-0020 DF11-SB-GP02-0-0050
Total Solids (%) 76.69 76.24 77.53 85.09 88.25 85.07 85.91 85.18 77.08 76.78 81.84 80.75 78.37 79.35 83.76 80.67 84.01 84.17 84.93 79.27 82.39 80.57 83.52 79.21 82.01 82.72 81.23 80.48 81.25
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Table VII. Total Percent Solids in Soil Samples Continued
Exygen ID
C0153338 C0153339 C0153340 CO153341 C 0 153342 C0153344 C0153345 C0153346 CO153347 C0153348 CO153349 C0153350 C0153351 C0153352 C0153353 CO153354 C0153356 C0153357 C0153358 C0153359 C0153360 C0153361
S a m p le D es c rip tio n D F 1 1-SB -G P 03-0-0000 D F 1 1-SB -G P 03-0-0005 D F 1 1-S B -G P 03-0-0010 DF11-SB-GP03-DB-0020 D F 11-S B -G P 0 3 -0 -00 2 0 D F 1 1 -S B -G P 0 3 -0 -00 5 0 DF11-SB-GP04-0-0000 D F 1 1-S B -G P 0 4-0 -0 0 05 DF11-SB-GP04-0-0010 D F 1 1-SB -G P 04-0-0020 D F 11-S B -G P 0 4 -0 -00 5 0 DF05-SB-GP01-0-0000 DF05-SB-GP01-0-0005 D F 05-S B -G P 0 1 -0 -00 1 0 DF05-SB-GP01-0-0020 D F 05-S B -G P 0 1 -0 -00 5 0 DF05-SB-GP02-0-0000 D F 0 5 -S B -G P 0 2 -0 -00 0 5 DF05-SB-GP02-0-0010 D F 05-S B -G P 0 2 -0 -00 2 0 D F 05-S B -G P 0 2 -0 -00 5 0 DF05-SB-GP02-DB-0050
Total Solids (%) 84.51 84.04 83.56 81.84 82.75 80.26 85.76 83.58 82.09 80.19 82.04 85.92 85.62 83.15 80.85 80.79 85.84 83.89 83.92 89.00 84.00 83.25
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FIGURES
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Figure 1. Typical Calibration Curve for PFBS in Methanol
03170GE Soil.rdb (PFBS): "Lineai" Regression (M / x" weighting): y * 98.8 x + 0.00180 (r 0.9941)
Area, counts
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Figure 2. Extracted Standards of PFBS in Methanol, 25 ng/L and 50 ng/L, Respectively
X a a U O t-1 PFBS (Standard) 299.0/93.9amu -tam pla 2o f 39 from 031306C.wfff A n a : 3692 counts H eigkt: 2.43a+002cp* R T: 0.545 min 0.54
Time, min
I XC031400-2 - PFBS (Standard) 298.0/88.0 amu - sample 3 of 38 from 031308C.wiff Area: 5004 counts Height: 3.25e+002 ops RT: 0.543 min
0.54
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Figure 3. PFBS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively
I Reagent Control PFBS (Unknown) 299. 0/99.0 amu - tam ple 9o f 38 from 031306C.wiff (peak uot found)
3.55
Oao). 00-
ft
2cS5? c
40
20-10. 0<
^30 2.40 J -,/A ry/vA.//i .
u.07 ^
5 14 6.08
7.25 7.05 8.66 0.72 11 27 11.70
11 12
Tim e, min
R eagent Spk A - P FB S (Q C ) 200.0/00.0 amu - sample 10 of 38 from 031300C.wiff Area: 5729 counts Height: 2.99e+002 ops R T : 0.544 min
0.54
13.56 14.07
17.81^
15.08
A
13 14 15 10 17
Tim e, min
Reagent Spk B - P FBS (Q C ) 209.0/99.0 amu - sample 11 of 38 from Q31300C.wiff Area: 52530 counts Height: 2.77e+003 ops R T : 0.558 min
0.50
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Exygen Study No.: P0001131
Figure 4. Chromatogram Representing a Soil Sample Analyzed for PFBS (Exygen ID: C0152904, Data Set: 031306C)
I C01529Q4 - PFBS (UtltHOWK) 299. M 9.tam u -sample 15o f 3* from 031306C.wiff A n a : 17294eoaats Haight: l i f e +003cp* RT: 0. 535mia
Intensity, cps
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Exygen Study No.: P0001131
Figure 5. Typical Calibration Curve for PFHS in Methanol
031300C P700-1131 Soil.rav2.rdb (PFHS): "U n ta i" Regression f1 />f weighting): y * 465 x + 0.00131 (r= 0.0004)
Area, counts
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Figure 6. Extracted Standards of PFHS in Methanol, 25 ng/L and 50 ng/L, Respectively
I XC03140S-1 - PFHS (Standard) 39S.m O.tam u -sam ple 2o f 3 from 031306C-wiff A n a : 12620counts H eight: 9S2a*0ncps itT : 9.40min
Area: 24021 counis Height: 2.03 +003 cps RT:0.41 min
S.41
Intensity, cps
Intensity, cps
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Figure 7.
PFHS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively
R eagiii t C o n tro l - PFHS (Unknown) 39910* 0.0 j m u -sam ple 9 o f 39 from 03130C.wlff fre a k n o t found) 8.22
Intensity, cps
Intensity, cps
I R*agnt Spk A - PFHS (Q C) 308.0/80.0 im u - sampl* 10 of 38 from 031306C.wiff A fta : 25403 counts Height: 2.15*+003 cps RT: 0.42 min 0.42
Tim*, min R*ag*nt Spk 8 - PFHS (Q C) 300.0/80.0 amu - sampl* 11 of 38 from 031306C.wiff A rta : 228685 counts Haight: 1.80+004 cps RT:0.41 min
0.41
Intensity, cps
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Figure 8. Chromatogram Representing a Soil Sample Analyzed for PFHS (Exygen ID: C0152904, Data Set: 031306CR)
C01S2904 - PFHS (Uakaow a) 399.0/00.0 amu -tam p / 17o f 69 from 031306CR.wiff A n a : 107331 counts H aight: #.8fe+003cp# RT: 9.26 atin 0.20
Intensity, cps
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Figure 9. Typical Calibration Curve for PFOS in Methanol
031406AR P780-1131 S oil REV1 .rdb (P F O S ): "L in o ji" R o g ro sio n f l ) X" wtighting): y 90.9 x + 0.000734 (r 0.9061)
Area, counts
C onci ntrotion. ng/L
1000
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Figure 10. Extracted Standards of PFOS in Methanol, 25 ng/L and 50 ng/L, Respectively
I XC0314M-1 - P FO S (Staadard) 419.0/8.$ ama -tam ple 2o f 39 from 031306C.wiff A rts : 10962coaatt H a ig tt: & 7S e+ 0 0 2 cpa R T: 11.8mia 11.84
Intensity, cps
A rts : 15182 counts Height: 1.12.+003 cps RT: 11.8 min
11.84
Intensity, ops
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Figure 11. PFOS in Methanol Reagent Blank, 50 ng/L Fortified Reagent Spk A, and 500 ng/L Fortified Reagent Spk B, Respectively
R tagant C ontrol P FO S (Unknown) -910/801fl ama -ta rn p it 9 o f 39from 031506C.wlff A n a : 231count* H eight: f.?6e+0O 7c/>r R T: 11.5min 11.62
Intensity, cps
Tima, min R aagantSpkA - P F O S (Q 0)400.0/80.0 amu-sampla 10 o f3 8 fromG31506C.wiff
Araa:4501 counts Haight: 3.07a+002 ops RT: 11.5 min
Intensity, cps
Raagant Spk B - PFOS (00)400.0/80.0 amu-sampla 11 of 38 from 031506C.wff A r j: 44637 counts Haight: 2.87a+003 ops RT: 11.5 min
11.52
Intensity, cps
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Exygen Study No.: P0001131
Figure 12. Chromatogram Representing a Soil Sample Analyzed for PFOS (Exygen ID: C0152909, Data Set: 031406AR)
Intensity, cps
I C0152909 - PFOS (Unknown) 499.M 0.0 amu -ta m p la 20 o f 51 from 03140tAR.wiff A rea: 59900 coaats H aight: 453e+003 cp* RT: 11.4m ia 11.44
4000 3500 3000 2500
2000 1500
1000
500-
O-l-------------1------------- 1 ' I I............. ............................ ....... 12 3 4 5 5 7 8
i. min
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APPENDIX A
Study Protocol P0001131
(Exygen Study No. P0001131) with Analytical Methods and
Protocol Amendments
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Exygen Study No.: P0001131
Exygen Protocol Number P0001131
STUDY PROTOCOL
Study Title: Analysis of Perfluorobutanesulfonate (PFBS),
Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Liver and Small
Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program
Exygen Protocol Number: P0001131
Performing Laboratory: Exygen Research 3058 Research Drive State College, PA 16801
Phone: (814) 272-1039
Sponsor Representative: M ichael A. Santoro Director o f Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374
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DISTRIBUTION:
1) Jaisimha Kesari, Study Director, Weston Solutions 2) John M. Flaherty, Principal Investigator, Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research Quality Assurance Unit
Exygen Research
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PROTOCOL APPROVAL
Study Title: Analysis o f Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate (PFHS), and Perfluorooctanesulfonate (PFOS) in Water, Soil, Sediment, Fish, Clams, Vegetation, Small Mammal Livers and Small Mammal Serum Using LC/MS/MS for the 3M Decatur Monitoring Program
Exygen Protocol Number: P0001131
APPROVALS
JaisimhalKsan, S Weston Solutions
Michael A. Santoro, Sponsor Representative 3M Comparfy
Date
President, Facility Management :ad, Quality Assurance Unit
Date
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TABLE OF CONTENTS
TITLE PAGE.............................................................................................................................................. 1 DISTRIBUTION......................................................................................................................................... 2 PROTOCOL APPROVAL...........................................................................................................................3 TABLE OF CONTENTS............................................................................................................................ 4 INTRODUCTION....................................................................................................................................... 5 TEST MATERIALS................................................................................................................................... 5 OBJECTIVE...............................................................................................................................................6 TESTING FACILITY................................................................................................................................. 6 STUDY DIRECTOR................................................................................................................................... 7 SPONSOR REPRESENTATIVE................................................................................................................. 7 PRINCIPAL INVESTIGATOR................................................................................................................... 7 PROPOSED EXPERIMENTAL START AND TERMINATION DATES................................................... 7 IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM.......................................................8 SAMPLE PROCUREMENT, RECEIPT AND RETENTION...................................................................... 8 SAMPLE IDENTIFICATION.....................................................................................................................9 ANALYTICAL PROCEDURE SUMMARY............................................................................................... 9 VERIFICATION OF ANALYTICAL PROCEDURE...................................................................................9 METHOD FOR CONTROL OF B U S .........................................................................................................11 STATISTICAL METHODS........................................................................................................................ 11 GLP STATEMENT.....................................................................................................................................11 REPORT.....................................................................................................................................................11 SAFETY AND HEALTH............................................................................................................................ 12 AMENDMENTS TO PROTOCOL..............................................................................................................13 DATA RECORD KEEPING....................................................................................................................... 13 QUALITY ASSURANCE........................................................................................................................... 14 RETENTION OF DATA AND ARCHIVING..............................................................................................14 APPENDIX I, ANALYTICAL METHODS................................................................................................. 15
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INTRODUCTION
The purpose o f this study is to perform analysis for perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum using LC/MS/MS for the 3M Decatur Monitoring Program.
The study will be audited for compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by the Quality Assurance Unit o f Exygen Research.
TEST MATERIALS
The test materials are perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) and are all supplied by 3M.
PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (CiF^SOj'K*) Lot Number: 101 Purity: 96.7% Transitions Monitored: 299 -> 99 Structure:
PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (C F u S O s^ ) Lot Number: SE036 Purity: 98.6%
Transitions Monitored: 3 9 9 -+ 8 0 Structure:
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PFOS Chemical Name: Periluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (C |Fi7S0 3 ~K'f) Lot Number: 217 Purity: 86.9% Transitions Monitored: 4 9 9 - 9 9 Structure:
OBJECTIVE
The purpose o f this study is to perform analysis for perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS) and perfluorooctanesulfonate (PFOS) in water, soil, sediment, fish, clams, vegetation, small mammal livers and small mammal serum for the 3M Decatur Monitoring Program using the current versions o f the following Exygen analytical methods:
V0001780: "Method o f Analysis for the Deteimination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS"
V0001781: "Method o f Analysis for the Deteimination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS"
V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS"
V0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS"
V0001784: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS"
V0001785: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS"
V0001786: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS"
TESTING FACILITY
Exygen Research 3058 Research Drive State College, PA 16801 Phone:(814)272-1039
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STUDY DIRECTOR
Jaisimha Kesari P.E., DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: (610) 701-3761 Fax:(610)701-7401 j .kesari@westonsolutions.com
SPONSOR REPRESENTATIVE
Michael A. Santoro 3M Company Director o f Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374
PRINCIPAL INVESTIGATOR
John M. Flaherty Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039 john.flaherty@exygen.com
PROPOSED EXPERIMENTAL START AND TERMINATION DATES
It is proposed that the analytical portion o f this study be conducted from October 01, 2004 to December 31, 2005. The actual experimental start and termination dates w ill be included in the final report.
