Document ZBrGRxxOxLL2JoQggBqpKMB2J
Project / Study Archive Request
I Shaded areas for Archivist use only I
Requestor Name
Kent Lindstrom
Phone
651 778.5352
Date
04105105
Request for:
Archival
~
0 Retrieval
Report
laFC I
I
LIMS number: E05-0210
I
Title/Summary: Interim Report #2 - Analysis of Surface Water 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 Executive Summary GrouD: 3M Decatur
~
Group Project Number: Exygen Protocol Number PO001131 Project Lead/ Study Director: Gary Hohenstein (data management), Mike Santoro (Sponsor), John Flaherty (Principal Investigator), Jaisimha Kesari P.E., DEE (Study Director, Weston Solutions, Inc.) Contract: NA
Contract Proiect #: NA
Analyte(s): PFBS, PFHS, PFOS
Test System (Sample Matrix): water
Project Type
Ix) Analysis
0Degradation
c]Duct Residue Analysis
0Ecotox
Method Validation
0NA 0Phys/Chem Properties
ResearchMethod Development
0 I
Regulatory Status:
Non-GLP
I
GLP Other:
m i a t i o T D a t e : 11/05/04 (Protocol Signed by Study Director), Analytical Start Date for
Interim Report: 02/14/05
Completion Date: Analytical Termination Date for Interim Report #2: 02/17/05
Signed by Study Director 03/23/05, Signed by Sponsor 03/30-31/05
Electronic Media:
Data CD:
FTIR Data Instrument Data 0None (raw data files archived at
Exygen)
Page 1 o f 2
Project / Study Archive Request
Archive Date
Scan Date
Key Words: Decatur Memorandum of Understanding, MOU, GLP, Protocol PO001131, PFBS, PFHS, PFOS, E05-0209, E05-0210, Interim Report #2
Comments:
Distribution Point Gary Hohenstein (Interim Report #2 PDF) Jai Kesari (Interim Report #2 PDF)
Date 04/05/05 04/05/05
Page 2 o f 2
INTERIM REPORT # 2-Analysis of Surface Water 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 REOUIREMENTS EPA TSCA Good LaboratoryPractice Standards40 CFR 792
STUDY DIRECTOR Jaisimha Kesari P.E., DEE
Weston Solutions, Inc. 1400 Weston Way
West Chester, PA 19380 Phone: 610-701-3761
INTERIM REPORT COMPLETION DATE March 22,2005
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: POOOl 131 Exygen StudyNumber: POOOl 131
Total Pages: 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
Exygen Study Number PO001131, entitled "Analysis of Perfluorobutanesulfonate(PFBS), Perfluorohexanesulfonate (F`FHS), 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 EPA TSCA Good Laboratory Practice Standards 40 CFR 792 by Exygen Research.
hn Flaherty
J
L n c i p a l Investigator
Exygen Research
Study Director Weston Solutions, Inc.
Michael A
F
Sponsor R esentative
3M Company
Exygen Research
& Date
Page 2 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen StudyNo.: POOOl 131
QUALITY ASSURANCE STATEMENT
Exygen Research's Quality Assurance Unit reviewed Exygen Study Number POOOl 131, 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 LCMSMS for the 3M Decatur Monitoring Program". All reviewed phases' 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
Date InsDected
Date Reported to Date Reported to
Principal
Exygen Date Reported to
Investigator Management Study Director
7. Raw Data Review 03/10,11/05 03/14/05
0311 8/05
0311 8/05
8. Interim Analytical Report Review
03/17/05
03/18/05
0311 8/05
0311 8/05
`Note: 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.
Exygen Research
Page 3 of 100
Interim Report #2-halysis of Surface Water Samples
Exygen Study No.: PO001131
CERTIFICATION OF AUTHENTICITY
This interim report, for Exygen Study Number POOO1131, 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:
O0L m f d 4
Exygen Research
3bdk
Date
Exygen Research Facility Management:
'Richard A. Gradzini
/
Date
President f -
Exygen Research
Study Directgr, Weston Solutions, Inc.
Weston Solutions, ~ n c . Sponsor Representative, 3M Company:
,
M a Michael A. Sant ro
P Director of Regulatory Affairs
Exygen Research
Date ' Page 4 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO00113I
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:
PO001131
EXYGEN STUDY NUMBER: PO00113I
TYPE OF STUDY:
Residue
SAMPLE MATRIX:
Surface Water
TEST SUBSTANCE:
Perfluorobutanesulfonate (PFBS), Perfluorohexanesulfonate(PFHS), and Perfluorooctanesulfonate (PFOS)
SPONSOR:
3M Company 3M Building 0236-01-B-10 St. Paul, MN 55144
STUDY DIRECTOR:
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:
02/14/05
Interim Analytical Termination Date: 02/17/05
Interim Report Completion Date: 03/22/05
Exygen Research
Page 5 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
PROJECT PERSONNEL
The Study Director for this project is Jaisimha Kesari at Weston Solutions, Inc. The following personnel fiom Exygen Research were associated with various phases of this interim portion of the study:
Name John Flaherty Karen Risha Paul Connolly Chrissy Edwards Mark Ammerman Amy Sheehan
Title
Vice President Scientist
Technical Lead-LCMS Technician
Sample Custodian Associate Scientist
Exygen Research
Page 6 of 100
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No .. PO001131
TABLE OF CONTENTS
Pane
TITLE PAGE ....................................................................................................................... 1 GOOD LABORATORYPRACTICE COMPLIANCESTATEMENT.............................. 2 QUALITY ASSURANCE STATEMENT .......................................................................... 3 CERTIFICATIONOF 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 ............................................................................................................... 11 3.0 INTRODUCTION....................................................................................................... 11 4.0 ANALYTICALTEST SAMPLES.............................................................................. 12 5.0 REFERENCE MATERIAL ........................................................................................ 12 6.0 DESCRIPTION OF ANALYTICALMETHOD ........................................................ 13
6.1. Extraction Procedure ............................................................................................... 13 6.2 Preparation of Standards and Fortification Solutions............................................... 13 6.3 Chromatography ....................................................................................................... 14 6.4 Instrument Sensitivity............................................................................................... 15 6.5 Description of LC/MS/MS Instrument and Operating Conditions........................... 15 6.6 Quantitation and Example Calculation ..................................................................... 16 7.0 EXPERIMENTAL DESIGN ...................................................................................... 17 8.0 RESULTS ................................................................................................................... 17 9.0 CONCLUSIONS......................................................................................................... 17 10.0 RETENTION OF DATA AND SAMPLES ............................................................. 17
Exygen Research
Page 7 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Table I.
Table II.
LIST OF TABLES
Pane
Summary of PFBS, PFHS and PFOS in Surface Water Samples...................19
Matrix Spike Recovery of PFBS, PFHS and PFOS in Surface Water
Samples.......................................................................................................... .20
Exygen Research
Page 8 of 100
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
LIST OF FIGURES
Pane Figure 1. Typical Calibration Curve for PFBS in Water................................................ 23
Figure 2. Extracted Standards of PFBS in Water, 50 ng/L and 100 ng/L,
Respectively................................................................................................... .24
Figure 3. PFBS in Reagent Water, 500 ng/L Fortified Reagent Water, and
5000 ng/L @F=lO) Fortified Reagent Water, Respectively......................... ..25
Figure 4. Chromatogram Representing a Surface Water Sample Analyzed
for PFBS (Exygen ID: C0052282, Data Set: 021405C) ................................. 26 Figure 5. Typical Calibration Curve for PFHS in Water ............................................... 27
Figure 6. Extracted Standards of PFHS in Water, 50 ng/L and 100 ng/L,
Respectively.. ................................................................................................. .28
Figure 7. PFHS in Reagent Water, 500 ng/L Fortified Reagent Water, and
5000 ng/L @F=lO) Fortified Reagent Water, Respectively.. ......................... 29
Figure 8. Chromatogram Representing a Surface Water Sample Analyzed
for PFHS (Exygen ID: C00522831, Data Set: 021405C) ............................... 30 Figure 9. Typical Calibration Curve for PFOS in Water ............................................... 31
Figure 10. Extracted Standards of PFOS in Water, 50 ng/L and 100 ng/L,
Respectively.. ................................................................................................. .32
Figure 11. PFOS in Reagent Water, 500 ng/L Fortified Reagent Water, and
5000 ng/L @F=lO) Fortified Reagent Water, Respectively.......................... .33
Figure 12. Chromatogram Representing a Surface Water Sample Analyzed
for PFOS (ExygenID: C0052282, Data Set: 021405C) ................................. 34
Exygen Research
Page 9 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOOl 131
LIST OF APPENDICES
PaRe
Appendix A Study Protocol POOOl 131 (Exygen Study No. POOOl 131)with Analytical Method: VOOOl780:"Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by
LC/-MS/MS"...............................................................................................35
Exygen Research
Page 10 of 100
Interim Report #2-halysis of Surface Water Samples
Exygen Study No.:POOOl131
1.0 SUMMARY
Exygen Research extracted and analyzed surface water samples for the determination of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) according to Exygen Method VOOOl780 (Appendix A).
The limit of quantitation for PFBS, PFHS and PFOS in water was 50 n g L and the limit of detection was 25 ng/L.
The PFBS, PFHS and PFOS residues found in surface water samples are summarized in Table 1.
The average percent recoveries L- standard deviations for PFBS, PFHS, PFOS in surface water samples were 139% 2 21%, 95% 2 7%, and 96% _+ 12%, respectively.
2.0 OBJECTIVE
The objective of the analytical part of this study was to determine levels of perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), and perfluorooctanesulfonate (PFOS) in surface water according to Protocol POOOl 131 (Appendix A).
3.0 INTRODUCTION
This report details the results of the analysis for the determination of PFBS, PFHS and PFOS in surface water using the analytical method entitled, "V0001780: Method of
Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by
LCMSMS."
The study was initiated on November 05, 2004, when the study director signed protocol number POOOl 131. The analytical start date for this interim report was February 14, 2005, and the analytical termination date for this interim report was February 17,2005.
Exygen Research
Page 11 of 100
Interim Report#2-Analysis of Surface Water Samples
Exygen StudyNo.: PO001 131
4.0 ANALYTICAL TEST SAMPLES
Forty-four surface water samples (Exygen ID COO52282-COO52299 and COO52310C0052335) were received on wet ice on December 04, 2004 from Charles Young at Weston Solutions, Inc. The samples were logged in by Exygen personnel and placed in refrigerated storage.
Sample log-in and chain of custody information will be kept at Exygen Research until the conclusion of the study; they will then be located in the raw data package associated with this study. 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 July, 06, 2000. 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
Exygen Inventory No.. SP0000252 SP0002401 SP0002694
Lot # 101 SE036 430180-1
Purity (%) 96.7 98.6 101.2
Expiration Date 12/04/06 10118/06 04/23/06
The molecular structures of PFBS, PFHS and PFOS are given below:
PFBS
Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (C4F$03X+)
Transitions Monitored: 299 + 99
Structure:
F
F
Exygen Research
Page 12 of 100
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight: 438 supplied as the potassium salt (CsF13S03-K')
Transitions Monitored: 399 + 80
Structure:
PFOS Chemical Name: Perfluorooctanesulfonate Molecular Weight: 538 supplied as the potassium salt (C8F17S03-K')
Transitions Monitored: 499 + 80
Structure:
6.0 DESCRIPTION OF ANALYTICAL METHOD
The analytical method "V0001780: Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" was used for this study.
6.1. Extraction Procedure
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 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.
Exygen Research
Page 13 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
6.2 Preparation of Standards and Fortification Solutions
Individual stock standard solutions of PFBS, PFHS and PFOS were prepared as specified in Exygen method V0001780. The stock standard solutions were prepared at a concentration of 100 pg/mL by dissolving 10 mg of each of the standards (corrected for purity and salt content) in methanol. From these solutions, mixed 1.O pg/mL fortification standard solutions were prepared by taking 1 mL of each of the appropriate stock solutions and bringing the volume up to 100 mL with methanol. By taking 10 mL of the mixed 1.0 pg/mL fortification standard and bringing the volume up to 100 mL with methanol, a mixed 0.1 pg/mL fortification standard was prepared. By taking 10 mL of the mixed 0.1 p g / d fortification standard and bringing the volume up to 100 mL with methanol, a mixed 0.01 pg/mL fortification standard was prepared.
A set of standards containing PFBS, PFHS and PFOS was prepared in water and processed through the extraction procedure, identical to samples. The following concentrations were prepared:
Conc. of Fort Fort
Solution
Volume
(ng/mL)'
(pL)
0
0
10
100
10
200
10
400
100
100
100
200
* 100
400
of PFBS, PFHS and PFOS
Volume of Fortified Sample
(mL)
40 40 40 40 40 40 40
Final Conc. of Calibration Std.
(ng/L) 0 25 50 100
250
500 1000
The stock standard solution and all fortification and calibration standard solutions were
stored in a refrigerator (4" f 2OC) when not in use. Documentation of standard
preparation will be kept at Exygen Research until the conclusion of the study; it will then be located in the raw data associated with this study.
6.3 Chromatography
- - - Quantification of PFBS, PFHS and PFOS was accomplished by L C N S N S electrospray.
The retention times of PFBS, PFHS and PFOS were 3.9mins, 10.4mins, and 12.6 mins, respectively. Peaks above the LOD were not detected in any of the reagent blank samples corresponding to the analyte retention time.
Exygen Research
Page 14 of 100
Interim Report #ZAnalysis of Surface Water Samples
Exygen StudyNo.:PO001131
6.4 Instrument Sensitivity
The smallest standard amount injected during the chromatographic run had a concentration of 25 ng/L of PFBS, PFHS and PFOS.
6.5 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.: 35' C Injection Vol.: 15 pL Mobile Phase (A): 2 mM Ammonium Acetate in water Mobile Phase (B): Methanol
Time (rnin)
%A
0.0
65
35
1.o
65
35
8.0
25
75
10.0
25
75
11.0
65
35
18.0
65
35
Total run time: -18 min
Flow Rate: 0.3 mL/min
Ions monitored:
Analvte
PFBS PFHS PFOS
Mode
negative negative negative
Transition Monitored 299 -+ 99 399 -+ 80
499 + 80
Approximate Retention Time
(min) -3.9 min.
-1 0.4 min. -12.6 min.
Exygen Research
Page 15 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
6.6 Quantitation and Example Calculation
Fifteen microliters of sample or calibration standard were injected into the L C M S M S . The peak area was measured and the standard curve was generated (using l/x fit weighted linear regression) by Analyst software using six concentrations of standards. The concentration was determined fiom the equations below.
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 calculated the percent recovery.
Equation 2: Recovery (%) =
(analyte found ( n a ) - analyte in control (ny./L)) x 100%
amount added (n&)
An example of a calculation using an actual sample follows (calculation is for PFOS
only):
Water sample Exygen ID: COO52282 Spk C (Set: 021405C), fortified at 500 ng/L
with where: peak area intercept slope dilution factor
- 84394
- 3070
-
162
-
1
ng/L PFOA added (fort level) = 500
amt in corresponding sample = 0 (Not Detected)
From equation 1:
Analyte found (ng/L) = 184394- 30701x 1
162
From equation 2: % Recovery
= 502.0ng/L
= (502.0 - 0 nn/L) x 100% 500
= 100%
Exygen Research
Page 16 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
7.0 EXPERIMENTAL DESIGN
For samples designated as field matrix spikes, PFBS, PFHS and PFOS were added at a known concentration to the bottles in the laboratory before being shipped to the field. The samples were filled to a 200 mL volumetric fill line in the field.
The samples were extracted in two sets. Each set consisted of one reagent blank, two reagent blanks fortified at known concentrations. One set contained five sample sites and the second set contained four sample sites, along with all the field blanks and field blank spikes collected for the surface water samples. For each site, a sample, a field duplicate and two matrix field spikes were collected. For each site, a laboratory duplicate was extracted and two laboratory matrix spikes were also extracted.
8.0 RESULTS
The PFBS, PFHS and PFOS found in the surface water samples are listed in Table I.
Fortification recoveries for PFBS, PFHS and PFOS in the water samples are detailed in Table 11. The average percent recoveries k standard deviations for PFBS, PFHS, PFOS in water samples were 139% f 21%, 95% f 7%, and 96% k 12%, respectively.
9.0 CONCLUSIONS
The surface water samples were successfully extracted and analyzed for PFBS, PFHS and
PFOS according to analytical method V0001780.
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 sponsor. 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
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Interim Report #2-Analysis of Surface Water Samples
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Exygen StudyNo.:PO001131
TABLES
Exygen Research
Page 18 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Table I.
