# 

GLP10

- 01 -02, Interim Report 3 5: Analysis of PFBS, PFHS, and PFOS in Groundw ater

Samples; Decatur Waste Water Treatment Plant (WWTP) Pump Test

– June 2012

Study Title

Analysis of Perfluorooctane Sulfonate (PFOS), Perfluorohexane Sulfonate (PFHS) and

Perflu orobutane Sulfonate (PFBS) in Groundwater, Soil and Sediment for the 3M Decatur Phase 3

Site

- Related Monitoring Program

Data Requirement

EPA TSCA Good Laboratory Practice Standards 40 CFR Part 792

Study Director

Jaisimha Kesari P.E., DEE

Weston Solution s, Inc.

1400 Weston Way

West Chester, PA 19380

Phone: 610

- 701

- 3761

Author

Susan Wolf

3M Environmental Laboratory

Interim Report Completion Date

Date of Last Signature

Performing Laboratory

3M Environmental Health and Safety Operations

Environmental Laboratory

3M Center, Bldg 260

- 05 -N- 17

St. Paul , MN 55144

Project Identification

GLP10

- 01

- 02 -35

Total Number of Pages

74

Testing Cert #2052.01 The testing reported herein meet the requireme nts of ANSI/ISO/IEC 17025:2005 “General Requirements for

the Competence of Testing and Calibration Laboratories”, in accordance with the A2LA Testing Certificate #

2052.01. Testing that complies with this International Standard also meets principles of IS O 9001:2000.

### This page has been reserved for specific country requirements.GLP10-01-02 ; Interi m Repor t 3 5

Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Sample s

Decatu r VWVT P Pum p Tes t - Jun e 201 2



### GL P COMPLIANC E STATEMEN T

Repor t Title : GLP10-01-02

, Interi m Repor t 35 : Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Samples ; Decatu r Wast e Wate r Treatmen t Plan t (VVWTP ) Pum p Tes t - Jun e 2012

.

Study : Analysi s o f Perfluorooctan e Sulfonat e (PFOS)

, Perfluorohexan e Sulfonat e (PFHS ) an d

Perfluorobutan e Sulfonat e (PFBS ) i n Groundwater

, Soi l an d Sedimen t fo r th e 3 M Decatu r Phas e 3

Site-Relate d Monitorin g Program

.

Thi s analytica l phas e wa s conducte d i n complianc e wit h Toxi c Substance s Contro l Ac t (TSCA ) Goo d

Laborator y Practic e (GLP ) Standards

, 4 0 CF R 792

, wit h th e exception s liste d below :

• Thes e ar e environmenta l sample s wher e ther e i s n o specifi c tes t substance

, n o specifi c tes t

syste m an d n o dosin g o

f a tes t system

.

• Th e referenc e substance s hav e no t bee n characterize d unde r th e GLP s an d th e stabilit y unde r storag e condition s a

t th e tes t sit e hav

e no t bee n determine d unde r GLPs

.

Gar y Mohensteii /

Sponso r Representativ e Dat e

Jaisimh a r^ari

, P.E.

, DEE

, Stud y Directo r

Pag e 3 o f 7 4GLP10-01-02 ; Interi m Repor t 3 5

Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Sample s

Decatu r VWVT

P Pum p Tes t - Jun e 201 2



### QUALIT Y ASSURANC E STATEMEN T

Repor t Title : GLP10-01-02

, Interi m Repor t 35 : Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Samples ; Decatu r Wast e Wate r Treatmen t Plan t (WWTP ) Pum p Tes t - Jun e 2012

.

Study : Analysi s o f Perfluorooctan e Sulfonat e (PFOS)

, Perfluorohexan e Sulfonat e (PFHS ) an d

Perfluorobutan e Sulfonat e (PFBS ) i n Groundwater

, Soi l an d Sedimen t fo r th e 3 M Decatu r Phas e 3

Site-Relate d Monitorin g Program

.

Thi s analytica l phas e wa s audite d b y th e 3 M Environmenta l Laborator y Qualit y Assuranc e Uni t (QAU)

,

a s indicate d i n th e followin g table

. Th e finding s wer e reporte d t o th e principa l investigato r (P.I.)

, laborator y managemen t an d stud y director

.

Inspectio n Date s Phas e Dat e Reporte d t o

Inspectio n Date s Phas e Testin g Facilit y Managemen t Stud y Directo r

08/09/1 2 Dat a an d Repor t 08/20/1 2 08/20/1 2



### QA U Representativ^j? ^ Dat e

Pag e 4 o f 7 4

# T ABLE OF 

# C ONTENTS

GLP Compliance Statement

...................................................................................................................... 3

Quality Assurance Statement

.................................................................................................................... 4

Table of Contents

................................................................................................ ....................................... 5

List of Tables

................................................................................................ .............................................. 6

1 Study Information

................................................................................................ ................................ 7

2 Summary

................................................................................................ ............................................. 8

3 Introduction

................................................................................................ .......................................... 8

4 Test & Control Substances

................................................................................................................ 9

5 Reference Substances

..................................................................................................................... 10

6 Test System

................................................................................................ ...................................... 11

7 Method Summary

............................................................................................................................. 11

7.1 Method

.............................................................................................................................. 11

7.2 Sample Collection

............................................................................................................. 11

7.3 Sample Preparation

.......................................................................................................... 11

7.4 Analysis

............................................................................................................................. 11

8 Analytical Results

.............................................................................................................................. 13

8.1 Calibration

......................................................................................................................... 13

8.2 System Suitability

............................................................................................................. 13

8.3 Limit of Quantitation (LOQ)

.............................................................................................. 13

8.4 Continuing Calib ration

...................................................................................................... 14

8.5 Blanks

................................................................................................ ................................ 14

8.6 Lab Control Spikes (LCSs)

.............................................................................................. 14

8.7 Ana lytical Method Uncertainty

......................................................................................... 16

8.8 Field Matrix Spikes (FMS)

................................................................................................ 16

### 8.9 Lab Matrix Spikes (LMS)

.................................................................................................. 17

9 Data Summary and Discussion

....................................................................................................... 17

10 Conclusion

................................................................................................ ......................................... 22

11 Data/Sample Retention

.................................................................................................................... 22

12 Attachments

................................................................................................ ...................................... 22

13 Signatures

................................................................................................ ......................................... 23



# L IST OF 

# T ABLES

Tab le 1. S ummarized PFBS, PFHS, and PFOS Results (WWTP Pump Test

– June 2012).

... 8

Table 2. Sample Description Key Code.

..................................................................................... 11

Table 3. Instrument Parameters.

................................................................................................ . 12

Table 4. Liquid Chromatography Conditions.

............................................................................. 12

Table 5. Mass Transitions.

........................................................................................................... 13

Table 6. Limit of Quantitation (LOQ).

.......................................................................................... 13

Table 7. Laboratory Control Spike Recovery.

............................................................................. 15

Table 8. Analytical Uncertainty.

................................................................................................... 16

Table 9. Field Matrix Spike.

......................................................................................................... 17

Table 10. Laboraotry Matrix Spike

.............................................................................................. 17

Table 11. DAL GW WW8R 120627

- 1000

................................................................................... 18

Table 12. DAL GW WW8R 120627

- 2130

................................................................................... 18

Table 13. DAL GW WW8R 120628

............................................................................................. 19

Table 14. DAL GW WW9R 120625

............................................................................................. 19

Table 15. DAL GW WW9R 120625

- 2400

.................................................................................. 20

Table 16. DAL GW WW9R 120626

............................................................................................ 20

Table 17. DAL GW TRIP01 120625

............................................................................................ 21

## 1 **Study Information**

Sponsor

3M Company

Sponsor Representati ve

Gary Hohenstein

3M EHS Operations

3M Building 224

- 5W

- 03

Saint Paul, MN 55144

- 1000

Phone: (651) 737

- 3570

Study Director

Jaisimha Kesari, P.E., DEE

Weston Solutions, Inc.

West Chester, PA 19380

Phone: (610) 701

- 3761

Fax: (610) 701

- 7401
j.kesari@westonsolutions.com

Study Location

Testing Facility

3M EHS Operations

3M Environmental Laboratory

Building 260

- 5N

- 17

St. Paul, MN 55144

Study Personnel

William K. Reagen, Ph.D., 3M Laboratory Manager

Cleston Lange, Ph.D., Principal Analytical Investigator. ( clange@mmm.com ); phone (651)

- 733

- 9860

Susan Wolf, 3M Analyst

Kevin Eich, Analyst

Kelly Ukes, Analyst

Study Dates

Study Initiation: March 8, 2010

Interim 35 Experimental Termination: August 3, 2012

Interim Report Completion: Date of Interim Report Signing

Location of Archives

All original raw data and the analytical report have been archived at the 3M Environmental

Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard

reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792

## 2 **Summary**

The 3M Environmental Laboratory received groundwater sa mples from a pump test conducted at two

monitoring wells at the 3M Decatur facility in Decatur, AL. A total of twenty sample bottles were received at the 3M Environmental Laboratory for perfluorooctane sulfonate (PFOS), perfluorohexane

sulfonate (PFHS) and perfluorobutane sulfonate (PFBS)

, and included duplicate groundwater samples

and a f ield matrix spike (FMS) sample from each sampling location. Samples also included one trip

blank containing Milli

- QTM water and a trip blank spike. All samples were logged into the laboratory

information management system (LIMS) under project GLP10

- 01

- 02

- 35. The groundwater samples

and trip blanks were received fr om Weston personnel on June 29, 2012. All of the samples wer e prepared and analyzed for PFBS, PFHS, and PFOS following 3 M Environmental Laboratory Method

ETS -8- 044.1 “Method of Analysis for the Determination of Perfluorinated Compounds in Water by

LC/MS/MS; Direct Inject ion Analysis”

.

The average and relative percent difference (RPD) for the PFBS, PFHS, and PFOS concentrations

detected in the primary and field duplicate ground water samples are summarized in Table 1. The trip

blank sample was below the lower limit of quantitation (LLOQ) for all analytes, indicating adequate

control of sample contamination during shipping and sample collections. The analytical method

uncertainties associated with the reported results are : PFBS + 19 %, PFHS + 17 % and PFOS + 22 %.

Table 1

. S ummarized PFBS, PFHS, and PFOS Results ( WWTP Pump Test

– June 2012 ).

PFBS PFHS PFOS

Sampl ing Location Avg. Conc

. (ng/mL) ± RPD Avg. Conc

. (ng/mL) ± RPD Avg. Conc

. (ng/mL) ± RPD



### DAL

- GW

- WW8R

- 120627

- 1000 575 ± 1.2 % 2610 ± 0 .0 % 5280 ± 0.38 %

DAL

- GW

- WW8R

- 120627

- 2130 489 ± 3.3 % 2280 ± 1. 3 % 455 0 ± 0.44 %

DAL

- GW

- WW8R

- 120628 45 3 ± 4.6 % 2070 ± 1.5 % 4170 ± 0.48 %

DAL

- GW

- WW9R

- 120625

- 1250 233 ± 5.2 % 129 0 ± 3.1 % 262 0 ± 3.8 %

DAL

- GW

- WW9R

- 120625

- 2400 248 ± 1.6 % 1340 ± 0.75 % 2610 ± 0.38 %

DAL

- GW

- WW9R

- 120626 254 ± 3.1 % 1330 ± 3.8 % 2550 ± 9.8 %

DAL

- GW

- TRIP01 Blank <0.100 <0.100 <0. 0928

(1) The analytical method uncertainties associated with the reported results are: PFBS ± 19 %, PFHS ± 17 %,

and PFOS ± 22 %.



## 3 **Introduction**

This analytical study was conducted as part of the Phase 3 En vironmental M onitoring and A ssessment

P rogram for the 3M facility located in Decatur, Alabama. The objective of the overall program is to gain

information regarding concentrations of perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate

(PFHS) and perfluorobutane sulfonate (PFBS)

, in various environmental media such as groundwater

, soils and sediments that are associated with and near the Decatur facility. This analytical study was

conducted to analyze groundwater samples collected from a pump test conducted at two on

- site

monitoring wells at the 3M Decatur facility in Decatur, AL for PFOS, PFHS and PFBS in an effort to

characterize regional groundwater conditions.

### The 3M Environmental Laboratory prepared sample bottles (250 mL high

- density polyethylene) which

were shipped to Decatur, AL Weston personnel prior to field sampling. Sample bottle sets for each

groundwater sampling location included a field sample, field sample duplicate, and a field spike sample.

Each empty container for groundwater samplings was marked with a “fill to here” line to produce a final

sample volume of 200 mL. Containers designated for field matrix samples were fortified with an

appropriate matrix spike solution containing PFBS (linear isomer ), PFHS (linear isomer ), and PFOS

(linear and branched isomers ) prior to being sent to the field for sample collection. All sample bottles

included the addition of 18 

### O 2 

### - PFBS, 13 

### C 3 

### - PFHS, and 13 

### C 8 

### - PFOS (internal standard) at a nominal

c oncentrati on of 1 ng/mL and 13 

### C 4 

### - PFOS (surrogate recovery standard) at a nominal concentration of

0.1 ng/mL.

Due to the level of PFBS, PFHS and PFOS detected in samples, and the need to dilute the samples

100

- fold and 200

- fold, the internal s tandards 18 

### O 2 

### - PFBS, 13 

### C 3 

### - PFHS, and 13 

### C 8 

### - PFOS were not used for

quantitation , nor could the surrogate recovery standard 13 

### C 4 

### - PFOS be quantitated. In addition, the field

matrix spike sample at 1000 ng/mL was not appropriate for all analytes at each sampling location.

Where necessary, a laboratory matrix spike was prepared. See section 8.8 of the report for field matrix

spike levels. See section 8.9 of the report for laboratory matrix spike leve ls.

Samples were prepared and analyzed according to the procedure defined in 3M Environmental

Laboratory method ETS -8- 044. 1 “ Method of Analysis for the Determination of Perfluorinated

Compounds In Water by LC/MS/MS; Direct Injection Analysis”.

Table 1 su mmarizes the average and relative percent difference (RPD) for the PFBS, PFHS, and

PFOS concentrations detected in the primary and field duplicate ground water samples and the trip

blank sample

. Tables 10 -15 summarize the individual sample results and the associated field matrix

spike recoveries. All results for the quality control samples prepared and analyzed with the samples

are reported and discussed elsewhere in this report.



## 4 **Test & Control Substances**

There was not a test substance or control substances in the classic sense of a GLP study. This study

was purely analytical in nature.

## 5 **Reference Substances**

Reference

Substance PFBS

(Linear) PFHS

(Linear) PFOS

( Linear + Branched)

Chemical Name Potassium Perfluorobutane

sulfonate Sodium Perfluorohexane

sulfonate Potassium

Perfluorooctane sulfonate

Chemical Formula 

### C 4 

### F 9 

### SO 3 - 

### K + 

### C 6 

### F 13 

### SO 3 - 

### Na + 

### C 8 

### F 17 

### SO 3 - 

### K +

Identifier NA L- PFHXS Br

- PFOSK

Use Target Analyte Reference

S tandard Target Analyte Reference

Standard Target Analyte Reference

Standard

Source 3M Wellington Wellington

Expiration Date 01/10/2017 03/25/2018 12/01/2014

Storage Conditions Frozen Frozen Frozen

Chemical Lot

Number 41

- 2600

- 8442 -5 LPFHxSAM08 brPFOSK11 11

TCR Number TCR

- 121 TCR08

- 0018 TCR11

- 0041

Physical Description White Powder Crystalline Liquid

Purity 96.7% 100% 99.9%

Reference

Substance PFOS (1)

(Linear + Branched)

Chemical Name Potassium Perfluorooctane

sulfonate

Chemical Formula 

### C 8 

### F 17 

### SO 3 - 

### K +

Identifier CAS # 2795

- 39 -3

Use FMS Reference Standard

Source S igma

- Aldrich

Expiration Date 0 2/ 0 4/2014

Storage Conditions Ambient

Chemical Lot

Number 1424328V

TCR Number TCR11

- 0028

Physical Description Whi te Powder

Purity 99.7%

(1) Due to the level of PFOS detected in the samples, the FMS reference standard for PFOS was not

utilized in this analysis.

## 6 **Test System** 

The test systems for this study are groundwater samples collected from wells located in Decatur, AL by

Weston Solutions, Inc. personnel. Samples for this study are “real world” samples, not dosed with a

specific lot of test substance.

Table 2 . Sample Description Key Code.