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IDENTIFICATION AP JUSTIFICATION OF THE TEST SYSTEM
The following are the test systems for this study: Water (groundwater and surface water) Soil Sediment Fish Clams Vegetation Small Mammal Liver Small Mammal Serum
The samples will be collected by Weston Solutions. The control samples will be purchased and prepared by the testing facility. Purchase and processing details for the control samples will be included in the final report associated with this study.
The test systems were chosen to access the environmental impact o f PFBS, PFHS and PFOS in the Decatur, Alabama area.
SAMPLE PROCUREMENT, RECEIPT AND RETENTION
Water, soil, sediment, fish, clam, vegetation, small mammal liver and small mammal serum samples will be received at Exygen directly from Weston Solutions. The details o f sample procurement for this study are outlined in the 3M work plan entitled "Phase 2 Work Plan for Sampling Environmental Media." The number and types o f samples collected will vary depending availability in the field. The total number o f samples received and analyzed for each matrix will be documented in the final report associated with this study.
Water, soil, and sediment samples will be used as received without further processing at Exygen. These samples will be stored refrigerated at 2C-8C. Fish, clam, vegetation and small mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These samples will be stored frozen at -10C. Small mammal whole blood samples will be centrifuged in the field at the time o f collection and the serum fraction will be used for the study. Small mammal serum will be stored frozen at 5 -10C.
The receipt and processing o f the samples will be documented in the final report and raw data associated with the study.
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^ SAMPLE IDENTIFICATION
Prior to analysis, each sample will be assigned a laboratory sample reference number. The reference number will be unique and will distinguish each laboratory sample that is processed throughout the analytical procedure. Chromatographic data will be identified by the laboratory sample reference number.
Sample storage conditions and locations will be documented throughout the study.
ANALYTICAL PROCEDURE SUMMARY
References: V0001780: "Method o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Water by LC/MS/MS" V0001781: `M ethod o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Soil by LC/MS/MS" V0001782: "Method o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Sediment by LC/MS/MS" V0001783: "Method o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Fish and Clams by LC/MS/MS" V0001784: "Method o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Vegetation by LC/MS/MS" V0001785: `M ethod o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Small Mammal Liver by LC/MS/MS" V0001786: `M ethod o f Analysis for the Determination o f Perfluorooctanoic
Acid (PFOA) in Small Mammal Serum by LC/MS/MS"
The above methods use analytical conditions capable o f separating the isomers o f PFBS, PFHS and PFOS. The final report will include the isomers summed into total PFBS, total PFHS, and total PFOS found.
VERIFICATION OF ANALYTICAL PROCEDURE
A laboratory control sample will be used for the preparation o f fortified control samples. The test substance will be made into solutions as per the method, and added to the matrices via a micropipette.
For water sampling, Exygen will supply one bottle per sample collected. The (**. bottles will be 500 mL precleaned Sci/Spec Premier wide mouth HDPE
bottles. These bottles have been routinely used for fluorochemical sample
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/-- collection at the testing facility and have been shown to be free o f PFBS, PFHS and PFOS. Samples will be added to each container to a volumetric fill line at 200 mL. A field duplicate, a low field spike and a high field spike of each sample will be collected. The low and high field spike bottles will contain PFBS, PFHS and PFOS as well as perfluorooctanoic acid (PFOA) and 1.2-13C perfluorooctanoic acid (13C PFOA). PFOA and l3C PFOA are included in the solutions used to spike the samples. The results for PFOA and >3C PFOA will not be reported in this study. Exygen will supply one field blank (control water) and two field blank spikes (control water fortified with PFBS, PFHS and PFOS at a low and high level) for every twenty samples collected. At the testing facility, each water sample (excluding field duplicates and field spikes) will be extracted in duplicate and will also be fortified at a low and high concentration with PFBS, PFHS and PFOS and processed through the described procedure to determine method accuracy and to check for bias.
For soil, sediment, clams, and vegetation, Exygen will supply one 500 mL precleaned Sci/Spec Premier wide mouth HDPE bottle per sample collected or a zip-seal bag. All containers/bags used for sample collection will be shipped to the sample location. Samples will be added to each container or bag in the field. At the testing facility, each sample will be extracted in duplicate and will also be fortified at a known concentration with PFBS, PFHS and PFOS at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias.
For small mammal liver, Exygen will supply a 50 mL polypropylene centrifuge tube. For small mammal serum, Exygen will supply a collection kit for each sample containing serum separator tubes (red top), vacutainers, needle holders and needles, transfer pipettes, and polypropylene tubes. At the testing facility, each liver and serum sample will be extracted in duplicate and will also be fortified at a known concentration with PFBS, PFHS and PFOS at both a low and high level and processed through the described procedure to determine method accuracy and to check for bias.
Low and high spiking levels for each matrix are defined below:
M atrix
Low Spiking Level
High Spiking Level
Water
500ng/L
5000 ng/L
Soil
4ng/g
40 ng/g
Sediment
4ng/g
40 ng/g
Fish
lOng/g
100 ng/g
Clams
10 ng/g
100 ng/g
Vegetation
10ng/g
100 ng/g
Small Mammal Liver
10ng/g
100 ng/g
Small Mammal Serum
10 ng/mL
100 ng/mL
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Recoveries are anticipated to be between 70% and 130% o f the fortified levels; however, the exact precision and accuracy will be determined by the analysis o f the quality control samples described above. A statement o f accuracy will be included in the final report.
METHOD FOR CONTROL OF BIAS
Control o f bias will be addressed by taking representative sub-samples from a homogeneous mixture o f each matrix from untreated control samples, and by analyzing at least two levels o f fortifications.
STATISTICAL METHODS
Statistics will be limited to those specified in the subject methods and to the calculation o f average recoveries, as applicable.
GLP STATEMENT
All aspects o f this study shall be performed and reported in compliance with EPA TSCA Good Laboratory Practice Standards 40 CFR 792. The final report or data package (supplied to the Sponsor) shall contain a statement that the study was conducted in compliance with current and applicable GLP standards and will outline any deviations in the study from those standards. This statement will be signed by the Study Director and Sponsor Representative.
REPORT
A final report will be prepared by the principal investigator or their designee at the conclusion o f the study. The report will include, but will not be limited to, the following: The name and address o f the Study Director, Sponsor Representative, and
o f the testing facility.
A statement o f GLP compliance (any related documentation, such as chain-of-custody records, must be in the study records).
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The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates o f study inspections and dates findings were reported to the Study Director and Management.
A description o f the exact analytical conditions employed in the study. If the subject method was followed exactly, it is necessary to include only a copy o f the analytical method. Any modifications to this method will be incorporated into the report. I f the method is photo-reduced, the project number and page number must be included on each page.
Description o f the instrumentation used and operating conditions.
All results from all sets analyzed. Control and fortified samples will be identified and the data table will include sample number and fortification level.
Representative chromatograms for each analyte in each matrix, including chromatograms o f a standard and a control sample, and a chromatogram at a fortification level. The location o f the analyte peaks will be clearly identified in all chromatograms.
All circumstances that may have affected the quality or integrity o f the data will be documented in the report.
Locations where raw data and the final report are to be archived.
Additions or corrections to the final report shall be in the form o f an amendment signed by the Study Director. The amendment shall clearly identify that part o f the report that is being altered and the reasons for the alterations. The amendment will be signed and dated by the Study Director and the Sponsor Representative.
All applicable requirements for reporting o f study results as per 40 CFR 792.185.
SAFETY AND HEALTH
Laboratory personnel w ill practice good sanitation and health habits.
Every reasonable precaution shall be taken to prevent inadvertent exposure of personnel and the environment to the test or reference substancefs).
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AMENDMENTS TO PROTOCOL
All significant changes to the analytical protocol outlined here will be expressed in writing, signed and dated by the Study Director and Sponsor Representative. Amendments usually will be issued prior to initiation o f study plan change. However, when a change is required without sufficient time for the issue o f a written amendment, that change may be effected verbally with supporting documentation signed and dated by the Study Director and followed with a written amendment as soon as possible. In this case, the effective date o f the written amendment will be die date o f the documented change. Copies o f the signed amendments will be appended to all distributed study plan copies. The original amendment will be maintained with the original study plan. Any deviations from the study plan or from the analytical method as provided will be documented and reported promptly to the Sponsor Representative.
DATA RECORD KEEPING
Records to be maintained include the following (as appropriate):
Sample tracking sheet(s) Sample receipt records, storage history, and chains o f custody History and preparation o f standards (stock, fortification, calibration) Description o f any modifications to the method Instrument run sheets, bench-sheets or logs Analytical data tables All chromatographic and instrumental conditions Sample extraction and analysis dates A complete listing o f study personnel, signatures and initials Chronological presentation o f all study correspondence Any other documentation necessary for the reconstruction o f the study
Chromatograms- All chromatograms will contain the following:
Sample identification, injection date, arrow or other indication o f the area o f interest, and injection number corresponding to the run.
Additionally, fortifications will include the amount o f analyte added and the sample number o f the sample that was fortified.
Analytical standard chromatograms will additionally include the concentration (e.g., pg/mL).
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As part o f the documentation the following sheets will be included in each analytical set: a run sheet listing the samples to be run in the set, and an instrument conditions sheet describing the instrument type and operating conditions.
QUALITY ASSURANCE
The QA Unit o f Exygen Research will inspect the study at intervals adequate to assure compliance with GLP's, and will report the findings o f audits to the Study Director, Exygen Management, and the Sponsor Representative.
RETENTION OF DATA AND ARCHIVING
All hard copy raw data, including, but not limited to, the original chromatograms, worksheets, correspondence, and results shall be included with the data package submitted to the Study Director. These will be archived with the original study plan, amendments, final report, and all pertinent information from the Sponsor.
The testing facility shall keep all electronic raw data and any instrument, equipment, and storage logs for the period o f time specified in 40 CFR 792.195. An exact copy o f the materials submitted to the study director will also be kept at Exygen Research.
Exygen will obtain permission from the study director before discarding or returning samples.
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APPENDIX I
ANALYTICAL METHODS
V0001780: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS"
V0001781 "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS"
V0001782 "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS"
V0001783 `M ethod o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS"
_ V0001784 `M ethod o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS"
V0001785 "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS"
V0001786: "Method o f Analysis for the Determination o f Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS"
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ANALYTICAL METHOD MethodNumber: V0001780
Method ofAnalysis for the Determinati of Perflaorooctanole Acid (PFOA) In Water by LC/MS/MS
Analytical TestingFacility:
Exygen Research 3058Research Drive
State College, PA 16801
Approved By:
C~Y^-X. ..L=XLi_________
Paul Connolly
'
Technical Leader, LC-MS, ExygenResearch
Date
Exygen Research
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Enygta lUiMich
Method Number VOOOI7BO
ANALYTICAL m ethod
MethodofAnalysis for theDeterminationofPerfluorooctanoic Acid (PFOA) in Water by LC/MS/MS
1.0 Scope
TUa method ia tobe employed for foe eolationand quantitation of perfluorooctanoic add by High Performance Liquid Chromatography coupled to a tandem Mass SpectrometricDetector(LC/MS/MS) in water.
2.0 Safety
2.1 Always observe safe laboratorypractices. 2.2 Consult theappropriate MSDS before handling anychemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At least 40mL oftest sample for extraction. 3 2 No sampleprocessing is needed for watersamples. 3J Samples stored refrigerated should be allowed to equilibrate to room
temperature. 3.4 All samplesmust be thoroughlymixed before being sampled for extraction 3.3 Any samples containing particles should be centrifuged at -3000 rpm for -5
minutes andthe supernatant used for the extraction. 3.6 Sample collection procedures will be specified in the sampling plan for this
project.
4.0 Reagent*andStandards
4.1 Water- HPLC grade 4.2 Methanol - HPLC grade 4.3 AmmoniumAcetate-A.C.S. Reagent Grade 4.4 Perfluorooctanoic Acid- Sigma-Aldrich
3.0 Instrument andEquipment
3.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume iiyoctor capable of injecting 3-200 pL toa tandemMaes Spectrometer (LC/MS/MS).
5.2 Adevice tocollect rawdata forpeak integration andquantitation. 3.3 Analyticalbalancecapableofreadingto 0.00001 g. 5.4 50mLdisposablepolypropylenecentrifogetube. 5.5 13mLdisposablepolypropylenecentrifoge tube. 5.6 Disposable micropipets (50-lOOuL, 100-200uL). 5.7 125-mLLDPE narrow-mouthbottles. 5.8 2 mLclearHPLCvial kit. 5.9 Disposable pipettes. 5.10 Autopipettes (100-1000pLand 10-100pL), with disposable tips. 5.11 Waters Sep Pak Vac 6 cc (Ig) IC18 SPEcartridges.
Pise2of ?
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ExygnXmarch
Method Number VOOOI78C
| ANALYTICAL METHOD |
MethodofAnalysis forthe DeterminationofPerfluorooctenolc Acid (PFOA) in Waterby LC/MS/MS
5.12 SPEvacuummanifold. 5.13 Ceotrifiigecapableofapinning 50mL polypropylenetubes at 3000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: FluophaaeRP (Keystone Scientific), 2.1 mm x SOmm, 5m (P/N: 82505*052130)
6.2 Temperature: 30C 6.3 MobilePhase(A): 2 mMAnunottiumAcetate in Water 6.4 MobilePhase(B) : Methanol 6.5 Gradient Program:
Timefmin) 0.0 1.0 8.0 20.0 22.5
HA 65 65 25 25 65
Flow Rate fmL/minl
35 0.3 35 0.3
75 0.3 75 0.3
35 0.3
6.6 InjectionVolume: 15pL (canbe increasedto as much ai 50 pL). 6.7 Quantitation: PeakArea-external standardcalibration curve. 6.8 RunTime: -2 3 minutes.