Summary of PFBS, PFHS and PFOS in Surface Water Samples
Exygen ID
COO52282 COO52282 Rep
COO52283 COO52286 COO52286 Rep COO52287 COO52290 COO52290 Rep COO52291 COO52294 COO52294 Rep COO52295 COO52298 COO52298 Rep COO52299 COO52312 COO52312 Rep COO52313 COO52316 COO52316 Rep COO52317 COO52322 COO52322 Rep COO52323 COO52326 COO52326 Rep COO52327 COO52330 COO52333 COO52320 COO52321
Client
C4 Sulfonate PFBS
Sample ID
Peffluorobutanwulfonat.
DL3-SW-LOCOOI-0-041201
ND
DL3-SW-LOC001-0-041201'
ND
DL3-SW-LOC001-0-041201 DuP
ND
DL2-SW-LOC001-0-041201
ND
DL2-SW-LOCOOI-0-041201'
ND
DL2-SW-LOC001-0-041201 D u ~
ND
DBC-SW-LOC001-0-041202
93.8
DBC-SW-LOCOOI-0-041202'
96.2
DBC-SW-LOC001-0-041202 DUp
97.5
DOU-SW-LOC001-0-041202
14000
DOU-SW-LOC001-0-041202'
14700
DOU-SW-LOCOOI-0-041202D u ~
12800
DDO-SW-LOC001-0-041202
ND
DDO-SW-LOCOOI-0-041202'
ND
DDO-SW-LOC001-0-041202 DuP
ND
DLI-SW-LOC001-0-041202
76.8
DLI-SW-LOC001-0-041202'
77.4
DLI-SW-LOCOOI-0-041202 Dup
77.4
DMC-SW-LOCOOI-0-041202
I07
DMC-SW-LOC001-0-041202'
97.6
DMC-SW-LOCOOI-0-041202D u ~
104
DAA-SW-LOC005-0-041202
2460
DAA-SW-LOCOO5-0-041202'
3270
DAA-SW-LOCOO5-0-041202 DUP
3100
DAA-SW-LOCOO2-0-041202
379
DAA-SW-LOCOO2-0-041202'
456
DAA-SW-LOC002-0-041202D u ~
432
Field Blank
ND
Field Blank
ND
DMC-SW-LOCOOI -2-041202
ND
DMC-SD-LOCOOI -2-041202
ND
Analyte Found (ppt, n@L) C6 Sulfonate PFHS
Perlluoroheunuulfonat.
ND ND ND ND ND ND 231 233 252 1620 1650 1600 ND ND ND 45.7 48.3 49.2 53.6 49.5
54.0
7980 7350 7880 375 433 445 ND ND ND ND
C8 Sulfonate PFOS
Perlluorooctanuulfonate
ND ND ND ND ND ND 3460 2880 3920 5710 5820 5640 ND ND ND 179 185 213 284 257 283 18800 18000 19600 2780 2930 3760 ND ND ND ND
'Laboratory Duplicate
ND = Not detected at or above 25 ng/L. NQ = Not quantifiable = Measuredconcentrationbetween25 ng/L and the Limitof
Quantitation(LOQ)which is 50 ngL.
Exygen Research
Page 19 of 100
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Interim Report #ZAnalysis of Surface Water Samples
Exygen Study No.: PO001131
Table 11. Matrix Spike Recovery of PFBS, PFHS and PFOS in Surface Water Samples
Sample Description DL3-SW-LOCO010441201 (CWSUU SpkC. 500 ndL Splke)
DL3-SW-LOC0010041201 (COOSUU Spk 0. WOO Wl. Splke)
DL3-SW-LOCOO14441201Low Spk (COOSUM. 500 ndL SpuU)
DL3-SW-LOCOOl0-041201 High Spk (CWSU8S. 5OOO n e t Spllu)
x Spikec
500 5oOo 500 5000
C4 SUlfOMte PFBS
W F o u n d Amount
in Sample Recovered Recovei
(npll)
(ngiL)
(54)
ND
748
150
ND
7090
142
ND
694
139
ND
7320
146
C6 Sulfonate PFHS
4ntFound Amount
In Sample Recovered Recover
(nm)
(%)
ND
485
97
ND
4560
91
ND
496
99
ND
4580
92
DLZ-SW-LOC0010-041201
(cwszznspk E. npnsplke)
500
ND
779
156
ND
516
103
DLZ-SW-LOC0010041201
(COOSZZM Spk F. WOO ndL S p W
5000
ND
7200
144
ND
4630
93
DLZ-SW-LOCOO10441201Low Spk
(CMSUU.500 npnSplb)
500
ND
734
147
ND
492
98
DLZ-SW-LOC0010-041201 High Spk
(cooaur. moo non spike)
5000
ND
7200
144
ND
4660
93
DBC-SW-LOCOOl0-041202 (CooOnO Spk0. W n#L Splke)
500
93.8
819
145
231
714
97
DBC-SW-LOCWl0-041202
(CooOnO Spk H. WOO npn Splke)
5oM)
93.8
7330
145
231
5060
97
DBC-SW-LOC0010-041202 Low Spk
(CM52n2.500ndL Spike)
500
93.8
742
130
231
667
87
DBC-SW-LOCOOl0-041202High Spk
(COOa?nl. 5OOO ndL Spllu)
5ooo
93.8
6830
135
231
4600
87
DOU-SW4OC0010-041202 (CooUnr Spk I. 500 npn Splke)
500
14000
12200
DOu-Sw-LOC0010-041202 (COO5ZZMSpk J. WOO n f l Splke)
so00
14000
14800
16"
DOU-SW-LOCO010041202LOWSpk
(CMa2w.500 ndL Splke)
500
14000
15200
DOU-SW-LOCW14-041202High Spk
(CWS22#7. 5OOO nwL Spike)
5Ooo 14000
14100
2"
1620
2130
102
1620
6160
91
1620
21 50
106
1620
5720
82
DDO-SW-LOC0014-041202 (COOS2288SpkK. 500 n#L SpuU)
500
ND
eo5
161
ND
482
96
DDO-SW-LOCO014441202
(CoosUrC SpkI..WOO n p n S p h )
5oOo
ND
7240
145
ND
4590
92
DW-SW-LOC0014-041Z02 Low Spk
(coo62alo. mn a spy.)
500
NO
794
159
ND
517
103
- DDOSW-LOCOO10041202Hlgh Spk
5oOo
ND
5310
106
ND
4680
94
*Samplemidue exweds hsplking level slgnlficently;therefore.an accuraterecoveryvalue cannot be calculated
**Pantwas removed hornVIE calwlatlonof average and standarddeviationa8 per email requestdated 0321105.
ND = Not detectedat OT above 25 ngn (PPI). NQ = Not quantifiaMe= Maaswed concentratlionbe(ween25 I@.(ppt) and !he Limit of
Quantllatlon ( L a ) which I8 50 I@.(ppt).
Note: Sin- thb summaryt8Ms show rounded naultr. recoveryvelmsmayvarysllghtly
from tho n l m s In the raw d8h.
C8 Sulfonate PFOS
MFound Amount In Sample Recovered Recovery
ND
502
100
ND
4450
89
ND
506
102
ND
3840
77
ND
490
98
ND
4680
94
ND
515
103
ND
5440
109
3460
3840
3460
7930
89
3460
3800
3460
7090
73
5710 5710 571 0 5710
5820
10300
92
6720
9350
73
ND
505
101
ND
4600
92
ND
517
103
ND
5ooo
100
Exygen Research
Page 20 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen StudyNo.: PO001131
Table 11. Matrix Spike Recovery of PFBS, PFHS and PFOS in Surface Water Samples Continued
Sample Description DLlSW-LOCO010041202
(COOPS12spk c. WO IldL S*)
DLlSW-LOC0010441202 (COOS2312Spk 0, Iwo ndL Spit.)
DLI-SW-LOCO010041202 Low Spk (CW52314. WO ndL Splka)
DLl-SWIOCOO10011202 High Spk (COO62115,Woo ndL Splka)
GG
Spiked
&f&k
500
5000
500
5000
C4 Sulfonate PFBS untFound Amount nSarnple Recovered Recover
C6 Sulfonate PFHS
h t F o u n d Amount
In Sample Recovered Recover!
C8 Sulfonate PFOS
4mtFound Amount
in Sample Recovered Recovery
(ng/L)
(ng/L)
(%)
76.8
849
154
45.7
556
111
179
707
106
76.8
7280
144
45.7
4810
96
179
4800
92
76.8
820
149
45.7
566
113
179
726
109
76.8
7970
158
45.7
4700
94
179
4100
78
DMCSWIOC0010-041202 (COO62311Spk E. 600 Spit.)
500
107
DMC-SWIOC0010011202
(COO62316Spk F, 6ooon@LSplka)
5000
107
DMC-SW-LOC0010-041202 Low Spi
(CWSZ311, 600ndL Spllu)
500
107
DMCSW-LOCOOl-0441202High Spl
(COO623lr.Woo n@ SpUu)
5000
107
893
1 57
53.6
556
100
284
7780
153
53.6
4740
94
284
786
136
53.6
544
98
284
6480
127
53.6
4560
90
284
764
96
51 30
97
807
105
3890
72
DAA-SW-LOCOO50011202 (COO62S22 Opk G. WO ng& Spit.)
DAA-SW-LOC0050441202 (COO62U2 spk H. Woo ng& Spit.)
DAA-SW-LOC0050041202Low Spk (CW52124.500ndL SplY
DAA-SW-LOCO050041202 Hbh Spl (COOPUS. ndL S p k )
500
5000 500 5000
2460 2460 2460 2460
4100
6840
88
3840
10800
1 67
7980
9270
7980
122M)
84
7980
8290
7980
11900
78
18800
23500
18800
23600
96
18800
20500
18800
20200
28"
DAA-SW-LOCOO2-0011202 (COOPUI Spk I,WO ndL Spit.)
DAA-SW-LOC0020041202 (COOPS26 Spk J. Woo m#L SpUu)
DAA-SW-LOC0024-041202Lav Spk (CW52SZl.600ndL Splka)
DAA-SW-LOCOO20011202HighSpk ICW52329. Woo n e t S P W
500 5000 500
5000
FleM Blank Low Spk (CW52SS1. WO ndL SpHu)
Fleld Blank High Spk (COUZUI. Woo ndL SpUu)
FIeld Blank Low Spk (cmezrw.noo ndL spwu)
Flold Bbnk High Spk
500
5ooo 500
5000
cilpc
379
1250
174
379
6430
121
379
762
77
379
7850
149
ND
594
119
ND
6070
121
ND
625
125
ND
6550
131
Average: 139 Standard Deviatlon: 21
375
891
103
375
5320
99
375
860
97
375
51 80
96
ND
458
92
ND
4670
93
ND
475
95
ND
4450
89
Average: 95 Standard Deviation: 7
2780 2780 2780 2780
3530
8310
111
3440
7740
99
ND
574
115
ND
5470
109
ND
541
108
ND
51W
102
Average: 96 Standard Deviatlon: 12
'Sample residue exceedsthe splkicq levelsQniflcanUy,lherefore.an armrate recoveryvalue cannolbe CalaMed
"Point was removBd fmnthe calculation of average and standard deviationas per m i l requestdated 03R1/05. ND = Not d m e d at m above 25 ngA (ppt). NQ = Not qwntlflable= Measured wnc~ntrationbe(ween25 n(lll(ppt) and the Llrnit of
Quanutation(LOQ) whkh k 50 ngA (ppt).
Not.: Slnw Ullssummry tab* shows rounded m u b . mwvoryvalues mynrySllght)y from tha valuos In the raw data.
Exygen Research
Page 21 of 100
Interim Report #2-halysis of Surface Water Samples
Exygen Study No.:PO001131
FIGURES
Exygen Research
Page 22 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 1. Typical Calibration Curve for PFBS in Water
021405C nvl.rdb (PFBS): "Unrar" Regression (Y /x"weighting): y = 52.7 x + 0.000802 (r = 0.9884)
0 .o
0
100
200
300
400
500
600
700
800
900
1000
Concentration. ng/mL
Exygen Research
Page 23 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Figure 2. Extracted Standards of PFBS in Water, 50 ng/L and 100 ng/L, Respectively
3.83
v)
P u
-%c(0
-Q1
c
2
4
6
8
10
12
14
16
I - - Time. min XCO21105-3, I00 nglL Standard PFBS (Standard)299.0190.0 amu sample 4 of 52 from 021405C.wiff Area: 5100 counts Height 105. cps RT: 4.00 min
- v) 100
a.
0
zi 50-
0)
c
-cc Q)
0
4.00
2
4
6
8
10
12
14
16
Time. min
Exygen Research
Page 24 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
Figure 3.
PFBS in Reagent Water, 500 ng/L Fortified Reagent Water, and 5000 ng/L @F=lO) Fortified Reagent Water, Respectively
n u
cQ)
0'
c
~
2
4
6
8
10
12
14
16
- Time. min
IReagent Spike A, 500 nglL- PFBS (QC)2QQ.O/QQ.0 amu - sample 10 of 52 from 021405C.wiff
Area: 26775 counts Height: 633. cps RT: 3.93 min
v)
E?
3.83
500
i5vc0) 0 0.83 ,I.#
5.866.437.20 8.729.55- 10.13 11:56,lI.Q5
c c
2
4
6
8
10
12
- Time, min
I - - Reagent Spike 8.5000 nglL. DF=lO PFBS (QC)2QQ.ORQ.0 amu sample 11 of 5 2 from Area: 25448 counts Height: 586. cps RT: 3.94 rnin
14.0a14.37 j 5 . 6 4
14
16
021405C.wiff
17.36
0)
0 P
- 500 Z iv)
c
-cc Q)
3.94
5.00
2
4
6
8.00,8.45
8
10
Time, min
11.71,12.1013.04 12
,13.72 14
15.4c15.77 17.55 16
Exygen Research
Page 25 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 4.
Chromatogram Representing a Surface Water Sample Analyzed for PFBS (Exygen ID: C0052282, Data Set: 021405C)
- COO52282 - PFBS (Unknown}299.0)99.0 amu sampfe 75 o f 5 2 from 021405C.wiF (peak nof b u n d )
- 180 0.58 - 140 - 120
100u
ziv)
c
a,
c c
- -60
40- 8 . 0 2
13.04
Exygen Research
Page 26 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 5. Typical Calibration Curve for PFHS in Water
021405C rev1.rdb (PFHS): "Linear" Regressionc1 I Y" weighting): y = 187 Y + -0.000613 (r = 0.99C14)
,2.O0e5
- 1 . 8 ~ 5
IBe5 -
1.4e5 -
- c v) 1 . 2 ~ 5
c
3
0
- 0
1.0&
m-
2(I) 8.0e4-
6.Oe4 -
4.0e4 -
0
I00
200
300
400
500
800
700
800
900
1000
Concentration, nglmL
Exygen Research
Page 27 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 6. Extracted Standards of PFHS in Water, 50 ng/L and 100 ng/L, Respectively
I - XEO27705-2, 50 ng/L Standard PFHS mandard] 39LO#LO alwu -sample 3 of 52 from 027405twiff Area: 8?27counts might: 69s c p RT:7a4min
10.42
I - Time, min XCO21105-3, 100 nglL Standard PFHS (Standard)399.0#0.0 amu -sample 4 of 52 from 021405C.wiff Area: 16628 counts Height: 1380. cps RT: 10.5 min
E(0 1000:
&v)
500-
- c
I=
0-
2.32
Time, min
11.39 12.47 ,13.40
16.78
Exygen Research
Page 28 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 7.
PFHS in Reagent Water, 500 n g L Fortified Reagent Water, and 5000 ng/L @F=lO) Fortified Reagent Water, Respectively
I - Reagent Cortro#A'FP @ea¬ found]
(UtAnown]39SWLLOamu -sample 9 of 52from 0214U5Cwifl
u)
8 --ZO
s.-
(0
c
0
-c 0C -)
2
'4
6
10.46
8
10
12
14
16
Time, min
I - Reagent Spike A, 500 nglL- PFHS (QC)399.0#0.0 amu sample 10 of 52 from 021405C.wiff
Area: 94857 counts Height: 7770. cps RT: 10.5 min
u)
Q u
10.46
i5cu)
c 0 Q)
c
2
4
6
8
10
12
14
16
- Time. min
I - - Reagent Spike 8,5000 nglL. DF=lO PFHS (QC)399.0#0.0 amu sample 11 of 52 from 021405C.wiff
Area: 80963 countr Height: 6530. cps RT: 10.4 min
u)
u P 5000
-63
c
Q)
0
c c
10.43
2
4
6
8
10
12
14
16
Time, min
Exygen Research
Page 29 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 8.
