# 

GLP10

- 01 -02, Interim Report 3 1: Analysis of PFBS, PFHS, and PFOS in Groundw ater

Samples from New On

- Site Wells

- Adjacent to the 3M Decatur Wastewater Treatment Plant

(WWTP) April 2012

Study Title

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

Perfluorobutane 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 K esari P.E., DEE

Weston Solutions, Inc.

1400 Weston Way

West Chester, PA 19380

Phone: 610

- 701

- 3761

Author

Susan Wolf

3M Environmental Laboratory

Interim Report Completion Date

Date of signing

Performing Laboratory

3M Environmental Health and Safety Oper ations

Environmental Laboratory

3M Center, Bldg 260

- 05 -N- 17

St. Paul , MN 55144

Project Identification

GLP10

- 01

- 02

- 31

Total Number of Pages

89

Testing Cert #2052.01 The testing reported herein meet the requirements of ANSI/ISO/I EC 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 ISO 9001:2000.

### This page has been reserved for specific country requirements

.GLP10-01-02 ; Interi m Repor t 3 1

Analysi s o f PFBS

, PFHS

, an d PFO S I n Groundwate r

Ne w On-sit e VWVT P Well s - Apri l 201 2



### GL P COMPLIANC E STATEMEN T

Repor t Title : GLP10-01-02

, Interi m Repor t 31 : Analysi s o

f PFBS

, PFHS

, an d PFO S i n Groundwate r Sample s fro m Ne

w On-Sit e Well s - Adjacen t t o th e 3 M Decatu r Wastewate r Treatmen t Plan t (WWTP )

Apri l 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 detemnine d unde r GLPs

.

Gar y Hohehsteirf

, Sponso r Representativ e Dat e

Jaisimh a Kesari

, P.E.

, DEE

, Stud y Directo r ' Dat e

Pag e 3 o f 8 9GLP10-01-02 ; Interi m Repor t 3 1

Analysi s o f PFBS

, PFHS

, an d PFO S I n Groundwate r

Ne w On-Sit e WWT P Well s - Apri l 201 2



### QUALIT Y ASSURANC E STATEMEN T

Repor t Title : GLP10-01-02

, Interi m Repor t 31 : Analysi s o f PFBS

, PFHS

, an d PFO S i n Groundwate r Sample s fro m Ne

w On-Sit e Well s - Adjacen t t o th e 3 M Decatu r Wastewate r Treatmen t Plan t (WWTP )

Apri l 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 R e porte d t o

Inspectio n Date s Phas e Testin g Facilit y

Managemen t Stud y Directo r

4/26-27/12

, 5/1/1 2 Dat a an d Repor t 5/4/1 2 5/4/1 2



### QA U Representativ e

Pag e 4 o f 8 9

# 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

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

8 Analytical Results

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

8.1 Calibration

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

8.2 System Suitability

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

8.3 Limit of Quantitation (LOQ)

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

8.4 Continuing Calibration

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

8.5 Blanks

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

8.6 Lab Control Spikes (LCSs)

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

8.7 Analytical Method Uncertainty

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

8.8 Field Matrix Spikes (FMS)

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

### 8.9 Lab Matrix Spikes (LMS)

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

9 Data Summary and Discussion

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

10 Co nclusion

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

11 Data/Sample Retention

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

12 Attachments

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

13 Sig natures

................................................................................................ ......................................... 24



# L IST OF 

# T ABLES

Table 1. S ummarized PFBS, PFHS, and PFOS Results (Residuum Monitoring Wells).

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

Table 2. Sample Description Key Code.

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

Table 3. Instrument Parameters.

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

Table 4. Liquid Chromatography Conditio ns.

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

Table 5. Mass Transitions.

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

Table 6. Limit of Quantitation (LOQ).

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

Table 7. Laboratory Control Spike Recovery.

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

Table 8. Analytical Uncertainty.

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

Table 9. Field Matrix Spike Levels.