The above conditions are intendedas a guide and maybe changed in order to optimize the HPLCsystem.
7.0 MS/MSSystem 7.1 Mode: ElectrosprayNegative MRMmode, monitoring 413 -* 369 nVz.
The above condition! are intended ai a guide and may be changed in order to optimizethe MSMS system.
8.0 Preparation ofSolutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water it prepared by adding 0.154 g of ammoniumacetateto 1000mLofwater.
Alternatevolumesmaybe prepared.
Ptye J wl
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ExygeaKasseidi
Method Number VOM1710
ANALYTICAL M ETHOD
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Acid (PFOA) in Water by LC/MS/MS
9.0 StandardPreparation
9.1 StandardStoek/FortificationSolution 9.1.1 Prepare a stock solution of-100 pg/mL of PFOA by weighing 10 mg ofanalytical standard (corrected for purity) and dilute to 100 mL with
methanol in a 125-mL LDPEbottle. 9.1.2 A 10 pg/raL fortification solutionof PFOA is prepared by bringing 10
mL ofthe 100 pgAnL solution to a final volume of 100 with methanol in a 125mL LDPEbottle. 9.1.3 A 1.0 pg/mLfortification solution ofPFOAi* prepared by bringing 10 mL offoe 10 pg/mLsolution to a final volume of 100 with methanol in a 125mLLDPEbottle. 9.1.4 A0.1 pg/mL fortification solution ofPFOAit prepared by bringing 10 mLoffoe 1.0pg/mLsolutionto a final volumeof 100with methanol in a 125mLLDPEbottle. 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 ina 125mLLDPEbottle. 9.1.6 The stodc andfortificationsolutions are to be stored in a refrigerator at approximately 4*C and are stable for a maximum period of 6 months from foe date of preparation.
9.2 StandardCalibrationSolutions
9.2.1 LC/MS/MS calibration standards are prepared in HPLC water. The calibration standards are processed through foe extraction procedure, identical to samples.
9 2 2 The following ii a typical example: additional concentrations may be preparedas needed.
Final
Concentration Fortification Volum s o f Concentration of Calibration
o f Fortification Votums Fortified Contini Calibration
Standard ID
Solution foob)
(pL>
Simule (mL) Standard (not)*
(example)
0 0 40
0 XCmrnddyy-0
10 100 40
25 XCmmddyy*l
to 200 40
50 XCmmddyy-2
10 400 40
100 XCmroddyy-3
too 100 40
250 XCmmddyy-4
too 200
40
500 XCmmddyy-5
100 400
40
1000
XCmmddVY-6
*The extractedconcentrationofthecalibration standard is equal to 8x its initial
concentration, duetothe concentrationofthe standard during the extraction (SPE).
XC* extractedcalibrationstandard.
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BxygtalUsaarcfa
Mbod Number VOOOI7SO
ANALYTICAL M ETHOD
MethodofAnalysis forthe DeterminationofPcrfluorooctiuoic Acid (PFOA) in Water by LC/MS/MS
9.2.3 A zoo itandarti aohition (reagent binde) mint be prepared with each
aet offtandartii extracted. 9.2.4 Store all extractedcalibration standards in 15-mL polypropylene tubes
at 2*C to 6*C, upto two weeks. 9.2.5 Alternate volumes andconcentrations ofstandards may be prepared as
needed.
10.0 B atch Set U p
10.1 Each batch of aamplea extracted (typically 20 or leu) must include at least one reagent control (method blank using HPLC water) and two reagent controls fortified at known concentrations (lab control spikel to venfv procedural recoveryforthebatch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified in diequality assurance plan for thisproject.
11.0 SampleExtraction
11.1 Measure 40 mLofsample or a portion ofsimple diluted to 40 mL with water into 50 mLpolypropylene centrifage tubes (fortify as needed replace lid and mix well).
11.2 Condition the Cis SPE cartridges (1 g. 6 mL) by pauing 10 mL methanol followedby5 mLofHPLCwater(~2 drop/sec). Do not let column run dry
11.3 Loadsampleon conditionedCis SPEcartridge. Discard eluate. 11.4 Elute with -5 mL 100S methanol. Collect 5 mL of eluate into graduated
15mLpolypropylenecentriftigetubes(final volume " SmL). 11.5 AnalyzesamplesusingelectrosprayLC/MS/MS.
12.0 Chromatography
12.1 Inject the same mount ofeach standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples.
12.2 StandardsofPFOAcorrespondingto at least five or more concentration levels must be Deluded 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. Aa an alternative, an entire set of 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 aet of extracted standards). In either esse, extracted
calibrationstandards mustbe the first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves are
generatedfordie analyte by linearregrauionusing l/x weighting ofpeak area
Pag S of 7
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exyysn Rm arch
Method Number V00017M
ANALYTICAL M ETHOD
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Acid (PFOA) in Waterby LC/MS/MS
venue calibrationstandard concentration using MaasLynx 3.3 (or equivalent)
software system. 13.5 Sample response should not exceed standard responses. Any samples that
exceedstandardresponses shouldbe Airther diluted and reanalyzed.
13.0 AcceptanceCriteria
13.1 Chromatogrammust 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 (he daughter ion(369amu) represents the lots ofcarbon 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 beobtainedand the entireset must be re-extracted.
I3J Recoveries of control spikes mid matrix apikes must be between 70-!30*/Gof their known values. If a control spike tails outside the acceptable limits, the entire set of sauries should be rt-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 Ore total number of extracted calibration standards that could be occluded must not exceed 20% of die total number of extracted standards injected.
13.5 The conviction coefficient (R) for calibration curves generated must be 20.992 (R2 20.985). 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 %withinan analytical nm. Ifretention time drift exceeds this limit within an analytical run thenthe 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 fee standard curve (linear regression parameters) generatedbythe MassLynx softwareprogram:
PFOAfound (ng/L) - (Peak area - intercept) x DF slope
DF- Actor bywhichthe final volume was diluted, ifnecessary.
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygaa RttMrcb
MMbodNumtwr VO00I780
1 AWALVTICALMETHOD
|
MethodofAulyda for theDeteranation ofPerfiuorooctanoic Acid (PFOA) in Water by LC/MS/MS
142 For samples fortified with known amounts of PFQA prior to extraction, use foefollowing equation to calculate foepercent recovery.
Reeovwy(H)-
[totalanalytefound(ng/L) - analytefoundincontrol(ngfc)l |QQ analyteadded(ng/L)
Exygen Research
Pag7 or? P age 22 o f 65
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Exygen Study No. : P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber V0001781
M ethod o f A nalyib for the Determ ination o f PerftooroocU noic Acid (PFOA) io Soil by LC /M S/M 5
Analytical TestingFacility:
Exygen Research
3058 ResearchDrive Stale College. PA 16801
Approved By:
T U . C-JlL
Paul Connunuolllvy
Technical Leader, LC-MS, ExygenResearch
Date
ohn Flaherty f Vice Prsidait, Operations, ExygenResearch
Dit
Exygen Research
Total Pifer 7
P a g e 23 o f 65
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
EHMin n k______________________________________MhodNnibrVOOOI7H
I ANALYTICAL METHOD
MethodofAnalysis fortheDeterminationofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
1.0 Scope This method is to be employed for the isolation and quantitation ofperfluorooctanoic acid by High Performance liquid Chromatography coupled to a tandem Mass Spectrometric Detector(LC/MS/MS) in soiL
2.0 Safety
2.1 Alwaysobserve safe laboratorypractices. 2.2 Consult tbs appropriate MSDS before handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At lean 15g oftestsample forextraction. 3.2 Nosampleprocessingis neededfor soil samples. 3.3 Samples stored reftigaratod should be allowed to equilibrate to room
temperature. 3.4 All samples must be thoroughlymixedbefore beingsampled forextraction. 3.5 Sample collection procedures will be specified in the sampling plan for this
project.
4.0 Rge"fr Standards
4.1 Water-HPLC grade A2 Methanol - HPLCgrade 4.3 AmmoniumAcetate-A.C.S. Reagent Grade 4.4 PerfluorooctanoicAcid- Sigma-Aldrich
5.0 InstrumentandEquipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume iqjector capable of injecting 5-200 pL connectedto a tandemMass Spectrometer (LC/MS/MS).
5.2 Adeviceto collect rawdata for peak integration and quantitation. 5.3 Analytical balance capable ofreadingto 0.00001 g. 5.4 50mLdisposable polypropylene centrifoge tubes. 5.5 15mLdisposablepolypropylene centrifoge tubes. 5.6 Disposablemicropipets (SO-lOOuL, 100-200uL). 5.7 125-mLLDPBnarrow-mouthbottles. $.8 2mLclear HPLCvial kit. 5.9 Disposable pipettes. 5.10 Autopipettes(100-1000jiLand 10-100 pL), with disposable tips. 5.11 Waters SepPak Vac 6 cc (Ig) tC18 SPEcartridges. 5.12 SPE vacuummanifold. 5.13 Ultrasonic bath.
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Exygen Study No.: P0001131
Exygen Protocol Number: POOOl 131
Exygen Hamich
Method N u o ta V0001781
I ANALYTICALMETHOD
|
MethodofAnalysis fordieDeterminationofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
5.14 Wrift-ectionihaker. 5.15 Centrifuge capableofspinning 50 mLpolypropylene tubes el 5000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: FluophaseRP (Keystone Scientific), 2.1 mm x 50 mm. 5jj (P/N: 82305-052130)
6.2 Temperature: 30*C 6.3 Mobile Phaae(A) : 2 mMAmmoniumAcetate in Water 6.4 Mobile Phaae(B) : Methanol 6.5 GradientProgram:
Tima fminl
0.0 1.0 8.0 20.0 22.5
%A 65 65 25 25 65
Flow Rate 2lb (mL/minl 35 0.3 35 0.3
75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: 15 pL(canbe increasedto as much as 50 jiL). 6.7 Quantitation: PeekAm - external standardcalibration curve. 6.8 RunTune: ~ 23minutes.
The above conditionsare intendedas a guide and maybe changed in order to optimize theHPLCsystem.
7.0 MS/MS System
7.1 Mode: BlectrosprayNegativeMRMmode, monitoring 413- 369 m/z for PFOA.
The above conditions ire intendedas guide and may be changed in order io optimizethe MSMS system.
8.0 PreparationofSolutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water it prepared by adding 0.154 g oT ammoniumacetate to 1000mLofwater.
Alternatevolumes maybeprepared.
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygee R w u rh
M bod Nttmbr V0001781
ANALYTICAL M ETHOD
MethodofAnalysisfor theDeterminationofPerftuorooctanoic Acid (PFOA) in Soil by LC/MS/MS
9.0 StandardPreparation
9.1 StandardStock/FortificationSolution 9.1.1 Prepare a node solutionof '100 pg/roL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125hiiL LDPEbottle. 9.1.2 A 10 pgfatL 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 125mLLOPEbottle. 9.1.3 A 1.0pg/raL fortificationsolution of PFOA is prepared by bringing 10 mLofthe 10pg/mL solution to a final volume of 100 with methanol in a 125 mLLDPEbottle. 9.1.4 A 0.1 p^mL fortificationsolutionofPFOAprepared by bringing 10 mL of)he 1.0pg/mLsolutionto a final volume of 100 with methanol in a 125mLLDPEbottle. 9.1.5 A 0.01 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of die 0.1 pg/mL solution to a final volume of 100 with methanolin a 125mLLDPEbottle. 9.1.6 Tbe stock and fortificationsolutions are to be stored in s refrigerator at approximately4*C and are stable for a maximum period of 6 months fromthedate ofpreparation.
9.2 StandardCalibrationSolutions
9.2.1 LC/MS/MS calibration standards are prepared in HPLC water The calibration standards are processed through tbe extraction procedure, identical to samples.
9.2.2 The following ia a typical example: additional concentrations may be prepared aaneeded.
Final
Conoentration Fortification Volume o f Concentration o f Calibration
e f Fortification Volume Fortified Control Calibration
Standard ID
Solution (neb)
(uL)
Samole (mL) Standard foot)4
(examcle)
0 0 40
0 XCmmddyy-0
10 100 40
25 XCmmddyy-1
10 200 40
so XCmmddyy-2
10 400 40
100 XCmmddyyO
100 100 40
250 XCmmddyy-4
100 200
40
500 XCmmddyy-5
100 400
40
1000
XCmmddYY-6
*The extractedcoaccntratioaofthecalibration standard is equal to 8* its initial
concentration, doeto thecoaccntratioa of the standard during the extraction (SPE). XC extractedcalibrationstandard.
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Exygen Study No.: P0001131
Exygen Protocol Number: POOOl 131
Exygao R m aicb
Method Nw*srV00I7Si
ANALYTICAL M ETHOD
]
MethodofAnalysis for theDeterminationofPerftuorooctanoic Acid (PFOA) in Soil by
LC/MS/MS
9.2.3 A zero standard solution (reagent blank) must be prepared with each set ofstandards extracted.
9.2.4 Store all extracted calibration standards in 15-mL polypropylene lubes at 2*Cto 6*C, upto two weeks.
9.2.$ 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 leu) must include at least mm reagent control (method blank using S mL of methanol) and two reagent controls fortified at known concentrations (lab control spike) to verify procedural recovery forthebatch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified inthequalityassuranceplan 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 andmix well).