String Number String Descriptor Example

**Example DAL** 

**- GW** 

**- WW9R** 

**- DB** 

**- 120625** 

**- 2400** 

**1 Sample Location**  **DAL =**  **Decatur , Alabama** 

**2 Sample Type**  **GW = Ground Water**

**3 Well ID**  **Example: WW9R** 

**4 Sample Type**  **0=primary sam ple** 

**DB =duplicate sample** 

**FM S =**  **field matrix spike** 

**5 Sampling Date**  **1 20635 = June 25, 2012** 

**6 Sampling Time**  **2400 = 24:00 or 12:00 am**

**Not all sampling locations included a**

**sampling time in the sample description**



## 7 **Method Summary**

7.1 Method

Analysis for all analytes was completed following 3M Environmental Laboratory method ETS -8- 044. 1

“ Method of Analysis for the Determination of Perfluorinated Compounds In Water by High Performance

Liquid Chromatography/Mass Spectrometry Direct Injection Analysis”.



## 7.2 Sample Co lle ction

Samples were collected in 250 mL Nalgene TM 

### (high

- density polyethylene) bottles prepared at the 3M

Environmental Laboratory. Sample bottles associated with GLP10

- 01

- 02

- 35 were returned to the

laboratory at ambient conditions on June 29 , 2012. Samples were stored refrigerated at the laboratory

after receipt. A set of laboratory prepared Trip Blank and a Trip Blank field matrix spike were sent with

the set of sample collection bottles.



## 7.3 Sample Preparation

Due to the level of PFBS, PFHS, and PFOS de tected in all sampling locations, the primary and field

duplicate samples were diluted 1:100 or 1:200. The 1:100 dilution was preapred by diluting 0.10 mL of

a well mixed sample with 9.9 mL of Milli Q water

. The 1:200 dilution was preapred by diluting 0.050 mL

of a well mixed sample with 9.9 mL of Milli Q water (dilution factor of 199).

The Trip Blank FMS sample was diluted 1:100 by diluting 0.10 mL of a well mixed sample with 9.9 mL of

Milli Q water. The Trip Blank sample was aliquoted and analyzed with out sample dilution.



## 7.4 Analysis

All study samples and quality control samples were analyzed for PFBS, PFHS, and PFOS using high

performance liquid chromatography/ tandem mass spectrometry (HPLC/MS/MS). Detailed instrument

parameters, the liquid chromatography gradient program, and the specific mass transitions analyzed are

### described in the raw data hard copies placed in the final data packet, and are briefly described below in

Table 3

, Table 4 and Table 5

.

Due to the nature of the sample, the wide range of concentrations found in the sample, and the

environmental occurrence of multiple isomers of the laboratory’s analytes of interest, the software used

for processing the analytical results is not able to consistently integrate the analytical peak, manua l

integration of the analytical peak is necessary. All manual integrations are performed following the

procedures outlined in method ETS

- 12

- 010. The consistency of the laboratory’s integration is ensured

through the training of laboratory personnel, the peer review process required for all manual integrations,

the review of manual integrations by the QAU, and where necessary the review of manual integrations

by laboratory management.

Table 3

. Instrument Parameters.

Instrument Na me ETS McCoy

Analytical Method Followed ETS

- 8

- 044.1

Analysis Date 7/20/2012 and 8/3/2012

Liquid Chromatograph Agilent 1290

Guard column Betasil C18 (4.6 mm X 100 mm), 5 μ

Analytical column Betasil C18 (4.6 mm X 100 mm), 5 μ

Injection Volume 5 μ L

Mass Spectrometer Applied Biosystems API 55 00

Ion Source Turbo Spray

Electrode Turbo ion electrode

Polarity Negative

Software Analyst 1.6.1



### Table 4 . Liquid Chromatography Conditions.

**Step**

**Number Total Time** 

**(min) Flow Rate** 

**(** μ

**L/min) Percent A** 

**(5 mM ammonium** 

**acetate :0.01% Acetic Acid**  **) Percent B** 

**( Acetonitrile**  **)**

0 0.00 750 90.0 10.0

1 0.50 750 90.0 10.0

2 4.00 750 70.0 30 .0

3 6.00 750 70.0 30.0

4 11.0 750 20.0 80.0

5 13.0 750 20.0 80.0

6 13.5 750 10.0 90.0

7 16.0 750 10.0 90.0

8 16.5 750 90.0 10.0

9 19.0 750 90.0 10.0

### Table 5. Mass Transitions.

**Analyte Mass Transition** 

**Q1/Q3 Reference Mate**  **rial**

**Structure**

**PFBS 299/80**  **Linear 299/99** 

**PFHS 399/80**  **Linear 399/99** 

**PFOS 499/80** 

**Linear +**  **Branched 499/99** 

**499/130**

Dwell time was 3 0 msec for each transition. The individual transitions were

summed to produce a “total ion chromatogram” (TIC), which was used for

quantitation.



## 8 **Analytical Results** 

8.1 Calibration

Samples were analyzed against an external standard calibration curve. Calibration standards were

prepared by spiking known amounts of the stock solutions containing PFBS, PFHS, and PFOS

(re ference standard containing both linear and branched isomers) into Milli Q water. Standards ranging

from 0.10 ng/mL to 75 ng/mL (nominal) were analyzed (0.10 mg/mL to 25 ng/mL for PFBS analysis on

8/3/120) . A quadratic, 1/x weighted, calibration curve of the peak area counts was used to fit the data

for each analyte. The data were not forced through zero during the fitting process. Calculating the

standard concentrations using the peak area confirmed accuracy of each curve point. Each curve point

was quantitated using the overall calibration curve and reviewed for accuracy. Method calibration

accuracy requirements of 100±25% (100±30% for the lowest curve point) were met for PFBS, PFHS,

and PFOS. The correlation coefficient (r) was greater than 0.995 for PFBS, PFHS, and PFOS.



## 8.2 System Suitability

A calibration standard was analyzed four times at the beginning of each analytical sequence to

demonstrate overall system suitability. The acceptance criteria of less than or equal to 5% relative

standard deviation (RSD) for peak area and retention time criteria of less than or equal to 2% RSD w as met for PFBS, PFHS and PFOS for each analysis.



## 8.3 Limit of Quantitation (LOQ)

The LOQ for this analysis is the lowest non

- zero calibration standard in the curve that mee ts linearity

and accuracy requirements and for which the area counts or area ratio are at least twice those of the

appropriate blanks. The LOQ for all analytes can be found in Table 6.

Table 6 . Limit of Quantitation (LOQ).

**Analys is Date**  **Dilution**

**Factor PFBS** 

**LOQ, ng/mL PFHS** 

**LOQ, ng/mL PFOS** 

**LOQ, ng/mL**

7/20/12 1 0.100 0.100 0.0928

7/20/12 100 10.0 10.0 9.28

7/20/12 199 19.9 19.9 18.5

8/3/12 100 10.0 NA NA

NA = Not Applicable

## 8.4 Continuing Calibration

During the course of each analytical sequence, continuing calibration verification samples (CCVs) were

analyzed to confirm that the instrument response and the initial calibration curve were still in control. All

CCVs met method criteria of 100% ± 25% for PFBS, PFHS and PFOS for each analysis

.



## 8.5 Blanks

Two types of blanks were prepared and analyzed with the samples: method procedural blanks and a trip

blank. Method procedural blank results were reviewed and used to evaluate method performance and

to determine the LOQ for PFBS, PFHS,and PFOS. The trip blank reflects the shipping and sample

collection conditions the sample bottles and samples experience.



## 8.6 Lab Control Spikes (LCSs)

Low, mid, and high lab control spikes were prepared and analyzed in triplicate. LCSs were prepared b y spiking known amounts of the analytes into 10 mL of Milli Q water to produce the desired concentration.

The method acceptance criteria, average of LCS at each level within 100% ± 20 % with an RSD ≤20%,

were met for PFBS

, PFHS, and PFOS with the following exception

.

7/20/12 Analysis: The low set of LCS samples for PFOS had an average recovery of 79.3%. A method

deviation is included with the raw data.

The following calculations w ere used to generate data in Table 7 for laboratory control spikes:

100% * ion Concentrat Spike ion Concentrat Calculated Recovery Percent LCS

=

100% *

recovery LCS average replicates LCS deviation standard RSD LCS%

=

### Table 7 . Laboratory Control Spike Recovery

.

ETS -8- 044.1

Analyzed 7/20/12 **PFBS PFHS** 

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery**  **Spiked**

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120720

- 1 0.200 0.199 99.6 0.200 0.189 94.6

LCS

- 120729

- 2 0.200 0.196 97.9 0.200 0.162 81.2

LCS

- 120729

- 3 0.200 0.210 105 0.200 0.188 94.0

Average ± %RSD 101 % ± 3.7 % 89.9 % ± 8.4 %

LCS

- 120729

- 4 2.00 1.89 94.3 2.00 1.95 97.6

LCS

- 120729

- 5 2.00 1.85 92.4 2.00 1.96 98.2

LCS

- 120720

- 6 2.00 1.94 97.2 2.00 1.90 94.9

Average ± %RSD 94.6 % ± 2. 6 % 96.9 % ± 1.8 %

LCS

- 120720

- 7 20.0 18.4 92.1 20.0 18.8 94.2

L CS

- 120720

- 8 20.0 19.5 97.4 20.0 20.3 101

LCS

- 120720

- 9 20.0 19.7 98.3 20.0 19.8 99.0

Average ± %RSD 95.9 ± 3.5 % 98.1 % ± 3. 6 %

ETS -8- 044.1

Analyzed 7/20/12 **PFOS (Linear and Branched)**

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculate**  **d**

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120720

- 1 0.186 0.168 90.5

LCS

- 120729

- 2 0.186 0.135 72.5

LCS

- 120729

- 3 0.186 0.140 75.0

Average ± %RSD 79.3 % (1) ± 12 %

LCS

- 120729

- 4 1.86 1.74 93.3

LCS

- 120729

- 5 1.86 1.66 89.4

LCS

- 120720

- 6 1.86 1.60 86.0

Averag e ± %RSD 89.7 % ± 4.1 %

LCS

- 120720

- 7 18.6 17.4 93.4

LCS

- 120720

- 8 18.6 18.6 99.7

LCS

- 120720

- 9 18.6 17.4 93.6

Average ± %RSD 95.6 % ± 3.7 %

(1) LCS did not meet acceptance criteria of 100 ± 20%.

### Table 7 continued. Laboratory Control Spike Recovery.

E TS -8- 044.1

Analyzed 8/03 /12 **PFBS**

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120803

- 1 0.200 0.225 112

LCS

- 120803

- 2 0.200 0.217 108

LCS

- 120803

- 3 0.200 0.220 110

Average ± %RSD 110 % ± 1.8 %

LCS

- 120803

- 4 2.00 2 .10 105

LCS

- 120803

- 5 2.00 2.00 100

LCS

- 120803

- 6 2.00 2.10 105

Average ± %RSD 103 % ± 2.8 %

LCS

- 120803

- 7 20.0 21.0 105

LCS

- 120803

- 8 20.0 22.4 112

LCS

- 120803

- 9 20.0 22.6 113

Average ± %RSD 110 % ± 4.0 %

(1) LCS did not meet acceptance criteria of 100 ± 20%.



## 8.7 Analytical Method Uncertainty

Analytical uncertainty is based on historical QC data that is control charted and used to evaluate

method accuracy and precision. The method uncertainty is calculated following ETS

- 12

- 012.2. The

standard deviation is calculated for the set of accuracy results (in %) obtained for the QC samples. The

expanded uncertainty is calculated by multiplying the standard deviation by a factor of 2, which

corresponds to a confidence level of 95%.

Table 8

. Analytical Uncertainty.

Analyte Standard Deviation Method Uncertainty

PFBS 9.39 ± 19 %

PFHS 8.7 7 ± 17 %

PFOS 10.8 ± 22 %



## 8.8 Field Matrix Spikes (FMS)

A field matrix spike was collected at each sampling point to verify that the analytical method is appli cable

to the collected matrix. Field matrix spikes were generated by adding a measured volume of field

sample to a container spiked by the laboratory with PFBS (linear), PFHS (linear), and PFOS (linear and

branched ) prior to shipping sample containers for sample collection. Field matrix spike recoveries within

method acceptance criteria of 100±30% confirm that “unknown” components in the sample matrix do not

significantly interfere with the extraction and analysis of the analytes of interest. Field matrix spike

concentrations must be 50% of the sample concentration to be considered an appropriate field spike.

The field matrix spike sample was not appropriate for all analytes at each sampling location. Where

necessary, a laboratory matrix spike was prepared. See section 8.9 of the report for laboratory matrix

spike levels. Field matrix spike recoveries are presented in section 9 of this report.

### T able 9 . Field Matrix Spike

.

Sampling Location PFBS,

ng/mL PFHS,

ng/mL PFOS,

ng/mL

All locations and Trip Blank 1002 9 94 1012

100% *

on Concentrat Spike Dup.) Sample Field & Sample Field: ion Concentrat Average FMS of ion Concentrat ( Sample Recovery FMS −

=



## 8.9 Lab Matrix Spikes (LMS)

Due to the high level of PFHS and PFOS detected in several sampling locations, a laboratory matrix

spike (LMS) sample was prepared to verify that the analytical method is applicable to the collected

matrix. Laboratory matrix spikes were generated by adding a measured volume of the target analytes to

an aliquot of the primary sample. T he laboratory matrix spike added was based on the o n- column

instrument concentration.

All sampling locations: the primary sample was diluted 1:200, to which a nominal concentration of 30

ng/mL was added for a LMS concentration of approximately 6000 ng/mL.

Table 10

. Laboraotry Mat rix Spike

Sampling Location PFBS,

ng/mL PFHS,

ng/mL PFOS,

ng/mL

All locations 6012 6006 5579



### Lab matrix spike recoveries within method acceptance criteria of 100±30% confirm that “unknown”

components in the sample matrix do not significantly interfere with the extraction and analysis of the

analytes of interest. Lab matrix spike concentrations must be 50% of the sample concentration to be

considered an appropriate field spike. Lab matrix spikes are presented in section 9 of this report.

The followin g calculation was used to calculate the lab matrix spike recovery in Section 9 of the report:

100% *

on Concentrat Spike Dup.) Sample Field & Sample Field: ion Concentrat Average LMS of ion Concentrat ( Sample Recovery LMS −

=



## 9 **Data Summary and Discussion**

The tables below summarize the sample results and field and/ or lab oratory matrix spike recoveries for

the sampling locations as well as the Trip Blank

. Results and average values are rounded to three

significant figures according to EPA rounding rules. Because of rounding, values may vary slightly from

those listed in the raw data. Field matrix spike and lab m atrix spike recoveries meeting the method

acceptance criteria of ± 30%, demonstrate that the method was appropriate for the given matrix and

their respective quantitative ranges

.

All QC elements of the method; field matrix spikes (FMSs) and laboratory matrix spikes (LMSs) met

method acceptance criteria.