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

Table 10. DAL GW TW1R 120405

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

Table 11. DAL GW TW2R 120405

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

Table 12. DAL GW TW3R 120405

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

Table 13. DAL GW TW4R 120405

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

Table 14. DAL GW TW5R 120405

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

Table 15. DAL GW TW6R 120405

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

Table 16. DAL GW TRIP01 120405

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

Table 17. Rinsea te Blank

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

## 1 **Study Information**

Sponsor

3M Company

Sponsor Representative

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

W eston 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

Chelsie Grochow, Analyst

Kelly Ukes, Analyst

Study Dates

St udy Initiation: March 8, 2010

Interim 31 Experimental Termination: April 28 , 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

Labo ratory 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 samples fro m six new on

- site temporary

residuum monitoring wells in the vicinity of the 3M Decatur facility Wastewater Treatment Plant

(WWTP) in Decatur, AL. A total of twenty

- eight sample bottles were received at the 3M Environmental

Laboratory for perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate (PFHS) and

pe rfluorobutane sulfonate (PFBS)

, and included duplicate groundwater samples and two field matrix

spike (FMS) samples from each sampling location. Samples also included one trip blank containing

Milli

- QTM water and appropriate trip blank spikes and one equipment rinseate blank

. The equipment

rinseate blank did not have FMS samples prepared for determination of PFOS, PFHS and PFBS

recovery. All samples were logged into the laboratory information management system (LIMS) under

project GLP10

- 01

- 02

- 31 . The groundwater samples and trip blanks were received from Weston

personnel on April 9 , 2012. All of the samples were 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 Injection Analysis”

.

The average measured PFBS, PFHS, and PFOS concentrations are summarized in Table 1. The trip

blank sample was belo w 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 + 12 %, PFHS + 12 % and PFOS + 35 %.

Table 1

. S ummarized PFBS, PFHS, and PFOS Results ( Resid uum Monitoring Wells , April

2012 ).

PFBS PFHS PFOS

Sampl ing Location Avg. Conc

. (ng/mL) ± RPD Avg. Conc

. (ng/mL) ± RPD Avg. Conc

. (ng/mL) ± RPD



### DAL GW TW1R 120405 33.8 ± 0.30% 57.3 ± 2.1% 29.0 ± 9.0%

DAL GW TW2R 120405 117 ± 5.1% 579 ± 5.5% 1090 ± 17%

DAL GW TW3R 120405 304 ± 7.6% 1310 ± 15% 3660 ± 17%

DAL GW TW4 R 120405 184 ± 4.9% 19.1 ± 1.0% 247 ± 2.4%

DAL GW TW5 R 120405 522 ± 4.4% 43.0 ± 1.6% 696 ± 2.3%

DAL GW TW6R 120405 156 ± 0.64% 46.7 ± 5.6% 281 ± 4.6%

DAL GW TW3R R inseate B lank <0. 250 <0. 250 <0.232

DAL

- GW

- TRIP01 Blank <0. 250 <0. 250 <0.232

The analytical method uncertainties associated with the reported results are: PFBS ± 12 %, PFHS ± 12 %, and

PFOS ± 35 %.



## 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 facil ity. This analytical study was

conducted to analyze groundwater samples collected from six new temporary on

- site residuum

### monitoring wells located adjacent to the 3M Decatur WWTP for PFOS, PFHS and PFBS in an effort to

characterize regional groundwater conditions.

The 3M Environmental Laboratory prepared sample containers (250 mL high

- density polyethylene

bottles) which were shipped to Decatur, AL Weston personnel prior to field sampling. Sample

containers for each sampling location included a field sample, field sample duplicate, and two field

matrix spike samples. Each empty container 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 (i nternal standard) at a nominal

concentrati 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 the samples, neither the internal

standards nor the surrogate recovery standard was utilized during sample analysis. Laboratory matrix

spikes were prepared for sampling locations TW2, TW3, TW4, TW5, and TW6 since the field matrix

spike levels were not appropriate for these sampling locations for at least one of the target analytes.

See section 8.9 of the report for laboratory matrix spike levels. See section 8.8 of the report for field

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

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 summarizes the average PFBS, PFHS, and PFOS concentrations for the duplicate surface

water samples collected and the trip blank sample

. Table s 10 -17 summarize the individual sample

results and the associated field matrix spike recoveries. All r esults 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 stu dy

was purely analytical in nature.