1U Add 5mLofmethanol andshake on a wrist action shaker for~15 minutes IU Transferthetubestoanultrasonic bath and sonicate for-15 minutes.
11.4 Bring the volume up to 40 mL with water in the $0 mL polypropylene
^ 11.5 centrifogetube. CentrifUgefor-10 minutes at ~3000 rpm. 11.6 Condition the Cts SPB cartridges (1 g. 6 mL) by passing 10 mL methanol followedby 5mL ofHPLC water(- 2 drop/iec). Do not let column run dry 11.7 Load (decant) the sample on the conditioned Cn SPE cartridge. Discard ehiate. 11.8 Elute with >5 mL 100% methanol. Collect 5 mL of eluste into graduated 1$mLpolypropylenecentrifoge tubes (final volume * 5 mL). 11.9 Analyze samples usingelectrospray LC/MS/MS.
12.0 Chromatography
12.1 Iqject the sane amount ofeach standard, sample and fortified sample into the LC/MS/MS system. A calibration standard must precede and follow all analyzed samples.
12.2 Standards ofPFOAcorrespondingto at least five or more concentration levels muet beincluded in an analyticalact.
12.3 An entire set of extracted calibration standards must be included at the beginning and it the end of a simple set Extracted standards must be interspersedbetweenevery 5*10samples. As an alternative, an entire set of
Page 5 of?
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExytmUeaettch
Method Number VQQQ17|
|
ANAIVTICAL METHOD .....
...
MethodofAnalysts for theDeterminationofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
extracted calibration standaids may be injected at the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to account for a second set ofextracted standards). In either case, extracted calibration standards must be the first and last infection in a sample set.
12.4 Use linear tiandard curves for quantitation. Linear standard curves are generated forfoe analyte by linearregression using 1/x weighting ofpeak area
versus calibration standard concentration using MaasLynx 3.3 (or equivalent)
software system. 12.5 Sample response should not exceed standard responses. Any samples that
exceedstandardresponses should be fortherdiluted and reanalyzed.
13.0 AcceptanceCriteria
13.1 Chromatogrammust show a peak of a daughter ion at 369 amu from a parent of 413 amu. The 413 amu parent corresponds to foe PFOA anion, while the daughter ion(369 amu) represents the loss ofcarbon dioxide.
13.2 Method blanks must not contain PFOA at levels greater than foe LOQ. If a blank contains PFOA at levels greater than SOng/L, then a new blank sample must be obtainedandthe entireset must be re-extracted.
13.3 Recoveries ofcontrol spikes and matrix spikes must be between 70-130*4 of their known values. If a control spike M s outride foe acceptable limits, the entire set of samples should be re-extracted. Any matrix epike 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
Bitot Test, may be excluded from foe calculation of foe calibration curve. However, foe total number of extracted calibration standards that could be excluded must not exceed MS of foe total number of extracted standards
injected. 13.5 The correlation coefficient (R) for calibration curves generated must be
20.992 (R3 20.985). If calibration remits foil 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
1 4 %withinanlyrical run. Ifretention timedrift exceeds this limit within ananalytical run thenthe set must bereanalyzed.
Paces of7
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExygeaRaHHcb
Method N uaber V000I7SI
1 ...... ANALYTICAL METHOD
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS
14.0 Calculations
14.1 Ute foe following equation to calculate foe amount of PFOA found (in ng/U baaed on peak area) wing the standard curve (linear regression parameters) generatedbyfoeMamLynxsoftware program:
PFOA found (ng/L)
xDF
DF" factorbywhichfoe final volumewas diluted, ifnecessary.
14.2 For samples fortified with known amounts of PFOA prior to extraction, use foefollowingequationto calculate thepercent recovery.
Recovery(%)
[totalanalytefound(ng/L) - analytefoundincontrol (ng/L)] analyteadded(ng/L)
14.3 Use the following equation to convert foe amount of PFOA found in ng/L to ng/g(ppb).
PFOAfound(ppb) * fPFOAfoundfaa/D x volumeextracted (0.04U1 sample weight (5 g)
14.4 Use the following equation to calculate foe amount of PFOA found in ppb baaedon dryweight
PFOAfound(ppb) dry weight PFOA found (ppb) x [100%/ total solida(%)]
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Exygen Study No. : P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber V000J782
Metta) ofAnalysis for Am Determination of Perfluorooctaaoic Add (PFOA) la Sediment by LC/MS/MS
Analytical TestingFacility:
Exygen Research 3058 ResearchDrive StateCollege, PA 16801
Approved B y.
f-jt.
Paul Connolly
(
Technical Leader, LC-MS, ExygenResearch
//>?
ohin Flaherty / VicePresident,Operations, ExygenResearch
___ l O k h M Dote Date
Exygen Research
Total Pages: 7
P age 30 o f 65
Page 99 of 136
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
NtabodNumber V00017B2
| ANALYTICAL M ETHOD
|
MethodofAnalytic forthe DeterminationofPerfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS
1.0 Scope
This method ii to he employed for die isolation andquantitation ofperfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass SpectrometricDetector (LC/MS/MS) in sediment
2.0 Saftty
2.1 Alwaysobsetvesafolabontoiy practices. 2.2 Consultthe appropriate MSDSbeforehandling anychemical for proper safety
precautions.
3.0 SampleRequirement
3.1 At least 30goftest sample forextraction. 3.2 Nosample processingis needed for sediment samples. 3.3 Samples stored refrigerated should be allowed to equilibrate to room
temperature. 3.4 All samplesmust be thoroughlymixed beforebeing sampled for extraction. 3.5 Sample collection procedures will be specified in the sampling plan for this
project
4.0 Reagentssod Standards
4.1 Water- HPLCgrade 4.2 Methanol- HPLCgrade 4.3 AceticAdd-Reagent grade 4.4 AmmoniumAcetate- AC-S. Reagent Grade 4.5 PerfluorooctanoicAdd -Sigma-Aldrich
5.0 Instrument andEquipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equippedwith a variable volume injector capable of injecting 5-200 pLconnectedto a tandemMass Spectrometer(LC/MS/MS).
5.2 Adeviceto collect rawdata forpeak integration and quantitation. 5.3 Analytical balance capableofreadingto 0.00001 g. 5.4 50mLdisposablepolypropylenecentrifoge tubes. 5.5 15mLttoposabiepolypropylenecentrifugetubes. 5.6 Disposablemicropipets (50*l00uL, 100-200uL). 5.7 125-mLLDPEnanow-mouth bottles. 5.8 2mLclearHPLCvial kit. 5.9 Disposablepipettes. 5.10 Autopipettes(100*1000 pL and 10*100pL), with disposable tips. 5.11 Waters Sep PakVac 6 cc (lg) tC18 SPEcartridges. 5.12 SPEvacuummanifold.
Pag 2 of 7
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Exygen Study No.: P0001131
Exygcn Protocol Number: P0001131
E i y f n Raaaicfa
Method Number VQ0017S2
| ANALYTICAL METHOD
|
MethodofAnalysis forthe Determination ofPerfluorooctanoic Acid (PFOA) in Sediment bv
LC/MS/MS
'
5.13 Voftexer. 5.14 Wrist-actionshaker. 5.15 CentriiUgecapable ofspinning50mLpolypropylene tubes at 3000 rpm.
6.0 ChromatographicSystem
6.1 Analytical Cohunn: FluophaseRP(Keystone Scientific) 2.1mm x 50 mm. 5m (P/N: 825054)52130)
6.2 Temperature: 30*C 6.3 Mobile PbSM(A): 2 mMAmmoniumAcetate in Water 6.4 Mobile Phase (B): Methanol 6.5 GradientProgram:
Tima fmin)
00 1.0 8.0 20.0 22.5
2kA 65 65 25 25 65
Flow Rate (mL/minl
35 0.3 35 0.3 75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: IS |iL(can be increasedto as much as 50 pt). 6.7 Quantitation: PeakAna - external standard calibration curve. 6.8 RunTime: ~ 23 minutes.
The above conditions an intendedas a guide and may be changed in order to optimizethe HPLCsystem.
7.0 MS/MSSystem 7.1 Mode: ElectzoaprayNegative MRMmode, monitoring 413 369 m/z for
Theaboveconditions are intended u a guide andmaybe changed inorder to optimize theMSMSsystem.
8.0 PreparationofSolutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of mmnniim acetateto 1000 mLofwater.
Page 3 o f7
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Exygen Study No.: P0001131
Exygen Protocol Number: POOO1131
BuygwRm w I i
Method Number VOOOI782
| ANALYTICAL METHOD |
MethodofAnalysis for the DeterminationofPerfluorooclanoic Acid (PFOA) in Sediment by LC/MS/MS
8.2 ExtractionSolutions
8.2.1 IS acetic add in water is prepared by adding 10 mL of acetic acid to 1000 mL of water.
Alternatevolumes maybeprepared.
9.0 Standard Preparation
9.1 StandardStock/FortificationSolution 9.1.1 Prepare a stock solution of -100 jig/mL of PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with
methanol in a 125-mLLDPEbottle. 9.1.2 A 10pg/mL fortificationsolution of PFOA i*prepared by bringing 10
mL of the 100 pgtaL solution to a final volume of 100 with methanol
in a 125mL LDPEbottle. 9.1.3 A 1.0 ng/mLfortificationsolution of PFOA is prepared by bringing 10
mL ofthe 10pgAnL solutionto a final volume of 100 with methanol in a 125mLLDPEbottle. 9.1.4 A0.1 gg/mLfortificationsolution ofPFOA is prepend by bringing 10 mLofthe 1.0jig/mLsolution to a final volume of 100with methanol in a 125mL LDPEbottle. 9.1.5 A 0.01 pgfaiL fortification solution of PFOA is prepared by bringing 10 mL of the 0.1 pgtaL solution to a final volume of 100 with
methanolin a 125 mLLDPEbottle. 9.1.0 The stock and fortification solutions are to be stored in a refrigerator at
approximately 4*Cand are stable for maximum period of 6 months fromthe date of preparation.
9 2 StandardCalibrationSolution!
9.2.1 LC/MS/MScalibrationstandards areprepared in methanol via dilution oftbeO.l pg/mLfortificationsolution.
9.2.2 The following is a typical example: additional concentrations may be
Concentration o f Fortification Solution fn rtn L )
100 100 100 10 5 2
Volume fm U 10 5 2 10 10 10
M uted to
ta U 100 100 100 100 100 100
Final Concentration
inarinL) 10.0 SO 2.0 1.0 o.s 0.2
Page 4 of 7
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Method Number VOOOl 7S2
| ANALYTICAL METHOD | MethodofAnalysis fbrtheDeterminationofPerfluorooctanoic Acid (PFOA) in Sediment by
LG/MS/MS
9.2.3 Store all calibration standards in 125-reL LDPE narrow-mouth bottles at 2*Clo6*C, upto sixmonths.
91.4 Alternate volumes endconcentrations of standards may be prepsred as needed.
10.0 Batch Set Up 10.1 Bech batch of samples extracted (typically 20 or lees) must include at least one untreated control and two untreated controls fortified at known concentrations (labcontrol spike) toverifyprocedural recovery forthe batch. 10.2 Requirements for field and laboratory duplicates and spikes will be specified in the qualityaasursneeplan forthisproject.
11.0 SampleExtraction 11.1 Weigh 3 g of sample into 30 mL polypropylene centrifoge tubes (fortify as needed, replace lid andmix well). 11.2 Add 33 mL of 1%acetic acid, cap, vortex and shake on a wrist action shaker for-60 minutes. 11.3 Centrifoge the tubes at ~3000rpm for-20 minutes. 11.4 Condition the Ci* SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followedby 20mLofHPLC water (~ 2 drop/sec). Do not let column rundry 11.3 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 30 mL centrifoge tube. Of, vortex and shake on a wrist action shaker for *-30 mhmte. 11.7 Centrifogethetubes at"3000rpmfor~20minutes. 11.8 Decantthemethanol ontothe same SPEcartridge. Collect the eluate. 11.9 Washthe columnwith 4 mLofmethanol. Collect the eluate and add itto the eluatecollectedin step 11.8. 11.10 Conditiona secondCu SPEcartridge (1 g, 6 mL) by passing 10mL methanol followedby 20mLofHPLCwater (~ 2 drop/eec). Do not let column rundry 11.11 Add the methanol to -200 mL of water and load on the second conditioned SPEcartridge. 11.12 Elute with ~3 mL 100K methanol. Collect 3 mL of eluate into graduated 13mLpolypropylenecentrifoge tubes (final volume " 3 mL). 11.13 Analyzesamples using electrosprayLC/MS/MS.
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
B iy|-- It-- sawh
M sttod Number V0001782
| ANALYTICAL METHOD
|
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Aeid (PFOA) in Sediment by LC/MS/MS
12.0 Chromatography
12.1 Inject the seme amount ofeach standard, sample and fortified sample into the
LC/MS/MS system. A calibration standard must precede and follow all analysed samples. 12.2 Standards ofPFOAcorrespondingto at least five or more concentration levels must be included inan analytical set. 12.3 An entire act of extracted calibration standards must be included at the beginning and at die end of a sample set Standards must be interspersed between every 5*10 sample. 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 Iasi inaction ina sample set 12.4 Use linear standard curves for quantitation. Linear standard curves arc generated forthe analytebylinearregressionusing 1/x weighting of peak area versus calibration standard concentration using MastLynx 3.3 (or equivalent)
software system. 12.3 Simple response should not exceed standard responses. Any samples that
exceedstandardresponses shouldbe ftnther diluted and reanalyzed.