### Table 11

. DAL GW WW8R 120627

- 1000

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 3 5

- 001 DAL

- GW

- WW8R

- 0

- 120627

- 1000 578 NA 2610 NA 5270 NA

GLP10

- 01

- 02

- 35

- 002 DAL

- GW

- WW8R

- DB

- 120627

- 1000 571 NA 2610 NA 5290 NA

GLP10

- 01

- 02

- 35

- 003 DAL

- GW

- WW8R

- FMS

- 120627

- 1000 1560 98.4 3600 NC 5940 NC

GLP10

- 01

- 02

- 35

- 001 ; LMS DAL

- GW

- WW8R

- 0

- 12 0627

- 1000 6760 103 8800 103 11300 108

**Average Concentration (ng/mL) ±**  **%RPD 575**  **ng/mL ±**  **1.2 %**  **2610 ng/mL**  **± 0.0**  **% 5280**  **ng/mL ±**  **0.38 %** 

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

Samples analyzed using a 1 :199 dilution factor



### Table 12

. DAL GW WW8R 120627

- 2130

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 004 D AL

- G W

- WW8R

- 0

- 120627

- 2130 497 NA 2290 NA 4540 NA

GLP10

- 01

- 02

- 35

- 005 DAL

- GW

- WW8R

- DB

- 120627

- 2130 481 NA 2260 NA 4560 NA

GLP10

- 0 1

- 02

- 35

- 006 DAL

- GW

- WW8R

- FMS

- 120627

- 2130 1400 90.9 3170 NC 5660 NC

GLP10

- 01

- 02

- 35

- 004; LMS DAL

- GW

- WW8R

- 0

- 120627

- 2130 6500 100 8010 95.5 10400 105

**Average Concentration (ng/mL) ±**  **%RPD 489**  **ng/mL ±**  **3.3 %**  **2280 ng/mL**  **± 1**  **.3 %**  **4550 ng/mL**  **± 0.44**  **%**

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

Samples analyzed using a 1:199 di lution factor

### Table 13

. DAL GW WW8R 120628

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 007 DAL

- GW

- WW8R

- 0

- 120628 46 3 NA 2050 NA 4160 NA

GLP10

- 01

- 02

- 35

- 008 D AL

- GW

- WW8R

- DB

- 120628 442 NA 2080 NA 4180 NA

GLP10

- 01

- 02

- 35

- 009 DAL

- GW

- WW8R

- FMS

- 120628 1400 94.6 3070 NC 4490 NC

GLP10

- 01

- 02

- 35

- 007; LMS DAL

- GW

- WW8R

- 0

- 120628 6780 105 8010 99.0 10400 112

**Average Concentration (ng/mL)**  **± %RPD**  **453 ng/mL**  **± 4.6**  **% 2070**  **ng/mL ±**  **1**

**. 5%**  **4170 ng/mL**  **± 0.48**  **%**

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

Samples analyzed using a 1:199 dilution factor



### Table 14

. DAL GW WW9R 120625

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 010 DAL

- GW

- WW9R

- 0

- 120625

- 1250 239 NA 1310 NA 2570 NA

GLP10

- 01

- 02

- 35

- 01 1 DAL

- GW

- WW9R

- DB

- 120625

- 1250 227 NA 1270 NA 2670 NA

GLP10

- 01

- 02

- 35

- 012 DAL

- GW

- WW9R

- FMS

- 120625

- 1250 1170 93.5 2170 88.5 3290 NC

GLP10

- 01

- 02

- 35

- 010; LMS DAL

- GW

- WW9R

- 0

- 120625

- 1250 6320 101 7150 97.6 8970 114

**Average Concentration (ng/mL) ±**  **%RPD 233**  **ng/mL ±**  **5.2 %**  **1290 ng/mL**  **± 3.1**  **% 2620**  **ng/mL ±**  **3.8 %** 

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

Samples analyzed using a 1:199 dilution factor

### Table 15

. DAL GW WW9R 1206 25

- 2400

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 013 DAL

- GW

- WW9R

- 0

- 120625

- 2400 250 NA 1330 NA 2600 NA

GLP10

- 01

- 02

- 35

- 014 DAL

- GW

- WW9R

- DB

- 120625

- 2400 246 NA 1340 NA 2610 NA

GLP10

- 01

- 02

- 35

- 015 DAL

- GW

- WW9R

- FMS

- 120625

- 2400 1210 96.0 2590 126 3710 NC

GLP10

- 01

- 02

- 35

- 013; LMS DAL

- GW

- WW9R

- 0

- 120625

- 2400 NA (1) NA (1) 7110 96.2 7970 96.2

**Average Concentration (ng/mL) ±**  **%RPD 248**  **ng/mL ±**  **1.6 %**  **134 0**  **ng/mL ±**  **0.75 %**  **2610 ng/mL**  **± 0.38**  **%**

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

PFHS and PFO S diluted 1:199 and reported from 7/20/12 analysis. PFBS diluted 1:100 and reported from 8/3/12 analysis.

(1 ) Sample was not analyzed.



### Table 16

. DAL GW WW9R 12062 6

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentr ation** 

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 016 DAL

- GW

- WW9R

- 0

- 120626 258 NA 1350 NA 2670 NA

GLP10

- 01

- 02

- 35

- 017 D AL

- GW

- WW9R

- DB

- 120626 250 NA 1300 NA 2420 NA

GLP10

- 01

- 02

- 35

- 018 DAL

- GW

- WW9R

- FMS

- 120626 1140 88.4 2180 86.0 3180 NC

GLP10

- 01

- 01

- 36

- 016; LMS DAL

- GW

- WW9R

- 0

- 120626 6050 96.4 7130 96.7 8460 106

**Average Concentration (ng/mL) ±**  **%RPD 254**  **ng/mL ±**  **3.1% 1330**  **ng/mL ±**  **3.8 %**  **2550 ng/mL**  **± 9.8%** 

NA = Not Applicable

NC = Not Calculated; Spike level was less than 0.5x the endogenous sample concentration.

Sample s analyzed using a 1:199 dilution factor

### Table 17 . DAL GW TRIP 01 120625

**PF BS**  **PF HS**  **PFOS**

**3M LIMS ID Description**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery**  **Concentration**

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 35

- 019 DAL

- GW

- TRIP01

- 0

- 120625 <0.100 NA <0.100 NA <0.0928 NA

GLP10

- 01

- 02

- 35

- 020 DAL

- GW

- TRIP01

- FMS

- 120625 947 94.5 939 94.5 785 77.6

NA = Not Applicable

FMS Sample analyz ed using a 1:100 dilution factor

## 10 **Conclusion**

Laboratory control spikes, field matrix spikes , and lab matrix spikes were used to determine the

analytical method accuracy and precision for PFBS, PFHS, and PFOS. A nalysis was successfully

completed following 3M Environme ntal Laboratory method ETS -8- 044. 1 described herein.



## 11 **Data/Sample Reten tion** 

All remaining samples and associated project data (hardcopy and electronic) will be archived according

to 3M Environmental Laboratory standard operating procedures.



## 12 **Attachments**

At tachment A: Protocol Amendment 35 (General Project Outline)

Attachment B: Representative Chromatograms and Calibration Curves

Attachment C: Analytical Method

- ETS -8- 044. 1

Attachmetn D: Protocl Amendment GLP10

- 01

- 02

- 35 DeviationGLP10-01-02 ; Interi m Repor t 3 5

Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Sample s

Decatu r VVWT

P Pum p Tes t - Jun e 201 2



## 13 Signatures

Clesto n Lange

, Ph.D.

, 3 M Principa l Analytica l Investigato r Dat e

Willia m K

. Reagen

, Ph.D.

, 3 M Environmenta l Laborator y Technica l Directo r Dat e

Gar y Hdhensljbin

, SAonso r Representativ e Dat e

Jaisimh a Kesari ; Sfild y Directo r

Pag e 2 3 o f 7 4

# A TTACHMENT 

# A: P ROTOCOL 

# A MENDMENT

# A TTACHMENT 

# B: R EPRESENTATIVE 

# S AMPLE 

# C HROMATOGRAMS AND



# C ALIBRATION 

# C URVE 

# ( S 

# )mc120720a.rdb (PFOS): "Quadratic" Regression ("1 / x" weighting): y = -51.6 x^2 + 4.98e+004 x + -649 (r = 0.9999)0 5 10 15 20 25 30 35 40 45 50 55 60 65Concentration, ng/mL0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e6

Area, counts*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:38:39 AMPage 1 of 1mc120720a.rdb (PFHS): "Quadratic" Regression ("1 / x" weighting): y = -24.9 x^2 + 3.46e+004 x + 859 (r = 0.9996)0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75Concentration, ng/mL0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e62.1e62.2e62.3e62.4e62.5e6

Area, counts*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:28:09 AMPage 1 of 1mc120720a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -65.9 x^2 + 7.56e+004 x + -478 (r = 0.9996)0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75Concentration, ng/mL0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e63.6e63.8e64.0e64.2e64.4e64.6e64.8e65.0e65.2e65.4e6

Area, counts*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:28:32 AMPage 1 of 1Sample Name: "mc120720a014" Sample ID: "11 012-74-10" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "5.0 ng/mL FC mix in Milli Q water" Annotation: ""Sample Index: 15Sample Type: StandardConcentration: 5.00 ng/mLCalculated Conc: 4.90 ng/mLAcq. Date: 7/20/2012Acq. Time: 3:23:14 PMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.0 minArea: 368723 countsHeight: 1.89e+005 cpsStart Time: 9.94 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e5

Intensity, cps10.01Sample Name: "mc120720a021" Sample ID: "Method Blank" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "Milli Q water TN12-0228" Annotation: ""Sample Index: 22Sample Type: UnknownConcentration: N/ACalculated Conc: 0.0108 ng/mLAcq. Date: 7/20/2012Acq. Time: 5:39:53 PMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.0 minArea: 335 countsHeight: 2.76e+002 cpsStart Time: 9.96 minEnd Time: 10.0 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min020406080100120140160180200220240260280300320340360380400420

Intensity, cps10.4711.9212.0110.7410.01 10.95 11.2112.4310.519.4911.4410.4110.398.56 9.30 9.819.077.99 10.198.428.777.70 9.688.41Sample Name: "mc120720a022" Sample ID: "GLP10-01-02-35-001" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 23Sample Type: UnknownConcentration: N/ACalculated Conc: 578. ng/mLAcq. Date: 7/20/2012Acq. Time: 5:59:24 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.0 minArea: 218612 countsHeight: 1.06e+005 cpsStart Time: 9.73 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.005000.001.00e41.50e42.00e42.50e43.00e43.50e44.00e44.50e45.00e45.50e46.00e46.50e47.00e47.50e48.00e48.50e49.00e49.50e41.00e51.05e5

Intensity, cps10.00Sample Name: "mc120720a025" Sample ID: "GLP10-01-02-35-001; LMS" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 26Sample Type: UnknownConcentration: N/ACalculated Conc: 6760. ng/mLAcq. Date: 7/20/2012Acq. Time: 6:57:57 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.99 minArea: 2479263 countsHeight: 1.12e+006 cpsStart Time: 9.73 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e61.10e6

Intensity, cps9.99*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:56 AMPage 1 of 12Sample Name: "mc120720a027" Sample ID: "GLP10-01-02-35-004" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 28Sample Type: UnknownConcentration: N/ACalculated Conc: 497. ng/mLAcq. Date: 7/20/2012Acq. Time: 7:36:59 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.99 minArea: 188092 countsHeight: 9.80e+004 cpsStart Time: 9.72 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e48.5e49.0e49.5e4

Intensity, cps9.99Sample Name: "mc120720a030" Sample ID: "GLP10-01-02-35-004; LMS" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 31Sample Type: UnknownConcentration: N/ACalculated Conc: 6500. ng/mLAcq. Date: 7/20/2012Acq. Time: 8:35:32 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.98 minArea: 2389338 countsHeight: 1.09e+006 cpsStart Time: 9.71 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e6

Intensity, cps9.98Sample Name: "mc120720a036" Sample ID: "GLP10-01-02-35-007" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120628 (1:200 dil)" Annotation: ""Sample Index: 37Sample Type: UnknownConcentration: N/ACalculated Conc: 463. ng/mLAcq. Date: 7/20/2012Acq. Time: 10:32:41 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.98 minArea: 175234 countsHeight: 8.39e+004 cpsStart Time: 9.72 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e4

Intensity, cps9.98Sample Name: "mc120720a041" Sample ID: "GLP10-01-02-35-010" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120625-1250 (1:200 dil)" Annotation: ""Sample Index: 42Sample Type: UnknownConcentration: N/ACalculated Conc: 239. ng/mLAcq. Date: 7/21/2012Acq. Time: 12:10:17 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.98 minArea: 90171 countsHeight: 3.93e+004 cpsStart Time: 9.72 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e43.0e43.2e43.4e43.6e43.8e4

Intensity, cps9.98*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:56 AMPage 2 of 12Sample Name: "mc120720a050" Sample ID: "GLP10-01-02-35-013" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120625-2400 (1:200 dil)" Annotation: ""Sample Index: 51Sample Type: UnknownConcentration: N/ACalculated Conc: 237. ng/mLAcq. Date: 7/21/2012Acq. Time: 3:06:02 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.98 minArea: 89552 countsHeight: 4.00e+004 cpsStart Time: 9.70 minEnd Time: 10.0 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e43.0e43.2e43.4e43.6e43.8e4

Intensity, cps9.98Sample Name: "mc120720a055" Sample ID: "GLP10-01-02-35-016" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 56Sample Type: UnknownConcentration: N/ACalculated Conc: 258. ng/mLAcq. Date: 7/21/2012Acq. Time: 4:43:41 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.97 minArea: 97575 countsHeight: 4.37e+004 cpsStart Time: 9.70 minEnd Time: 10.0 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e43.0e43.2e43.4e43.6e43.8e44.0e44.2e4

Intensity, cps9.97Sample Name: "mc120720a057" Sample ID: "GLP10-01-02-35-018" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-FMS-120626 (1:200 dil)" Annotation: ""Sample Index: 58Sample Type: UnknownConcentration: N/ACalculated Conc: 1140. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:22:44 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.97 minArea: 431768 countsHeight: 2.15e+005 cpsStart Time: 9.69 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e52.0e52.1e5

Intensity, cps9.97Sample Name: "mc120720a058" Sample ID: "GLP10-01-01-36-016; LMS" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 59Sample Type: UnknownConcentration: N/ACalculated Conc: 6050. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:42:16 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.97 minArea: 2228465 countsHeight: 1.05e+006 cpsStart Time: 9.68 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e6

Intensity, cps9.97*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:56 AMPage 3 of 12Sample Name: "mc120720a066" Sample ID: "LCS-120719-7" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "20ppb LCS" Annotation: ""Sample Index: 67Sample Type: QCConcentration: 20.0 ng/mLCalculated Conc: 18.4 ng/mLAcq. Date: 7/21/2012Acq. Time: 8:18:31 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 9.96 minArea: 1370575 countsHeight: 6.49e+005 cpsStart Time: 9.89 minEnd Time: 10.1 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps9.96Sample Name: "mc120720a069" Sample ID: "LCS-120719-4" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "2ppb LCS" Annotation: ""Sample Index: 70Sample Type: QCConcentration: 2.00 ng/mLCalculated Conc: 1.89 ng/mLAcq. Date: 7/21/2012Acq. Time: 9:17:07 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 9.96 minArea: 141944 countsHeight: 6.51e+004 cpsStart Time: 9.90 minEnd Time: 10.0 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e4

Intensity, cps9.96Sample Name: "mc120720a072" Sample ID: "LCS-120719-1" File: "mc120720a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "0.2ppb LCS" Annotation: ""Sample Index: 73Sample Type: QCConcentration: 0.200 ng/mLCalculated Conc: 0.199 ng/mLAcq. Date: 7/21/2012Acq. Time: 10:15:44 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 10.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 9.96 minArea: 14585 countsHeight: 6.86e+003 cpsStart Time: 9.92 minEnd Time: 10.0 min8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5Time, min0500100015002000250030003500400045005000550060006500

Intensity, cps9.9610.4411.928.827.88Sample Name: "mc120720a014" Sample ID: "11 012-74-10" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "5.0 ng/mL FC mix in Milli Q water" Annotation: ""Sample Index: 15Sample Type: StandardConcentration: 5.00 ng/mLCalculated Conc: 4.96 ng/mLAcq. Date: 7/20/2012Acq. Time: 3:23:14 PMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 10.9 minArea: 171950 countsHeight: 7.79e+004 cpsStart Time: 10.9 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e4

Intensity, cps10.94*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:56 AMPage 4 of 12Sample Name: "mc120720a021" Sample ID: "Method Blank" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "Milli Q water TN12-0228" Annotation: ""Sample Index: 22Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 7/20/2012Acq. Time: 5:39:53 PMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.9 minArea: 303 countsHeight: 2.40e+002 cpsStart Time: 10.9 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0200400600800100012001400160018002000220024002600

Intensity, cps13.0511.4410.92 13.319.15 12.4211.64 12.1110.55Sample Name: "mc120720a022" Sample ID: "GLP10-01-02-35-001" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 23Sample Type: UnknownConcentration: N/ACalculated Conc: 2610. ng/mLAcq. Date: 7/20/2012Acq. Time: 5:59:24 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 449550 countsHeight: 1.88e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e5

Intensity, cps10.9210.82Sample Name: "mc120720a025" Sample ID: "GLP10-01-02-35-001; LMS" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 26Sample Type: UnknownConcentration: N/ACalculated Conc: 8800. ng/mLAcq. Date: 7/20/2012Acq. Time: 6:57:57 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 1474424 countsHeight: 6.81e+005 cpsStart Time: 10.6 minEnd Time: 11.1 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e5

Intensity, cps10.91Sample Name: "mc120720a027" Sample ID: "GLP10-01-02-35-004" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 28Sample Type: UnknownConcentration: N/ACalculated Conc: 2290. ng/mLAcq. Date: 7/20/2012Acq. Time: 7:36:59 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 395133 countsHeight: 1.54e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e5