Chromatogram Representing a Surface Water Sample Analyzed for PFHS (Exygen ID: C00522831, Data Set: 021405C)
140
120
100
u)
Q
is0 80 cu)
-cc Q)
60
40
20
0
10.47
2
4
6
8
10
12
14
16
Time, min
Exygen Research
Page 30 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
Figure 9. Typical Calibration Curve for PFOS in Water
021405C rev1.rdb (PFOS): "Linear" Regression Cl I x" weighting): y = 182 x + 3.07e403 (r = 0.9978)
1.8e5 ,
c v)
c
J 0
u
m-
Concentration, nglmL
Exygen Research
Page 31 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO00113I
Figure 10. Extracted Standards of PFOS in Water, 50 ng/L and 100 ng/L, Respectively
I - - XCO27705-2, 50 np/L Standard PFOS (Standard) 49%0/80.0 ~ I N I J samph 3of 52 from 0274flJCwiff Area: 9937counts might: 505. cpr RT: 726 min
12.60
I - - Time. min XCO21105-3. 100 nglL Standard PFOS (Standard)49.0#0.0 amu sample 4 of 52 from 021405C.wiff Area: 16106 counts Height: 925. cps RT: 12.6 min
I
v1 P
u
& 500-
c0 )
-cQ)
4-
0-
1.80
1 3 . 6 44~.01
Exygen Research
Page 32 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 11. PFOS in Reagent Water, 500 n g L Fortified Reagent Water, and 5000 ng/L @F=lO) Fortified Reagent Water, Respectively
v)
B -
12.64
i5 - _ _ cv)
3.55 4.06 6.31
8.71, 10.14,10.66
c 0
c
2
4
6
8
10
12
14
16
- Time, min
I - R e a g e n t Spike A. 500 n g l L - P F O S (QC)4QQ.0#0.0 a m u sample 10 of 52 from 021405C.wiff
Area: 78386 counts H e i g h t 5080. cps R T : 12.6 m i n
v)
P
- 5000
i5v)
cc 0 c Q)
12.62
12.18,
I
2
4
6
8
10
12
14
16
- Time, min
I - - R e a g e n t Spike 8.5000 nglL. D F 4 0 P F O S (QC)4W.0#0.0 a m u sample I 1 of 52 from 021405C.wiff
Area: 72672 counts H e i g h t 4660.cps R T : 12.6 m i n
v)
Q u
&0 )
c
Q)
0
-c c
12.82 12.16.
2
4
6
8
10
12
14
16
Time, min
Exygen Research
Page 33 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Figure 12. Chromatogram Representing a Surface Water Sample Analyzed for PFOS (Exygen ID: C0052282, Data Set: 021405C)
I - COO52282 PFOS (1Rrk~owAI4J980/8LOamu =sample75 of 52 from 027405Lwiff Area: 3595 counts Might: 885 cpt R E f27 min
12.68
80
I
.56
70 -
60 -
50 -
40-
12.17.
30-
20 -
10
I 13.74 ~ L A4.18
0
2
4
8
8
10
12
14
I6
Time, min
Exygen Research
Page 34 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO00113I
APPENDIX A
Study Protocol PO001131
(Exygen Study No. P0001131) with Analytical Method: VOOO1780:66Methodof
Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water
by LCIMSIMS"
Exygen Research
Page 35 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOOl131
Exygen Protocol Number: PO001 13 I
STUDY PROTOCOL
Study Title: Analysis of Perfluorobutanesulfonate,(PFBS),
Periluorohexanesulfonate(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: POOOl 131
Performine Laboratorv:
Exygen Research
3058 Research Drive
State College, PA 16801
Phone: (814)272-1039
SDonsor Rcmesentative: Michael A. Santon,
Director of Regulatory Affairs 3M Building 0236-01-B-10
St. Paul, MN 55144 Phone: (651)733-6374
Exygen Research
Page 36 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
ExygenProtocol Number: PO00I I3 1
DISTRIBUTION:
1) JaisimhaKesari, Study Director, Weston Solutions 2) John M. Flaherty, PrincipalInvestigator,Exygen Research 3) Michael A. Santoro, Sponsor Representative, 3M Company 4) Exygen Research QualityAssurance Unit
Exygen Research
Page 37 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOOl131
- PROTOCOL APPROVAL
Exygcn Protocol Number: PO001 13 1
Study Title: Analysis of 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:POOOl 131
APPROVALS
-
I
Weston Solutions
P
Michael A. Sj&toro, Sponsor Representative 3M Compady
Fohn M. Flaherty, hincipal Investigator
/ Exygcn Research
Exygcn R e s e w
Clr
ent. Facility Management
i/ N 0
Date
fl-w-w#
Date
P-
Exygen Research
Page 38 of 100
~
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number: PO001131
TABLE OF CONTENTS
TITLE PAGE ....................................................................................................................
.I
DISTRIBUTION.................................................
.......
..................................... 2
PROTOCOLAPPROVAL................................................................................................................................ 3
TABLE OF CONTENTS...................................................................................................................
INTRODUCTION..........................................................................................................
TEST MATERIALS.
................................................................................................ 5
OBJECTIVE..................................................................................................................................................... 6
TESTING FACILITY..............................................
STUDY DIREaOR......... .................................................................................................................. 7
SPONSOR REPRESENTATIVE..................
PRINCIPAL INVESTIGATOR........................................................................................................................ 7
- PROPOSED EXPERIMENTAL START AND TERMINATTON DATES ... IDENTIFICATION AND JUSTIFICATION OF THE TEST SYSTEM ............... SAMPLE PROCUREMENT, RECEIPT AND RETENTION ............ SAMPLE IDENTIFICATION........................................................................ ANALYTICALPROCEDURESUMMARY ................................................................................................... 9
VERIFICATION OF ANALYTICALPROCEDURE...........
METHOD FOR CONTROL OF BIAS .............................................
STATISTICAL METHODS ............................................................................................................................. I I
GLP STATEMENT ................................
REPORT ........................................................................................................................................................... 1 1
SAFETY AND HEALTH .......................
...........
AMENDMENTSTOPROTOCOL........ ............................................................................................ 13
DATA RECORD KEEPING ............................................................................................................................ 13
QUALITY ASSURANCE ............
.............. ...........................................
...........14
RETENTIONOF DATA AND ARCHIVING ................................................................................................. I4
APPENDIX I, ANALYTICAL METHODS..................................................................................................... I5
Page 4 of 65 -
Exygen Research
Page 39 of 100
~
~~
~~~
~
Interim Report #2-Analysis of Surface Water Samples
~~
Exygen Study No.:PO001131
Exygen Protocol Number: PO00113I
m
INTRODUCTION
The purpose of 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 of Exygen Research.
TEST MATERIALS
The test materials are pertluorobutanesulfonate (PFBS), perfluorohexanesulfonate(PFHS) and perfluorooctanesulfonate (PFOS) and are all supplied by 3M.
PFBS Chemical Name: Perfluorobutanesulfonate Molecular Weight: 338 supplied as the potassium salt (C4F&OiK+)
Lot Number: 101 Purity: 96.7%
Transitions Monitored: 299 + 99
structure:
PFHS Chemical Name: Perfluorohexanesulfonate Molecular Weight:438 supplied as the potassium salt (CaF&3O,X+) Lot Number: SE036
Purity: 98.6%
Transitions Monitored: 399 + 80
Structure: FFF
~
Page 5 of 6S
Exygen Research
Page 40 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1131
Exygen Protocol Number: POOO I I > I
PFOS ChemicalName: Perfluomctanesulfonate Molecular Weight: 538 supplied as the potassium salt (C,IF~~SO~X+) Lot Number: 217 Purity: 86.9%
Transitions Monitored: 499 + 99
Structure:
F
F
F
F
F
FFFF
OBJECTIVE
The purpose of 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
n
current versions of the following Exygen analytical methods:
V0001780: V0001781: V0001782: V0001783: V0001784 VOOOl785: V0001786
"Method of Analysis for the Determination of Pertluorooctanoic Acid (PFOA) in Water by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA)in Soil by LCM/Ms" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA)in Sedimentby LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic
Acid (PFOA) in Fish and Clams by LC/MS/MS" "Method of Analysis for the Determination of Pertluorooctanoic
Acid (PFOA) in Vegetation by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Liver by LC/MS/MS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LCIMS/MS'
TESTING FACILITY
Exygen Research 3058 ResearchDrive State College, PA 16801 Phone: (814) 272-1039
Page 6 of 65
Exygen Research
Page 41 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
- STUDY DIRECTOR
JaisimhaKesari P.E.. DEE Weston Solutions, Inc. 1400 Weston Way West Chester, PA 19380 Phone: (610) 701-3761
Fa:(610) 701-7401
j .kesari@westonsolutions.com
Exygen ProtocolNumber:PO001131
SPONSOR REPRESENTATIVE
Michael A. Santom 3M Company Director of Regulatory Affairs 3M Building 0236-01-B-10 St. Paul, MN 55144 Phone: (651) 733-6374
*
PRINCIPAL INVESTIGATOR
John M. Plaherty Exygen Research 3058 Research Drive State College, PA 16801 Phone: (814) 272-1039
john.flaherty@exygen.com
PROPOSEDEXPERIMENTALSTARTAND TERMINATION DATES
It is proposed that the analytical portion of this study be conducted from October 01, 2004 to December 31. 2005. The actual experimental start and
termination dates will be included in the final report.
Exygen Research
Page 42 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number: PO001131
c4
IDENTIFICATIONAND JUSTIFICATION OF THE TEST SYSTEM
The following are the test systems for this study: Water (groundwaterand surfacewater) 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 repon associated with this study.
The test systems were chosen to access the environmental impact of 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 of sampleprocurement for this study are outlined in the
3M work plan entitled "Phase 2 Work Plan for Sampling Environmental
Media" The number and types of samples collected will vary depending availability in the field. The total number of 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 2OC-8"C. Fish, clam, vegetation and mall mammal liver samples will be processed according to the appropriate analytical method (see Appendix I). These
samples will be stored frozen at 5 -10C. Small mammal whole blood
samples will be centrifuged in the field at the time of collection and the serum fraction will be used for the study. Small mammal serum will be stored
frozen at 5 -1OOC.
The receipt and processing of the samples will be documented in the final
c.4
report and raw data associatedwith the study.
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Interim Report #2-Analysis of Surface Water Samples
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Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 13 I
/".
SAMPLE DEN"EICATION
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 VO001781: V0001782: V0001783: VOOO1784: V0001785: V0001786:
"Method of Analysis for the Determination of Perfluorooctanoic
Acid (PFOA)inWater by LCIMSIMS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LCIMSIMS"
"Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA)inFish and Clams by LCIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic
Acid (PFOA)inSmall Mammal Liver by LC/MS/MS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS"
The above methods use analytical conditions capable of separating the
isomers of PFBS,PFHS and PFOS. The final report will include the isomers
summed into total PFBS, total PFHS, and total PFOS found.
VERIFICATIONOF ANALYTICAL PRC)ICEBURE
A laboratory control sample will be used for the preparation of 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 ScVSpec Premier wide mouth HDPE bottles. These bottles have been routinely used for fluorochemical sample
Pagr Y of 65
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: Po001 If I
collection at the testing facility and have been shown to be free of PFBS, PFHS and PFOS. Sampleswill 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 perfluorooctanoicacid (PFOA) and
1.2-13C perfluorooctanoic acid (% PFOA). PFOA and I3C PFOA are
included in the solutions used to spike the samples. The results for PFOA and I3C 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 ScVSpec Premier wide mouth HDPE bottle per sample collected or
a zip-seal bag. All containershags 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 concentrationwith PFBS, PFHS and PFOS at
P-
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 centrihge 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 concentrationwith 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.
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 131
Recoveries are anticipated to be between 70% and 130% of the fortified levels; however, the exact precision and accuracy will be determined by the analysis of the quality control samples described above. A statement of accuracy will be included in the final report.
METHOD FOR CONTROL OF BIAS
Control of bias will be addressedby taking representativesub-samples from a homogeneous mixture of each matrix from untreated control samples, and by analyzing at least two levels of fortifications.
STATISTICAL METHODS
Statistics will be limited to those specified in the subject methods and to the calculation of average recoveries, as applicable.
GLP STATEMENT
P-
All aspects of 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 fmm 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 of the study. The report will include, but will not be limited to, the following:
The name and address of the Study Director, Sponsor Representative, and of the testing facility.
a A statement of GLP compliance (any related documentation. such as chainaf-custody records, must be in the studyrecords).
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Exygen StudyNo.: PO001131
Exygcn Protocol Number:PO001131
The signed and dated statement by the Exygen Research Quality Assurance Unit regarding dates of study inspections and dates findings were reported to the Study Director and Management.
A description of the exact analytical conditions employed in the study. If the subject method was followed exactly. it is necessary to include only a copy of the analytical method. Any modifications to this method will be incorporated into the report. If the method is photo-reduced, the project number and page number must be included on each page.
Descriptionof the instrumentationused and operatingconditions.
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 chromatogramsof a standard and a control sample, and a chromatogram at a fortification level. The location of the analyte peaks will be clearly identified in ail chromatograms.
All circumstances that may have affected the quality or integrity of the data will be documentedin 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 of an amendment signed by the Study Director. The amendment shall clearly identify that part of 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.
0 All applicable requirements for reporting of study results as per 40 CFR 792.185.
SAFETYANDEIEALTH
Laboratorypersonnel will practice good sanitationand health habits.
Every reasonable precaution shall be taken to prevent inadvertent
exposure of personnel and the environment to the test or reference
substance@).
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Exygen Study No.: PO001131
Exygen Protocol Number:Po00I 13 1
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. Amendmentsusually will be issued prior to initiation of study plan change. However, when a change is required without sufficient time for the issue of 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 of the written amendment will be the date of the documented change. Copies of the signed amendmentswill 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) Samplereceipt records, storage history, and chains of custody
0 History and preparation of standards(stock, fortification,calibration) Descriptionof any modificationsto the method
0 Instrumentrun sheets, bench-sheetsor logs 0 Analytical data tables
All chromatographicand instrumentalconditions 0 Sample extraction and analysis dates
A complete listing of studypersonnel, signaturesand initials Chronologicalpresentationof all study correspondence 0 Any other documentationnecessary for the reconstruction of the study
Chromatograms-All chromatogramswill contain the following:
Sample identification, injection date, arrow or other indication of the area of interest, and injection number correspondingto the run.
Additionally, fortifications will include the amount of analyte added and the sample number of the sample that was fortified.
e
Analytical standard chromatograms will additionally include the
concentration (e.g., pg/mL).
Page 13 of (IS
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number: PO001 I3 1
As part of the documentationthe followingsheets 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 of Exygen Research will inspect the study at intervals adequate to assure compliance with GLP's. and will report the fmdings of audits to the Study Director, Exygen Management, and the Sponsor Representative.
RETENTION OF DATA A N D 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 of time specified in 40 CFR 792.195. An exact copy of the materials submitted to the study director will also be kept at Exygen Research.
Exygen will obtain permission !?om the study director before discarding or returning samples.
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Exygen StudyNo.: PO001131
Exygen Protocol Number: PO001131
APPENDIX I
ANALYTICAL METHODS
V0001780 V0001781: V0001782: V0001783: VOOO1784: V0001785: V0001786:
"Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Water by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Soil by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Sediment by LC/MS/MS" "Method of Analysis for the Determination of Perfluorooctanoic Acid QFOA) in Fish and Clams by LCIMSIMS"
"Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Vegetation by L.CIMSIMS" "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA)in Small Mammal Liver by LCMWMS' "Method of Analysis for the Determination of Perfluorooctanoic Acid (PFOA) in Small Mammal Serum by LC/MS/MS"
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
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Exygen Study No.: PO001131
Exygen Protocol Number: PO00I13 I
ANALYTICAL METHOD MahodNumbcr: V0001780
Mytiul ratingFacility
Exygcn R-h 3058 RaarchDrive SUte COUege,PA 16801
Vice Rcridau,Operations, E x y w R a d
Exygen Research
ToulPagu: 7
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
ExygcnProtocolNumber:PO001I3 1
r 1 exyg8noaurrsll
MlcbodNuakr VWOl7W
A%ALWICAI. METHOD Method of My&for the DclermiinUion of Pcrfluoroocwic Acid (PFOA) in Water by
LclMslMS
I .o seeps
n*mabod is to be emplopl fm the iMl.ti00 md quantitationof perfluomcwaic
acid by High
Liquld Chrwutopngby coupled to a lvldm Mass
S p m m m i c DctWorW X b W M S )in w.tor.
2.0 SrfW
2.I Alwaya observe n f c labomtorypnclicca 2.2 Con& thc qppdate MSDS bcfon lmdling MY chanicd for pmpcr safety
proclldim.
3.0 Sample Rcq-
3.1 At b t 4OmL oft& umple for~xcnccion. 3.2 No ample pmceuiyIs npcdod torwua umplo. 3.3 Smplm n o d nhgmlcd lhovld be .Ilowed lo Cquilibruc IO room
MlpBuuro.