13.0 AcceptanceCriteria
13.J
mint th f*
a d ^ g H--*ifm *
a parent
of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the
daughterion(309amu) representsthelossofcarbon dioxide.
13.2 Method blanks must not oontain PFOA it 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 obtainedand the entireset 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 foils outside the acceptable limits, the
entire set of sanies should be re-extracted. Any matrix spike outside 70
130% should be evaluated by foe 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 foe calculation of foe calibration curve.
However, foe total wmlw of extracted calibration standards that could be
excluded must not exceed 20% of foe total number of extracted standards
injected.
13.3 The correlation coefficient (R) for calibration curves generated must be
0.992 (R* 0.985). If calibration results foil outside these limits, then
appropriate steps must be taken to adjust instrument operation, and the
standards orthe relevant set of samples shouldbe reanalyzed.
Pss*4of 7
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Exygen Study No.: P0001131
Exygen Protocol Number: P001131
Method Number VOOOI7I2
1 ANALYTICAL METHOD
|
MethodofAnalyse forthe Detenninaton ofParfluorooctanoic Acid (PPOA) in Sediment by LC/MS/MS
13.6 Retention times between standards and samples must not drift more than
4 % within an analytical run. Ifretention time drift exceeds this limit within an analytical run thenthe set must bereanalyzed.
14.0 Calculations 14.1 Use the followingequationto calculatethe amount of PFOA found (in ng/mL. based on peak area) using the standard curve (linear regression parameters) generatedby the Maas Lynxsoftwareprogram:
PFOAfound (ng/mL) - (Peak area - intercept) x OF slope
OF factorbywhich the final volumewas diluted, ifneceesary.
14.2 For samples fortified with known amounts of PFOA prior to extraction, use the followingequationto calculate the percent recovery.
Recovery(%)-
[ total analytefound(ng/mL) - analytefoundincontrol(ng/mL)] analyteadded(ng/mL)
14.3 Use the followingequationto convert die amount ofPFOA found in ng/mL to ntf (PPb).
PFOA found(ppb) - rpun* (ni/mU x foul volume 1} mL)l sample weight (3 g)
14.4 Use foe following equation (if necessary) to calculate the amount of PFOA foundin ppb basedondryweight.
PFOAfound(ppb) dry weight **PFOAfound(ppb) x (100%/ total solids(%l]
Pese 7 of 7
P age 36 o f 65
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Page 105 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber: V0001783
Method ofAaafyait for die Determlaatioo ofFerflaorooctanolc Add (PFOA) in Fisb aid Clam by LC/MS/MS
Analytical TottingFacility:
Exygen Retouch
3058 Retouch Drive State College, PA 16801
Approved By:
V - l f - J L ____
Paul Connolly
1
Technical Leader, LC-MS, ExygenReeearch
Dele
Flaherty Vice President, Operations, ExygenRetearch
Date
Exygen Research
Total Paget: 8
P a g e 37 o f 65
Page 106 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No. : P0001131
Exygen Protocol Number: P0001131
Exygea Xcmrch
Method Number VOOQ17S3
I ANALYTICAL METHOD
|
MethodofAnalysis fortheDeterminationofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
1.0 Scope
Tbit method is to be employed forthe isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Mass SpcctromotricDetector(LC/MS/MS) in fish andclams.
2.0 Safety
2.2 Alwaysobservesafe laboratorypractices. 2.2 Consult the appropriate MSDS beforehandling any chemical for proper safety
precautions.
3.0 SampleRequirement
3.1 At least 20 g oftest sample forextractioa 3.2 Samples should be processed before extraction. Place the frozen sample in a
foodprocessor and homogenize with dry ice. Place the samples in containers and 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 tire sampling plan for this project
Reagents andStandards
4.1 Water-HPLC grade 4.2 Acetonitrile- HPLCgrade 4.3 Carbon(120-400mesh)- Reagent grade 4.4 Methanol - HPLC grade 4.5 Silicagel (60-200mesh)- Reagent grade 4.4 Ftorisil (60-100mesh)- Reagent grade 4.7 SupercleanLC-NHj - Reagent grade 4.8 1-Octanol - HPLCgrade 4.9 L-Ascorbicacid- Reagent grade 4.10 DimethyidichknosiUme- Reagent grade 4.11 Toluene- Reagent grade 4.12 AmmoniumAcetate- AC.S. ReagentGrade 4.13 Perfluorooctanoic Acid- Sigma-AJdrich
3.0 Instrumentand Equipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume iqjeetor capable of injecting 5*200 pLconnectedto a tandemMass Spectrometer(LC/MS/MS).
S 2 A devioeto collect rawdata forpeak integration and quantitation. 5.3 Analytical balance capableofreadingto 0.00001 g.
Par tors
P a ge 38 o f 65
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Page 107 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number P001131
ExytMRMMNh
M ethod Number VOOOI78J
[ ANALYTICAL m ethod MethodofAnalyiii fortheDeterminationofPerfluorooctanoic Add (PFOA) in Fish and Clamsby LOMS/MS
5.4 Rotaryevaporator.
5.5 Tusumizar. 5.6 125mLpear-shaped flasks. 5.7 50mLdisposablepolypropylenecentriftige tubes.
5.8 15mLdisposablepolypropylene centrifuge tubes. 5.9 Disposablemicropipets (50-100uL, 100-200uL). 5.10 12^mL LDPEnarrow-mouthbottles.
5.11 2 mLclearHPLCvial lot. 5.12 Disposablepipettes. 5.13 Autopipettes (100*1000pLand 10-100 pL), with disposable tips.
5.14 SPEtubes (20mL) (Supeko cat. no. N057I77). 5.15 Wrist actiontaker. 5.16 Centrifugecapableoftptanwg SOmLpolypropylene tubes at 2000 rpm.
6.0 Chromatographic System
6.1 Analytical Column: Fhiopbsse RP {Keystone Scientific). 2.1 mmx 50 mm. 5m (P/N: 82505-052130)
6.2 Temperature: 30*C 6.3 Mobile Phase (A) : 2 mMAmmoniumAcetate in Water 6.4 MobilePhase(B) : Methanol 6.3 GradientProgram:
Ttmafminl
0.0 1.0 8.0 20.0 22.5
2i A 65 65 25
25 65
Flow Rate fmLftniiri
35 0.3 35 0.3 75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: 1$ pL (canbe increased to as much as SOpL). 6.7 Quantitation: PeakArea- external standardcalibration curve. 6.8 RunTime: - 2 3 minutes.
The aboveconditions are intended as a guide and may be changed in order to optimize the HPLC system.
PattJofS
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Exygen Study No. : P0001131
Exygen Protocol Number: P0001131
Exygen K-- ech
MethodNuocer VQQ0J7S3
A NA LYTICAL M ETH O D
MethodofAnalysis forthe Determination ofPerfluorooctinoic Acid (PFOA) in Fish and Clams byLC/MS/MS
7.0 MS/MS System
7.1 Mode; ElectroprayNegative MRMmode, monitoring 413- 369 m/z for PFOA.
The aboveconditions are intendedas a guide and may be changed inorder to optimize theMSMSsyctem.
8.0 PreparationofSolution!
8.1 MobilePhase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of ammoniumacetateto 1000mLofwater.
8J Extraction Solutions
8.2.1 2%ascorbic add inmethanol is prepared by dissolving 2 g ofascorbic acidm 100mLofmethanol.
8.2.2 30%Dimethykiichlorosilane in toluene is prepared by bringing 3 mL ofdimethyldichloroatianeto a final volume of 10mL with toluene.
Alternatevolumesmaybe prepared.
9.0 StandardPreparation
9.1 StandardStock/Fortifioation Solution
9.1.1 Prepare a Hook solution of-100 >tg/mLof PFOA by weighing 10 mg of analytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mLLDPEbottle.
9.1.2 A 1.0 p^mL fortification solution of PFOA is prepared by bringing 1 mL ofdie 100 pgrinL solution to a final volume of 100 with methanol ina 125mL LDPEbottle.
9.1.3 A0.1 pgffaLfortificationsolutionofPFOA is prepared by bringing 10 mLoftbe l.Ojig/raL solution to a final volume of 100 with methanol in a 12S mLLDPEbottle.
9.1.4 A 0.01 pgfaiL 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 ina 125mL LDPE bottle.
9.1.5 The stockand fortificationsolutions are to be stored in a refrigerator at approximately 4*C and are stable for a maximum period of 6 months fromthedate ofprepaidon.
Page 4 of S
P age 40 o f 65
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Page 109 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExygeaResearch
MsthodNumberVQOO1783
1 ... ANALYTICAL M E T H O D
MethodofAnalysis fortheDeterminationofPerfhiorooctanoic Acid (PFOA) in Fith and Clamaby LC/MS/MS
9.2 Studied Calibration Solutions
9.2.1 LC/MS/MS calibrationstandards are prepared in methanol via dilution ofdie 1.0pgfaL fortificationsolution.
9.2.2 The foliowing is a typical example: additional concentrations may be preparedat needed.
Concentration
Fine)
ofFortification Volume Dilutedto Concentration
Solution(ut/mL) (mL)
(mL)
(ua/mL)
1JH 5.0 100
0.05
1.0 2.5 100
0.025
1.0 1.0 100
0.01
0.05 10 100
0.005
0.025 10
too
0.0025
0.1 10 100
0.001
0.005 10
100
0.0005
9.2.3 Store all calibrationstandards in 125-mL LDPE narrow-mouth bottles
at 2*Cto 6*C, up to sixmonths.
9.2.4 Alternate volumes and concentrations ofstandards may be prepared u
needed.
10.0 BatchSet Up
10.1 Each batch of samples extracted (typically 20 or leas) must include at least one untreated control and two untreated controls fortified at known concentrations(lab control spike)to verilyprocedural recovery forthe batch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified in thequalityassurancepin forthis project.
11.0 Sample Extraction
11.1 Weigh 5 g of frozen sample into 50 mL polypropylene centrifuge tubes (fortifyas needed, replace lidand mix well).
11.2 Add30 mLofacetonitrile andshake on a wrist action shaker for-15 minutes L1.3 Place the tubes in a freezer for-1 hour. 11.4 Pack andconditiontheSPEtubes and silsnizethe pear-shaped flasks. 11.5 Pack the 20 mL SPE tubes in sequence with 2 g Qoriril, 2 g silica gel, 2 g
carbon, and I 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 foe 125 mL pear-shaped flasks by rinsing with the 30% dimethyklichlorosilane in toluene solution. Rinse foe flask with toluene once, followed by methanol (time tiroes). Dry the flasks completely before use, either byair-dryingorwitha streamofnitrogen.
Page 5 of a
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Page 110 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExygraRssMttli
MftbodNumber V000PU
A N A LYTIC A L M ETHO D
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Acid (PFOA) in Fish and Clams by LC/MS/MS
11.7 Cemriftige the 50 mL polypropylene tubes containing sample at -2000 rpm for-10 minutes.
11.8 Decant the extrect on to a conditioned SPE column fitted inside the mouth of the pear-shaped flask. Collect the eluate in the 12S mL silanized pear-shape
flask. 11.9 Add 10 mL of acetonitrile to the sample in the 50 mL centrifuge tube.
Homogenize the frozen fiuphaee using a tissumizer for -30 seconds and rinse thetiseumizerwifo-10 mLofacetonitrileinto the tube. 11.10 Shakethe sample againfor -10 minutes on a wrist-action shaker.
11.11 Placethe tubes in a freezer for - 1hour more. 11.12 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 ipm
for-10 minutes.
11.13 Decant the extract onto the seme SPE column. Collect the eluate into the same pear-shaped flask and combine with foe eluent from the initial
extraction.
11.14 Pass 20 mL ofacetonitrile throughthe SPE column and combine the eluate in
the samepear-ehapedflask. 11.15 Add 3-4 drops of i-octanol to foe extract in foe pear-shaped flask and
evaporate at reducedpressure usinga rotaryevaporator(at < 40C).
11.16 Make foe final volume, by adding 2 mL of 2% ascorbic acid in methanol to foepear-shapedflaskand swirl to mix/diisolve.
11.17 Transferthe extracts loHPLCvials usingdisposable pipis.
11.18 Analyze samplesusing electrospny LC/MS/MS.
12.0 Chromatography
12.1 Inject foe tame amount of each standard, sample and fortified sample into (he LC/MS/MS system. A calibration standard must precede and follow alt analyzed samples.
12.2 Standards ofPFOAcorrespondingto at least five or more concentration levels must be includedin an aruriyticai set
12.3 An entire set ofcalibration standards must be included at the beginning and at
foe end of a sample set. Standards must be interspersed between every 5-10 simples. 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 ease, calibration standards must be the first end lest injection in u sample set 12.4 Use linear standard curves for quantitation. Linear standard curves are generated forthe analyte bylinearregression using 1/xweighting ofpeak area versus calibration standardconcentration using MassLynx 3.3 (or equivalent) software system.
Page6ofS
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Page 111 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Bxyya to n reh
Method Namber V0001783
analytical method
MethodofAnalysis lor the DeterminationofPerfluorooctmoic Acid (PFOA) in Fish end Clams by LOMS/MS
12.5 Sample response should not exceed standard responses. Any samples that exceedstandardresponses shouldbe Anther diluted and reanalyzed.
13.0 AcceptanceCriteria
13.1 Chromatogrammust 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(369amu) representstheloes ofcarbon dioxide.
13.2 Method blanks must not contain PFOA at levels greater than die LOQ. If a blank contains PFOA at levels greater than 0.5 ppb, then a new blank sample must be obtained andthe entire set mustbe re-extracted.