Intensity, cps10.9110.82*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:56 AMPage 5 of 12Sample Name: "mc120720a030" Sample ID: "GLP10-01-02-35-004; LMS" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 31Sample Type: UnknownConcentration: N/ACalculated Conc: 8010. ng/mLAcq. Date: 7/20/2012Acq. Time: 8:35:32 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 1346882 countsHeight: 6.31e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps10.91Sample Name: "mc120720a036" Sample ID: "GLP10-01-02-35-007" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW8R-0-120628 (1:200 dil)" Annotation: ""Sample Index: 37Sample Type: UnknownConcentration: N/ACalculated Conc: 2050. ng/mLAcq. Date: 7/20/2012Acq. Time: 10:32:41 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 353858 countsHeight: 1.36e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e5

Intensity, cps10.9010.80Sample Name: "mc120720a041" Sample ID: "GLP10-01-02-35-010" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120625-1250 (1:200 dil)" Annotation: ""Sample Index: 42Sample Type: UnknownConcentration: N/ACalculated Conc: 1310. ng/mLAcq. Date: 7/21/2012Acq. Time: 12:10:17 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 227444 countsHeight: 9.35e+004 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e48.5e49.0e4

Intensity, cps10.9010.80Sample Name: "mc120720a050" Sample ID: "GLP10-01-02-35-013" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120625-2400 (1:200 dil)" Annotation: ""Sample Index: 51Sample Type: UnknownConcentration: N/ACalculated Conc: 1330. ng/mLAcq. Date: 7/21/2012Acq. Time: 3:06:02 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 231736 countsHeight: 9.36e+004 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e48.5e49.0e4

Intensity, cps10.8910.79*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 6 of 12Sample Name: "mc120720a055" Sample ID: "GLP10-01-02-35-016" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 56Sample Type: UnknownConcentration: N/ACalculated Conc: 1350. ng/mLAcq. Date: 7/21/2012Acq. Time: 4:43:41 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 233560 countsHeight: 9.62e+004 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e48.5e49.0e49.5e4

Intensity, cps10.8910.79Sample Name: "mc120720a057" Sample ID: "GLP10-01-02-35-018" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW9R-FMS-120626 (1:200 dil)" Annotation: ""Sample Index: 58Sample Type: UnknownConcentration: N/ACalculated Conc: 2180. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:22:44 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 376341 countsHeight: 1.69e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e5

Intensity, cps10.8910.79Sample Name: "mc120720a058" Sample ID: "GLP10-01-01-36-016; LMS" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 59Sample Type: UnknownConcentration: N/ACalculated Conc: 7130. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:42:16 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 10.9 minArea: 1201801 countsHeight: 5.90e+005 cpsStart Time: 10.6 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e5

Intensity, cps10.89Sample Name: "mc120720a066" Sample ID: "LCS-120719-7" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "20ppb LCS" Annotation: ""Sample Index: 67Sample Type: QCConcentration: 20.0 ng/mLCalculated Conc: 18.8 ng/mLAcq. Date: 7/21/2012Acq. Time: 8:18:31 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.9 minArea: 643954 countsHeight: 3.09e+005 cpsStart Time: 10.8 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e5

Intensity, cps10.88*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 7 of 12Sample Name: "mc120720a069" Sample ID: "LCS-120719-4" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "2ppb LCS" Annotation: ""Sample Index: 70Sample Type: QCConcentration: 2.00 ng/mLCalculated Conc: 1.95 ng/mLAcq. Date: 7/21/2012Acq. Time: 9:17:07 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.9 minArea: 68284 countsHeight: 3.18e+004 cpsStart Time: 10.8 minEnd Time: 11.0 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e43.0e4

Intensity, cps10.8812.96Sample Name: "mc120720a072" Sample ID: "LCS-120719-1" File: "mc120720a.wiff"Peak Name: "PFHS" Mass(es): "399.000/80.000 Da,399.000/99.000 Da"Comment: "0.2ppb LCS" Annotation: ""Sample Index: 73Sample Type: QCConcentration: 0.200 ng/mLCalculated Conc: 0.189 ng/mLAcq. Date: 7/21/2012Acq. Time: 10:15:44 AMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.0 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 10.9 minArea: 7403 countsHeight: 4.04e+003 cpsStart Time: 10.8 minEnd Time: 10.9 min8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0Time, min02004006008001000120014001600180020002200240026002800300032003400360038004000

Intensity, cps10.8812.9711.3413.44Sample Name: "mc120720a014" Sample ID: "11 012-74-10" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "5.0 ng/mL FC mix in Milli Q water" Annotation: ""Sample Index: 15Sample Type: StandardConcentration: 4.64 ng/mLCalculated Conc: 4.50 ng/mLAcq. Date: 7/20/2012Acq. Time: 3:23:14 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 222193 countsHeight: 7.37e+004 cpsStart Time: 11.4 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e4

Intensity, cps11.7011.5612.33Sample Name: "mc120720a021" Sample ID: "Method Blank" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "Milli Q water TN12-0228" Annotation: ""Sample Index: 22Sample Type: UnknownConcentration: N/ACalculated Conc: 0.0204 ng/mLAcq. Date: 7/20/2012Acq. Time: 5:39:53 PMModified: NoProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 11.7 minArea: 368 countsHeight: 3.34e+002 cpsStart Time: 11.7 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min02004006008001000120014001600180020002200240026002800300032003400360038004000420044004600

Intensity, cps12.2912.3812.1912.6813.0713.2413.6811.69*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 8 of 12Sample Name: "mc120720a022" Sample ID: "GLP10-01-02-35-001" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 23Sample Type: UnknownConcentration: N/ACalculated Conc: 5270. ng/mLAcq. Date: 7/20/2012Acq. Time: 5:59:24 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 1282070 countsHeight: 2.64e+005 cpsStart Time: 11.3 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e52.0e52.1e52.2e52.3e52.4e52.5e52.6e5

Intensity, cps11.6811.5411.42Sample Name: "mc120720a025" Sample ID: "GLP10-01-02-35-001; LMS" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW8R-0-120627-1000 (1:200 dil)" Annotation: ""Sample Index: 26Sample Type: UnknownConcentration: N/ACalculated Conc: 11300. ng/mLAcq. Date: 7/20/2012Acq. Time: 6:57:57 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 2648963 countsHeight: 7.33e+005 cpsStart Time: 11.3 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e5

Intensity, cps11.6711.53Sample Name: "mc120720a027" Sample ID: "GLP10-01-02-35-004" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 28Sample Type: UnknownConcentration: N/ACalculated Conc: 4540. ng/mLAcq. Date: 7/20/2012Acq. Time: 7:36:59 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 1108850 countsHeight: 2.25e+005 cpsStart Time: 11.3 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e52.0e52.1e52.2e5

Intensity, cps11.6711.5211.40Sample Name: "mc120720a030" Sample ID: "GLP10-01-02-35-004; LMS" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW8R-0-120627-2130 (1:200 dil)" Annotation: ""Sample Index: 31Sample Type: UnknownConcentration: N/ACalculated Conc: 10400. ng/mLAcq. Date: 7/20/2012Acq. Time: 8:35:32 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 2456945 countsHeight: 6.99e+005 cpsStart Time: 11.3 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e5

Intensity, cps11.6611.52*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 9 of 12Sample Name: "mc120720a036" Sample ID: "GLP10-01-02-35-007" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW8R-0-120628 (1:200 dil)" Annotation: ""Sample Index: 37Sample Type: UnknownConcentration: N/ACalculated Conc: 4160. ng/mLAcq. Date: 7/20/2012Acq. Time: 10:32:41 PMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 1018661 countsHeight: 2.22e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e52.0e52.1e52.2e5

Intensity, cps11.6511.51Sample Name: "mc120720a041" Sample ID: "GLP10-01-02-35-010" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW9R-0-120625-1250 (1:200 dil)" Annotation: ""Sample Index: 42Sample Type: UnknownConcentration: N/ACalculated Conc: 2570. ng/mLAcq. Date: 7/21/2012Acq. Time: 12:10:17 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.7 minArea: 633565 countsHeight: 1.16e+005 cpsStart Time: 11.3 minEnd Time: 11.8 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.005000.001.00e41.50e42.00e42.50e43.00e43.50e44.00e44.50e45.00e45.50e46.00e46.50e47.00e47.50e48.00e48.50e49.00e49.50e41.00e51.05e51.10e51.15e5

Intensity, cps11.6511.5011.38Sample Name: "mc120720a050" Sample ID: "GLP10-01-02-35-013" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW9R-0-120625-2400 (1:200 dil)" Annotation: ""Sample Index: 51Sample Type: UnknownConcentration: N/ACalculated Conc: 2600. ng/mLAcq. Date: 7/21/2012Acq. Time: 3:06:02 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 642210 countsHeight: 1.19e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.005000.001.00e41.50e42.00e42.50e43.00e43.50e44.00e44.50e45.00e45.50e46.00e46.50e47.00e47.50e48.00e48.50e49.00e49.50e41.00e51.05e51.10e51.15e5

Intensity, cps11.6411.4911.37Sample Name: "mc120720a055" Sample ID: "GLP10-01-02-35-016" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 56Sample Type: UnknownConcentration: N/ACalculated Conc: 2670. ng/mLAcq. Date: 7/21/2012Acq. Time: 4:43:41 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 658491 countsHeight: 1.31e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.005000.001.00e41.50e42.00e42.50e43.00e43.50e44.00e44.50e45.00e45.50e46.00e46.50e47.00e47.50e48.00e48.50e49.00e49.50e41.00e51.05e51.10e51.15e51.20e51.25e5

Intensity, cps11.6411.4911.38*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 10 of 12Sample Name: "mc120720a057" Sample ID: "GLP10-01-02-35-018" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW9R-FMS-120626 (1:200 dil)" Annotation: ""Sample Index: 58Sample Type: UnknownConcentration: N/ACalculated Conc: 3180. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:22:44 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 782368 countsHeight: 1.72e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e5

Intensity, cps11.6311.4911.38Sample Name: "mc120720a058" Sample ID: "GLP10-01-01-36-016; LMS" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "DAL-GW-WW9R-0-120626 (1:200 dil)" Annotation: ""Sample Index: 59Sample Type: UnknownConcentration: N/ACalculated Conc: 8460. ng/mLAcq. Date: 7/21/2012Acq. Time: 5:42:16 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 2012804 countsHeight: 5.45e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e53.2e53.4e53.6e53.8e54.0e54.2e54.4e54.6e54.8e55.0e55.2e5

Intensity, cps11.6311.49Sample Name: "mc120720a066" Sample ID: "LCS-120719-7" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "20ppb LCS" Annotation: ""Sample Index: 67Sample Type: QCConcentration: 18.6 ng/mLCalculated Conc: 17.4 ng/mLAcq. Date: 7/21/2012Acq. Time: 8:18:31 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 849171 countsHeight: 2.69e+005 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e52.0e52.1e52.2e52.3e52.4e52.5e52.6e5

Intensity, cps11.6311.52Sample Name: "mc120720a069" Sample ID: "LCS-120719-4" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "2ppb LCS" Annotation: ""Sample Index: 70Sample Type: QCConcentration: 1.86 ng/mLCalculated Conc: 1.74 ng/mLAcq. Date: 7/21/2012Acq. Time: 9:17:07 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 85622 countsHeight: 2.94e+004 cpsStart Time: 11.3 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e4

Intensity, cps11.6211.5112.23*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 11 of 12Sample Name: "mc120720a072" Sample ID: "LCS-120719-1" File: "mc120720a.wiff"Peak Name: "PFOS" Mass(es): "499.000/80.000 Da,499.000/99.000 Da,499.000/130.000 Da"Comment: "0.2ppb LCS" Annotation: ""Sample Index: 73Sample Type: QCConcentration: 0.186 ng/mLCalculated Conc: 0.168 ng/mLAcq. Date: 7/21/2012Acq. Time: 10:15:44 AMModified: YesProc. Algorithm: Specify Parameters - MQ IIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 0 pointsRT Window: 30.0 secExpected RT: 11.7 minUse Relative RT: NoInt. Type: ManualRetention Time: 11.6 minArea: 7735 countsHeight: 3.00e+003 cpsStart Time: 11.4 minEnd Time: 11.7 min9.5 10.0 10.5 11.0 11.5 12.0 12.5 13.0 13.5 14.0Time, min020040060080010001200140016001800200022002400260028003000320034003600380040004200

Intensity, cps12.2211.6313.2312.1312.8213.3111.5113.9013.61*ETS-McCoyResults Name: mc120720a.rdbPrinting Date: Wednesday, August 08, 2012Data printed by STWPrinting Time: 8:43:57 AMPage 12 of 12mc120803a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -87.3 x^2 + 8.59e+004 x + -511 (r = 0.9998)0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 16.0 17.0 18.0 19.0 20.0 21.0 22.0 23.0 24.0 25.0Concentration, ng/mL0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e62.1e62.2e6

Area, counts*ETS McCoyResults Name: mc120803a.rdbPrinting Time: 9:10:14 AMPrinting Date: Monday, August 06, 2012*Data printed by KJUPage 1 of 1Sample Name: "mc120803a015" Sample ID: "11 012-74-9" File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "2.5 ng/mL FC mix in Milli Q water" Annotation: ""Sample Index: 15Sample Type: StandardConcentration: 2.50 ng/mLCalculated Conc: 2.58 ng/mLAcq. Date: 8/3/2012Acq. Time: 2:39:11 PMModified: NoProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ValleyRetention Time: 9.84 minArea: 220928 countsHeight: 8.31e+004 cpsStart Time: 9.73 minEnd Time: 9.95 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e4

Intensity, cps9.849.80Sample Name: "mc120803a021" Sample ID: "Method Blank" File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "Milli Q water TN12-0248" Annotation: ""Sample Index: 21Sample Type: UnknownConcentration: N/ACalculated Conc: 0.00767 ng/mLAcq. Date: 8/3/2012Acq. Time: 4:36:22 PMModified: NoProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ValleyRetention Time: 9.82 minArea: 147 countsHeight: 3.56e+002 cpsStart Time: 9.82 minEnd Time: 9.83 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min010020030040050060070080090010001100120013001400150016001700180019002000210022002300

Intensity, cps10.3311.6910.3511.7311.6411.8210.4111.8510.4610.0510.5111.0411.4711.9010.579.9011.1612.237.658.609.828.899.598.227.927.449.38Sample Name: "mc120803a022" Sample ID: "GLP10-01-02-35-013 " File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "DAL-GW-WW9R-0-120625-2400 (1:100 dil)" Annotation: ""Sample Index: 22Sample Type: UnknownConcentration: N/ACalculated Conc: 250. ng/mLAcq. Date: 8/3/2012Acq. Time: 4:55:55 PMModified: YesProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ManualRetention Time: 9.83 minArea: 213565 countsHeight: 7.64e+004 cpsStart Time: 9.41 minEnd Time: 9.92 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e4

Intensity, cps9.83Sample Name: "mc120803a028" Sample ID: "LCS-120803-1" File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "0.2ppb LCS" Annotation: ""Sample Index: 28Sample Type: QCConcentration: 0.200 ng/mLCalculated Conc: 0.225 ng/mLAcq. Date: 8/3/2012Acq. Time: 6:53:08 PMModified: NoProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ValleyRetention Time: 9.83 minArea: 18799 countsHeight: 7.05e+003 cpsStart Time: 9.75 minEnd Time: 9.90 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min05001000150020002500300035004000450050005500600065007000

Intensity, cps9.8311.6711.6411.7110.3310.3411.7510.3911.8911.5211.0110.5412.0610.079.737.497.748.857.799.178.66Printing Date: Wednesday, August 08, 2012Data printed by STWPage 1 of 2Sample Name: "mc120803a031" Sample ID: "LCS-120803-4" File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "2.0ppb LCS" Annotation: ""Sample Index: 31Sample Type: QCConcentration: 2.00 ng/mLCalculated Conc: 2.10 ng/mLAcq. Date: 8/3/2012Acq. Time: 7:51:46 PMModified: NoProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ValleyRetention Time: 9.83 minArea: 179608 countsHeight: 6.90e+004 cpsStart Time: 9.75 minEnd Time: 9.90 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e4

Intensity, cps9.83Sample Name: "mc120803a034" Sample ID: "LCS-120803-7" File: "mc120803a.wiff"Peak Name: "PFBS" Mass(es): "299.000/80.000 Da,299.000/99.000 Da"Comment: "20ppb LCS" Annotation: ""Sample Index: 34Sample Type: QCConcentration: 20.0 ng/mLCalculated Conc: 21.0 ng/mLAcq. Date: 8/3/2012Acq. Time: 8:50:24 PMModified: NoProc. Algorithm: IntelliQuan - IQA IIMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 5 pointsRT Window: 30.0 secExpected RT: 9.84 minUse Relative RT: NoInt. Type: ValleyRetention Time: 9.83 minArea: 1765631 countsHeight: 6.48e+005 cpsStart Time: 9.72 minEnd Time: 9.95 min7.5 8.0 8.5 9.0 9.5 10.0 10.5 11.0 11.5 12.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps9.83Printing Date: Wednesday, August 08, 2012Data printed by STWPage 2 of 2