3.4 AU wpplu mustbe thomu&hly mixedbefor0 being rpmplcd for cxuaclion 3.5 Any umplor conUinin( pmiclec should be ccnlrifugod at -3000 rpm for -5
minufa ud therupanrtrnt wed tor tho cxm~tion. 3.6 Sunplc collcotioll pmcedurer will bc ~pccifidin the impling plan for this
proj=r
4.0 ~ e o u M d S w l d U d c
4.1 WW-HPLCgndC 4.2 Me(lumol-HPLCpdo
- 4.3 AmmonivmA m A.C.S. R-Wt Gndc - 4.4 PeffluomoctlnoicAcid Sipu-AkIrich
5.0 I W I U W l t U K l Q ~ l
5.1 A high pdomurm liquid clmuutngrqh crprblc of pumpin@ up to 2 rolvcau equippod with a wilblc volume injector upable of injecting 5-200 pL OOllllQdod to a uadcm M.u Spcctrommcr (LCJMWS).
5.2 A dcviscto collect nw drt.for peak inkgalion .adqusntilalion. 5.3 ruulyciulM.Oc0crprslaofm84ingto o.oooo1 g.
5.4 5 0 m L ~ o p o l y p r o p y l ~ c ~ t r i f l r g c t u b a . 5.5 15 mLdirpoublopolypmpylaE d f u g e tubw.
5.6 Diqonbkmicmpipu(50-100uL.IWZohrL). 5.7 125-mL LDPE nUmW-~Oumbonlo.
5.8 2 mLC l e m HPLC vw kit
5.9 Dirpomblcpipdtca. 5.10 A u t o p ~ ( l O O - 1 o O O p L m dlOlOOpL). wilhdi8poaabletipc. 5.11 W.unSopPtVu6~((ly)ClUSPEculridgoc.
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
ExygenProtocol Number: P o 0I 131
I .AhirLYllCAL METHOD Mabodof Andy& br th@Dc4rd&on of PdumoocuDoic Acid (PFOA)in Wiler by
5.12 SPBvlcuwarmrdfbld 5.13 CODIlifigocapableof lpinning50mLpol~mpylmc~ b awt 3(xxI rpm.
6.0 chmrmtoplphic syctm
6.1 M y c i u l Column:Fluophuc RP (Kcyttono Scientific). 2.I mm x 50 mm. Jp (PM: 82505052130)
6.2 Trmpcn(urr: 30.C 6.3 Mobile pturc (A) : 2 mM Ammonium Acetate in Water 6.4 Mobiiphuo(B): Maha01 6.5 cindimth~opnm:
& & ? M y 0.0
1 a
65
8.0
25
20.0
25
22.5
65
Flow Rate
';ja
lrellrmnl 0.3
35
0.3
75
0.3
15
0.3
35
0.3
6.6 6.7 6.8
- - InjectionVolume: 15)rL(cmk imrmscd
Qu~tiutionP: wk Arcr cxtcrml Run T i e : 23 minulsl.
to LI much M 50 ~1). crlibmlion curve.
Ths.bovceoaditiool~iara&d u r guidesnd maybe changed inorder to
oplimilc the mu:Iyrce3n
7.0 Ms/Ms syllan
7.1 Mode: Elo*mopnyNeprtiveMRMmodc,manitonng413+369m/z.
The above cnadicioar M incosdod u guide rad m y be changed in order 10 OptMiSo the MSMS 8yntem.
8.0 Prqnntion of Solvriolu 8.1 Mobilapbuo
8.1.1 2 mM .mmotlium rotrcD in wrtar is p n p d by d i n g 0.154y of
lmmoaiumlcancto1ooomLofw.tn.
Altamue volvmcl rmy be prepared.
I
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Exygen Study No.: POOO1131
Exygen Protocol Number: POOO1131
I . ANALyTIc.u METHOD Method of Analyds tkrho DatamiDuionof Perflumwauroic Acid (PFOA) in Wuor by Lc/Ms/MS
9.1 Slud.rdStock/pdfiutionSoh*on
9.1.1 Rcp.rr a dock solution of-100 j~gftnLof PFOA by weighing IO my
of &@wl ctmdpd (mrrcstcdfor purily) and dilute IO IW mL wilh
mdhpIol in a 1 2 5 d LDPB bottle. 9.13 A 10 pl/mL forliflcltion rolutionofPFOA i8 pnpued by brinying IO
mL Of UIE I00 p w roiution to a Anrt volume of 100 wilh methanol
in Ii u m~LDPB bonk.
9.1.3 A 1.0p@nL fotthiuiion solutionof PFOA i8 pmpad by bringing I O
mLof tho IO pB/mLrolutionIO. find volum of 100 with m c r h d m
125 mL LDPE botllo.
9.1.4 A 0.1 p#mL fod6c.tionsolutiw of PFOA ir prspMd by bringing IO
mL oftbo 1.0 p g h L mltltion to a fuul volumeof 100 wilh me(h.nol in
a 125 mL LDPR boltla
9.1.5 A 0.01 pgftnL fdfisrcion rolulion of PFOA u preparedby bringing
IO mL of h 0.1 p#mL rolution IO a f i ~ vl olwnc of IW with
&I
in a 125 mL LDPE bottle.
9.1.6 'Ibs dock md fortifidon rolutionau e IO be awed in a refrigerator PI
lpprorimuoly C C and Mulblo for a nuximum pond of 6 months
hthe &lo ofpnpusion.
9.2 SlaMhrd cdibntioosolutioa,
92.1 L4XWMS a l i t d o n staudds m mated in HPLC wcr. The ulibntion nudpdc M pmcrued through the artnetion pmecdure. idcntiul la umplor.
9.2.2 'Ibs fobwing u a rrpial oxunplo: dditiooll oonocnmtionsmay be propvodu needed
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Exygen StudyNo.:PO001 131
ExygcnProtocolNumber: PO001131
'B a p l . * u F h ANALYTICUMElllOD
Mahod of Analysis fortbe Dctaminuiin ofMw-ic WIMS
MnbodNvmbn VWOl 780
I Acid (PFOA) in Witcr by
9.2.3 A zuu slmjud wlU(i0a (t'wp11 blmlr)
with each
#( of ImdpbartMcd
9.2.4 ScOnd ahaclcdultbntion UauidIin 15-mL polypmpylenc Nber
n 2% m 6'C, up m two weeks.
9.2.5 Allanale vohrwr and c o a e no~f Nndudr may bo prepwed as
needed.
10.0 8.tcbsaq
10.1 Erhbatch of umpla cxlractcd (typically 20 or l a s ) mwl include at leu1 one r a g a 1 con(r0l (mahad blank wing HPLC water) and IWO rrr$rnl conrrols fdfied 81 b r m conccnlrrtionr (lab control spike) IO verily procodunlrooovayfortheb.lch.
10.2 R c p ~ forlfid~d md labanlory duplicster and cpika will be specified inthc qualityusurawc plm for thisp m j a
11.0 Slmplc Emution
I 1.1 Mauuro 40mL of umple or a partionof umplc diluted to 40 mL with water into 50 mL palyppopylmccenhifipeluku (fasiify u needed, replace lid and
mix well).
11.2 Condition the QI SPE
(1 g, 6 mL) by passing IO mL methanol
followedby 5 mL of HPLCWItDT (- 2 dmp/roc). Do 1101 let column rundry
- 11.3 Lod~~pleoncaditionedG~SPEcutridgDe.i ~ ~ u d ~ l ~ a t ~ .
11.4 Elute with -5 mL 100% mabaol. Collat 5 mL of eluate into grdualcd
15 mLpO1ypmpylac w * e tuba(mvolume 5 mL). I 1.5 M y z s rvnpla Udng clcamrpny mms.
12.0 c b m d a p p h y
12.1 hjat ulc y l ~ pclount ofucb # t d d ,slmplc and forlificd umplc into the
LCIMSIMS
A cdibnlion Itud.rd musl prscedc and follow all
dyrd umplor. 12.2 sondvdr of PFOA comapadugto N lw five or more concamtion levels
mudbo iruhddin krluuiylicdtu.
12.3 An & sat of ntnclcd a l r ~ o aoud.rdr must be included a1 the beginning mui SIthc ad of a vmplc set. Extnclcd rlandudr mu1 hc
intmpad brmocllway 5-10 umpla. AI m dtarutivc. UI entire sa of
atnclcd ulibntion Imdpb m y bo ir\jccred st the beginning of a re1
followod by Qmrtcd ulibntion rulrdvdr intcnpcncd cvsry 5-10 ample5
O(I ICCW for a d sel of artncted rlududs). In cilhcr cue. e x u u l a l
calibration19adubmultbe thc fint md I.rt injection in a umplc HI. 12.4 Use lineu r(lld.rd cyva far qumtit.tion. Linur rundud curves an!
g dfwthe d y u by Linmmgrmion using LIx weighting of peak area
pW.y.lo<i
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Exygen StudyNo.: POOO113 1
Exygcn Protocol Number: POOOI I3 1
13.1 ~ g m tlnumt show ips*of I) daughter ion al 369 m u Pmm a pareni of 413 MU.The 413 m u pMnt concrpondr Io the PFOA anion, while Ihc daughter ion (369mu) rcprslsntithe louof carbon dioxide.
132 Mahod blmka mum MIcontain PFOA at lcvcli p i e r t h th~e LOQ. If I blank containr FTOA al Iwch @'cue#chm 50 nglL, ulm Inew blank tamplr muUbc obumodmdthe mtire wt m w bc rc-oxmctcd
13.3 Rceovcria of oon(r0l s p k md mmix spikes mum be between 70-13090 of h i r horn vdwr. i t a conhol $pike f8Ih outside tho acceptablelimits. the d i m wt of smpln should bc re-cxtnctcd. Any mafrix spike outside 70-
1% sould be evdwtad by the d y i t to determine if re-extraction is
wyM1cd.
13.4 Any cdibrdion rtlodud Iovldto bc i#fUi~tiuolutlier by using the Huge Ermr Toy may be acludd from the Dllculation of the cllibdon curve. H m e r , tho toul numbs of umctcd odibntion standards chol could be cxcludod muct not aoood 20% of the total numbs of oxmcted ItMdlrds
iqicctd.
13.5 The ramehtion owfkicat (R) lor alibntioa wya gmmtd must be
M.!BZ (R* M.995). If ulilnation raultl fdI outride these limits. then ippaoPiUc Uopl m w be taken to adjust inrlrument operation, and lhs daodmd~or the nlwmllaof umplu shouldbe rswlyzed. 13.6 Retention timol bawm sundudi d sunplw must not dritl more thnn
f 4 K within an larlfiul run. If rctcntiontime drift cxcccdathia limit within u1uulfial tun then tbc l a m w be 4 y L C d .
14.0 clkulatioru
14.1 Ute Ute followin# ap&m to alou*to thc amount of PFOA found (in ne/L.
bucd on ma) tuing the a d a d lye (linwregression pamelers)
~
~ by tbocM m Lydnx roltwuc pmgmm:
- PFOA Iound (ng5)
x DF slope
-DF hnor by which thc Brulvolumew u diluted,if nwcsury.
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Exygen Study No.: POOO1 131
Exygen Protocol Number:POOOI I3I
Mscbod of -8
1 AIyALnlCAL MLTtIOD
forcbo DatnmidmofPufluoroo*lmic Acid (PFOA) in Water by
14.2 F a umplr fortitied with known MOYDU of PFOA prior to cxtmtion. use
tho R-vcry
-followingcpurtion10 (W
Ulculalc
cbo
perm11 r
c
c
o
v
q
.
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Exygen Study No.: PO001131
Exygen Protocol Number: PO001 131
ANALYTICAL METHOD
MdwdNumkr: VW1781
MItbcd of AnWL for tbr Ihtrrmln81baof Perlluorooct.nok Add (PFOA) In Soil by LcIM9/M8
Anrlpicrl rating Pseility:
exyscDR-h
1058 Rh-
Drive
Sutc College. PA 16801
Appmvcd By:
Ppvl colmouv
\
iolulo4
Date
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Exygen StudyNo.: PO00113I
Exygco Protocol Number: PO001131
L 1 ANALYTICAL METHOD Mahod of AdyU for thc Dbonni~dmof Pafluoroocmoic Acid (PFOA) in Soil by LcIMs/MS
1.0 stops
Thir mochod uto ba anplopd for thc h l u i n and qudtation of pduomocunoic acid by High Pcrformmcc Liquid Chromuopnphy coupled to a Undcm Mass
SpcCaDmceiCDac*or (LcmsMs)in mil.
2.0 safely
2.1 Alwaycobravsufe*bontorypncricor. 2.2 C o d t Uw .ppmpriltoMSDS before butdlinn MYchemical for omwr saliiv
prrC8UtiOUI.
3.0 Sample Rquirancnt
3.1 At I d 15 gof tcaumple for ormotion. 3.2 No umple pmudngi8 Mododh r mil umplu. 3.3 Sample# itorsd mhipor.lod lbwld ba &wed to cguilibntc to room
ccmpatura.
3.4 All m p k r mud k Ihoroughly mixed bafom bringu m p l d for cxmnion. 3.5 Sample collection pmcdwu will bc apccificd in Ihs umpling plan for this
pmjoct.
4.0 Ragml~lodSU&&
4.1 W.O-HPLC&
4.2 Mcthulol-KPLCgde
4.3 4.4
- ML!IUAc*rts-AC.S.RarWdGndo
PCrRuomocuDoicAcid SipM-Aldrich
5.0 ~ a l t t 4 l I i p m o n t
5.1 A higb portornusee liquid c4uumUopph capable of pumping up 10 2 mlvmn qui@ with a VlrLble wlvmc injectorupabk of injecting 5-200
pL conwoted to 1UadmMua Spsaromaq(LUMS/MS). 5.2 A devicem colloctraw data forPSJC intogruton mdqumtitation.
5.3 A d y t i C d bJmcsClpoblOO f I d i l l 8 100.OoM)I 0. 5.4 50 mL dispouble polypmpylcnc~ ~ n h i k tgueka. 5.5 15 mL dispouble polypmpylcnc oclffikge tu&.
5.6 Dispoubla micmpipcu (5&1oouL. 100-200uL).
5.7 1 2 5 d LDPBnumw-mouthbottlcs. 5.8 ZmLClovHPLCvillkit.
5.9 Dirpoublcpipdta 5.10 Aulopipoasl(lM)-10~LtIO-1W~Awitbdicpoubletipl
3.11 Wl(ar Sop W Va76 oc(1.g) CIS SPE u m i d g a . 5.12 S P E v W ~ d f o l d .
5.13 ULmconicb.th.
Exygen Research
Page 59 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn ProtocolNumber: PO001131
1 ANALYTICAL METHOD Mahod ofAMIyru for the Datamidmof Pe&mmo&mic
1 Acid PFOA.)in Soil by
5.14 WhU-rCCimr & a h . 5.15 Contrifigeaplble ofspinning 50 mL polypmpylcncN b a u So00 rpm.
6.0 ~ I O g r ~ i lsipCtm
6.1 Myiiul Cahunn:FluophroRP (Keynon~Scientific). 2.1 nun x 50 mm. 5p (PM 82505452130)
6.2 Tsmpruurs: 3WC 6.3 Mobile Pturs(A) : 2 mMAmmonium Acstlre in Warn 6.4 Mobile P h w (e) : Matlunol 6.1 GndierrlPmgwn:
Tinrclminlu
0.0
6.3
1.0
65
8.0
25
20.0
25
22.5
65
Flow Rate
Y ' " " "0.3
35
0.3
15
0.3
15
0.3
35
0.3
6.6 6.7 6.0
- - Isjoaion Voluma: 15 pL (abo inrraredto u much u 50 pL).
Quuuiwion: P a tA m extmnd mrdudcdibntion DUTYO. Run Tim: 23 minluor
The above mndirionr Mintoldodu
Optimimtba~~
d n u y b c changed in order lo
7.0 MslMsSyitaa
7.1 Mode Elcctmpny NegativeMRM made, monitoring413 -b 369 d z for
PFOA.
The &ow d t i o n r arc intmdrd u a pido ud m y bs changed in order IO optimize thc MSMS syltcm.
8.0 prrpuuion of solutimu 6.1 MobilewuO
8.1.1 2 m~ muonium wmta in rmor is pmpmd by adding 0.154 p or ammoniumICC(UC IO 1000mL of watcr.
AlIanue voluma nuy bo pcpuod.
~.
Exygen Research
Page 25 of 65
Page 60 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen ProtocolNumber: PO001 131
Iurn-
MIlhod Nvnrbr v~o1781
ANALYTICAL MCTHOD
Mabod of kulylia fortbc DC(ermi0pmof P c c f h w c M o i c Acid (PFOA) in Soil by
LC/Ms/MS
9.0 Sund.rdRepm'Uim
9.1 sund.~ rdSto~ cWP.onilcuMnsolution
9.1.1 Rcpyc I stock solution of -100 pgmL of PPOA by weighing 10 ms
of d y l i c d atandud (eo-
for purity) and dilutc 10 100 mL with
m h l in 125-mL LDPE bottle.