13.3 Recoveries of oontrol spikes and matrix spikea must be between 70-130% of their known values. If oontrol spike Adis outside the acceptable limits, the entire set oftemplet shouldbe 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 exceed20%ofthe total numberofstandards injected.
13.5 The correlation coefficient (R) for calibration curves generated must be 20.992 (R2 20.985). If calibration results foil outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standardsor the relevant set ofsamples shouldbe reanalyzed.
13.6 Retention times between standards and samples must not drift more than i 4 %withinan analytical nm. Ifretention time drift exceeds this limit within an analytical nmthen the set must be reanalyzed.
14.0 Calculations
14.1 Usefoe followingequationto calculate the amount of PFOA found(in ng/mL, based on peak area) using foe standard curve (linear regression parameters) generatedby foeMass Lynx software program:
PFOAfound(ngfaiL) - (Peak area - intercept) slope
14.2 Ueefoe followingequationto convert foe amount ofPFOA found in ng/mL to n*/g(ppb).
PFOAfoundftmM- fPPOAfound
find volume fmLl DF1
ample weight (g)
DF* factorbywhichthe final volume was diluted, if necessary.
P l|t 7 of*
P a g e 43 o f 65
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exyfgq fceacaich
Method Number V000I783
[ ANALYTIC.!! A1ETUOD MethodofAnalysis for the Determination oPerfluorooctanoic Acid(PFOA) inFishand Clamsby LC/MS/MS
14.3 For samples fortified with known amounts of PFOA prior to extraction, use the followingequationto calculate thepercent recovery.
Recovery(%) -
[ totalanalytefound(ng/g) analytefoundincontrol (ng/g)] j]D analyteadded(ng/g)
Exygen Research
Pag* t o f 8 Page 44 o f 65
Page 113 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No. : P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber V0001784
Method ofAnalysis for the Determination ef Peruorooetanolc Add (PPOA) in Vegetation by LC/MS/MS
Analytical TestingFacility:
Exygen Research 3058 Research Drive State College, PA 16801
Approved By:
'S - 1 c J i ,
Paul Connolly
'
Technical Leader, LC-MS, ExygenResearch
n /s n f/U /________
J o h n Flaherty ^ VicePresident, Operations, Exygen Research
__ la ji& M Date Dat
Exygen Research
Total Pigea: 7
P a g e 45 o f 65
Page 114 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exjrgan R e m rc h
M ethod Number VOOOI784
ANALYTICAL METHOD
MethodorAnalysis for the DeterminationofPerfluorooctaooic Acid (PFOA) in Ve^etauon by LC/MS/MS
1.0 Scope
This method is to be employed forthe isolation and quantitation of pcrfluorooctanoic add by High Performance Liquid Chromatography coupled to s tandem Mass Spcctromctric Detector (LC/MS/MS) in vegetation.
2.0 Safety
2.1 Alwaysobserve safe laboratorypractices. 2.2 Consult foeappropriate MSDSbefore handling any chemical for proper safety
precautions.
3.0 Simple Requirement
3.1 At least 20g ofteet sample forextraction. 3.2 Samples should be processed before extraction. Place the frozen sample in s
foodprocessor and homogenize with dry ice. Place the samples in containers and 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 bo specified in the sampling plan for this project
4.0 Reagents and Standards
4.1 Water -HPLC grade 4.2 Acetonitrile-HPLC grade 4.3 Carbon(120-400mesh) - Reagent grade 4.4 Methanol-HPLC grade 4.3 Silicagel (60-200mesh) - Reagent grade 4.6 Florilil (60-100mesh) - Reagent grade 4.7 Superclean LC-NHj - Reagent grade 4.8 1-Octanol-HPLC grade 4.9 L-Ascorbic add - Reagent grade 4.10 Dimethyldichloioailane-Reagent grade 4.11 Toluene - Reagent grade 4.12 AmmoniumAcetate-A.C.S. Reagent Grade 4.13 PerfhiorooctanoicAcid- Sigma-Aldrich
3.0 Instrument andEquipment
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 pLoonnectedto a tandemMass Spectrometer(LC/MS/MS).
5.2 Adevicetocollect rawdata forpeak integration and quantitation. 5J Analyticalbalancecapableofreadingto 0.00001 g.
Page 2 of 7
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygwi Umweh
Method N u o to V0Q017S4
ANALYTICAL METHOD
Method ofAnalysis for the Detomination ofPerfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS
5.4 Rotaryevaporator.
5.5 125mLpear-thaped flasks. 5.6 SOmLdi^wiAle polypropyloecenbiiSifeUbw. 5.7 15mLdisposablepolypropylenecentrifuge tubes. 5.6 Disposablemicropipets (30-lPOuL. 100200uL).
5.9 125-mLLDPEnarrow-mouthbottle*. 5.10 2 mLclear HPLCvie] kit. 5.11 Disposablepipettes.
5.12 Aulopipettes (100-1000 pLand 10*100pL), with dispouble tip*.
5.13 SPEtube*(20mL) (Supelcoeat. no. N057177). 5.14 Wriet actionshaker. 5.15 Centrifbgecapableofapitming 50mLpolypropylene tube*at 2000 rpm.
6.0 ChromatographicSystem
6.1 Analytical Column: FhtophaseRP(Keystone Scientific), 2.1 mm x 50 mm. 5p (P/N: 82505-052130)
6.2 Temperature: 3Q*C 6.3 Mobile Phase(A): 2 mMAmmoniumAulite in Water 6.4 Mobile Phase(B): Methanol 6.5 Gradient Program:
Time fmitri
0.0 1.0 8.0 20.0 22.5
SIA 65 65
25 25 65
Flow Rate
2i fmL/min> 35 0.3 35 0.3 75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: 15pL(can be increased to aa much as 50 pL). 6.7 Quantitation: PeakArea- external atandard calibration curve. 6.8 RunTime: -2 3 minutes.
The above conditions are intended aaa guide end maybe changed in order to optimizethe HPLCsystem.
7.0 MS/MS System
7.1 Mode: ElectrosprayNegative MRMmode, monitoring 413 - 369 m/z for PFOA.
Page 3 of?
P age 47 o f 65
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Page 116 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
E xy|ta HcMAfch
Mrthod Number V00UI784
ANALYTICAL METHOD
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Add (PFOA) in Vegetation by LC/MS/MS
The aboveconditions are intendedas a guide and maybe changed in order to optimize the MSMS system.
8.0 PreparationofSolutions
8.1 MobilePhase
8.1.1 2 mM anunoahim testae in water is prepared by adding 0.154 g of ammoniumacetateto 1000mL ofwater.
82 Extraction Solution*
8.2.1 2% aaooriric add inmethanol if prepared bydissolving 2 g of ascorbic add in 100mLofmethanoL
8.2.2 30% Dimethyldichlorofilane in toluene is prepared by bringing 3 mL ofdimetiiyldicMoiodlaneto a final volume of 10mLwith toluene.
Alternate volumes may be prepared.
9.0 Standard Preparation
9.1 StandardStock/FortificationSolution
9.1.1 Prepare a stock solutionof--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-mLLDPEbottle.
9.1.2 A 1.0 jtgfaiL fortificationsolution of PFOA it prepared by bringing I mL of the 100 jigfaiLsolution to a final volume of 100 with methanol ina 125mL LDPEbottle.
9.1.3 A0.1 wtaL fortificationsolutionofPFOA ipreparedby bringing 10 mLoftite 1.0pg/mL solutionto a final volumeof 100 with methanol in a 125mL LDPEbottle.
9.1.4 A 0.01 pg/mL fortification solution of PFOA is prepared by bringing 10 mL of the 0.1 pgAnL solution to a final volume of 100 with Ina 125 mLLDPEbottle.
9.1.5 The stockand fortificationsolutions are to be stored in a refrigerator at approximately4*C and are stable for a maximum period of 6 months fromthedate ofpreparation.
9.2 StandardCalibrationSolutions
9.2.1 LC/MS/MS calibration standards are prepared tn methanol via dilution oftite 1.0 pgfaiLfortificationsolution.
Page 4 of `
P age 48 o f 65
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Page 117 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exy|*oJUmrch
MethodNumber VIXXH784
I .......
ANaLYIICAL METHOD
MethodofAnalysis for tbe Determination ofPerfluorooctmoic Add (PFOA) in Vegetation
by LC/MS/MS
9 2 .2 The following is a typical example: additional concentrations may be
Coocotraticn
Final
ofFortification Volume Dilutedto Concentration
SolutionfuafaiL) (mL)
fmL)
WmL)
1.0 5.0 100
0.05
1.0 2.5 100
0.025
1.0 1.0 100
0.01
0.05 10 100
0.005
0.025 10
100
0.0025
0.1 10 100
0.001
0.005 10
too
0.0005
9.2.3 Store all calibration standards in 125-mL LDPE narrow-mouth bottles
at 2*Cto 6*C, up to sixmonths.
9 2 .4 Alternate volumesand concentration*ofstandards may be prepared as
needed.
10.0 BatchSet Up
10.1 Each batch of simples extracted (typically 20 or less) must include it least one untreated control and two untreated controls fortified at known concentrations (labcontrol spike) to verifyprocedural recovery for the batch.
10.2 Requirements for field and laboratory duplicates and spikes will be specified inthe qualityassurance plan forthis project.
11.0 SampleExtaction
11.1 Weigh 5 g of frozen sample into 50 mL polypropylene centrifuge tubes (fortifyas needed, replace lid andmix well).
11.2 Add30mL ofacetonitrileand shake on a wrist action shaker for-15 minutes. 11.3 Centrifuge the 50 mL polypropylene tubes containing sample at -2000 rpm
for-10 minutes. 11.4 Packandconditiontbe SPBtubesand tUanize thepear-shaped flasks.
11.5 Pack the 20 mL SPE tubes in sequence with 2 g florisii, 2 g silica gel. 2 g carbon, and 1 g LC-NHj. Condition die columns with 20 mL of methanol, than 20 mL of acetonitrile. Discard all washes. Do not allow tbe column to
dry. 11.6 Silanize the 125 mL pear-shaped flasks by rinsing with the 30%
dimethyldichlOToeUanein toluene solution. Rinse die flaskwith toluene once, followed by methanol (three times). Dry the flasks completely before use, eitherbyair-dryingor with a streamofnitrogen. 11.7 Decant the extract on to a conditioned SPE column fitted inside the mouth of the pear-shaped flask. Collect tbe eluate in die 125 mL sihuiized pear-shape flask.
P age 49 o f 65
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Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygea tUsearch
Me&od Number V000I7M
ANA LYTICAL m e t h o d
]
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Acid (PFOA) inVegetation
by LC/MS/MS
11.8 Add20mLofacetonitrileto thesample in the 50mL centrifijge tube. 11.9 Shake the ample again for~10minuteson a wrist-action shaker.
11.10 Ccntrifoge the 50 mL polypropylene tubes containing sample at -2000 rpm for-5 minutes.
11.11 Decant the extract onto the skdc SPE column. Collect the eluate into the wwe peanehaped flask and combine with the eluent from the initial
extraction. 11.12 Repeat itepa 11.8through 11.11 again. 11.13 Add 3-4 drops of 1-octanoi to the extract in the pear-ahaped flask and
evaporate at re&icedpressure usinga rotaryevaporator (at < 40C).
11.14 Make the final volume, by adding 2 mL of 2% ascorbic acid in methanol to thepear-shapedflaskid swirlto mix/disaolve.
11.13 Transferthe extractto HPLCvials usingdisposable pipets.
11.16 Analyze sample*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
analyzedsamples. 12.2 Standards ofPFOAcorrespondingto at least five or more concentration levels
must be includedinan 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 attracted calibration standards may be injected a the beginning of a set followed by extracted calibration standards interspersed every 5-10 samples (to weewinf for a second set of extracted standards). In either case, extracted calibrationstandards must be die first and last injection in a sample set.
12.4 Use linear standard curves for quantitation. Linear standard curves are generated for theanalyte by linearregression using 1/xweighting ofpeak area versus calibration standard concentration using MsssLynx 3.3 (or equivalent)
software system. 12.5 Sample response should not exceed standard responses. Any samples that
exceedstandardresponses shouldbe further diluted and reanalyzed.
13.0 AcceptanceCriteria
13.1 Chromatopam must show a peak of a daughter ion it 369 amu from a parent of 413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughterion(369 amu) represents the loss ofcarbon dioxide.
Page6o ff
P age 50 o f65
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Page 119 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study N o.: P0001131
Exygen Protocol Number: P0001131
BiypaltMafch
MethodNumber VOOQI784
ANALYTICAL m ethod
MethodofAnalysis for theDeterminationofPerfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS
13.2 Method bUnka must not contain PFOA at levels greater than the LOQ. If a blank contains PFOAat level greater than 0.5 ppb, thee a new blank sample moat be obtainedandtheentiresetmust bere-extracted.
13.3 Recoveries of control spikes and matrix spikes must be between 70-130*4 of their known values. If a control spike falls outside the acceptable limits, the entire set ofsamples shouldbe 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 exceed20%ofthetotal number ofstandards injected.
13.5 The correlation coefficient (R) for calibration curves generated must be
0,992 (R* k0.983). If calibration results fall outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standards or therelevant set ofsamplesshouldbe reanalyzed. 13.6 Retention tirw--between standards and samples must not drift more than 4 % withinan analytical nm. Ifretention timedrift exceeds this limit within an analytical runthenthe set must be reanalyzed.