# A TTACHMENT 

# C: A NALYTICAL 

# M ETHOD 

# ( S 

# )

### ETS -8- 044. 1 Page 2 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis 

## 1 Scope and Application

This method describes the direct injection analysis of perfluorinated compounds (PFCs) from

water matrices using high

- performance liquid chromatography tandem mass spectrometry

(HPLC/MS/MS). The method is generally applicable but not limited to the measurement of

perfluoroalkyl sulfonamides and perfluorinated alkyl acids (PFAAs) such as perfluorosulfonic

acids (PFSAs) and perfluorocarboxylic acids (PFCAs) (Table 1). Water samples containing

heavy particulate may require preparation by an alternate method such as ETS -8- 154

“ Determination of Perfluorinated Acids, Alcohols, Amides, and Sulfonates In Water By Solid

Phase Extraction and High Performance Liquid Chromatography/Mass Spectrometry ”. The

method is applicable to both external standard and internal standard calibration 1



### .

Table 1. Representative Tar get Analytes

**Acronym Analyte**  **Chemical Abstract Services**

**Registry Number (CASRN)**

PFBA (C4 Acid) Perfluorobutanoic acid 375

- 22

- 4

PFPeA (C5 Acid) Perfluoropentanoic acid 2706

- 90

- 3

PFHxA (C6 Acid) Perfluorohexanoic acid 307

- 24

- 4

PFHpA (C7 Acid) Perfluoroheptanoic acid 375

- 85

- 9

PFOA (C8 Acid) Perfluorooctanoic acid 335

- 67

- 1

PFNA (C9 Acid) Perfluorononanoic acid 375

- 95

- 1

PFDA (C10 Acid) Perfluorodecanoic acid 335

- 76

- 2

PFUnA (C11 Acid) Perfluoroundecanoic acid 2058

- 94

- 8

PFDoA (C12 Acid) Perfluorododecanoic acid 307

- 55

- 1

PF TrD A (C1 3 Acid) Perfluorotridecanoic acid 72629

- 94

- 8

PFBS (C4 Sulfonate) Perfluorobutanesulfonic acid 375

- 73

- 5

PFHS (C6 Sulfonate) Perfluorohexanesulfonic acid 355

- 46

- 4

PFOS (C8 Sulfonate) Perfluorooctanesulfonic acid 1763

- 23

- 1

FBSA (C4 Sulfonamide Perfluorobutanesulfonamide 30334

- 69

- 1

FOSA (C8 Sulfonamide) Pefluorooctanesulfonamide 754

- 91

- 6



### The Minimum Reporting Level (MRL) is the Limit of Quantitation (LOQ) that meets Data

Quality Objectives (DQOs) that are developed based on the intended use of this method.

Method Flexibility

– This is a performance

- based method and may be generally applied to the

determination of perfluorinated compounds in water matrices when analysis batch quality

c ontrol ( QC ) criteria are met 2



### . Each set of samples are prepared in an analysis batch with

calibration standards, LCS s, blanks, and continuing calibration check standards analyzed on

the same instrument during a time period that begins and ends with the analysis of the

appropriate continuing calibration check standards. The laboratory is permitted to modify the

LC column, mobile phase composition, LC conditions, and MS/MS conditions. Method

modifications should be considered to improve method performance or to meet data quality

objectives for the study. In all cases where method modifications are implemented, the batch

1 The method is supported by validation with internal standard calibration for C4

- C13 PFCAs, C4, C6, and C8 PFSAs,

and C8 perfluoroalkane sulfonamide in laboratory control samples under 3M method validation E11

- 0667.

2 Guidance for establishing method QC Criteria based on a.) FDA May 2001, “Guidance for Industry, Bioanalytical

Method Vali dation”, b.) EPA Method 537, and c.) European Commission: Guidance for Generating and Reporting

Methods of Analysis in Support of Pre

- registration Data Requirements for Annex II (Part A, section 4) and Annex III

(Part A,section 5) of Directive 91/414, SANCO/3029/99 rev. 4 (11/07/00).

### ETS -8- 044. 1 Page 3 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis analytical QCs ( section 9) must be completed and pass QC acceptance criteria ( section 13 ) if

the data from the analytical batch are to be reported

.



## 2 Method Summary

Water samples are analyzed as neat aqueous sample or as solvent diluted aqueous samples

by direct injection using LC/ MS/MS. Samples containing heavy particulate may not be

suitable for analysis by this method. Samples containing suspended particulate should be

centrifuge d or filtered prior to removing a sample aliquot or diluting with solvent

. The water

sample is mixe d well prior to removing an aliquot or diluting, if necessary, with ASTM Type I

water, HPLC water, other suitable water, or solvent (methanol).

Quantitation is by stable isotope internal standard calibration in laboratory reagent water. All

perfluorinat ed compounds ( PFC s) target analyte concentrations of perfluorosulfonic acids

(PFSAs) and perfluorocarboxylic acids (PFCAs) are reported as anions and corrected for

their salt or free acid forms. Alternatively, quantitation may be performed by external

st andard calibration.

This is a performance

- based method. Method uncertainty for each target analyte is

determined for each analytical batch using multiple laboratory control spikes at multiple

concentrations. This method also requires that the precision and accuracy for each sample

be determined using field matrix spikes to verify that the method is applicable to each sample

matrix.

Calibration standards for PFUnA, PFDoA, PFTrDA, and FOSA have been found to be

un stable for more than 2 days in 100% water. Samples requiring analysis for these

compounds by this method should be diluted 1:1 with methanol and analyzed against a

calibration curve prepared in 1:1 synthetic groundwater:MeOH.



## 3 Definitions

3.1 Analysis Batch

A set of study samples that are prepared with calibration standards, laboratory control

samples, and procedural blanks , and analyzed on the same instrument during a time period

that begins and ends with the analysis of the appropriate continuing calibration check

standards.



## 3.2 Analytical Sample

A portion of a laboratory sample prepared for analysis.



## 3.3 Calibration Standard

A solution prepared by spiking a known volume of the Working Standard (WS) into a

predete rmined amount of ASTM Type I, HPLC grade water, or other suitable water (i.e.

matrix water), and analyzed according to this method. Calibration standards are used to

calibrate the instrument response with respect to analyte concentration.



## 3.4 Laboratory Duplicate Sample (LDS, or Lab Dup)

A laboratory duplicate sample is a separate aliquot of a sample taken in the analytical

laboratory that is analyzed separately with identical procedures. Analysis of LDSs compared

to that of the first aliquot give a measure of the precision associated with laboratory

procedures, but not with sample collection, preservation, or storage procedures.

### ETS -8- 044. 1 Page 4 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis 

## 3.5 Field Blank (FB)/Trip Blank (TB)

ASTM Type I, HPLC grade water, or other suitable water, placed in a sample container in the

laboratory and treated as a sample in all respects, including exposure to samp ling site

conditions, storage, preservation and all analytical procedures. The purpose of the TB is to

determine if test substances or other interferences are present in the field environment. This

sample is also referred to as a Trip Blank.



## 3.6 Field Duplicate Sample (FDS, Field Dup)

A sample collected in duplicate at the same time from the same location as the sample. The

FDS is handled under identical circumstances and treated exactly the same throughout field

and laboratory procedures. Analysis of the FDS compared to that of the first sample gives a

measure of the precision associated with sample collection, preservation and storage, as well

as with laboratory procedures.



## 3.7 Field Matrix Spike (FMS)

A sample to which known quantities of the target analyte s , ISs and SRSs are added to the

sample bottle in the laboratory before the bottles are sent to the field for collection of aqueous

samples. A known, specific volume of sample must be added to the sample container

without rinsing. This may be accomplished by making a “fill to this level” line on the outside of

the sample container. The FMS is analyzed to ascertain if any matrix effects, interferences,

or stability issues may complicate the interpretation of the sample analysis.



## 3.8 Trip Blank Matrix Spike ( T BMS )

An aliquot of ASTM Type I, HPLC grade water, or other suitable water, to which known

quantities of the target analytes , ISs and SRSs are added in the laboratory prior to the

shipment of the collection bottles. The TBMS is analyzed exactly like a study sample to help

determine if the method is in control and whether a loss of analyte or analytical bias could be

attributed to sample holding time, sample storage and/or shipment issues. A low and high

TBMS are appropriate when expected sample concentrations are not known or may vary.



## 3.9 Internal Standard (IS)

A compound added to each study sample, calibration standard, laboratory control samples,

and procedural blanks at a consistent level (typically around 1 ng/mL). T he internal

standard (s) are stable isotope labeled versions of the target analyte s . The area count ratio of

the target analyte to the internal standard is used for calibration. Surrogate ISs are applied

when stable isotope ISs of target analytes are unavailable. A surrogate IS is not necessar ily

a stable isotope labeled version of the target analyte, but is treated as an internal standard for

quantitation.



## 3.10 Laboratory Control Sample (LCS)

An aliquot of control matrix to which known quantities of the target analytes , ISs and SRSs

(when applicabl e) are added in the laboratory at the time when samples are aliquotted . At

least three levels (two levels for SRSs) in triplicate are included, one generally at the low end

of the calibration curve and one near the mid range and the upper end of the curve. The

LCSs are analyzed exactly like a laboratory sample to determine whether the stability of the

standards . LCSs should be prepared each day samples are aliquoted

.



## 3.11 Laboratory Matrix Spike (LMS)

A laboratory matrix spike is an aliquot of a sample to which known quantities of target

analytes , ISs and SRSs (when applicable) are added in the laboratory. The LMS is analyzed

exactly like a laboratory sample to determine whether the sample matrix contributes bias to

the analytical results. The endogenous concentrations of the analytes in the sample matrix

must be determined in a separate aliquot and the measured values in the LMS corrected for

these concentrations. LMSs are optional for analysis of aqueous samples.

### ETS -8- 044. 1 Page 5 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis 

## 3.12 Laboratory Sample

A portion or aliquot of a sample received from the field for testing.



## 3.13 Limit of Quantitation (LOQ)

The lower limit of quantitation (LLOQ) for an analytical batch is the lowest concentration that

can be reliably quantitated within the specified limits of precision and accuracy. The LLOQ is

generally selected as the lowest non

- zero standard in the calibration curve that meets method

acceptance criteria. The LLOQ for each target analyte is established for each analysis batch

as the lowest calibration standard with area counts at least twice that of the average area counts of the procedural blanks.

The upper limit of quantitation (ULOQ) for an analytical batch is the highest concentration that

can be reliably quantitated within the specified limits of precision and accuracy. The highest

standard in the calibration curve that meets method acceptance criteria is defined as the

ULOQ.



## 3.14 Method /Procedural Blank

An aliquot of control matrix that is treated exactly like a laboratory sample including exposure

to all glassware, equipment, solvents, and reagents that are used with other laboratory

samples. The method blank is used to determine if test substances or other interferences are

present in the laboratory environment, the reagents, or the apparatus.



## 3.15 Sample

A sample is an aliquot removed from a larger quantity of material intended to represent the

original source material.



## 3.16 Stock Standard Solution (SSS)

A concentrated solution of a single

- analyte prepared in the laboratory with an assayed

reference compound.



## 3.17 Surrogate Internal Standard

An IS that is not necessarily a stable isotopically labeled target analyte, but is treated as an

internal standard for quantitation. Surrogate ISs are used when isotopically labeled

counter parts of the target analyte are not commercially or re adily available.



## 3.18 Surrogate Recovery Standard (SRS)

An isotopically labeled standard , not used as an internal standard, that is added to each

sample and appropriate QC sample as a means to evaluate the method performance for a

chemical class of compounds (e .g., PFSAs, PFCAs)

.



## 3.19 Working Standard (WS)

A solution of several analytes prepared in the laboratory from SSSs and diluted as needed to

prepare calibration standards and other required analyte solutions.



## 4 Warnings and Cautions

4.1 Health and Safety

The acute and chronic toxicity of the standards for this method have not been precisely

determined; however, each should be treated as a potential health hazard. The analyst

should wear gloves, a lab coat, and safety glasses to prevent exposure to chemicals that

might be present.

### ETS -8- 044. 1 Page 6 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis The laboratory is responsible for maintaining a safe work environment and a current

awareness of local regulations regarding the handling of the chemicals used in this method.

A reference file of material safety data sheets (MSDS) should be available to all personnel

involved in these analyses.



## 4.2 Cautions

The analyst must be familiar with the laboratory equipment and potential hazards including,

but not limited to, the use of solvents, pressurized gas and solvent lines, high voltage, and

vacuum systems. Refer to the appropriate equipment procedure or operator manual for

additional information and cautions.



## 5 Interferences

During sample preparation and analysis, major potential contaminant sources are reagents

and glassware. All materials used in the analyses shall be demonstrated to be free from

interferences under conditions of analysis by running method blanks.

Parts and supplies that contain Teflon® should be avoided or minimized due to the possibility

of interference and/or contamination. These may include, but are not limited to: wash bottles,

Teflon ® 

### lined caps, autovial caps, HPLC parts, etc.

The use of disposable micropipettes or pipettes to aliquot standard solutions is recommended

to make calibration standards and matrix spikes.



## 6 Instrumen tation, Supplies, and Materials

6.1 Instrumentation

Analytical balance capable of reading to 0.0001g

HPLC/MS/MS or HPLC/MS system, as described in Section 10

.



## 6.2 Supplies and Materials

Sample collection bottles

— HDPE (e.g., NalgeneTM) wide

- mouth bottles with screw cap.

Note: Do not use fluorinated or Teflon® bottles or lined caps.

Coolers or boxes for sample shipment.

15

- mL and 50

- mL disposable polypropylene centrifuge tubes.

Class A pipettes and volumetric flasks, various.

2 mL HPLC auto vial s

Disposable pipettes, polypropylene or glass as appropriate

Centrifuge capable of spinning 15

- mL and 50

- mL polypropylene tubes at 3000 rpm.



## 7 Reagents and Standards

Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent

performance may be ach ieved using chemicals obtained from other suppliers. Do not use a

lesser grade of chemical than those listed.



## 7.1 Chemicals

Water

– Milli

- Q, HPLC grade, or other suitably appropriate sources

Calcium Acetate

- A.C.S. Reagent Grade

### ETS -8- 044. 1 Page 7 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis Magnesium Acetate

- A.C.S. Re agent Grade

Methanol

– HPLC grade

Ammonium Acetate

– A.C.S. Reagent Grade



## 7.2 Representative Target Analytes, ISs, and SRSs

PFBA, Heptafluorobutyric Acid, (C 4 

### Perfluorinated Acid)

PFPeA , Nonafluoropentanoic Acid (C 5 

### Perfluorinated Acid)

PFHxA, Perfluorohexanoi c Acid (C 6 

### Perfluorinated Acid)

PFHpA, Tridecafluoroheptanoic Acid, (C 7 

### Perfluorinated Acid)

PFOA, Ammonium perfluorooctanoate, (C 8 

### Perfluorinated Acid)

PFNA, Heptadecafluorononanoic Acid, (C 9 

### Perfluorinated Acid)

PFDA, Nonadecafluorodecanoic Acid (C 10 

### Per fluorinated Acid)

PFUnA, Perfluoroundecanoic Acid, (C 11 

### Perfluorinated Acid)

PFDoA, Perfluorododecanoic Acid, (C 12 

### Perfluorinated Acid)

PFTrDA, Perfluorotridecanoic Acid, (C 13 

### Perfluorinated Acid)

FBSA, Perfluorobutanesulfonamide

FOSA, Perfluorooctanesulfo nylamide

PFBS, Potassium Perfluorobutane sulfonate

PFHS, Perfluorohexanesulfonate

PFOS, Potassium perfluorooctanesulfonate

PFOA [1,2, 3, 4

- 13 

### C], 13 

### C 4 

### - isotopically labeled perfluorooctanoic acid (SRS)

PFOS [1,2, 3, 4

- 13 

### C], 13 

### C 4 

### - isotopically labeled Perfluorooctanesulfonate (SRS)

PFUnA [1,2

- 13 

### C], 13 

### C 2 

### - isotopically labeled Perfluoroundecanoic acid (SRS)

A custom mix of ISs in a methanolic solution containing ([1,2,3,4

- 13 

### C 4 

### ]PFBA, [1,2

- 13 

### C 2 

### ]PFHxA, [1,2,3,4 ,5,6,7,8

- 13 

### C 8 

### ]PFOA, [1,2,3,4,5 ,6,7,8,9

- 13 

### C 9 

### ]PFN A, [1,2

- 13 

### C 2 

### ]PFDA,

[1,2 ,3,4,5,6,7

- 13 

### C 7 

### ]PFUnA, [1,2

- 13 

### C 2 

### ]PFDoA, [1,2 ,3

- 13 

### C 3 

### ]PFHS, [1,2,3,4 ,5,6,7,8

- 13 

### C 8 

### ]PFOS

, and [1,2,3,4,5,6,7,8

- 13 

### C 8 

### ]PFOSA (Wellington Laboratories, Guelph, ON) in combination with

added ([1,2,3,4 ,5

- 13 

### C 5 

### ]PFPe A

, ([1,2,3,4

- 13 

### C 4 

### ]PFHpA, and [ 18 

### O 2 

### ]PFBS can be used to prepare
a stock IS solution. Alternatively, individual stable isotope ISs can be used to prepare a stock

IS mixture.