9.1.2 A 10 pphnL fonificuionaolutionof PFOA is prcparcd by bringing IO mL of mC 100 p#mL rolulion to I find volume of 100 with methmnl
in a 125mL LDPE botcle.
9.I .3 A I .O @IL fortificuionrolutionof PFOA is pmparcd by bnnginy IO
mL of the 10 p#mL solution to find volume of I00 with mthinulii1
i1.25 mL LDPE bottle.
9.1.4 A 0.1 pg/mL forbflution solutimof PFOA upnp.rod by bringing I O
mLof~l.OpplmLsol~mto~flnrlvolumcof100withmoh.noltn
I 115 mLLDP8 bottle.
9.1.5 A 0.01 pvmL Wication ldutionof PFOA ia preparedby bringing
10 mL of tho 0.1 p@nL solution to 8 find volume of 100 with
mahmlin 125 mL LDPE M e .
9.1.6 The uock md fonifiwtlw r ~ l u t i M o ~IObs stored in I refrigeratorat approxhwely 4.C Ird PO r table for a maximum H o d o f 6 months
ftoDntboduo ofpmpuuioo.
9.2 St.ladud CdibnIion SolutiW
9.2.1 Lc/MS/MS c d i b d m roadudr MpropMd in WLC water Thc cdibntion rundvdr M prcrouod thmugh Lo axtnction pmsedure. idcntial to pmpla.
9.2.2 Ibo followi u 8 typical ex.ntplo: additional wnccnmlions may be PrepSrrdUMcdrd.
Fhl
Comenmioa Fmifia~ion V o l u of ccnornlntionof ofFatifiollion Volumc FdficdCmmol Cdibntion
Qlibnlion StandardU)
SalutioaIPpb) (rL)
0
0
Samole (mL) 4a
SmndvdbP0' (eaample)
0
xcmmddyy-0
IO
100
40
IO
200
40
23
xcmmidyy. I
so
xcmmddyy-2
10
4GQ
40
I00
xCmmddyy.3
100
IGQ
40
250
XCmmddyy.4
100
200
40
100
4GQ
40
500
xcmmddyy-5
loo0
xcmmddw.6
xc *Tbccmraual-
'onof tbo alibntiw d u d ia equal to Bx its initial
-kmion, dw to tbo -tim
of thc Nndud duiung Le utrution (SPE).
.Icxtmtedcllibntimu1Ddpd
c-
Exygen Research
Page 26 of 65
Page 61 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygen Protocol Number: PO001 13 1
9.2.3 A ?.anmadud solution (magonI blank) Mof .unduboxmcted.
with each
9.2.4 Ston dl oxfnucd calibration rtadrrdr in 1 5 4 . polypmpylmi u k u ZTIO6T,up 10 two w&.
9.2.5 Alt~nusvoluma ad coDCCILblti~ol~f standardsmay bo prepand as
Moded.
10.0 BUchSctUp
10.1 E d buch of vmplm exfnucd (typically 20 or less) mu81 include pt least OM magcu~mI tml (method bhnk using 5 mL of methanol) md two rcaycni contmli Mfld U kmwn consmmtiolu (lab control spike) to verily paoeodunlrocavEy forIhcb.tcb.
10.2 Rcquirwcllu for Bcld ad llbontoryduplicates and spike will bo specified io the qualityuutmwp h for ch* p j e c t .
11.1 Weigh 5 g of umplo into 50 mL p~lypropylsnscatribge d e s (fortify as needed, replace lid Pdmix MII).
113 Add 5 mLofmahn01ad chlrc on a m k t d o n Ibakcr for45 minute. 11.3 h k t h o t u b u t o YI ulhumicbUh aad Moicato for -15 minutes. 11.4 Bring tbe volnnc up IO 40 mL with WUUin tbe SO mL polypropylene
cmtrifuge(uk. 11.5 C m k i f u g o f o r - l O m i u - 3 W p .
11.6 C d t i w tho Q t SPE d d g a (I p 6 mL) by pwsing 10 mL muhlnol f0llOWd by 5 of WLC WUOT (- 2 drogrD2). Do no1 IC( column Nn dry
11.7 Lord (decurt) the sample 011 Ihc wdtiond CISSPE cnmidge. Discard
- tlurte.
11.8 Elutc with -5 mL 100% mcthulol. C o l l a 5 mL of cluue into graduaid I5 mL polypopyloatWUriIbgotubm (ruulvolume 5 mL).
11.9 AnrlyrcUmplm wingo l m ~ Lcy/Ms/Ms.
___
Exygen Research
Page 27 nj 6s ._
Page 62 of 100
~~
~
~
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
ExygcnProtocolNumber: PO001 131
byp. Rup*
M n b o d h h b a VOOO1181
I 1 ANALYTICAL METHOD Method of Mpii for theD e t m h n'onof PerlluomoctPnoicAcid (PFOA) in Soil by
LClMsMS
12.4
12.5
13.1 IhwW0gmmudlbool rprk ofrd.uOboosion 81 369 m u from I plrcnt of413 mu. T b 413 MU pmnt carapoldr 10 tho PWA mion. while the dwgh(a ion (369 8mu) repmsds tho lorr of carbondioxide.
13.2 Method b W mwt no1 cowrin PFOA 8l levels pala (h.n the MQ. If a b M : MU~IU PPOA 8l levdl lportorIllan50 I@, then 8 new blank rnmpls mwl bo obtliaod md themtin lam w be rwxmclod.
13.3 RocoVri~o~fumkol spikes d mmiX rpikw m W bo botwocn 70-130'6 of the hwMl vrlucr If a mntrol rpika Wlr outside Cho acceptablelimits. the mtLs IC( of nmph sbould be tbextrrted. Any mlhix spike outside 7013oY. should be onlurtd by the d p t m dacrminC if mxlraction is
WpMtod
13.4 b y Ulibntion S d a d buDd to be r nrrinieal outlior by wing the Huge Enor Tea m y be excluded fnnn thc olloulrtionof the wlibntion curve. Howowr. thc toul numbs of ox!nctcd ulibntion nrnddr thu could be excluded mud MaIaed 20% of tbc total number of cxwaclod aandanlr
injcasd.
13.5 Ihc mrroluion cwtficient (R) for calibration c u l ~ e sgeneraled n m l he 20.992 (R' 20.985). If wlibruion rcrvlti fall outiide thac limits. then appropriate rwps mud be ultm to ldjw inrcrument operation. und the lundudr or the mlcnnt (01of umph should be rsrrulyud.
13.6 RotaUion limes I#cwoon d.ad.rdr a d umplu murc not drift more than f 4 K witbin an mulyticd NIL IfraoatiOn time drift ex& this limit within o uulyiiulmffimtha rctmurtbenuulyrcd.
Exygen Research
Page 28 of65
__
Page 63 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn ProtocolNumber: PO001131
.."
14.0 C W m
14.1 UM Ihs following cquuioo to dcuW the mount of PFOA found (io ny/L. b u d OIL peak ucr) u a b the amnjxd culw (lincu wgrcuion parameters) pdcd by the Mu8 Lynx tofhvw program:
14.2 P a r w p l c l forcifiod with known unountr of PFOA pnor IO exuu-tion. UK Ihc followingq u i m u)&Ul* thc proson( neovory.
wg - 14.3 UM tbc following quation to Comra tho mount of PFOA found in ng/L IO W).
PFOA found @pb)
umpls weight (5 g)
14.4 Use the foUowing cqutim to dcu*re the mount of PFOA found in pph
busd0ndryW~L
-__
Exygen Research
Page 29 of 65
-
Page 64 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygen ProtocolNumber: Po001I3 1
ANALYTICAL METHOD
M a b O d N m k V0001782
Method of Andpb fW tba Dccermhmlonof ?trflncroo*.noic Acid (PFOA) In
WmMI by Lmms
Analytical Testing F=ility
Exyw Raouch 3058 Rcvuch Lhivc Sbta College, PA 16801
Exygen Research
Page 65 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO00113I
ExygcnProtocolNumber: PO001 131
1.0 scope
This mclhod b tobo OmpbyOaforthc hlrtion andquititation of~ u ~ o o c m o i c rid by High Pafnrmsw Liquid chmrmtognphy coupled to s Undem Mass
Spectromekic Dctmof (LG'hWMS) in dimeat.
2.0 way
2.1 Ahvl~obsuwufo~laMtorypnctica.
2.2 COaRJI tbc .ppmpriueMSDS b o d D h~ ndllnsMYcbemisd for proper safely
pluxutioM.
3.0 SunplcRequuaneut
3.1 Atlart30gOfMumpIeforcxtnc~~. 3.2 No #amplep-iag is mcdedfor rcdimontsamplsr 3.3 Sunplcc rtaod rrbiporucd should be d l o w d to quilibntc io mom
m.
3.4 All umplc~m u be thoroughly mixedbefom being umplcd for extraction. 3.5 Sunplc collation prcculw n U be rpsciGed in thc sampling plan for this
pmj=t.
4.0 RwgentIMdshududI
4.1 W l r a - H P L C M
- .
4.2 4.3
- U e t h o l - H P U : ~ O
A~eticA d m t plao
4.5
5.0 INillll 5.1
5.2 5.3
54 5.5 5.6 5.7 5.8 5.9 5.10 5.1 I 9.12
Exygen Research
Page 31 of 65
_.__ -
Page 66 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001131
,-
tMabod of kUl*
I ANALYTICAL hIETHOD
for tho Dctormirution of Pufluomoctmic Acid (PFOA)in Sedimentby
Lc/Ms/MS
5.13 VoPfp(01. 5.14 Wrillrcciocl &&r. 5.15 Cptmll\lpcupableof spinning50 mLpolvpropylcnctuba at 3000 'pm.
6.0 chroml(0Snphic Sflm
6.1 Analytical C o l m phlophw RP (KoySouc Scientific). 2.1 mm x 50 mm. 5p (PN 82505-052130)
6.2 'Tanpmluw.3w 6.3 Mobile P h w (A) : 2 mM Ammonium Aoauc io Wa(a 6.4 MobilOPhw(8): Mabrmol 6.5 GndisntFTo~:
ZaanunA
0.0
65
1.0
65
8.0
25
20.0
25
21.5
65
Flow Rue
ULmlLmin)
35
0.3
35
0.3
75
0.3
75
0.3
35
0.3
- 6.6 w&tioaVolumo: 15 pL.(cmbe iaaawdtoas much u 50 pL),
6.7 Qucncitltion: Pmk AIM -cx(anrl mndvdulibmion curve. 6.8 Run Time: 23 minuto*
The rbovs wnditiw Miatcndedu 8 guideud m y bechangedin ordor 10
optimi?al&HpLccyrtcm.
7.0 Ms/Ms syrtan
7.1 Mode: ElsWlpny Negative MRM mode, m o n i w 413 +369 d z for
PFOA.
Tba aboved t i o m MWaded ua pliC ud my bo changed in order to optiinh m0 MSMS sptam.
8.0 PmmIuion of s o w
8.1.1 2 mM rnmonivm .cb.tc in wuer l a prepprod by adding 0.154 & or ammonium lootpteu)lo00mL of wuu.
Exygen Research
Page 32 of 63
Page 67 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen StudyNo.: POOO1131
ExygcnProtocol Number: POOOI I3 1
I ANALYnCALMETHOD
'Method of M y l i r for the Dctcrmidon of PcrtluorooaMoic Acid (PFOA) in Scdimml by Lc/Ms/Ms
8.2 ExtractionSoltuio~~
8.2.1 1% r c t i c .eidin wUa ir PrOpMd by adding 10 mL of acclic acid 10 lo00 mL ofrvlta.
A l l a ~ u kvolumol m y bo prqwed.
9.0 StmdUdRopMtion 9.I Stladpd StodJFoltific.tion Solution 9.1.1 Repuc a m c k solution of -100 p#mL of PFOA by weighink IO mg of Uulvliulrtladpd (eoncctd for p.wily.) and dilutc 0 100 mL with m&l m I12s-mLLDPEW e . 9.1.2 A IO po/mL fonitlcuion solutionof PFOA ic pmpMd by bringinp 10 mL of tho IM) pghdsolution to a W volume of I00 with mslhnnul in a 125 mL LDPE bonb. 9.1.3 A 1.0p@. forliftation solvtionof PFOA it prepared by bringing I O mLof the 10 p#mL solutionto a tid volume of 100 with methmolin a 125 mL U)PE botllc. 9.1.4 A 0.1 pl/mL IOrtMutionsolutionof PFOA iIprcplred by bringmy IO mLof~I~1.e0 p&L ~ l u t i o nIOIfhrrl volumaof 100 with mclhmol in a 12s mL LDPE w e . 9.1.5 A 0.01 p@. fortiKcltion solvtiOn of PFOA u prepucd by brinying IO mL of thc 0.1 p@L solution to L 6 4 volume of 100 with mgbaolh a 125 mLLDPE bonle. 9.1.6 Tho mock urd fortifiuton solutimu p'o to be otorod in a refrigeralorI qpmximucly 4.C and am mble for a muhum period of 6 months fmm thr btsofprrp.ntion.
9.2
Exygen Research
-.-
Page 33 of 65 -
Page 68 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 13 I
.-
LCIMSIMS
9.2.3 Storo1cllibnrions l m d d 8 in 125-mLLDPE numw-mouth bolllcr .I2 T 106% up tosix moatha.
93.4 Altnrrrte voluma mi wnwntrahuof standuds may be propprcd as &
10.0 BlGehSdVp
10.1 PIOh btcb of umplm omrctrd (typicdly 20 or la)mum includo I( lure
OM unrortod warn1 urd two untreated ooncmlr fortified .( known
EOOEOO1Rtlauga control spprc) 10 vod@ proculunlrocovcry for the batch. 101 9uiruimb lor 5sld lad hbmtorydupliuta md spike8 will bo rrmifiod
I thoqualityvnvuroplan kthir pvjst.
11.1 11.2 11.3 11.4 11.5 11.6
11.7 11.8 11.9 11.10 11.11 11.12
woigh 5 g of umplo into 50 mL polypnwylcne m u i h g e tuba (fonify ;IS needed, mpllce Lid mid mix woll).
Add 35 mL of 1% rcdic acid.up.vortex Md hake on 8 wrist action shaker
for-6ominu~
chuifugotha bham -3000 rpm Cos-20 minutor.
cwditiw tbo CU SPE currldga,(1 g 6 mL) by pouing 10 mL methanol
followsdby 20mL of WLC Ws (- 2 dmplm). Do not let COlUmn W dry Load (docm()tho umplo an tho condition4 C,I SPE h d p . Discard
Chute. Add 20 mL of m-1
to tbo & a t left in tho bllom of the SO mL
dhp minutea
Cap, VOM lad shake an 8 Wrsi d o n rh.lcr for -30
cdtinlge Ibcbhau -3000 rpnfa -20 minutor
we). Decanttho mdunolomtha umaSPE urtridge. Collsotthe eluate.
W u h tho solunm with 4 mLof moUumL Colloa tho oluatc and dd it IO the
dust8wlktcd ia app I 1.8. CODditiana rscoad CIISPE m d g e (I 6 mL) by w i n g 10mL methanol
f o l M by 20 mLof HPLCwlter (- 2
Do not let column w dry
Md mS -1
to -200 mL of warn urd kd on tho w n d conditioned
SPE amidla
auto wicb -5 mL 100% molluaol. CoUoct 5 ml of eluate into grnduatcd
- .
.
Exygen Research
--
Poge 34 of 6s
Page 69 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number: PO00113I
12.1 Ir\iect mCumo ullountof each Mandud, -le and fonificd sample into L e
Lc/Ms/MS sptan. A ulibntion U u h d must pmcdc and follow a11
4vrrd12.2 Shadmh ofPpOA conslpoDdingto at lsut five or more conccnmtion levels
muat be iaclvdsd io od y t i u l set.
12.3 An Q~LC(01 of ortwtod cdibnrioa Uanduda must be included DI the
beginning and u the ad of a umplc M. Stmdud, mum be intersped
M w m way 5-10 umplm. Aa m dtomativc. m oatire sc1 of calibnuun
m y be injaded .( tho beginning of a set followed by cdibniion
-aanduda ilrrcnparod way 5-10 -la (to .ccoynt for raond act or
-).
In d t k UIC. Cdibntion #.rdUdr mua be tbe firat Md IS1
hjccticmin a ample w.
12.4 UM linear ctlldrrd CUVQ for quutituion. Linear 1t~dSrdcurves arc
for m0
by Liaclrmgmsion wing I/x weightingof peak area
ullbntiond u d sonecntntl'onusing MurLynx 3.3 (or quivalcnll
12.5
13.1 Chmnntopmmudrhow a @ of. daughter ion 81 369 m u hwn 8 p a n 1 of 413 amu. The 413 .mu p m t compondr to thc PFOA mion. while the
daughterion (369unu)repwuUthe lossof cubon dioxide. 13.2 Method bluJD mmt not cmmin PPOA u Iweb &mtachn the LOQ. If a
blmk cuntdna PFOA It Iwoh pule# than 0.2 ag/mL, than a new blank
ample mustbe obtainedmdtbo m h Mmust be rwxtnctcd.