14.0 Calculation! 14.1 Use die followingequation to calculate die amount of PFOA found (in ng/mL, based on peak area) using the standard curve (linear regression parameters) generatedby theMass Lynx softwareprogram:
PFOAfound(ng/mL) - (Peak area - intercept) slope
14.2 Uaedie followingequationto convert the amount ofPFOA found in ng/mL to ng/g(ppb).
PFOA found(ppb) - [PFOAfound Ina/mLl x final volume (mL) x DFi sample weight (g)
DF- factor bywhichthe final volume was diluted, ifnecessary.
14.3 For samples fortified with known amounts of PFOA prior to extraction, use die followingequationto calculate thepercent recovery.
Recovery(%)
( totalanalytefound(ng/g) - analytefoundincontrol(ng/g)j analyteadded(ng/g)
Page 7 o t'*
P age S I o f 65
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Page 120 o f 136
Interim Report #21 - Analysis of Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber V0001785
Method ofAnalysis for the Determination ofPerfloorooctaoolc Add (PFOA) in Smell Mammal Liver by LC/MS/MS
Anelytiea] Testing Facility:
Exygen Research 3058 ResearchDrive State College, PA 16801
Approved By:
C--iL|------
Paul Connolly
I
Technical Leader, LC-MS, ExygenResearch
q/ m
Jo6hnFFUlaherty /f 'VViceProesitdent, Operations, Exygeo Research
___1>UA Date Date
Exygen Research
TotalPages: 7
P age 52 o f 65
Page 121 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No. : P0001131
Exygen Protocol Number: POOO1131
ExygaoJUmich
Method Number V00017I5
| ANALYTICAL METHOD..........
MethodofAnalysts fix1(beDetermination ofPerfluorooctiDoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS
1.0 Scope
Thie method is to be employed for the isolation and quantitation ofperfluorooctanoic acid by High Performance liquid Chromatography coupled to a tandem Maas SpectrometricDetector (LC/MS/MS) inemail mammal liver.
2.0 Safoty
2.1 Alwaysobserve safe laboratorypractices. 2.2 Consult the appropriate MSDSbefore handling any chemical for proper safety
precautions.
3.0 Sample Requirement
3.1 At least Sg ofteet 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 and leave open in frozen storage overnight to allow for carbon dioxide sublimation. Seal and place the samples in frozen storage until time of analysis. Alternately, if there it an insufficient amount of sample (-less than 5 g), then noprocessingis necessaryand the sample can be used as supplied 3.3 Sample collection procedures will be specified in the sampling plan for this project
4.0 Reagents andStamfords
4.1 Water-HPLC grade 4.2 Methanol- HPLCgrade 4.3 Acetonitrile-HPLC grade 4.4 AmmoniumAcetate- ACS. Reagent Grade 4.5 PerfluorooctanoicAdd - Sigma-Aldrich
3.0 Instrument andEquipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume iqjedor capable of injecting 5-200 pLconnectedto a tandemMesa Spectrometer (LC/MS/MS).
3.2 Adevicetocollect rawdata forpeak integration and quantitation. 5.3 Analytical balance capableofreading to 0.00001 g. 3.4 50mL disposablepolypropylenecentrifrigetubes. 5.3 15 mLdisposable polypropyleneceturiftige tubes. 5.6 Disposablemicropipets (50-100uL, 100-200uL). 5.7 125-mLLDP6narrow-mouthbottles. 5.8 2 mLdear HPLCvial kit
PSe2on
P age 53 o f 65
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: POOOl 131
EnyynR iW Kgh
Method Nuobor VOOOI7K5
I ANALYTICALMETHOD
MethodofAnalysis forthe DeterminationofPerfluorooctanoic Add (FFOA) in Small Mammal Liver by LC/MS/MS
3.9 Disposablepipettes. 3.10 Autopipettet (100*1000 pL aod 10*100pL), wilb disposable tips. 3.1! Walen Sep PakVac 6 oc(lg) tC18 SPEcartridgea. 3.12 SPEvacuummanifold.
5.13 Tissuemizcr. 5.14 Writt-action shaker. 5.15 Centrifuge capableofepinning 15mL polypropylene tubes at 3000 rpm.
0.0 Chromatographic System
6.1 Analytical Column: Fluophasa RP (Keystone Scientific), 2.1 mm x 50mm. 5p (P/N: 82505-052130)
62 Temperature: 30*C 6.3 MobilePhase (A) : 2mMAmmoniumAcetate in Water
MobilePhase(B): Methanol Gradient Program:
Timafmml
0.0 1.0
8.0 20.0 22.5
%A
65 65
25 25 65
FlowRate (mL/minl
35 0.3 35 0.3
75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: 15 pL (canbe increased to as much as 50 gL). 6.7 Quantitation: PeakArea- external standardcalibration curve. 6.8 RunTime: -23 minutes.
Theaboveconditions are intended as a guide and maybe changed in order to optimize the HPLCsystem.
7.0 MS/MSSystem
7.1 Mode: Electiospray NegativeMRMmode, monitoring 413 - 369 m/z for PFOA.
The above conditions ire intended as a guide and may be changed in order to optimizethe MSMS system.
pt*3on
Page 54 o f 65
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Page 123 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExygnlUaMKh
MttbodNunbcr VOOOI7S3
I AN/O.YTICAL METHOD
MethodofAnalysUibrtheDetenniiuuioQofPerfluorooctanoic Acid (PFOA) in Small
M ammal Liver by LC/MS/M S
8.0 PreparationofSolutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water it prepared by adding 0.154 g of ammoniumacetate to 1000mLof water.
Alternatevolumes maybe prepared.
9.0 StandardPreparation
9.1 StandardStock/FortificationSolution
9.1.1 Prepare a stock solution of ~100 pg/mL of PFOA by weighing 10 mg ofanalytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mLLDPEbottle.
9.1.2 A 1.0 ng /m L fortification solution of PFOA is prepared by bringing I mL ofthe 100 jt^mL solution to a final volume of 100 with methanol in a 125mLLOPE bottle.
9.1.3 A0.1 pg/mL fortificationaolutionofPFOA ia prepared by bringing 10 mLofthe 1.0pgfaL solutionto a final volume of 100 with methanol in a 125mLLDPE bottle.
9.1.4 The stockand fortificationsolutions are to be stored in a refrigerator at approximately 4C and e stable for a maximum period of 6 months Gromthedate ofpreparation.
9.2 StandardCalibrationSolutions
9.2.1 LC/MS/MScalibration standards are prepared in methanol via dilution of the0.1 pg/mLfortificationsolution.
9.2.2 The following is a typical example: additional concentrations may be prepared aa needed.
Concentration
Fine!
ofFortification Volume Dilutedto Concentration
SolutionfartiiL) (mL)
(mL)
(na/mL)
100 5.0 100
5.0
100 2.0 100
2.0
100 1.0 100
1.0
5.0 10 100
0.5
2.0 10 100
0.2
U 10 100
0.1
9.2.3 Stiue all calibration standards in 125-mL LDPE narrow-mouth bottles
at 2*Cto6"C, upto sixmonths.
9.2.4 Alternate volumes and concentrations of standards may be prepared as
needed.
PsgeSof?
P age 55 o f 65
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Page 124 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exypea R o ta rd i
Metbod Number V0001785
| A N A LYTICAL M ETH O D
MethodofAnalysis for theDeterminationofPerfluorooctwoic Acid (PFOA) in Small Mammal Liverby LC/MS/MS
!0.0 BatchSet Up
10.1 Bach batch of samples extracted (typically 20 or less) must include at least one untreated control and two untreated controls fortified at known concentrations(labcontrol spike)to verifyprocedural recovery for the batch.
10.2 Requirements for field a d laboratory duplicates and spikes will be specified infoe qualityassuranceplan forthis project.
11.0 Simple Extraction
11.1 Weigh 1 g of sample into a 50 raL polypropylene centrifuge tubes (fortify as needed, replace lid and mix well). Note that alternate weights of liver may be measured dependingon thesample size available foruse.
It .2 Addwaterto the sample for a final volumeof 10mL. 1U Homogenizesampleusing a tissuemizer for -1 minute. 11.4 Transfer 1 mL of foe sample using a disposable pipette into a IS mL
disposable centrifuge tube. 11.5 Add5 mLofacetonitrileandShakefor**20minutes on a wrist-action shaker. 11.6 Centrifuge foetubes at -3000 rpmfor *-5minutes. 11.7 Decant the supernatant into a SOmL diipoiable centrifiige robe and add 35
mLofwatar. 11.8 Condition foe C SPE cartridges (1 g, 6 mL) by passing 10 mL methanol
followedby 5mLofHPLCwater (~ 2 drop/sec). Do not let column rundry 11.9 Loadthe sampleon conditionedCi SPEcartridge. Discard eluate. 11.10 Elute with ~2 mL of methanol. Collect 2 mL of eluate info a graduated
IS mLpolypropylene eentrifogetube (final volume 2 mL). 11.11 Analyze sampleusingelectrosprayLC/MS/MS.
12.0 Chromatography
12.1 Inject the same amountof each standard, sample and fortified sample into the LC/MS/MS aystam. A calibration standard must precede and follow all analyzedsamples.
12.2 Standards ofPPOAcorrespondingto at toast five or more concentrationlevels mustbe includedin an analytical set
12.3 An entire eatofcalibrationstandards must be included at the beginning and at the end of a sample set. Standards must be interspersed between every 5-lu samples. As an alternative mi entire set of calibration standards may be iiyected at the beginning of a set followed by calibration standards intervened every $-10samples (to account for a second eel of standards). In either case, calibration standards must be die 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 linearregression using llx weightingofpeak area
Page S of?
P age 56 o f 65
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Page 125 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exypoft--ch
MahodNumberY000178S
I ....................... ANALYTICAL METHOD
MethodofAnalysis fortheDeterminationofPerfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS
versus cahbratioa standard concentration using MassLynx 3.3 (or equivalent) software system. 12.3 Simple response should not exceed standard responses. Any samples that exceedstandardresponses shouldbe Antherdiluted and reanalyzed.
13.0 Acceptance Criteria
13.1 Chromatogrammust show a peak of a daughter ion at 369 amu from a parent of413 amu. The 413 amu parent corresponds to the PFOA anion, while the daughter ion(369amu) represents die loss of carbon dioxide.
13.2 Method blanks must not contain PFOA at levels greet'than the LOQ. If a blank contains PFOA at levels greater dun 10 ng/g, then e new blank sample must beobtainedaid the entireset must bere-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 die 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 ro-extrsetion is warranted.
13.4 Any cahbratioa 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 exceed20%ofdie total numberof standards injected.
13.3 The correlation coefficient (R) for calibration curves generated must be 20.992 (R3 20.983). If calibration results foil outride these limits, then appropriate steps mutt be taken to adjust instrument operation, and the standards or therelevant set ofsamples should be reanalyzed.
13.6 Retention tunes between standards and simples must not drift more than 4 %withinananalytical run. Ifretention time drift exceeds this limit within ananalytical nmthenthe set mutt be reanalyzed.
14.0 Calculations
14.1 Use the followingequation tocalculatethe amount ofPFOA found (in ng/mL, based on peak area) using the standvd curve (linear regression parameters) generatedby dieMass Lynx softwareprogram:
PFOA found(ng/mL)-(Peak area -intercept! x DF x aliquot factor lope
DF factorby which the final volume was diluted, if necessary. Aliquot factor" 10
Paga S o n
P age 57 o f 65
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Page 126 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygen Renewh
Method Number V00017IJ
I ANALYTICAL method
MethodofAnalysts forthe DetanmnationofPeriluorooctanoic Acid (PFOA) in Small Mammal Liverby LC/MS/MS
14.2 For samples fortified with known amounts of PFOA prior to extraction, use tite followingequationto calculate thepercent recovery.
Recovery(%) -
[totalanalytefound(ng/mL) - analytefoundincontrol (ng/mL)] analyteadded(ng/mL)
14.3 Use the followingequationto convert the amount of PFOA found in ng/mL 10 nrttppb).
pfoa found(ppb) [PFOAfound(m /M.) x fimi YQlamfl (aUl sample weight (g)
Exygen Research
Page 7 o f 7 P age 58 o f 65
Page 127 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study N o.: P0001131
Exygen Protocol Number: P0001131
ANALYTICAL METHOD MethodNumber V0001786
Method ofAnalyala for the Determination of Perfieorooctaoolc Add (PFOA) in Small Mammal Serum by LC/MS/MS
Analytical TeetingFacility:
Exygen Research 3058 Research Drive
State College, FA 16801
Approved By:
c jUL
Paul Connolly
I
Technical Leader, LC-MS, Exygen Research
a/#? 4 / /
^JohnaFFllaahherty s ' Vice President, Operations, ExygenResearch
___ U l i f e M Date
/' J r
Due
Exygen Research
Total Pagca: 7
P age 59 o f 65
Page 128 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
ExygenReMuch
Method Number V000]716
I ANALVTICAt M IT H O P
MetbodofAnalysis fortheDeterminationofPerfluorooctanoic Acid (PFOA) in Small Mammal Serumby LC/MS/MS
1.0 Scope
This method ia to be employed for the isolation and quantitation of perfluorooctanoic acid by High Performance Liquid Chromatography coupled to a tandem Maes Speetrometric Detector(LC/MS/MS) in email mammal scrum.
2.0 Safety
2.1 Always obeervesafe laboratorypractices. 2.2 Consult foeappropriateMSDS before h*ndl"g any chemical for proper safety
precautions.