Other ISs can be applied.



## 7.3 Reagent Preparation

2 mM A mmonium acetate solution (Analysis)

— Weigh 0.3 g of A mmonium acetate and

dissolve in 2.0 L of reagent water.

Synthetic Groundwater (containing 25 ppm Ca and Mg)

– Weigh 0.61 g of Calcium Acetate

and 0.92 g of Magnesium Acetate and dissolve in 6.0 L of reagent water.

Note: Alternative volumes may be prepared as long as the ratios of the solvent to solute ratios

are maintained.

### ETS -8- 044. 1 Page 8 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis 

## 7.4 Stock Standard Solution (SSS) and Working Standard Solution

Preparation

The following standard preparation procedure serves as an example. Weighed amounts and

final volumes may be changed to suit the needs of a particular study. For example, μL

volumes may be spiked into volumetric flasks when diluting stock solutions to appropriate

levels.

100 μg/mL target analyte SSSs

— Weigh out 10 mg of analytical standard **(corrected for**

**percen t salt**  **, acid [ETS -4-**  **031] and purity**  ) and dilute to 100 mL with methanol or other

suitable solvent, in a 100 mL volumetric flask. Transfer to a 125 mL LDPE bottle or other

suitable container. Prepare a separate solution for each analyte. Expiration dates and

storage conditions of stock solutions should be assigned in accordance with laboratory

standard operating procedure. An example of purity and salt correction is given below for

PFOS.



### salt of weight moclecular anion of weight molecular factor correction salt

=

9275.0 538 499 factor correction )salt (K PFOS

== +



### 10 mg C 8 

### F 17 

### S0 3 - 

### K + 

### with purity 90% = 8.35 mg C 8 

### F 17 

### S0 3 – 

### (10 mg*0.90*0.9275=8.35 mg)

10 μg/mL (10,000 ng/mL) mixed working standard

— Add 5.0 mL each of the 100 μg/mL

SSSs to a 50 mL volumetric flask and bring up to volume with solvent.

1 μg/mL (1,000 ng/mL) mixed working stan dard

— Add 0.5 mL of the 100 μg/mL SSSs to a

50 mL volumetric flask and bring up to volume with solvent.

0.1 μg/mL (100 ng/mL) mixed standard

— Add 0.05 mL of the 100 μg/mL SSSs to a 50 mL

volumetric flask and bring up to volume with solvent.

Storage Conditi ons

— Store all SSSs and working standards in accordance with laboratory

standard operating procedure or in a refrigerator at 4°±2°C for a maximum period of 6 months

from the date of preparation.



## 7.5 Calibration Standards

Calibration can be performed by IS or external calibration. Using the working standards

described above, prepare calibration solutions in ASTM Type I water, HPLC water, other

suitable water, or a mixture of solvent and water using the information in Table 2 as a

guideline. Note: Volumes of wa ter or water/solvent mixtures and working standards may be

adjusted to meet the data quality objectives addressed in the general project outline.

Calibration levels other than those listed below can be prepared as needed.

For the quantitation of PFOA and PFOS, reference materials of certified mixed linear and

branched isomer are preferred. Alternately, reference materials of primarily linear isomers of

PFOA and/or PFOS may be used, however, when quantitating with predominantly linear

reference standards, additional LCS samples containing both linear and branched isomers of

PFOA and PFOS are required 3



### .

7.5.1 Internal Standard (IS) and Surrogate Recovery Standard (SRS)

For IS calibration, stable isotope internal standards of each target analyte or appropriate

surrogate ISs should be spiked at the same level in all calibration standards. Once the

calibration standards have been prepared as stated above in Section 7.5, all calibration

standards are spiked with a separate internal standard spiking solution. Typically the

3 A report summarizing an assessment of the use of reference standards containing certified linear and branched isomers of

PFOA/PFOS can be found in 3M report E11

- 0560.

### ETS -8- 044. 1 Page 9 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis concentration of the internal standard is consistent with the internal standard concentration

expected in the samples being prepared, usually 1 ng/mL. The concentration of the internal

standard spiking solution is typically 2 μg/ mL. A separate zero point or method blank is

typically prepared at the same time as the calibration standards, using the same solution

used to prepare the standards (ASTM Type I water, HPLC water, other suitable water, or a

solvent/water mixture), and is spiked with the internal standard at the same concentration as

the calibration curve, typically at 1 ng/mL.

If the sample s being analzyed w ere pre

- spiked with SRSs , the calibration curve prepared in

Section 7.5 is spiked with a separate SRS spiking solution. Typically, the sample bottles are

spiked with a SRS at 0.1 ng/mL. The final calibration curve must consist of at least six

calibration points after analysis. The following table provides an example of spike

concentrations and volumes used to achieve a multi

- point extracted calibration curve with

internal standard and surrogate standard.

Table 1 lists recommended stable isotope internal standards for several PFSA and PFCA

target compounds. A custom mix of isotopically labeled target analytes in a methanolic

solution containing ([1,2,3,4

- 13 

### C 4 

### ]PFBA, [1,2

- 13 

### C 2 

### ]PFHxA, [1,2,3,4,5,6,7,8

- 13 

### C 8 

### ]PFOA,

[1,2,3,4,5,6,7,8,9

- 13 

### C 9 

### ]PFNA, [1,2,3,4,5,6

- 13 

### C 6 

### ]PFDA, [1,2,3,4,5,6,7

- 13 

### C 7 

### ]PFUnA, [1,2

- 13 

### C 2 

### ]PFDoA, [ 1,2,3

- 13 

### C 3 

### ]PFHS, [1,2,3,4,5,6,7,8

- 13 

### C 8 

### ]PFOS, and [1,2,3,4,5,6,7,8

- 13 

### C 8 

### ]FOSA

(Wellington Laboratories, Guelph, ON) in combination with added ([1,2,3,4 ,5

- 13 

### C 5 

### ]PFPe A

, ([1,2,3,4

- 13 

### C 4 

### ]PFHp A , and [ 18 

### O 2 

### ]PFBS can be used to prepare a stock IS solution.

Alternative sources of certified stable isotope labeled target analytes are applicable.

Alternatively, individual stable isotope ISs can be used to prepare a stock IS mixture. The

table below lists the recommended stable isotope ISs and SRSs applied in the method.

Other stable isotope ISs and SRSs of target analytes not listed in the table may be used if

supported by validation and/or analysis batch QCs meeting method acceptance criteria (e.g.,

[ 13 

### C 2 

### ]- PFOA) . The same internal standard should be used for a given analyte throughout the

entire project/study. Note: some of the compounds listed below are appropriate to use as

surrogate ISs when a stable isotope IS of a target analyte is not available. Generally,

surrogate isotopically labeled PFCAs are used for PFCAs, and surrogate i sotopically labeled

PFSA s are used for PFSAs.

Table 2 provides examples of spike concentrations and volumes used to achieve a multi

- point

calibration curve with ISs and SRSs

.

### ETS -8- 044. 1 Page 10 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS;

Direct Injection Analysis Table 1 . Stable Isotope PFCAs and PFSAs used for ISs and SRSs

Compound Name Synonym or Acronym Analytical Purpose Reference Standard

Source

13 C 4 - Perfluorobutanoic acid [1,2,3,4

- 13 C 4 ]PFBA IS for PFBA Wellington Labs

(Mix or Individual)

13 C 4 - Perfluoro pentanoic acid [1,2,3,4 ,5

- 13 C 5 ]PFPe A IS for PFPeA Wellington Labs

(Mix or Individual)

13 C 2 - Perfluorohexanoic acid [1,2

- 13 C 2 ]PFHxA IS for PFHxA Wellington Labs

(Mix or Individual)

13 C 4 - Perfluoro hept anoic acid [1,2,3,4

- 13 C 4 ]PFHp A IS for PF HpA Wellington Labs

(Mix or Individual)

13 C 8 - Perfluorooctanoic acid [1,2,3,4,5,6,7,8

- 13 C 8 ]PFOA IS for PFOA and [1,2,3,4

13 C 4 ]PFOA Wellington Labs

(Mix or Individual)

13 C 9 - Perfluorononanoic acid [1,2,3,4,5,6,7,8,9

- 13 C 9 ]PFNA IS for PFNA Wellington Labs

(Mix or Individual)

13 C 6 - Perfluorodecanoic acid [1,2,3,4,5,6

- 13 C 6 ]PFDA IS for PFDA Wellington Labs

(Mix or Individual)

13 C 7 - Perfluoroundecanoic acid [1,2,3,4,5,6,7

- 13 C 7 ]PFUnA IS for PFUnA Wellington Labs

(Mix or Individual)

13 C 2 - Perfluorododecanoic acid [1,2

- 13 C 2 ]PFDoA IS for PFDoA, *PFTA Wellington Labs

(Mix or Individual)

18 O 2 - Ammonium Perfluorobutane sulfonate [ 18 O 2 ]PFBS IS for PFBS RTI International

(Individual)

13 C 3 - Ammonium Perfluorohexane sulfonate [ 1,2,3

- 13 C 3 ]PFHS IS for PFHS Wellington Labs

(Mix or Individual)

13 C 8 - Sodium Perfluorooctane sulfonate [1,2,3,4,5,6,7,8

- 13 C 8 ]PFOS IS for PFOS and

PFOS[1,2,3,4 13 C 4 ], Wellington Labs

(Mix or Individual)

13 C 8 - Perfluorooctanesulfonamide [1,2,3,4,5,6,7,8

- 13 C 8 ]FOSA IS for FOSA Wellington Labs (mix)

RTI International

(Individual)

13 C 4 - Perfluorooctanoic acid [1,2,3,4

- 13 C 4 ]PFOA SRS for all PFCAs : C4

- C8 Wellington

13 C 2 - Perfluoroundecanoic acid [1,2

- 13 C 2 ]PFUnA SRS for all PFCAs C9

- C13 Wellington

13 C 8 - Perfluorooctane sulfonate [1,2,3,4

- 13 C 4 ]PFOS SRS for all PFSA s: C4,

C6, and C8 Wellington

### ETS -8- 044. 1 Page 11 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS; Direct Injection Analysis Table 2 . Example Preparation of Calibration Curve with ISs and SRSs

**Sample Description Concentration** 

**of WS, μ**  **g/mL Volume of** 

**WS,** μ **L Volume of IS** 

**(2 μ**  **g/mL),** μ **L Concentration of** 

**Surrogate, μ**  **g/mL Volume of** 

**Surrogate,** μ **L Volume of ASTM Type I**  **Water, or**

**other suitable solvent**  **(1) , mL** 

0.025 ng/mL curve point 0.10 25 50 0.20 12.5 100

0.030 ng/mL curve point 0.10 30 50 0.20 15 100

0.04 ng/mL curve point 0.10 40 50 0.20 20 100

0.05 ng/mL curve point 0.10 50 50 0.20 25 100

0.1 ng/mL curve point 0.10 100 50 0.20 50 100

0.25 ng/mL curve point 0.10 250 50 0.20 125 100

0.5 ng/mL curve point 1.0 50 50 0.20 250 100

1 ng/mL curve point 1.0 100 50 0.20 500 100

2.5 ng/mL curve point 10.0 25 50 10.0 25 100

5.0 ng/mL curve point 10.0 50 50 10.0 50 100

10.0 ng/mL curve point 10.0 100 50 10.0 100 100

25.0 ng/mL curve point 10.0 250 50 10.0 NA 100

50.0 ng/mL curve point 10.0 500 50 10.0 NA 100

75.0 ng/mL curve point 10.0 750 50 10.0 NA 100

100 ng/mL curve point 10.0 1000 50 10.0 NA 100

N/A

- Not Applicable

(1) Samples requiring analysis for PFUnA, PFDoA, PFTrDA, and FOSA should be analyzed against a calibration curve prepared in 1:1 synthetic groundwa ter:MeOH.

### ETS -8- 044. 1 Page 12 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis 

## 8 Sample Collection and Bottle Preparation

Sample collection bottles are prepared by 3M Environmental Laboratory (or subcontract supplier) personnel for

shipment at ambient temperature to the collection site. Typically, four separate collection bottles are

associated with a single collection site: sample, field duplicate sample, low field matrix spike, and high field

matrix spike. Alternatively, the sample and field duplicate sample may contain SRSs in lieu of additional target

analyte low field matrix spike and target analyte high field matrix spike samples. Depending on the scope of

the project, additional replicates of the field sample and field matrix spikes may be added. Also, it is not

uncommon for additional mid

- level field matrix spikes to be collected if the expected sample concentrations are

truly unknown or could span a large concentration range.

High

- density polyethylene (HDPE) wide

- mouth Nalgene bottles are used for the sample collection containers.

(Volumes of the bottles may vary depending on how much sample is required to meet data quality objectives.)

Sample collection volumes are project specific and based on data quality objectives. The Nalgene bottles do

not require any pretreatment prior to use. Typically, placement of a sample bottle volumetric “fill to here” line is

done by using a sample bottle marker template. Alternatively, bottles may be weighed prior to bottle

preparation and weighed again after samples have been collected.

All bottles should be clearly labeled to indicate its intended use as a sample, field sample duplicate, low field

matrix spike, high field matrix spike, sample/SRS field matrix spike, field duplicate sample/SRS field matrix

spike, trip blank, or trip blank matrix spike. If each location has different designated spike levels, the label

should also clearly indicate the sample location designation. Generally, a set of bottles for a given collection

site are prepared then grouped together in plastic bags for organizational purposes. For each sample

collection event, at least one set of trip blank and trip blank matrix spikes are prepared.

Bottle preparation should be documented in a Note to File or on a sample preparation worksheet and should

include the following information: date prepared, total number of bottles prepared, number of sample sites, the

standard identification numbers and spike volumes used to prepare spiked bottles, the “fill to here” volume, and

any other pertinent information needed for reconstructibility of the data. The Note to File will be included in the

final data package for the project.

Samples are collected in the field and shipped to the laboratory at ambient temperature.



## 8.1 Field Matrix Spike Sample (FMS)

Field matrix spike samples are a requirement of the method. A FMS sample is defined as a QC sample to

which known quantities of appropriate target analytes are added to the sample bottle in the field or in the

laboratory before the bottles are sent to the field. The sample and field duplicate sample may contain

appropriate SRSs in lieu of target analyte FMS samples. Sample quantities are determined vol umetrically or

gravimetrically. A known, specific volume or weight of sample is added to the sample container without

rinsing. Volumetric sample measurements may be acquired by a laboratory applied “fill to this level” line on the

outside of the sample container. Target analyte FMS samples should be spiked at approximately 0.5

- 10 times

the expected analyte concentration in the sample. If the expected range of analyte concentrations is unknown,

multiple spikes at varying levels may be prepared to increase the likelihood that a spike at an appropriate level

is made. Typically a low and a high target analyte spike are prepared for each sampling location. In those

instances where SRSs are to be used in lieu of target analyte FMS samples, the sample and field duplicate

sample are spiked at approximately 2

- 5 times the target LOQ. The FMS is analyzed to ascertain if matrix

effects or sample holding time contributes bias to the analytical results. For the sample bottles designated for

matrix spikes, an appropriate volume of matrix spiking solution is added to the empty bottle prior to sampling.