## 5 **Reference Substances**

Reference

Substance PFBS

(Linear) PFHS

(Linear)

Chemical Name Potassium Perfluorobutane

sulfonate Sodium Perfluorohexane

sulfonate

Chemical Formula 

### C 4 

### F 9 

### SO 3 - 

### K + 

### C 6 

### F 13 

### SO 3 - 

### Na +

Identifier NA L- PFHXS

Use Target Analyte Reference

Standard Target Analyte

Reference Standard

Source 3M Wellington

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

Storage Conditions Frozen Frozen

Chemical Lot

Number 41

- 2600

- 8442 -5 LPFHxSAM08

TCR Number TCR

- 121 TCR08

- 0018

Physical Description White Powder Crystalline

Purity 96.7% 100%

Reference

Substance PFOS

(Linear + Branched) PFOS

(Linear + Branched)

Chemical Name Potassium Perfluorooctane

sulfonate Potassium

Perfluorooctane sulfonate

Chemical Formula 

### C 8 

### F 17 

### SO 3 - 

### K + 

### C 8 

### F 17 

### SO 3 - 

### K +

Identifier Br

- PFOSK CAS # 2795

- 39 -3

Use Target Analyte Reference

Standard FMS Reference Standard

Source Wellington Sigma

- Aldrich

Expiration Date 03/17/2014; 12/01/2014 0 2/ 0 4/2014

Storage Conditions Frozen Ambient

Chemical Lot

Number brPFOSK0708;

br PFOSK1111 1424328V

TCR Number TCR11

- 0010; TCR11

- 0041 TCR11

- 0028

Physical Description Liquid White Powder

Purity 99.9% 99.7%

## 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** 

**- TW1R** 

**- 0** 

**- 120405** 

**1 Sa**  **mple Location DAL**  **= Decatur**  **, Alabama**

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

**3 Well ID**  **Example: TW1R** 

**4 Sample Type**  **0=primary sample**

**DB =duplicate sample** 

**LS = low spike**

**HS = high spike**

**5 Sampling Date**  **1 20405** 

**– April 5, 2012** 



## 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 Sa mple Colle 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

- 31 were returned to the

laboratory at ambient conditions on April 9 , 201 2 . Samples were stored refrigerated at the laboratory

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

the sample collection bottles.



## 7.3 Sample Preparation

All samples were initially diluted 1:10 by diluting 1 mL of a well mixed sample with 9 mL of Milli Q water

and analyzed on 4/13/12.

Sampling locations TW2, TW3, TW4, TW5, and TW6 were re

- analyzed and included the preparation of

a laboraotry matrix spike with each sample set. Location TW2R was diluted 1:50 by diluting 0.2 mL of

the well mixed sample with 9.8 mL of Milli Q water and included a 1000 ppb LMS for PFHS and PFOS.

Location TW3R was diluted 1:100 by diluting 0.1 mL of the well mixed samp le with 9.9 mL of Milli Q

water and included a 2000 ppb LMS and 5000ppb LMS for PFBS, PFHS, and PFOS. Locations TW4R,

TW5R and TW6R were diluted 1:10 by diluting 1.0 mL of the well mixed sample with 9.0 mL of Milli Q

water. Loc a tion TW4R included a 200 ppb LMS for PFOS. Loc a tion TW5R included a 500 ppb LMS for

PFBS and PFOS. Loc a tion TW6R included a 200 ppb LMS for PFOS.

## 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.

Table 3

. Instrument Parameters.

Instrument Name ETS Ginger ETS Buster

Analytical Method Followed ETS

- 8

- 044.1 ETS

- 8

- 044.1

Analysis Date 4/13/12 4/16/12 , 4/27/12

Liquid Chromatograph Agilent 1100 Agilent 1100

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

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

Injection Volume 2 or 10 μ L 2 or 10 μ L

Mass Spectrometer Applied Biosystems API 50 00 Applied Biosystems API 4000

Ion Source Turbo Spray Turbo Spray

Electrode Turbo ion electrode Turbo ion electrode

Polarity Negative Negative

Software Analyst 1. 4 .2 Analyst 1. 4 .2



### Table 4 . Liquid Chromatography Conditions.

**Step**

**Number Total Time** 

**(min) Flow Rate** 

**(** μ

**L/min) Per**  **cent A**

**(2 mM ammonium acetate) Percent B** 

**( m**  **ethanol)**

ETS

- 8

- 044. 1

0 0 .0 750 97.0 3 .0

1 2 .0 750 97 .0 3 .0

2 14.5 750 5. 0 9 5 .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



### Table 5. Mass Transitions.

**Analyte Mass**  **Transition**

**Q1/Q3 Reference Material** 

**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 50 or 75 msec for each transition. The individual transitions were