3.0 SampleRequirement
3.1 At least l mLofteet sample forextraction. 32 No sample procceeing is needed for serum samples. However, frozen serum
ample#must to allowedtocompletelythaw to roomtemperature before use. 3J Sample collection proeedurea will be specified in the sampling plan for (his
project
4.0 Reagents and Standards
4.1 Water-HPLC grade 4.2 Methanol-HPLC grade 4.3 Acetonitrile - HPLCgrade 4.4 AmmoniumAoetate-A.CS. Reagent Grade 4.5 PerfluorooctanoicAcid- Sigma-Aldrich
5.0 Instrument andEquipment
5.1 A high performance liquid chromatograph capable of pumping up to 2 solvents equipped with a variable volume uyoctor capable of injecting 5-200 pLconnectedto a tandemMesa Spectrometer (LC/MS/MS).
5.2 Adevice to collect rawdata forpeak integration and quantitation. 5.3 Analytical balance capableofreadingto 0.00001 g. 5.4 50mLdiqioiablepolypropylenecentrifoge tubes. 5.5 15 mLdisposablepolypropylenecentrifogetube. 5.6 Disposable micropipets (50-100uL, 10Q-200uL). 5.7 125-mLLDPEnarrow-mouthbottles. 5.8 2 mLclearHPLCvial lat. 5.9 Disposablepipettas. 5.10 Autopipettes(100-1000pL and 10-100pL), with disposable tipi. 5.11 Waters Sep Pak Vac 6 cc (1|) tCl8 SPE cartridges. 5.12 SPEvacuummanifold. 5.13 Vortexer.
onPage 2
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P001131
Method Number VOOO1786
A1WYT1CAL METHOD MethodofAnalysis tor theDeterminationofPeriluorooctanoic Acid (PFOA) in Small
Mammal Serumby LC/MS/MS
5.14 Wrist-actionshaker. 5.15 Centrifugecapableofspinning 15 mLpolypropylene tubes at 3000 rpm.
6.0 ChromatographicSystem
6.1 Analytical Column: Ftuophase RP (Keystone Scientific), 2.1 nun x 50 mm. 5p (P/N: 82505-052130)
6.2 Temperature: 30*C 6.3 Mobile Phase (A) : 2 mMAmmoniumAcetatein Water
Mobile Phase(B) : Methanol Gradient Program:
Time (mart
0.0 1.0 8.0 20.0 22.5
&A 65 65 25 25
65
Row Rate %B (mL/minN
35 0.3
3S 0.3 75 0.3 75 0.3 35 0.3
6.6 InjectionVolume: 15pL (canbe increseedto as much as 50 pL). 6.7 Quantitation: PeakAna - external standard calibration curve. 6.8 RunTime: ~ 23minutes.
The above conditions are intendedas a guide and may be changed in order to optimize theHPLCsystem.
7.0 MS/MSSystem
7.1 Mode: EloctrosprayNegative MRMmode, monitoring 413 -* 369 m/z for PFOA.
The above conditions are intended u a guide and may be changed in order to optimize the MSMS system.
8.0 PreparationofSolutions 8.1 Mobile Phase
8.1.1 2 mM ammonium acetate in water is prepared by adding 0.154 g of wmaniumacetate to 1000mLofwater.
Alternatevolumesmaybe prepared.
Page 3 of 7
P age 61 o f 65
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Page 130 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: POOO1131
Bxygta R w r e h
kteihod Number VOOOI786
I ANALYTICAL METHOD
MethodofAnalyiii fintheDetermmtlion ofPorfluoroocttnoic Acid (PFOA) in Small Mammal Serumby LC/MS/MS
9.0 StandardPreparation
9.1 StandardStock/FoitificstionSolution 9.1.1 Prepare a Mocksolution of -100 |ig/mL of PFOA by weighing 10 mg ofanalytical standard (corrected for purity) and dilute to 100 mL with methanol in a 125-mLLDPE bottle. 9.1.2 A 1.0 pgteiL fortification solution of PFOA is prepared by bringing 1 mL of the 100 pgfooLsolution to a final volume of 100 with methanol
ina 125mLLDPEbottle. 9.1.3 A0.1 ugfoiLfortification solutionof PFOA is prepared by bringing 10
mLofthe 1.0p^mL solutionto a final volume of 100with methanol in a 125mLLDPEbottle. 9.1.4 The stockand fortificationsolutions are to be stored in a refrigerator at approximately 4*C and are stable for a maximum period of 6 months fromthe dateofpreparation.
9.2 StandardCalibrationSolutions
9.2.1 LC/MS/MScalibrationstandards are prepared in methanol via dilution ofthe 0.1 MffaL fortificationsolution.
9 2 .2 The following is s typical example: additional concentrations may be prepared as seeded.
Concentration
Final
ofFortification Volume Dilutedto Concentration
Solution(uteri.) (mL) 100 5.0
(mL) 100
(ntteiL) 5.0
100 2.0 100 100 1.0 too 5.0 10 100
2.0 1.0 0.5
2.0 10 100
0.2
1.0 10 100
0.1
9.2.3 Store all calibration standards in 125*mL LDPE narrow-mouth bottles
at 2Cto 6*0, up tosix months.
9.2.4 Alternate volisne* and concentrations of standards may be prepared as
needed.
10.0 BatchSet 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 strike) to verifyprocedural recovery for the batch.
10.2 Requirements for field sad laboratory duplicates and spikes will be specified in the qualifyassuranceplan for this project.
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Page 131 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygsa Ri o rcb
M ated NumberV00017S6
I ANALYTICAL METHOD
Method ofAnalysis forfoeDetetminttion ofPerftuorooctanoic Acid (PFOA) inSmall Mammal Seramby LC/MS/MS
11.0 Sample Extraction
11.1 Measure 1mLofsample into a 50 mL polypropyhme centrifuge tubes (fortify u needed, replace lid and mix well). Note dial alternate volumes of serum maybe measureddependingon thesample size available for use.
11.2 Addwaterto thesample for a final volume of20 mL. Cap tightly 11.3 Vortex fbr-l minute. 11.4 Transfer 1 mL of die sample using a disposable pipette into a 15 mL
disposable centrifiigetube. 11.5 Add 5 mLofacetonitrile andshake for-20 minutes on ewrist-action shaker. 11.6 CentriAigethe tubesat -3000 rpm for ~5 minutes.
11.7 Decant the supernatant into a 50 mL disposable centrifiige tube and add 35 mL of water.
11.8 Condition foe Cu SPE cartridges (1 g, 6 mL) by passing 10 mL methanol followedby5 mLofHPLCwater (- 2 drop/aee). Do not let column run dry
11.9 Loadthe sample onconditionedCu SPE cartridge. Discard eluate.
11.10 Elute with -2 mL of methanol. Collect 2 mL of eluate into a graduated 15mLpolypropylenecentrifiigetube (final volume * 2 mL).
11.tl Analyzesamples usingelectrospray LC/MS/MS.
12.0 Chromatography
12.1 hgect tbe same amount ofeach 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 correspondingto at least five or more concentration levels
mustbe includedin at analytical set. 12.3 An attire set ofcalibrationstandards must be included a 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 a foe beginning of a set followed by calibration standards intaspersed every 5-10ssmples (to account for s second set ofstandards). In
either case, calibration standards must be foe first and last injection in a sample set. 12.4 Use linear standard curves for quantitation. Linear standard curves 8re generated fortheanalyte by linear regression using I/x weighting ofpeak area versus calibration standard concentration using MassLynx 3.3 (ex'equivalent) software system. 12.5 Sample reeponao should not exceed standard responses. Any samples that exceedstandardresponses shouldbe Antherdiluted and reanalyzed.
Page 5 of 7
P age 63 o f 65
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Page 132 of 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
Exygen Rteeaitb
Method Number V000I7S6
I ANALYTlCALMltTHOP
MethodofAnalytic fixthe Detenninstion ofPerfluoiooctanoic Acid (PFOA) in Small Mammal Seramby LC/MS/MS
]
13.0 AcceptanceCriteria
13.1 Chromatogrammutt chowa peak of a daughter ion at 369 amu tram a parent of 413 amu. The 413 amu parent correiponds to the PFOA anion, while tlw delimiterion(369amu) repreeeatadie loaaofcarbon dioxide.
13.2 Method blank! muct not contain PFOA at level! greater than the LOQ. if a blank contains PFOA at level! greater than 10 ng/mL, then a new blank camplemuctbe obtainedand theentire 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 apike falls outside the acceptable limits, the entire set of camples 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 wing the Huge Error Test, may be excluded from the calculation of the calibration curve However, foe total number of calibration standards that could be excluded must not exceed20%ofthe total numberofstandards injected.
13.5 The correlation coefficient (R) for calibration curves generated must be 0.992 (R* 0.965). If calibration results foil outside these limits, then appropriate steps must be taken to adjust instrument operation, and the standardsor foerelevant set ofsamples shouldbe 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 ran then the set must be reanalyzed.
14.0 Calculations
14.1 Use the followingequationto calculate the amount ofPFOA found (in ng/mL. baaed on peak area) using the standard curve (linear regression parameters! generatedbyfoeMast Lynx softwareprogram:
PFOAfound(ng/mL) /Peak gca - intercept! x DF x aliquot factor slope
DF foctorbywhichfoe final volume was diluted, if necessary. Aliquot factor 20
14.2 For samples fortified with known amounts of PFOA prior to extraction, use foe followingequationto calculatethepercent recovery.
Recovery(%) -
[total analytefound(ng/mL) - analytefoundincontrol(ng/mL)] analyteaddod(ng/mL)
Page 6 o f 7
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Page 133 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
Exygen Protocol Number: P0001131
E x y y n R m irch
Mtfbod Number VOOO1786
I ANALITICA!. METHOD
Method ofAntlyui fortheDetennination ofPerfluoroocUnolc Acid (PFOA) in Small Mammal Seramby LC/MS/MS
14.3 Uie die followingequation to convert foe amount of PFOA found in ng/mL to ppb.
PFOA found fppb) - fPFQA found faaAnLI X final volume fml.ll sample volume (mL)
Exygen Research
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Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
3058 Research Drive
Phone: 814-272-1039
S tate College, PA 16801
Fax: 814-231-1580
PROTOCOL AMENDMENT
Amendment Number 1 Effective D ate: 0 1/19/05 Exygen Study Number P0001131 Client Study N um ber
P a g e lo M
D ES C R IPTIO N O F AM ENDED S E C TIO N 1) Analytical Procedure Sum m ary V0001780:Section 9.1 2 ) Verification o f Analytical Procedure
NOffo
AMENDED TO 1) Add to Section 9.1: Section 9 .1 .6 , Alternate weights o f standards m ay be used to prepare alternate concentrations o f stock solutions as necessary. A lternate levels of fortification solutions m ay also be prepared. 2 ) Low and high spiking levels o f the analytes fo r each matrix m ay be altered depending on sam ple size available for extraction and/or to cover analyte concentrations expected in the sam ples.
R A T IO N A L E 1) Higher concentrations o f standards need to be prepared in order to spike the sam ple bottles at higher levels. 2 ) The sam ple size available fo r small mammal Iv e r and serum w as sm aller than expected. Spiking at the pre-determ lned levels in the protocol puts the spiked concentration lower than the detection lim it Also, th e analyte levels in th e ground w ater sam ples are expected to greatly exceed the pre-determ lned spiking levels listed in the protocol. W hen the levels in the sam ples greatly exceed the spiking levels, an accurate recovery value cannot be calculated fo r the Q C sam ple. Higher spiking levels in the bottles will cover the analyte concentrations expected in the w ater sam ples.
IMPACT QN STUDY The LOQ Is 100 ng/g for a 0.1 g sam ple o f small mam mal liver and is 1000 ng/m L for a 0.01 mL sam ple o f sm all m am mal serum . Higher levels of spiking for the w ater sam ples will ensure that more Q C recovery data can be used.
LIBRARY ID. W000122S4` . ..
''
ADMINISTRATIVE FORM
Exygen Research
Page 135 o f 136
Interim Report #21 - Analysis o f Soil Samples
Exygen Study No.: P0001131
3058 Research Drive
Phone: 814-272-1039
S tate College, PA 16801
Fax: 814-231 -1580
Amendment Number Effective Date: Exygen Study Number
PROTOCOL AMENDMENT 2
03/07/05 P0001131 Client Study Num ber
Page 1 of 1 None
D ES C R IPTIO N O F AM ENDED S E C TIO N 1) Report, page 11 o f 65 2 ) T est M aterials, page 6 o f 65: PFO S transition monitored 499 -> 99.
AMENDED TO 1) Instead o f one final report, interim reports will be issued.
2 ) PFO S transition monitored m ay also be 4 9 9 -> 80.
RATIO NALE
1) D ue to the excessive sizes o f fhe data sets, interim reports w ll be issued to allow the
client to receive data in a tim elier m a n e *
e *h l*r 4t)ex
2 ) The A P I 4 0 0 0 LC /M S /M S systems detect the 4 9 9 -> 80 PFO S transition with greater
sensitivity than the 4 9 9 > 99 transition.
IM PACT O N S TU D Y 1 ) T he client will be able to receive and review the data m ore quickly. 2 ) The 4 9 9 -> 80 transition can be detected with greater sensitivity; therefore, giving better chromatography.
LIBRARY ID: VD001226-8-
Exygen QAU Review JJ OsJesfe ADMINISTRATIVE FORM
Exygen Research
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