The volume of spike solution added should produce the desired final concentration of target analytes once the

bottle is filled with sample to the “fill to here line”. The matrix spiking solution(s) should be prepared in a

suitable solvent and contain all of the appropriate target analytes, ISs, and SRSs. The target analyte matrix

spiking solution is often the same as the working standards used to create the calibration standards. An

example of a bottle spike is given below.

“Fill to here” volume = 200 mL (A 250 mL Nalgene bottle is used)

Desired Field Spike Concentration = 0.25 ng/mL

### ETS -8- 044. 1 Page 13 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis 500 μ L of a 0.1 μ g/mL spiking solution (containing the target analytes) is added to the bottle and the bottle cap

promptly sealed.



## 8.2 Internal Standard and Surrogate Recovery Standard

If analysis of a surrogate recovery standard (SRS) is included in the project objectives, an appropriate volume

of a surrogate standard solution is added to all the bottles prior to sampling and SPE. Typically sample bottles

are spiked with surrogate recovery standards at a final desired spike concentration of 0.1 ng/mL.

If quantitation by internal standard (IS) is included in the project objective, an appropriate volume of internal

standard solution is added to all the bottles prior to sampling and SPE . Typically sample bottles are spiked

with internal standard at a final desired spike concentration of 1 ng/mL.

For the trip blank, the SRS spike and IS spike i s added to the bottle and then ASTM Type I water (HPLC grade

reagent water or other suitable water may used) is added to the “fill to here” line. The bottle is capped and

sealing tape may be placed around the outer edge of the cap. Trip blank matrix spikes are prepared by adding

the appropriate volume of target analyte spiking solution, IS, and SRS spiking solutions and filling the bottle to

the desired volume with the appropriate water and capping and sealing the cap.



## 9 Quality Control and Data Quality Objectives

9.1 Data Quality Objectives

This method and required quality control samples is designed to generate data accurate to ±30% with a

targeted LOQ of 0.025 ng/mL. Any deviations from the quality control measures spelled out below will be

documented in the raw data and footnoted in the final report.



## 9.2 Method/Procedural Blanks

The method/procedural blank is zero point calibration standard (which includes ISs) analyzed in a regular basis

with each analysis batch. At a minimum, method blanks are analyzed prior to instrument calibration, prior to

the analysis of CCV samples, after every 10 sample injections, and at the end of the analytical run.

The mean area count or area ratios when using internal standard calibration, for each analyte in the method

blanks m ust be less than 50% of the area count counts or area ratios when using internal standard calibration,

of the LOQ standard. The standard deviation of the area counts, or area ratios when using internal standard

calibration, of these method blanks should be calculated. A specific %RSD acceptance criteria is not specified

but is assessed on an analytical batch basis. If the mean area counts or area ratios when using internal

standard calibration, of the method blanks exceed 50% of the LOQ standard, then the LOQ must be raised to

the first standard level in the curve that meets criteria. Method blanks may be eliminated if technical

justification can be provided (e.g. the procedural blank was analyzed after an unexpectedly high level sample).

If any procedural blanks are removed from the LOQ determination, document in the raw data and report as

appropriate. Laboratory Sample Replicates / Field Duplicate Sample

Typically, samples are collected in duplicates in the field. The relative percent difference (RPD) of duplicate

samples should be ≤20% for the precision of sample preparation and analysis to be considered in control.

Replicate samples not meeting the ≤20% RPD criteria are flagged and reported as outside of QC acceptance

criteria.



## 9.3 Laboratory Matrix S pikes (LMSs)

LMSs may be performed in lieu of FMSs if FMSs have previously been performed for the sample matrix.

Additionally, LMSs may be performed in lieu of FMSs for a sample matrix if the FMS levels were not

appropriate for determining spike recoveries relative to endogenous levels of target analytes and appropriate

SRSs. Generally, each sample location represents a different sample and sample matrix. LMSs are prepared

for each sample and analyzed to determine the matrix effect on spike recovery efficiency of each target analyte

and appropriate SRSs. LMSs should be prepared at a minimum of one level and in duplicate. LMS

concentrations should be prepared at approximately 0.5

- 10 times the endogenous concentration or

approximately 4

- 10 times the LOQ concentration of each target analyte.

### ETS -8- 044. 1 Page 14 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis Lab matrix spike recoveries should fall within ± 30% of expected values. Sample data with LMS recovery

outside of ±30% but within ±50% of the expected value are flagged and reported as outside of QC acceptance

criter ia. Data with LMS recovery outside of ±50% of the expected value are reported as NR, where NR is

defined as “Not Reportable” data outside of QC acceptance criteria.



## 9.4 Lab Control Sample

Lab control spikes are prepared for each analysis batch to determine method accuracy and precision. LCSs

should be prepared at three levels in triplicate for each target analyte and at a minimum of two levels in

triplicate for appropriate SRSs. Low lab control spikes should be prepared at a concentration in the range of

a pproximately four to ten times higher than the targeted lower LOQ, the mid lab control spikes should be

prepared at a concentration near the mid

- point of the calibration curve and the high lab control spikes at

approximately 80% of the upper LOQ. For each target analyte and SRSs, the percent relative standard

deviation (method precision) for each control spike level must be less than or equal to 20% and the average

recovery (method accuracy) for each control spike level must be 80

- 120%. Sample data for target analytes

outside of the laboratory control spike acceptance criteria will be handled as follows:

If the average recovery of a spiking level falls outside method acceptance, but at least 67% (6 out of 9 ) o f LCS

samples are within 20 % of their respective nominal value (33% of the QC samples, not all replicates at the

same concentration, may be outside 20% of nominal value), the average recovery will be flagged as outside

method acceptance criteria. All LCS samples will be control charted as per ETS -4- 02 6. If the average

recovery of one of the spiking levels exceeded the analytical method uncertainty as determined by ETS

- 12

- 012, that analytical batch uncertainty will be expanded for that particular study.

If more than 67% of the LCS samples fail to meet method acceptance criteria, the data will not be reported.

Calibration standards consisting of mixed branched and linear isomer PFOS/PFOA are preferred. However,

for PFOS/PFOA target analytes, if the calibration standards are comprised of predominantly linear isomers

only, at least one level of triplicate LCSs should be prepared using PFOS/PFOA which contains a mix of linear

and branched isomers. These LCSs will be used to demonstrate quantitative equivalency (or quantitative bias)

of the isomeric mix when using a predominantly linear standard for calibration. The mixed linear and branched

isomer PFOS/PFOA LCSs recoveries should fall within ± 30% of expected values. Alternatively, in lieu of

mixed branched and linear isomer PFOS/PFOA LCSs, mixed branched and linear isomer PFOS/PFOA

TBMSs may be applied to demonstrate method accuracy and precision.



## 9.5 Field Matrix Spikes (FMSs) / Surrogate Recovery Standards (SRSs)

FMSs are prepared for each sampling location and analyzed to determine the matrix effect and sample holding

time on the spike recovery of each target analyte and/or appropriate SRS. Generally, each sample location

represents a different sample and sample matrix.

FMSs are QC samples to which known quantities of appropriate target analytes are added to the sample bottle

in the laboratory before the bottles are sent to the field. Typically a low and a high target analyte FMS are

prepared for each sampling location. The sample and field duplicate sample may contain appropriate SRSs in

lieu of target analyte low field matrix spike and target analyte high field matrix spike samples.

Field matrix spike method acceptance criteria are recoveries within ±30% of the expected value. If FMS

recovery (target analyte or SRS spike) is outside of ±30% of the expected value or could not be assessed

because the FMS (target analyte) was spiked at an inappropriate level, the sample result is reported as follows:

1.) If target analyte FMS recovery could not be assessed because the FMS's were at an inappropriate level,

then Laboratory Matrix Spikes (LMS) may be substituted. If LMS recoveries are within ±30% the data are

reportable and flagged to indicate that the FMS spikes levels were inappropriate.

2.) If multiple target analyte FMS's were prepared on a sample and the closest FMS level to the reported

sample meets the ±30% acceptance criteria but additional FMS's are outside the ±30% acceptance range, the

data are reportable and flagged to indicate that while there were failing FMS’s, the uncertainty will not be

expanded since the most appropriate spike level passed.

### ETS -8- 044. 1 Page 15 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis 3.) If the target analyte FMS recoveries are outside of the ±30% acceptance range but at least 30 acceptable

historical reportable FMS sample results are available, the data may be reported but flagged with an expanded

uncertainty and as not meeting FMS criteria.

4.) Sample data with FMS recovery outside of ±30% but within ±50% of the expected value are flagged and

reported as outside of QC acceptance criteria with an expanded uncertainty.

5.) If FMS recovery is outside of ±50%, the sample result is reported as NR, where NR is defined as “Not

Reportable” due to noncompliant QC results.

The targeted fortification levels should be at least 50% of the endogenous level and less than 10 times the

e ndogenous level to be used without justification to determine the statement of accuracy for analytical results.

Note: It is possible for bottles utilized for Field Matrix Spike samples to be under

- filled or over

- filled during

sample collection. Since this scenario will effect the actual concentration of the FMS sample (surrogate and

internal standard concentrations will also be effected, if used), it is important that any obvious under

- filling or

over

- filling of sample bottles be documented in the data package and taken into account in the FMS , ISs, or

SRSs recovery calculations. Samples over

- filled or under

- filled by more than 10% will be require recalculation

of the FMS, ISs, and SRS true values.

The average of the sample and the field duplicate should be used to calculate the recovery.



## 10 Procedures

10.1 Water Sample Preparation

This method is applicable to water samples. Samples containing heavy particulate may not be suitable for

analysis by this method. Samples containing suspended particulate should be centrifuge prior to removing a

sample aliquot, or filtered.

• Thoroughly mix sample before removing a n aliquot and placing in a labeled autovial.

• Dilute sample, if necessary, with ASTM Type I water , HPLC water, other suitable water , or solvent

(methanol)

.

• L ab control spikes are prepared for each analysis batch to determine method accuracy and precision.

LCSs should be prepared at three levels in triplicate for each target analyte and at a minimum of two

levels in triplicate for appropriate SRSs. Low lab control spikes should be prepared at a concentration

in the range of approximately four to ten times higher than the targeted lower LOQ, the mid lab control

spikes should be prepared at a concentration near the mid

- point of the calibration curve and the high

lab control spikes at approximately 80% of the upper LOQ. For IS quantitation, stable isotope internal

standards of each target analyte or appropriate surrogate ISs should be spiked at the same level as the

samples being analyzed, in all LCSs

.

• If LCSs are being prepared us ing synthetic groundwater, allow the LCSs samples to equilibrate for a

minimum of 4 hours before aliquoting for analysis or diluting with solvent (methanol).



## 11 Sample Analysis
- LC/MS/MS

11.1 Instrument Setup

Note: In this example, an Applied Biosystems Sciex API 4000 (API 5000 or API 5500) Tandem Mass

Spectrometer (LC/MS/MS) is used. Other brands/models of LC/MS/MS instruments as well as single

quadrupole mass spectrometers (LC/MS) may be used as long as the method acceptance criteria are met.

Brand names, suppliers, part numbers, and models are for illustrative purposes only. Equivalent performance

may be achieved using apparatus and materials other than those specified here, but demonstration of

equivalent performance that meets the requirements of this method is the responsibility of the laboratory. The

operator must optimize and document the equipment and settings used.

### ETS -8- 044. 1 Page 16 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis Establish the LC/MS/MS system and operating conditions equivalent to the following:

Mass Spec: Applied Biosystems API 4000, API 5000, or API 5500

Ion Source: Turbo Ion Spray (ABS)

Mode: Electrospray Negative

Scan Type: MRM (Multiple Reaction Monitoring)

Computer: Dell DHM

Software: Windows 2000 or Windows XP, Analyst 1.4.2 or higher versi ons

HPLC: Agilent Series 1100,1200, or 1290

Agilent Quaternary Pump

Agilent Vacuum Degasser

Agilent Autosampler

Agilent Column Oven

Note: One or more C18 HPLC analytical columns (2.1 mm x 100 mm, 5 μ m or 2.1 mm x 50 mm, 5 μ m) may be

attached on

- line after the purge valve and before the sample injection port to retard and separate any residue

contaminants that may be in the mobile phase and/or HPLC system.

HPLC Column: Betasil C18, 4.6mm x 100mm, 5 μ m (ThermoElectron Corporation)

Column Temperature: 35 ° C

Inje ction Volume: 5μL

Mobile Phase (A): 2mM Ammonium Acetate in ASTM Type I water (See 7.3)

Mobile Phase (B): Methanol

Table 3

. Liquid Chromatography Gradient Program

.

**Step**

**Number Total Time** 

**(min) Flow Rate** 

**(** μ

**L/min) Percent A** 

**(2 mM ammonium** 

**acetate) Percent B** 

**(Methanol)**

0 0 750 97.0 3.0

1 2.0 750 97.0 3.0

2 14.5 750 5.0 95.0

3 15.5 750 5.0 95.0

4 16.5 750 97.0 3.0

5 20.0 750 97.0 3.0



### Note: Other HPLC gradients may be used as long as the method criteria and project data quality objectives are

met.

It may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with

different dimensions (e.g. 2.1mm x 30mm) and columns from different manufacturers (Keystone Betasil C18

etc.) may be used.

### ETS -8- 044. 1 Page 17 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis Table 4 Suggested MRM Transitions for Target Analytes, Surrogates, and Internal Standards

Analyte Analyte Description Mass Transition

Q1 (amu) Mass Transition

Q3 (amu)

PFBA (C4 Acid) Target 213 169

PFPeA (C5 Acid) Target 263 219

PFHxA (C6 Acid) Target 313 269, 119

PFHpA (C7 Acid) Target 363 319, 169

PFOA (C8 Acid) Target 413 369, 219, 169

PFNA (C9 Acid) Target 463 419, 169, 219

PFDA (C10 Acid) Target 513 469, 269, 219

PFUnA (C11 Acid) Target 563 519, 269, 219

PFDoA (C12 Acid) Target 613 569, 169, 319

PFTA (C13 Acid) Target 663 619, 369, 319

FBSA (C4 Sulfonamide) Target 298 78

FOSA (C8 Sulfonamide) Target 498 78

PFBS (C4 Sulfonate) Target 299 99, 80

PFHS (C6 Sulfonate) Target 399 99, 80

PFOS (C8 Sulfonate) Target 499 80, 99, 130

[1,2,3,4

- 13 C 4 ]PFBA IS for PFBA 217 172

[1,2 ,3,4,5

- 13 C 5 ]PFPe A IS for PFPeA 268 223

[1,2

- 13 C 2 ]PFHxA IS for PFHxA 315 270

[1,2,3,4

- 13 C 4 ]PFHpA IS for PFHpA 367 322

[1,2,3,4,5,6,7,8

- 13 C 8 ]PFOA IS for PFOA 421 376

[1,2,3,4,5,6,7,8,9

- 13 C 9 ]PFNA IS for PFNA 472 427

[1,2,3,4,5,6

- 13 C 6 ]PFDA IS for PFDA 519 474

[1,2,3,4,5,6,7

- 13 C 7 ]PFUnA IS for PFUnA 570 525

[1,2

- 13 C 2 ]PFDoA IS for PFDoA and PFTA 615 570

[ 18 O 2 ]PFBS IS for PFBS 303 84

[ 1,2,3

- 13 C 3 ]PFHS IS for PFHS 402 80

[1,2,3,4

- 13 C 4 ]PFOS IS for PFOS 503 80

[1,2,3,4,5,6,7,8

- 13 C 8 ]FOSA IS for FOSA 507 80

[1,2,3,4

- 13 C 4 ]PFOA Surrogate (C4

- C8 Acids) 417 372

[1,2,3,4

- 13 C 4 ]PFOS Surrogate(Sulfonates, FOSA) 503 80

[1,2

- 13 C 2 ]PFUnA Surrogate (C9

- C13 Acids) 565 520



### Multiple transitions for monitoring the analytes is an option. The use of one daughter ion is acceptable if data

sensitivity and selectivity is achieved and provided that retention time criteria are met to assure adequate

specificity. While the daughter ions may be chosen at the discretion of the analyst, mass transition 99 is

suggested for PFOS. Quantitation may be performed using the total ion chromatogram (TIC, or summed

MRMs) for a given analyte. For example, the PFOA TIC would sum all three of the monitored transitions. Use

of the suggested primary ion is recommended. Retention times may vary slightly, on a day

- to

- day basis,

depending on the batch of mobile phase and the gradient, column, guard column(s) used etc. Drift in retention

times is acce ptable within an analytical run, as long as the drift continues through the entire analysis and the

standards are interspersed throughout the analytical run.