13.3 Rooovsirr of umml @a d murix c p k m w be bmvcen 70-130% of ChL kmwn v h w . If caatml+ hlL ou(ddc tha vccptlble limitr. the
CntLO set of c~plashould be fo-extmcled. Any matrix spike ouuidc 70.
130% cbould be ovrhuted by the d y l l to detormim if rnxtraclion is
WYMtQd
13.4 Any c9b.tioa ctlldrrd found to bo a wiaiul outlier by using the Huge EnOD Tea,may be excluded fmm cbc cdculuion of tho calibration curve.
Howow, tbo !atd number of cxm*cd cllibntion rundudt that muld be
cxclvdod must llol o r d 20% Of 1& roW number Of exblclcd rIMdudS
yoflsd.
13.5 Tho correlation wetticioat (R) for dibnrioa GWO( gonmtcd must be
ZO.992 (R' ZO.985). If cdibntion -Its hll outside there limits. then ' acpl numl bo ukm to adjust lnsmunenl operation. and Ihc
z
r the mlovant(01of umplcl shouldbe reuulyzed.
~~
Page 35 of65
Exygen Research
Page 70 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
Exygen Protocol Number: PO001 13 1
Ery(mwh
~NnmbuV~llU
1 i ANALYTICAL METBOD
Mahod of Anrlyc* forthoDO(enninrti0nofPafhwmocMoic Acid PFOA) in Sediment by
LUA4SiMS
13.6 Rncntion timol bawooa standud# urd mmpla mwt 1101 drift more lhan i4 W within MlnrlyticdNO. Ifmtmtiontimodrift u d r thir limit within
Mmrlytied NO ma tksarmuf bcralulyzod
14.1 UM tho followingeprution to c & u W thc mount of PFOA found (in nplmL.
b a d on p k a m ) uing tho Nndud CWD (linear ngrersion pmmnen) gmaatad by Q Mur Lyax mltwuopmgrm:
PPOA bud(nghL)*
x DF
DF
-
factor
by
which
the
5114volume
wu
slop dilutod. if n
c
c
w
s
u
y
14.2 For 8mpla M6cd with ~JOI W .mount# of PFOA prior to exlction. use
R
-
thc vw
-following (K)
opualionto
u
l
c
u
l
r
t
c
thc
pacrnt
r
o
s
o
v
a
y
.
14.3 Uw thc followingquation to convwl thc mount of PFOA found in n#mL 10
@6 W). PPOA fouud
@
p
-b)
-
umple wei@ (5 s)
- 14.4 Uw tbo following epwica (if ~)(c.ou.ryt)o ulcuhte the mouol of PFOA found inppbbucd011 dry WeighL PFOA douad @pb) dry weight PFOA found (ppb)x [100% I IMal soli&(%.)]
Exygen Research
Page 71 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001131
ANALYTICAL METHOD M s m o d N ~ m b tV~OOO1183
Methodo f h d y ~fbortbe Deterdm.lb. of PerfluomoctrnolrAcld (PPOA) Io Fisb
and
by LCIMGMS
M y l i e d ToslingFacility
Exygw~Rsrarch 3058 ReJcarch Drive SUtc College.PA 16801
Approved By
--
Exygen Research
Page 37 of65
Page 72 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Numbcr: PO001 13 1
1.0 SMpC
Tbir methodis to be 0mployOa for the icolatiw rad qwntitationof portluorooctanoic u i d by Hi@ Pefomura Liquid Chmnufognphy coupled to a undem Mass Spoctmmcrnc Dctfftor (Lc/Ms/Ms) in fuh ud cluns.
2.0 srfcty
2.1 NWyO b r a V C U f C hbCdOlyPdCCD. 2.2 ConsulttheappropriateMSDS kfmltuujling my chemical for proper safety
prouutionr.
3.0 Sample RcquircmcDt
3.1 AtlowZOgoftsltrrmplaf~cx~tion. 3.2 Sunpls cbwld k proccslsd b&ro extnOtirn. Placethe fromsample in a
food procolsor .ndhomo+ with dry ice, Pllea the smplu in containers ud leave opm in ftozcn slonge ovanight to allow for urbon dioxide sublirmsion scll md pllea fhc nmpla in from storage until time o f
dyU.
3.3 Sunplo oaUkcion procuhm dll be rpbcitied in tho smnplinp plan for this pmjm.
4.0 h g m t # d S I & d #
4.1 W.tor-HPLC&
4.2 Acc(ocriDile-HPLC
- 4.3 Cuban (12o-400meah) went pride
4.4 Mothnol-HPLC@&
- 4.5 Silica gel (60-200msrh) h g m t prds
4.6 Fl&il(6O-l00mah)- h g o n t g d o
- 4.7 svpcrdomLC-MIa Ragult @e
4.8 I-oanol-HPu:~
- 4.9 L-Ararbic acid b g m t @e
4.10 Dimethyldichbnuihc-hgupaot @e
4.11 T ~ l ~ - R o y a n ( @ &
4.12 4.13
- AmwniumAcohte-A.C.S.ReWIItGndc
Pcrflmmo*ooic Acid Sismr-Aldfich
5.0 hshumcrUudEquipnotl(
5.1 A high pedonIuacc liquid cbmrmtomph upahlo of pumping up lo 2 colvcnta qippcd with L wilblc volume injectorcapable of injecting 5-2W pL EwDc*odto 1ludmMu,s m e m(LcIMs/Ms).
5.2 A d e v h to oollct nw dru Impeak intagdon ud qurnliution 5.3 Amlyticd balance capable ofreadinsto o.oooo1 g.
-
-
Exygen Research
Page 38 q/ 6j
Page 73 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number: PO00113I
..
I Method of AMI*#
I ANAL.\TICAL METHOD
lorthc DocorminnioOof P4uomooUnoic Acid (PFOA) in Fish and C h i by LC/Ms/Ms
5.4 5.5 5.6 5.7 5.8 5.9 5.10 5.11 5.12 5.13 5.14 5.15 5.16
6.0 C l n o m u o ~ h i c S ~
6.1 Aa8iytic.l Column:Fluophro RP ( K o ~ MScientific),2.1 mm x 50 m.511
(PM:82505-052130)
6.2 Tmperalw: 3WC
6.3 Mobile phrc (A) : 2 mM Ammonium
in Water
6.4 MobilePhuc(B): MeuvnOl
6.5 GndiontPmpm
Y lhdnlbl
0.0
I .o
65
8.0
25
20.0
25
22.5
65
Flow Rue
Y - 0.3
35
0.3
75
0.3
75
0.3
35
0.3
6.6 6.7 6.8
- - InjectionVolume: I IpL (CUI bo incmucdto 81 much 81 50 pL).
Q u n t i t h n : Puk A m artarulatandud calibration curve. Run Time: 23 minutes.
Tho .bowconditioarM intmdedu guide.odm y be c h g d inorder IO optimize mC HPLC iylto3a.
Exygen Research
Page 74 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen StudyNo.: PO001131
Exygen Protocol Number: PO001131
I I ANALYTICAL METHOD Methodof Aarlpu far cho DaorminrtimofPsrthuKoo*moic Acid @KIA) in Ftah and Clam bv Lc/Ms/MS
7.0 MSlMSSptcm
7.1 Modo: ElacUoqmyNa@ve MRM mode,monitoring413 + 369 rmz for
PFOA.
Tk above oanditiom M intudd u iguide .Odm y be changed in orda to optimizd cho MSMS aytam.
8.0 PrqNntioIlof S O l U t i ~ 8.1 Mobilephuo
8.1.1 2 mM mnrnonium roarto in wuer u prapucd by dding 0.154 g of d m u 8 m t o t o 1000mLofwatm.
83 Bx~mSoMool
8.1.1 2% .rabiorid in mcthmml u px~aradby dinolviq 2 g of ascorbic .cidinlWnL.ofmah.ool.
8.2.2 30% Dme(hyldicbkrmi1am in tolum ir prcpvtd by bringing 3 mL of dimtbyidichlormilMe to s hrulvoluma of 10 mL with toluene.
Altmula voluma m y be prrparad.
9.0 StududRCprntion 9.1 sundud smd;lFonifia(iM %h*W
9.1.1 Ropur 1nacL soluriono f -100 )rp/mLof PFOA by weighing 10 m y of dylMMDdud(conootd for purity) nnd dilute to I 0 0 mL with mnbrmol in a 125mL LDPE bottle.
9.13 A 1.0 p&L M k U i o n wlution o f PFOA it prepared by bringing I mL of tbo 100 p#mL solution to ifINI voluma of 100 with mcthwl in s 125 mL D P E botlla.
9.1.3 A 0.1 )r@Lfortificationlolutioa of PPOA is p~oplrcdby bringing IO mLoftha 1.0p@Lsolutionto~fINIvolwneof100withmerh~nolin s 121mL LOPE bottlc.
9.1.4 A 0.01 pvmL fortifiation wlvtim of PFOA is pnp.rsd by bringing 10 mL of UU 0.1 pghnL solution to s fiad volume of 100 with mahrnol in 1 125 mL LDPE bonk.
9.1.I) Tho stock f d u t i o n mlutiom M to be mod in imfrigerwr 81 ~ppmximtdyC C ad M atable for a maximum puiod of 6 months fmm tbc dale of plop.ntion.
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO00113 1
Exygen Protocol Number: Po001 13 1
I 1 ANAJATKAL METHOD MscbDd ofAdyais fatbrDaamirutionofPamcomaounoicAcid (PFOA) in Fish and Clamby LCIMSIMS
9.2 SundudC d i i i a r Solutioru
9.2.1 LCIMSIMS calibrationstmula& Mpropand in melhmol via dilution of thc 1.0pphnt foriific.tion solution.
9.21 Tim following ir a lvpicdcamplc: ditionrl concmlnrionsmay bc prcpmduMcdod
Frml
ofFatiflatica WJlicaOlJmL)
1 .o I .o I A
Volunm
(mL1
5.0 2.)
I .o
Dilutcdto (mL)
I00 I00
io0
Caummtion bO/mL)
0.05 0.025 0.01
0.01
IO
I00
0.00)
0.025
IO
100
0.0025
0. I
IO
100
0.00s
IO
I00
0.001 0.0oOs
9.2.3 S m dI alik.tion s t r r d u b in IZknL LDPE narrow-mouth bollla u 2.C to6.C. up to *x montlu.
9.2.4 Altsrmtc volumes mdcenccntmtiiolllof 8Undardi m y bo pnprnd 8s
noodad.
10.0 BatchSolVp
11.0 Sample Extnction
11.1 Woigh 5 g of hrs umple into 50 mL polypropylenc cnrnfugc lubes
(foriifyu Dcodsd. rsplwlid md mix well).
11.2 Md30mLof.celoniuilcvdlhrLoonam*lrlion~afor-l5 minutes
11.3 P b I h e t u ~ i n r ~ S f o r - l b o u r .
11.4 Pack d d l i w tbrSPE blbclUdailUJL0cho pcu-lhrpcd a&$.
11.5 Pack chc 20 mL SPE h b a in quart with 2 g Ooriril. 2 8 ailica gel. 2 g
arbon, md I g LC-NHI.Condition thc cohrrmu with 20 mL of mcthmol,
e. choll M Id,O f rolonihik.
dl wuhor. DO MI dlOW the CdUmn IO
11.6 S i b UIO I25 mL par-&@ flulrr by rinsing with tho 30%
dimatbyldiehlo~lamin toluene solution. Rime thc lluk with lolumc once.
followcd by muhano1 (Ibrea lima). Dry lho flukr completely boron use.
Oilbeby lir-drying 01 with a slmm o f n i t ~ ~ g a ~
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 13 1
I 1 .ANAL%l'lCAL WETHOD
Mcthodof AMIylia for the D*onninuion ofPerflwmocMoicAcid (PFOA)in Fish and Clam by LC/MsIMS
11.7 11.8
1 I .9
11.10 11.11 11.12 11.13
11.14 11.1s 11.16 11.17 11.18
Ccairihge UU SO mL polypaop~merubdl contlininp umple u -2000 rpm
for-lOl.ninUm. Doolnt ulo ectnct OII to ICODditionedSPE column fitted inside the moulhof the psrr-ahpsd flulr. Colla4 the eluate in the I25 mL rilanizcd pcar-rhrpe
fluk.
Md 10 mL of lcstonitdle to UU umplc in the 50 mL centrifuge tube. Hrnnoomitothe bowr fat phro tu4 a t i u u a ~ fhor -30 rccondr and nnsc tbc riuumiwwith -10mL ofacetonihileinto the tube. sb*otho sampleqpin h-lOminutea on Iwrist-actionrh.ltor.
Plroa IhorubdlmImcza for- I burmole.
Cmirifuga tbs 50 mL polypopykw htbw contlininp m p l c at -2000 rpm
fa -10 miouta.
Deunl mC exm* onto UIC smw SPB mlumn. Collect the eluuc into the
umo *shaped fluk ud com!&w with tho eluent h m the initial athutioa Pur 10mL oflootonitri* Ihmu& thc SPE column and combine the eluate in
thCWCpdUplflUk.
Md 3-4
of loctlool to tho extncl in Iho pcu-shaped flask and
wlpomeU nducad plaruro using 8 mWyovqmator (It C 4OOC).
M.lrs tbc ffDll volume, by Nldin#1 mL of 2% usorbic wid in methanol to
thc pur-shaped fluk md rarirl to mix/d*lolw.
~futbca~MWLcVilla~~blepipar. ~~crvnplauaingd~yLC/MS/MS.
12.0 CbrolDltopnphy
12.1 Iqa i the u m c ~ m o uo~f tcrh slmdd, umple and f d t i e d m p l e into the LLYMSMS ayltcm A Jlkrtirm 8laodd mlt precr.de ud follow all
12.2
~YrOdHmpla.
'
StddsofPPOAcxmwp&q
ta u I&
five or mom concmrmion levels
murt k includrdin M uulytiul set.
12.3 An entirea& of cdibntion r u n d d r murt be included u the beginning and ai
the a d of iample et. Smduda mum bc intmponod betwan wny 5. IO
aamplac. AI M dlarutivo. M dlltiro set of calibration stpndardr may be
i n j d U chc beginning of I 101 followed by calibration rtandards
ilurrpaedevery 5-10 lmtplu(to .cEwDt for iwcond rc( of alurdudr). In
utba uro. ulibntion st#ndda mum k the tint ud lut injectm in Y
umple sat 12.4 Uaa l i i IWUIUII RYVOT for qumtilation. Linear atandud curves are
gcnaatcdfor UU d y i e by liaaulegmaion u*lp Ilx weightingof puk m a vmua dibnrion mad.rd amcantnliontutu MurLym 3.3 (or equivalent)
lomvMrynan
~ a 6 ogr a
Page 42 of 65
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..
Exygen Research
Page 77 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
.
- .
Exygen Protocol Number:PO00113 1
1 ' k m - b
M.WNnmbn VOW1783
ANALYTICAL METHOD Mahod of AfulyluforthcDUumidonof Pafluomocunoic Acid (PFOA)in Fish and
C l M S by LuhWMs
12.5 Svlrplo rrrponw rhould DOI o x e d rundud rrrponrcr. Any ramplea thri ucsod dudud nrponsurbould be Mcrdilutod and reanalyzed
13.0 AscoptncoCritair
13.1 ChmmuopmnUIW rbow i)p d of 8 dau@a ion u 369 MU from a pnrenl
of 413 MU. 7hc 413 m u p n a t cmrrrpondrlo tho PFOA anion. while the
duybtorica(369 mu) ropmenbtho IOU of cubm dioxide.
e 13.2 Mahod b l a h mud not contain PFOA U lovelr p a l a than tho LOQ If a
blank c a d m PFOA 0 Iovdr
thn 0.5 ppb. them a new blank simple
mustboobhinod ud the mtim mmustbomox-tcd.
13.3 R a w u i r ofcam1 apikea md murix @a mud bo bowan 70430% of
tho$kmwn vrlua. If r 04ohOl pika falls ouuido tho acccplablc limits, the
a l hm orsullplw huldbcm.cxtnc~.
13.4 h y ulibntia,dlldud found to be a stuLtic.I outlier by using tho Huge
Enor Tclt may bo acludod from mS calculuion of tho calibration curve
Hmrdvor, tha total wrrkr of caliWon rturd.rd,vut could bo excluded
mw~aQlccoduW.ofthctotlln~ofwdudcyjoded.
13.5 Tha uurohtion c ~ ! t i ~ (iR~) tf a ulibntion curvu gcnomtd must be
20.992 (R' 20.985). If ulibntion d t a Bu ouUido lbuc limits. then
c p p r o p h M p s mud be Wrm lo lajwt insmnml oporuion. and the
~athcr~of~lusbouldbcd~.
13.6 hention timol betwoe ttmiuds md umplor mum 1101 driA mom than
f 4 Y. withinMuuIjlic81run. If ntmtion time drifl oxccodr this limit wilhiii UIwlytiul run then thc sa mustbo m y r a d .
14.0 Cdoul.tiolu
14.1 Usethe following qualion to cdcuW the mnouot of PFOA found (in ng/mL. b u d on peak aun) win8 the Nndvdcutw flinur ropeuion panmeten)
- gmmted by h o Mur Lynx soflware program: PFOA found ( a m ) dope
14.2 UM1&folMog oplutionto 0ollvQt tha pwuot of PPOA found inng/mL to ns/s W).
- PFOA bund(ppb)
m p l e weight (g)
-DF fvtcrby which the 6n.I volume vu diluted. ifncecwIy.
-
-
-
Page 43 of65
Exygen Research
Page 78 of 100
Interim Report #2-halysis of Surface Water Samples
Exygen Study No.: PO001131
Fxygen Protocol Number:PO001131
1 &\AL\'llC.U JlETtlOD Method of A ~ I y r i rfor cba D M e i d d o n of PahrwxUmo,c Acid (PFOA) in Fish urd
Exygen Research
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Page 79 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygea Pmtocol Number: PO001131
-
-
ANALYTICAL METHOD
MabodNumbcl: V0001784
__ .-Exygen Research
T o u l P a p ~ :7
Page 45 0f 65
_-
Page 80 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number:PO00I 131
I r y p n Irurcll
Mnhod Numbcr V W I 7114
1 &YAL$TlCAL METHOD
!
1
M W of ADllpufor the Daamhtion of Pcrfluomoctlaoic Acid QFOA) in Vcgelation
bv LCIMSIMS
I .o
2.0
3.0
4.0
5.0
-.
Exygen Research
Page 46 of63
Page 81 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
P.
Exygen ProtocolNumbcr:PO001131
5.4 5.5 5.6
5.7 5.8
5.9 5.10 5.11 5.12 5.13 5.14 5.15
6.0 CtnormllopnphicSflom
6.1 M y t i c a l Column:Fluophw RP (Kcytlonc Scientific). 2.1 nun x 50 mm. 5b1 ( P N 82505-052130)
6.2 Tanprrturr:3O.C 6.3 Mobilo FUM(A) : 2 mM knmoniumAc*rte in W W 6.4 MobibPhua(B): Mellur~ol 6.5 GndieaIPmpm:
Flow Rue
Y " " " " P-
IiauLQmu
0.0
65
0.3
1 .o
65
35
0.3
8.0
25
75
0.3
20.0
25
75
0.3
22.5
65
35
0.3
6.6 6.7
6.8
- - wcaVolumo: 15 pL (ormbe inauscd lou much u 50 pL).
Quntilltiolr:Palr AIU cxtmul ataidudwlibntion curve. Run Tim: 23 mlnucsl.
Tho&ova condiditionrME itltoDdodu 8 guide and may be changed inordcr to optimizO~HPLCayr(0m.
Exygen Research
Page 82 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1 131
Exygcn Protocol Number:POOOI13 1
Iiaygc. RpuI*
LMabod ofAnd&
W N u m b c r VOa11784
I A k A L Y T l W >lETHOD
for thc Darpmiion ofPortlwrwcmnoic Acid (PFOA) in Vegetation bv LC/MsIMS
8.0 PIsplrUionofSotutiom 8.1 Mobilophur
8.1.1 2 mM d u m acetate in wr(a b prspued by ldding 0.154 8 of .mmoniumuaueto 1wo d of water.
8.2 Extnction Solutiom
8.2.1 2% amubic acid in mocllurol isprepredby dissolving2 g of ucorbic
.Oidin1WmLOf~Ol.
8.2.2 30% Ddyldichlo!t4lrw in toluac L pmplnd by bringing 3 mL
o f d i m a h y l d ie~toa8ollvolumoof IOmLwithwlucnc.
Altsmrtc VOmW ay be proprrod.
9.0 sund.rdRcpontion 9.1 S M d v d StocwponjfiortionSolution
9.1.1 R c p r r r ~ n o c k a o l u t i o n o f - l W ~ ~ o f P F bOyAweighing 10my
of vvlytiul Nndpd(conoctodfor purity) and dilute to I W mL wilh
&I
io a I 2 5 y d LDPE bottle.
9.1.2 A 1.0 @L fau&uionaolution of PPOA is PnpMd by bringing I
mL of the 100 p#/mL rohlion to a LLnrl volume of 100 with methanol
in a 125 mL LDPE bottle. 9.1.3 A 0.1 pg/mL forciflcltion solutionof PFOA is prep& by bringing IO
mL of tho 1.0 p g h d mlution to a find volume of I00with mchrml in
L 12.5 mL LDPE botlle.
9.1.4 A 0.01 pg/mL fortihutioa solution of PFOA is p r e p a d by bringiny
10 mL of tho 0.1 &nL solution to a hrul volume of 100 with mglyDolina I25 mL LDPB boltla 9.1.5 Thc ltocL .ndlOrti6utioaaolutloiu Mto be atorcd in 8 nbiDorptor(It
q p m x h l y 4 T lad arc stable for a nuximum period of 6 months
from the due of prrplrrcion
9.2 sc.ldudCdbntionSolvtiom
9.2.1 LCIMSIMS ulibntion mbnduds YI prepuod in methmml via dilution oftho1.0pphnLlortifiutionsolutioa
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygen Protocol Number:PO001131
ExypnD*ush
I hielhod of And*
MnhodNumbn VOOOl784
I MALYllCAL lllllTLlOD
la tho Daamiarton of PCmuomocunOicAcid QFOA) in Vegetation
by L c / M s / M S
9.2.2 The blbwing ir L tvpiul cxmple: ddilionrl arncentntionr may k
preplrodUosdod
Q '.' OIFatificltia
Sohmbn(rx/KlL)
I .o
1.0 1.0 0.W 0.025
PI
Vdumc (mL) 5.0 2.5 1.0
IO
IO IO
Dilukdto (mL) 100 IW IW
la0
100 IM
Find caKmm(ion
[UdlllL) 0.05 0.025 0.01 0.005
0.0015 O MI
10.1 Ea91buoh o f pmpfcr cxaraed (cypkrlly 20 or lm) mwt urlude ai leasf one ull(l0ltod conlml rad fuo unlmtal wnuola fortified ai known concealntioru (lab umbul pike) lo vaify pmoodunl mcovory for the balch
101 Ryimumb la dcld d Lbantorydupliurcr md spikes will be rpecilied in thc qvllity UNMWplan for thispmject.
11.0 Sample Exm*ion
11.1 Weigh 5 g of horn wnple into SO mL polypmpyleoc centrifuge iubes (forti@u oaeded,replacelid rad mix well).
11.2 Add 30mL of lcctonitrilsand Ib*oon a wrid d o n shakerfor -15 minula. 11.3 Cenbifuge the SO mL polypropylenetuba containing sample at -2000 rpin
fa-10 minuta. 11.4 Pack ud Mnditim tho SPR tubcsradaimthcpur-dupuJ!lads. 11.5 Pack the 20 mL SPE hrbcl in mqnmw with 2 g floriril. 2 g silica pel. 2 g
c&on, Od I g LC-MIz. Gmdition Iho columna with 20 mL of mclhurol. ulsn 20 mL of W(0nitrilr. Diacud all wuhu. Do not allow the column to
dy.
11.6 S i k h UK 125 mL peu-shqd f l u k ~by nnring with the 30%
dimethyldichlomsIb intolwaowlution. Rinw h e fluk with toluene once. followed by methnol (tbnr timw). Dry the tL*r complstely before use, either by lir-dryiop Q wiL a strean of ni(r0gm. 11.7 Deumth e x m on lo L wnditionedSPB column fitted inside thc mouth 01' chs pop-asedauk. & l l a the elwte in ulc 125 mL silanired pcu-rhqx nuk.
~ --
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
~
~
~~
Exygen Study No.: PO001 131
ExygenProtocol Number: PO001131
13.1 Chmmtogmmmud rbmv ape.& ofadaughtor ion at 369 M U from a parent of 413 MU. Tbo 413 MU puslt SMOrpandr to thc PFOA anion. while Ihr duuphccrion(369MU) rrprrronuthe IOU of cuboo dioxide.
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._
Exygen Research
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Page 85 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1131
Exygen Protocol Number: POOO 1 I3 I
13.2 13.3 13.4 13.5 13.6 14.0 C&ul.tiau 14.1
14.2
14.3
~~
_.
Exygen Research
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Page 86 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1131
Exygen Protocol Number: POOO 1I3 I
Appmvd By:
ToulPrpot: 7
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InterimReport #2-halysis of SurfaceWater Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001131
1.0 smpe
This mahod u to k cmpr0)d for the ivrlition ud qmtitation of psmuomoctanolc acid by High PafomuDoc Liquid Chnalogmphy couplcd lo a tandem MMS Spccmnmic D*caor (Lc/Ms/Ms)in anrlld liver.
2.0 Su%y
2.1 Almyrokavcnfch~bontocypncticcr
2.2 W t the lpprqniltoMSDSW o n h a d i n g m y c h m i u l for proper safety
prsuutiolU.
3.0 SlmplcRoquiranaU
3.1 Allcut 5poftoltrmnpkforextrrtion.
3.2 Sunplor should ba proceurdkfm extraction. Place the fromsample in a
food paaror md hmno-
with dry ice. Place the a m p l a in containers
md law opm in hzut strmp ovani& b dow for &o dioxide
rubliiuion SUI ud pLvc the umplu in frozen r t o ~ euntil lime of
~ a p iA~lta.natdy, if thm is M b d l i c i u u unounf of sample (-Ius than
5 0). thenM pnnx&ng is n c ~ o u l ymd lhc samplewn be used as supplied
3.3 Sample collsCtim proccduru will be lpccifid in the sampling plan for this
pmjcct.
4.0 R w n d S l u d u b
4.1 W*-HPLCgdc
4.2 Mclhnol-WLCH
4.3 Acdonibih-HPLC@
4.4 4.5
- knmollirrmAcebte-Ac.s.RuJcmtond0
paflumooaam'e Acid S i m A l b i c h
v, 5.0 ~ t m d ! 4 p i p m m U
5.1 A high p0"mYlcf liquid
@IC
of pumping up lo 2
mlwDtr cqwppcdwtb 1 wiablc volume qOc(0r -IC
of injectin8 5.2W
pL wmos10d101lmdem M m SF.o?wMy(LC/MSmrS). 5.2 A dovim0 collestm&la br peak urteprulon mdquanliwion.
5.3 rwyliul blllnce Clplbl. of MdinpLo D . m I p.
5.4 SO mL dirponblcpolypmpyknc mbiip tubas.
5.5 15 mLdilpoublcpolypmpyloaoCcmtriiptubes.
5.6 ~ l e m i u o p i p a (t5C1oopL, 100-2ooUL).
5.7 125-td LDPE m W - m Q u t t hbollla.
5.6 2 m L c l w HPU:vid kit.
Page 53 0x65
Exygen Research
Page 88 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1131
Exygen Protocol Numbcr: POOOI 13 1
.- Ah&Y"ICAL A f m f O D
I J Mocbod ofAmlyru fcf thc DaOrmida~of POmuomod.noicAcid (PFOA) in Small
5,9 L)iqyablapipcua. 5.10 Auloprpata (1001ooO pL md 10-100 pL). with dilpouble tips 5.1I W u a S q PJr V r 6 w (Io) tCl8 SPE Cuaidgu. 5.12 SPEmwnmm~ilobl 5.13 Tirn#miur. 5.14 Wd-aclion alukci. 5.15 C d h g E clplbk O f o p b I I i q 15 mlpOlyPmpylUilENbOr U 3000 Qm.
6.0 QuomuosnphicSyitcm
6.1 Aml)tiul Column:pluophuRp W ~ IScWientibc), 2.1 mm x 50 mm. Sit
( P N 82505452130) 6.2 Tmpauuro: 30% 6.3 Mobile pbuc(A) : 2 mM Ammonium Acmcc in W.(n 6.4 Mobilephpe(B): Mathawl 6.5 O n d i e n t R o ~ u ~ ~ :
flow Ruc
9 UImlLSliol
0.0
35
0.3
1 .o
65
35
0.3
1.0
25
75
0.3
20.0
25
15
0.3
22.5
65
35
0.3
6.6 6.7 6.8
- - lnjdmVohunc: 15 pL (ank inmuod to u much u 50 #L).
Quatit.tion: Peak A m o x t a d lurdud dibratioocurve. RUB Time: 23 min.ocSr.
Thor b o v 0 W t i o 1m~mirrtadcdu rguida md my be chlagd in orderto
0plLnizothcHPLcry.tom.
7.1 Modo: E1-y PFOA.
NegativehfRM modc,monitoniq413 -b 369 d z for
'Tbe above d t i w u uc intaaded u L @de MII may be changed in order IO optimizethe MSMS lynan.
Exygen Research
Page 54 of6S
Page 89 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.:PO001131
ExygenProtocolNumber: PO001I 3 1
I .1 A X d A T I C M METHOD Mslbod o f d y d r forb DomminuioaofPlsfluomo*norc Acid (PFOA) in Small MMrmlLiverby LC/Ms/MS
8.0 RopMtiooofsolu~
8.1 Mobihphro
8.1.1 2 mM d m &-URIiCn water ia prcpucd by adding 0.154 y of` d u mrewe IOIO00 mLof water.
Altmu(o voluma mly bo PISpMd
9.0 SundudPmpamion
9.1 S M StawForliflUtimSolution 9.1.1 Rspma no* solution of -100 pghnLof PFOA by weighing IO mp ofmuly(ialrtPdud(corndodf~purity)~uldllutoelO0mL with mclhmol ina 12-mL LDPE W e . 9.13 A 1.0 p & L fortificmtion solution of PFOA is prcprrod by bnnginy I mL of tho 100 p g / d aolutioo (0 a find volume of IO0 with mahmol m L 125 mL LDPE boltlo. 9.1.3 A 0.1 p@L fortification wolution of PFOA is prspucdby brinpinp IO mL of h e 1.0pg/mL aolution to a find volumeof IO0with mrhrnol in 8 12) mL LDPE boule. 9.1.4 l l w nodr ad loltltlcuionaoluti~Mlobaalored in I~fril~cruoPIr a p p x h l d y 4`c md arc atable ku 8 nuximum period of 6 months h m (he date ofpqurtioa.
9.2 SurdudWbrm'onSolutiom
9.2.1 Lc/Ms/Msc r l r i o n ltlndpdr am proprod in m h o 1 via dilution of chs 0.1p&L aaciec.tionsolution
9 2 3 l l w followinl~ia a typical orunplc:rdditiod wnccnbuionr may he pcpucdunoodcd.
...
..
IW
1.0
IW
2.0
IW
1 A
100
I .o
51)
10
IW
os
1_ 0_
LO
I.W~~
0.1
1.0
IO
LOO
0.1
93.3 Ston dl UlibntiOOarrdudr in 125-mL LDPE numw-mouth buttler
rt 2.C to 6% up (0 ah montlu.
9.2.4 Altcrmtcvohuna md concSnm(i0ru of rt.nd.rda may bc p r e p a d YI
mdcd.
P.*e 4 O f 7
/-
Page 55 of 65
__
Exygen Research
Page 90 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 13 I
UY8- I*mrsL
Metbod ~ u m b ovr woiias
I AhdU.YTICALMETHOD
I
MathodofMyda fortho t ) e t m h b'Mof Pafhuvoocunoic Acid (PFOA) in Small
Munnml Uvcr by L C l M y M S
10.0 BltehSocUp
10.1 E r h bw1 of umplu oxmctal (typically 20 or Ius) mwt include at least 00. unhmtad control ud two untmted conaols foriificd at known c o ~ (Iw &canbul pike)to Wify pmcedunl raovery for the batch
10.2 Rcqummb far field md L.bontwydupliuta .nd spikes will be spzcificd inthc qualityUIP~Y~OPIOUI for thli project.
11.0 Sunplo Extnctiw
11.1
11.2 11.3 11.4
11.5 11.6 11.7
11.8
I I .9 11.10
11.1 I
Wmgb 1 g of umple into i50 mL polypropylene sentnfugc tubes (fomfy u nocded. replro lid md mix well). Nok that Jtcmite weightsof liver mny be
mDuuroddcpding Mthc umpbsits ivrilable for UM.