## 11.2 Calibration Curve

Quantitation is by internal standard or external standard calibration. Calibration standards may be prepared in

ASTM Type I, HPLC water, other suitable water, or a solvent/water mixture. If internal standard calibration

does not meet calibration acceptance criteria, external calibration can be applied. See Table 1 for

### ETS -8- 044. 1 Page 18 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis recom mended application of available internal standards. Quantitation of PFOA and PFOS is by summed

analyte

- specific mass transitions.

Analyze the standard curve prior to each set of samples. If internal standards were added to the calibration

standards area ratios are used to generate the calibration curve. The standard curve may be plotted using a

linear regression (y = mx + b), weighted 1/x or unweighted, or by quadratic fi t (y = ax 2 

### + bx + c), weighted 1/x or

unweighted

, using suitable software

. The mathematical method used to calculate the calibration curve should

be applied consistently throughout a study. Any change should be thoroughly documented in the raw data.

High and/or low points may be excluded from the calibration curves to provide a better fit over the range

appropriate to the data or because they did not meet the pre

- determined acceptance criteria. Low

- level curve

points should also be excluded if their area counts (or area ratio if quantitating by IS) are not at least twice that

of the ave rage area counts (or area ratio if quantitating by IS) of method and/or solvent blanks. The coefficient

of determination (r 2 

### ) value for the calibration curve must be greater than or equal to 0.990 (or a correlation

coefficient (r) of 0.995). Each point in the curve must be within ± 25% of the theoretical concentration with the

exception of the LLOQ, which may be within ± 30%. Justification for exclusion of calibration curve points will be

noted in the raw data. A minimum of 6 points will be used to construct the calibration curve.

If the calibration curve does not meet acceptance criteria, perform routine maintenance or prepare a new

standard curve (if necessary) and reanalyze.



## 11.3 Continuing Calibration Verification (CCV)

Continuing calibration verifications (CCV) are analyzed to verify the accuracy of the calibration curve. Analyze

a mid

- range calibration standard, one of the same standards used to construct the calibration curve, at a

minimum after every tenth sample, not including solvent blanks, with a minimum of one per sample set.

Calibration verification injections must be within ± 25% to be considered acceptable. The calibration curve and

the last passing CCV will then bracket acceptable samples. Multiple CCV levels may be used. Samples must

be bracketed by passing CCVs or the calibration curve and a passing CCV to be reportable.



## 11.4 System Suitability

A minimum of three system suitability samples should be injected at the beginning of each analytical run, prior

to th e analysis of the calibration curve. Typically these samples are at a concentration near the mid

- level of the

calibration curve and are repeated injections from one autosampler vial. It is suggested that the system

suitability injections have area counts or area ratios when using internal standard calibration, with a target RSD

of ≤5% and a target retention time RSD of ≤2%. There is no defined acceptability limit on these results as the

%RSD value is dependent on the number of MRM transitions being monitored in the LC/MS/MS run or time

period. Ultimately, any effects on these parameters for the System Suitability samples will also be evident on

all standards and QC samples analyzed as part of the analysis batch. Any effect of system suitability is

incorp orated within QC acceptance criteria. 4



## 11.5 Sample Analysis and QCs

For each analysis batch, the instrument analysis run sequence should include an initial calibration curve,

samples, FDSs, interspersed blanks, interspersed CCVs, appropriate QCs (i.e., LCSs, LMSs, FMSs, TBMSs,

and TBs), and a final CCV or calibration curve bracketing samples and appropriate QCs

Inject the same volume (between 5

- 100μL) of each standard, analytical sample and blank into the instrument

(unless an on

- instrument sample dilution is desired).

Samples containing analytes that are quantitated above the concentration of the highest standard in the curve

should be further diluted and reanalyzed.

4 3M Environmental Laboratory study E08

- 0096 evaluated the effect on these results as a function of the number of MRMs being monitored.

### ETS -8- 044. 1 Page 19 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis 

## 12 Data Analysis and Calculations

The chromatography analysis software will typically calculate the amount of target analyte in the sample

extracts using the established calibration curve.

Calculate the percent recovery of the LCS using the following equation:



### 100%*

)

mL ng (ion Concentrat Spike )

mL ng (ion Concentrat LCS

recovery LCS%

=

Calculate the percent recovery of the LMS using the following equation:

100%*

)
mL ng ( ion Concentrat Spike )
mL ng ( Sample of ion Concentrat -)
mL ng ( ion Concentrat LMS

recovery % LMS

=



### For samples fortified with known amounts of analyte prior to extraction, use the following equation to calculate

the percent recovery.



### 100 (ng/mL) added Analyte (ng/mL) sample in found analyte Average- (ng/mL) found analyte Total Recovery × =



## 13 Analysis Batch Method Performance Criteria

Any method performance parameters that are not achieved must be considered in the evaluation of the data.

Nonconformance to any specified parameters must be described and discussed in the final report if the

Technical Manager (non

- GLP study) or Study Director (GLP study) chooses to report the data.

If criteria listed in this method performance section are not met, maintenance may be performed on the system

and samples reanalyzed, or other actions taken as appropriate. Document all actions in the raw data.

If data are to be reported when performance criteria have not been met, the data must be footnoted on tables

and discussed in the text of the report.



## 13.1 System Suitability

– Analysis Batch

A minimum of three system suitability samples should be injected at the beginning of each analytical run.

These samples are run prior to the calibration curve. It is suggested that the system suitability injections have

area counts with a target RSD of ≤5% and a target retention time RSD of ≤2%. There is no defined

acceptability limit on these results as the %RSDs are dependent on the number of MRM transitions being

monitored in the LC/MS/MS run or time period. Any effect of system suitability is incorporated in the QC

acceptance criteria.



## 13.2 Calibration and Limit of Quantitation (LOQ)

– Analysis Batch

Calibration Curve: The coefficient of determination (r 2 

### ) value for the calibration curve must be greater than or

equal to 0 .990 corresponding to a correlation coefficient (r) = 0.995. Each point in the curve must be within

± 25% of the theoretical concentration with the exception of the LLOQ, which may be within ± 30%.

CCV Performance: The calibration standards that are interspersed throughout the analytical sequence are

evaluated as continuing calibration verifications in addition to being part of the calibration curve. The accuracy

### ETS -8- 044. 1 Page 20 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis of each curve point must be within 25% of the theoretical value ( within 30% for lowest curve point ) . Samples

that are bracketed by CCVs not meeting these criteria must be reanalyzed.

Limits of Quantitation (LOQ): The lower LOQ (LLOQ) is the lowest non

- zero active standard in the

calibration curve; the peak area of the LLOQ must be at least 2X that of the average area counts for all

prepared procedural blank(s). By definition, the measured value of the LLOQ must be within 30% of the

theoretical value.

Demonstration of Specificity: Specificity is demonstrated by chromatographic retention time (within 4% of

standard) and the mass spectral response of unique ions.



## 13.3 Blanks

– Method/Procedural Blanks and Trip

Method/ Procedural Blanks: Multiple procedural blanks should be interspersed throughout the analysis batch

and the analytical sequence. At a minimum, method blanks are analyzed prior to instrument calibration, prior

to the analysis of CCV samples, after every 10 sample injections, and at the end of the analytical run.

The mean area counts (or area ratios when using IS calibration) for each analyte must be less than 50% of the

area count of the LOQ standard. If the area counts of the procedural blanks exceed 50% of the LOQ standard,

then the LOQ must be raised to the first standard level that meets criteria.

Trip Blank: A t rip blank of ASTM Type I water (or lab equivalent) is prepared in a sample container in the

laboratory and treated as a sample, including exposure to shipping, sampling site conditions, storage,

preservation and all analytical procedures. The trip blanks results for each analyte are included with the

repo rted sample results.



## 13.4 Data Accuracy and Precision

– Analysis Batch

Lab Control Spikes: The average recovery at each LCS level for each target analyte and appropriate SRS

should be within 80

- 120% and the percent relative standard deviation of the recoveries must be less than or

equal to 20%. If the average recovery of a spiking level falls outside method acceptance, but at least 67% (6

out of 9 ) o f LCS samples are within 20 % of their respective nominal value (33% of the QC samples, not all

replicates at the same concentration, may be outside 20% of nominal value), the average recovery will be

flagged as outside method acceptance criteria. All LCS samples will be control charted as per ETS

- 12

- 012. If

the average recovery of one of the spiking levels exceeded the analytical method uncertainty as determined by

ETS

- 12

- 012, that analytical batch uncertainty will be expanded for that particular study. The average recovery

at each LCS level for mixed branched/linear isomer PFOA and PFOS should be within 70

- 130% and the

percent relative standard deviation of the recoveries must be less than or equal to 20%.

Field Duplicates: The relative percent difference (RPD) of duplicate samples should be less than 20% for the

precision of sample preparation and analysis to be considered in control. Replicate samples not meeting the

20% RPD criteria are flagged and reported as outside of QC acceptance criteria.

Field Matrix Spikes: FMS acceptance criteria are recoveries within ±30% of the expected value for each

target analyte and appropriate SRS. Sample data with FMS recovery outside of ±30% but within ±50% of the

expected value are flagged and reported as outside of QC acceptance criteria. Data with FMS recovery

outside of ±50% of the expected value are reported as NR, where NR is defined as “Not Reportable” data

outside of QC acceptance criteria. If FMS recovery could not be assessed because FMSs were at an

inappropriate level, then Laboratory Matrix Spikes (LMSs) may be substituted. If LMS recoveries are within

±30% for each target analyte and SRSs the data are reportable but flagged as not meeting the FMS method

acceptance criteria.



## 13.5 Analytical Method Uncertainty

Analytical method uncertainty for each target analyte and SRS is determined with control charted historica l

analysis batch LCS data for the method and reported with each analysis batch. 5 

### Uncertainty determinations

5 Method uncertainty based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression of

Uncert ainty in Measurement). Method application demonstrated in ETS

- 12

- 012, citing references: a.) EURACHEM/CITAC Guide,

“Quantifying Uncertainty in Analytical Measurement,” Second Edition; Editors: S.L.R. Ellison, M. Rosslein, and A. Williams

. b.)Georgian, Thomas, “Estimation of Laboratory Analytical Uncertainty Using Laboratory Control Samples,” Environmental Testing &

### ETS -8- 044. 1 Page 21 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis are based on INTERNATIONAL ANS/ISO/IED STANDARD 17025 reference (GUM, Guide to the Expression

of Uncertainty in Measurement) and described in ETS -12- 012 . At least thirty data points are required for

determining analytical method uncertainty. The method uncertainty is defined as 2x the standard deviation of

the percent recoveries of the pooled lab control spikes. While all LCS data points are control charted, only the

most recent fifty data points are used for determining the method uncertainty.

When less than thirty LCS data points have been generated for a given analyte, the analysis batch LCSs are

used to determine the data uncertainty. If FMSs meet the ±30% recovery criteria at a level appropriate to the

endogenous level, and the LCS meet the ±20% recovery criteria, then the uncertainty of the data is determined

as within 100±20%.

Analysis batch sample data with FMS recovery outside of ±30% but within ±50% of the expected value are

flagged and reported as outside of QC acceptance criteria with expanded uncertainties. Data with FMS

recovery outside of ±50% of the expected value are reported as NR, where NR is defined as “Not Reportable”

data outside of QC acceptance criteria. If FMS recovery could not be assessed because FMSs were at an

inappropriate level, then Laboratory Matrix Spikes (LMSs) may be substituted. If LMS recoveries are within

±30% for each target analyte and appropriate SRSs the data are reportable but flagged as not meeting the

FMS method acceptance criteria with uncertainties of ±30%. If FMS do not meet the ±30% recovery criteria,

and historical FMS data does not exist, the analytical uncertainty is evaluated on a sample

- by -s ample basis,

the data may be reported with expanded uncertainty and are flagged.



## 13.6 Quantitation of PFOA/PFOS

– Analysis Batch

Calibration standards consisting of mixed branched and linear isomer PFOS/PFOA are preferred.

Quantitation is performed by integrating the linear and branched isomers together. Alternately, the linear and

branched isomers can be integrated separately, applying the appropriate true value to each calibration curve

po int for each isomer. The LCS and samples are then quantitated by integrating the linear and branched

isomers separately (requires separate analytical results files) and quantitating the resulting peak against the

linear or branched calibration curve

. The results from both integrations are then summed to produce the final

result. Integrating the linear and branched isomers separately may be helpful for those samples where the

linear/branched ratios do not closely match those of the reference standards.

Ho wever, for PFOS/PFOA target analytes, if the calibration standards are comprised of predominantly linear

isomers only t he method requires the addition of LCS s of mixed branched/linear isomer PFOS/PFOA. The

purpose of including these LCSs is to demonstrate quantitative equivalency (or quantitative bias) of the

isomeric mix when using a predominantly linear PFOS or PFOA standard for calibration. Alternatively, in lieu

of mixed branched and linear isomer PFOS/PFOA LCSs, mixed branched and linear isomer PFOS/ PFOA

TBMSs may be applied to demonstrate method accuracy and precision.

An alternate method of quantitation can be performed whereby only the linear isomer of PFOS/PFOA is

integrated and used for generating the calibration curve. The LCS and samples are then quantitated by

integrating the linear and branched isomers separately (requires separate analytical results files) and

quantitating the resulting peak against the linear calibration curve. The results from both integrations are then

summed to produce the final result. Integrating the linear and branched isomers separately reduces the on

column concentration for those samples that contain both linear and branched isomers of PFOA/PFOS. This

ensures that the concentration detected is within the a range of the calibration curve that is comparable

regardless of whether the calibration curve was generated using predominantly linear isomers of PFOS/PFOA

or linear plus branched isomers of PFOS/PFOA.



## 14 Pollution Prevention and Waste Management

Waste generated when performing this method will be disposed of appropriately. The original samples will be

archived at the 3M Environmental Laboratory in accordance with internal procedures.

Analysis, November/December 2000. c.)Taylor, B.N. and CE. Kuyatt, NIST Technical Note 1297, 1994 Edition: “Guidelines for

Evaluating and Expressing the Uncertainty of NIST Measurement Results.”d.)Adams, T.M., “A2LA Guide for the Estimation of

Measurement Uncertainty in Testing”, July 2002.

### ETS -8- 044. 1 Page 22 of 22

Method of Analysis for the Determination of Perfluorinated Compounds in Wate r by LC/MS/MS; Direct Injection

Analysis 

## 15 Records

Each data package generated for a study must include all supporting information for reconstruction of the data.

Information for the data package must include, but is not limited to the following items: study or project

number, sample and standard prep sheets/records, instrument run log (instrument batch records, instrument

ac quisition method, summary pages), instrument results files, chromatograms, calibration curves, and data

calculations.



## 16 Affected Documents

None.



## 17 Revisions

Revision

Number Summary of Changes

1 Section 1

. Included the use of internal standard calibration by this method.

Section 2. Included the use of internal standard calibration by this method. Included the

use of a solvent/water mixture when analyzing for PFUnA, PFDoA, PFTrDA, and FOSA.

Section 3

. Added definitions for internal standard, surrogate internal standard, and

surrogate recovery standard.

Section 6.Removed the details regarding the instrument parameters to section 10 of the

method

. Section 7. Updated reference standards to include internal standards and surrogates.

Changed concentration levels for working standards and included the use of internal

standards and surrogates.

Section 8. Inserted a new section on sample bottle preparation.

Se ction 9 Quality Control. This section was previously section 10 in ETS -8- 044.0. Updated

QC criteria to be consistent with method ETS -8- 154.4.

Section 10 Procedures . This section was previously section 8 (Sample Handling) in E TS -8-

044.0. Added detail regarding the preparation of LCSs. Included the use of methanol as a

dilution solvent.

Section 11 Samp le Analysis

. This section was previously section 10 in ETS -8- 044.0.

Included the details regarding the instrument parameters.

Section 12 Data Analysis and Calculations. This section was previously section 11 in ETS

8- 044.0. Removed the equation for calculating the analytes concentration, indicating that

this is done by the instrument software.

Section 13 Method Performance. This section was previously section 12 in ETS -8- 044.0.

Updated QC criteria to be consistent with ETS -8- 154.4. Added information on the

determination of analytical method uncertainty and quantitation of PFOA/PFOS.

Section 14 Pollution Prevention. This section was previously section 13 in ETS -8- 044.0.

Section 15 Records. This section was previously section 14 in ETS -8- 044.0.

Section 16 Affected Documents. This section was previously section 15 in ETS -8- 044.0.

Section 17 Revisions. This section was previously section 16 in ETS

- 8

- 044.0.

# A TTACHMENT 

# D: D EVIATION 

# ( S 

# )