Add watertothe umple fora 6 n d volumeof 10mL. Homogah umplo wing Ltinwmizor for -1 minuts. TMIQ I mL of thc =PIC wing 8 dispouble pipene into a I5 mL
dicporr6k raaui- arbc. Add 5 mLof raoaitrilo ud duke b r -20 minutuon 8 wrist-actionshaker calrifuptho tubelu -3000 rpm tix -5 minutel. DDunt the mpaualmt into I 50 mL dispouble conaifigc Nbe md add 35 mL o f W W .
Conditica tho G g WE d d g a ( I g, 6 mL) by pauing 10 mL methanol Pollowed by 5 mL of HPU:water (- 2 WMCW)1.101 let columnrundry
- Lod thc&lo and t i o n c d C. SPE &ridge. Discad elurte.
Elvtc with -2 mL of m c t h w l Collat 2 mL of elute into a gmduted
15 mLpolypmpyIa18 c s a w lute(find volume 2 mL).
A M l p umplol *eloctrmpry LCIMSIMS.
12.1 lnjoct tho umc lDlo(ID1of orb Itladpd.rrmpk .adfortified umple into the
LUMSMs 8 w . A C d h U h d d must precede md follow all
d y r e d umplu. 12.2 SuodwbofPFOA
' to at IOU( five M mom cowontntionlevels
lnUdbe illCludedhlUI UldYdCdSo2
12.3 An entire IOI of ulikuion stauldI must be included at the begnningand ni
thc ad of L umplc Io(. Standvdr must be mlcnprncd between every 5-10
umplu. As M d ~ t i v c M , cntirc $01 of ulibntion sianndudr may hc
idcct6d L( the be@nninp of L Io( followed by cslibnuon standards
iDlarpoMd way 5-10 rvnplu (to account Tor L ward set of rtandardr) In cilhu cue. ulibntion wdudr m w be the first md IMI injection in 1
umplc ret.
12.4 Urn liner mndd culv~for qurntitrtion. Liocu sundud curves arc
gcnmtcd fortho d y l o by litmrregmuionwing Ihr weighting ofpeak uea
P-
__.__-
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Exygen Research
Page 91 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn ProtocolNumber: PO00113I
13.1 chrrmutoopn mwt lbow a porlr of (Id~ughtuion u 369 m u from a parent
of 413 mu. TbO 413 m u p ~ l lwlnwpod#lo tho PFOA mion. while the b o (369 .mu)repmenutho loor of cubon dioxide.
13.2 Mabodb b k a mud nol contain PFOA u levels greaterthan lhe LOQ. If II blU* corctlinr PFOA It lcvohgoaertbvl IO nu& lhcn new blank sample mudbc obtainednd tbs&M nuutbo n-oxtracted.
13.3 Rocowim of control @kea and llutrix spikea must br bewm 70-13W.of thir know values. If a oolllml rpiko Llh Wuide the wccpublc limits. he oath I* of mnple~cbould bo ~ w l m f t o d . Any maoh spike ouuidc 7033W dmdd bo evaluated by tho uulytl to determine if re-extnction is
W#mtltd.
13.4 Any ulibntim mndvd bud to bo l Uatistiul wllicr by wing the Huge Enor T a m y bo excluded fmm tbs uloulrtion of thc ulibntion curve Howcvn, tbs l0t.l numbs of dbr8tion rlndub I b r could bc excluded mudnot a d 20% of ulo totalnumbwof rtad.rdr injcctod.
13.5 lb conolltioD cocflicioat (R) TOC d i i c w c l gemrated must be 20.992 Ca' 20.98S). If mlibntioon m u l l s 1.11 outside tbcsc limits. then mum k ULea to djusi hdmnncnt Dpcntion. and the rurdpdr of tho nlcvlatmI of mmplorrhovld k rurulyzcd.
13.6 RdoDtion tMol bdwmn Nndmda md u m p l ~m~wi MI drin mom than
f 4 K within UI d y l j c d m. If roccOtion lime drift cxcccdr lhia limil within
U I d ~ C Mdltbmthc M nIud bo d y r e d .
14.0 Calcul.tiw
14.1 Use Uw C o w oquUionto mIcu*Icthc uwunt of PFOA found (in np/mL.
bated 011 peak ua)wing thc d u d ~ulye(lincu repusion parmc~en~
genmbd by t h o b Lym mfhrn prom:
- PFOA f d (nphnL)
x DF x aliquot fwor dopc
-DF Ewrby wiicb tho Mvolume wu diluted.if necsulry.
Auquothocar-lo
Exygen Research
Page S7 of 6s -.
Page 92 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
Exygcn Protocol Number: PO001 131
EayaeuRsanuch
M8lbod Numbcr VooOI7IJ
I 1 ANALY 1ICAL METHOD MethodofA d p b for thcDanminuiono fP e r f l q i c Acid (pFOA) in SmII
MaunulL i v r by w/Ms/Ms
-.
14.2 For umplr falifld with bmwn uIIouII1(o f PFOA prior IO extraction. use thc rollowiugOpUrtiaD(0u*u*tc tho porsclllncovcry.
14.3 Use tbc Mowingquatiion(0 oonvmthe mount of PFOA found in n@mL IO lyls W).
PFOA bund@pb)-
urnplc weight (g)
Exygen Research
Pl'C 7 or7
Page Sa of 65
Page 93 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
n
Exygcn Protocol Number:PO001131
ANALYTICAL. METHOD MolhodNumber: VoOo1786
AnalyticalT d n g Facility:
Exygcn Raomh 3058 Rcrcpch Drive State Collage. PA 16801
._
.. .... . _ _
Page 59 of 65 ._
Exygen Research
Page 94 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygcn Protocol Number: PO001 13 I
1 ' E.--
M*bodNumkr VW1786
ANALkllCAL METHOD Mahod of AnRlylhfor IhcD c k d n a h of Porfluomocpnoic Acid (PFOA) in Small
Munmd Saum by LcIMs/Ms
-
I .o stops
Thir mahod u to bs ompbM lor tho &OW md qurmtitati~of pcrfluomcctanoic wid by Hi@ P e f o m u ~ ~L~iqeuid clmmalography coupled to a tandem Mass
Spostmmctn'c Dclfftor (Lc/Ms/MS) in mdI manma1serum.
2.0 sflay
2. I Always oboosvcd e labontorypncticcr. 2.2 Consult L o .ppmPriakMSDS beforehudling any chemical for pmpcr safety
prsclutionr.
3.0 Smplc RoquLaamt
3.1 AtlloutImLoftwlaunploforo1o3h1otion.
3.2 No umple pmooUiag u noodod fm
umplcl. Howcvpr. b u n worn
lmrplclmult to llkwd toEomplotely b w IO mom rosnpcn?uobefore w.
3 3 Smplo solladonpmcedvror will k lpoci5ed in the umpllnn plan for this
4.0
5.0
Exygen Research
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Page 95 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: POOO1131
Exygen Protocol Number: POOOI I3 I
EanuEOurh
MelkdNlllllkrVW17Yb
I 1 Alhi.Yl1CAL. HETHOD M c l W o f A n r l y d : I m t b c D a m n i n u l o n o f P a n ~ i c A c i d (PF0A)inSmull
Mrmnul Scrvm by LC/MS/MS
_.
5.14 wrirt-.ctionItlnka. 5.15 Ccntrifug EIp.blcafspilmiq IS mL polypmpylme tubs at 3000 pm.
6.0 Cluumuo~icSyrtcm
6.1 Arulyticdcolumn:Pluophw RP (Koyrtons Scientific).2.I mm x 50 mm. 5p
(PM: 62505052l30) 6.2 Tompenturo: 3o.c
6.3 Mobik phvo (A): 2 mMAmmonium Ambte in Warn 6.4 MobilcPbuc(B): Mahrmol
6.5 GndicnlRogrm:
vI .o
8.0 20.0
22.5
u 65
65
25 25
65
Flow &IC
ZLBImlLminl
35
0.3
35
0.3
15
0.3
75
0.3
35
0.3
6.6 lDjoaion Volume: 15 pL (CUI be incrwcd to u much u 50 pL)
6.7 6.8
- Quaatitstion: Puk A m - exmud dlndud crlibntioncurve.
run Tim^: 23 min~ler
The aboveconditioluMirUcndcd8a 8 guide md may be c h p e d in order to OptimiEO the HPLC :yrten.
7.0 MS/MSSyUmn
7.1 Mode: Elweorpny Nquive MRMmode, monitoring4 I3 +369 m/z for PPOA.
Tlw above oooditiorv am in(0Sldced n I @ awl may be changed in order lo optimicthsMSMS tynan.
8.0 PrcgMtioo of solutiolv 8.1 M o b i l c ~
8.1.1 2 mM -Nw u-aaIe In w m i: prepared by adding 0.154 g of ammonium .cct.tcto loo0mL o fw w .
AltomnovahlmeJrmyboproplJDd
Exygen Research
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~~
Interim Report #'L-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygen Protocol Number:PO00I 131
m-
M I t h o d N d r VooO176b
1 J ANALYTICAL ,MISTHOD Methodof Analylir f o r t h D e e a of P~uoroOamoicAcid (PFOA) in Small
Mlmmal Serum by LCiMsIMS
9.0 Stlrrd.rdRsplntion
9.1 Suobrd Stawponificuion Solution 9.1.1 Pmpuc a atock solutionof -100 p&/mLof PFOA by weighing IO nig Of Wldyt*J a ( u d u d (-td foJ purity) Md dilute to 100 mL w l h mcthrnol in 8 125-mLLDPE bonle. 9.1.2 A 1.0 &TIL forciflw'on solution of PFOA is prop8redby bringing I mL of tho 100 pdmL aolution IO L 6nnl volume of L O O with w h o l in 8 125 mL LOPE W e . 9.1.3 A 0.1 p g h d fortifiulioocolution of PFOA hprepared by bringins IO mLofcbc1.0~~mLsollbionto~hnrlvolumclo0f0wiUlmcrhlnolin L 125 mL LDPe botue. 9.1.4 Tbo rtocL d folti6UtioacolutimPO to bo s t d in a refripemtorat lppmxiMcoly4.C urd uc atable for L m i m u m period of 6 months borncbcdueof prepmtior~
9.2 Stpdvd Cdibruioa SolvtiOrV
92.1 LCIMSIMSollibntionatmlmda uc prrpMdin m*hmol via dilution of (ho 0.1 rphaL W6c.tion M~UI~OII.
92.2 'lbfollowing u L typical ample: additional coocurtruionr may be
PrOpMd U MCdOd.
Caumbmion
Final
ofFmifKuiia Volume Dilutedo Concmmrioo
Solutim(r&nL) (mL)
(mL)
(ndtnL)
IW
5.0
IW
5.0
100
2.0
100
2.0
100
1 .o
IW
I .o
5.0
IO
100
0.5
2.0
10
100
0.2
1.0
IO
1W
0 .I
9.2.3 Store all ulibntions t d w J 8 in 125-mL LDPE nurowmouth bottles
u ZOCto 6T,up tor*.monthr.
9.2.4 AltanUe wluma ud coaccatntiocuof C t m d d may be preparediu
nseded.
Exygen Research
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Page 97 of 100
Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001 131
Exygen Protocol Number:PO001131
Manmal Senun by LCMS/Ms
11.0
11.1 M-
1 mL ofrrmplc into i50 mL polypmpylcac cenvifu8c tubu (fortify
u oecdod.rcp*cc lid and mix well). NOCS th.1 allmite volumes of serunl
m ybe mMllodw a g on the umple lise availableLr we.
11.2 Add wuor (0 tbe umple faIhlvolumeof 20 mL Cap tightly
11.3 Voltcr for4 minuto.
11.4 'hasfax 1 mL of tho umple wing a dilpoublo pipate into a I 5 mL
dispmlblcCoatriRy0~.
11.5 Add 5 mLof loawihile Ild chkc for -20 minutea Ma wrist-action rh.kcr.
11.6 Cavifuga tbetubor It-3000 rpm for-5 minuter.
11.7 DoMt tbo supmml~Iinto L SO mL dirpoublc ccoeifuge lube and add 35
mL of w e .
11.8 Condition tbc CISWE cutridgw ( I g, 6 mL) by ppsrins IO mL mnhanol
followedby S mL ofHPLC w&r (- 2 dmp/ls). Do no1 Is(column N n dry
11.9
tbt rrmolcon cwditioncdCIISPEcMxidnc. Dixud eluue.
12.0
12.1 Inject the umc uDouo(of each ttmd.rQ irmple and fortitid sample into the LClMslMS ryasm. A clllbntion amdud mwt pmodc and follow all
~yzodumplcl.
111 Sluxldsof PFOA
' m u lout ftvoor mora OOrSe~VUiOnlevels
mustbo included i n n mlytiulset.
12.3 An &set dulibnth urndds mutt be included.Ithe beguuung and ai
the end of 1 umple set. S l d w & m u be inlmpmed bchucm every 5- II1
umplcl. h UI lltanuivo, 111 onlirc KI of calibration standards may be q k t s d U mS beginning O f 1 MI followcd by cdibrauon rtandnnlb k&mpomd cvay 5-10umplos (to lffount for a w n d set of rundprds). In oicba cuo, &Won rt;lodub mutt bo the fint and lmt injection m a
U m p b 8eI.
12.4 UWl i i r(udud CU~YDI for quvltitatiim Linear ~(ud.rdcurves are gamated Tor the d y c by~ l i n a regmuion wing Ih weightingof peak area v m w dibntioa ltudvd concamtionwing MwLynx 3.3 (or equivalent)
wmvpo lystmn.
12.5 Sunplc rorporvc lbould not exceed rtudud M~OIUCS. Any samples ihai n c d almdvd rclponrcllbouldbe further diluted ud rcuulyzed.
Exygen Research
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Interim Report #2-Analysis of Surface Water Samples
Exygen Study No.: PO001131
ExygmProtocol Number: PO001131
' ' E a r n I*rusb
MIkdNumkr VMwII786
ANALYTICAL MYTAOD
Method of M y d r Emthc DacrminrtionofPOmuomocmmicAcid (PFOA) in Small Mumnd Scnunby Lc/Ms/Ms
13.1 ChasulopnmInUd cbow L POJ:of 8 daughter ion at 369 mnu lrom a pucnr
of413 mu. Tba 413 MU pMDt comrpoadt to h e PFOA anion. while IIK
h g h t a ion(369 MU)npclmtlthe Ion of urbondioxide.
13.1 Mnhodb h k a rnw not camin PFOA u level8 greater &an the Lop. If a
blank con& PFOA U level8 pats thn 10 WmL. than a new blink
umple must be Dbtllnedmdtheeatire wl must be Ic-oxv.sIcd.
13.3 RccoWic* of Foatml @or md lruhix spikes muct be Ween70-130./0 of
thcir known vdua. Ita conhDl spike f8lls outrids cho acceptable limits. the
&set of umpIa lhovldbe m-ahctcd. Any murix spike outside 70-
130% b u l d be wrhutod by tbo uulyn to dammine if mexulcrion is
WmULud. 13.4 Any mlibntioll lilldvdbund to be I UUi8ticrl o u t b by wing the Huge
h Tclc. may be cxchdal h tbo ukuluion of the calibration curve
Howsvcr. tbc tow numkr of dbntion amdvdr tbU could be excluded
must mlo x c d 20% oflbctow n u m bof aMdudr injcctcd.
13.5 The avnlrrion eOemciCm (R) for calibration WCIgenerued must bc
20.997. (R' 20.965). If cllibntion rsrvlu fdl outside thew limits. then
lppmprirtc aapr muat be taka to mljajya iiumunent operation. and rhc
8tMdads O r h NIWMrsIt of m p l u rhouldbe m r l y r d . 13.6 Ramtion h a bdwm iuadudr Md umplo8 musl not drift more lhdii
t 4 K witbin Muulyticll M.If romtion time drifI exceeds this limit within
c
Mwlytical tunthenthe Ict must be rcuulyrcd.
14.1 Use the bUowkgsqpdioalo UkuW the mountofPFOA found (in n@mL.
bwd at p d ana) wing the ecurve ( l i m regression panmetem
~Cnontodby the Maw Lynx wAwm pm-:
PFOA found (n&L) =
x DF I aliquoc factor
- - slope
DF Mor by which tho &ul wlvmow u diluted. if (LDOO~UI~. Miqwt frCa 20
14.2
. .....-. ..
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Interim Report #2-Analysis of Surface Water Samples
Exygen StudyNo.: POOO1131
Exygcn Protocol Number:POOOI I3 I
ai^. rumucb
MnhodNunhr VOW17116
- ANALYTICALMETHOD
I J Mclhodof M y a i i for tho Dnorminuionof PSmuomoctanaicAcid P F O N in Small
MMmd Serumby LcIMs/MS
14.3 UK tho fo1bwing eqvlrion 10 m v Q t Um @noun1of PFOA found in nlymL IO
PPb.
PFOA
fouud
@pb)
-
nmplc wlumc (mL)
--
- - ... -
- -._
Exygen Research
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Page 100 of 100