summed to pro duce 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 soluti on containing PFBS, PFHS, PFOS (reference

standard for PFOS containing both linear and branched isomers) into Milli Q water. Standards ranging

from 0.0250 ng/mL to 100 ng/mL (nominal) were analyzed. Low and or high curve points may have

been disabled to meet method criteria. 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 all analytes. The correlation coefficient (r) was greater than 0.995 for all analytes

. 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 all analytes. The

correlation coefficient (r) was gr eater than 0.995 for PFBS, PFHS and PFOS for each analysis

.



## 8.2 System Suitability

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

demo nstrate 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 meets 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).

**Analysis Date Dilution** 

**Factor PFBS** 

**LOQ, ng/mL PFHS** 

**LOQ, ng/mL PFOS** 

**LOQ, ng/mL**

4/13/12 10 0.250 0.250 NA

50 1.25 1.25 NA

4/16/12 10 0.250 0.250 0.23 2

50 1.25 1.25 1.16

100 2.50 2.50 2.32

4/27/12 10 10.0 9.98 9.28

50 50.0 49.9 46.4

100 100 99.8 92.8

250 250 250 232

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

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

blank, and an equipment rinseate 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 bla nk

reflects the shipping and sample collection conditions the sample bottles and samples experience. The

equipment rinseate blank is an aqueous sample that reflect the efficiency of equipment cleaning in the

field between different sample collections and are proof of no cross contamination of samples from the

equipment.



## 8.6 Lab Control Spikes (LCSs)

Low, mid, and high lab control spikes were prepared for the target analytes and analyzed in triplicate.

LCSs were prepared by spiking known amounts of the analyte into Milli

- Q water to produce the desired

concentration. The spiked water samples were then prepared and analyzed in the same manner as the

samples. The method acceptance criteria, average of LCS at each level should be within 100% ± 20%

with an RSD ≤20%

, was met for all analytes except for the low level LCSs for PFOS analyzed on

4/16/12 which had an average recovery of 75.2%

. All LCS samples were used in the determination of

the analytical method uncertainty on section 8.7. A method deviation is in cluded in Attachment D

.

The following calculations were 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

Analyze d 4/13 /12 **PFBS PFHS** 

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

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

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120411

- 1 0.498 0.479 96.2 0.497 0.472 95.0

LCS

- 120411

- 2 0.498 0.466 93 .7 0.497 0.464 93.3

LCS

- 120411

- 3 0.498 0.476 95.6 0.497 0.469 94.3

Average ± %RSD 95.2 % ± 1.4 % 94.2 % ± 0.91 %

LCS

- 120411

- 4 4.98 4.69 94.1 4.97 4.73 95.2

LCS

- 120411

- 5 4.98 4.77 95.8 4.97 4.76 95.7

LCS

- 120411

- 6 4.98 4.71 94.5 4.97 4.69 94.4

Average ± %R SD 94.8 % ± 0.94 % 95.1 % ± 0.69 %

LCS

- 120411

- 7 29.9 NA (1) NA (1) 29.9 NA (1) NA (1)

LCS

- 120411

- 8 29.9 NA (1) NA (1) 29.9 NA (1) NA (1)

LCS

- 120411

- 9 29.9 NA (1) NA (1) 29.9 NA (1) NA (1)

Average ± %RSD NA NA

NA = Not Applicable

(1) LCS spiked concentrat ion exceeded the calibration range.

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

### Table 7 continued. Laboratory Control Spike Recovery

.

ETS -8- 044.1

Analyzed 4/1 6 /12 **PFBS PFHS** 

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

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

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120411

- 1 0.498 0.505 101 0.497 0.489 98.3

LCS

- 120411

- 2 0.498 0.511 103 0.497 0.491 98.9

LCS

- 120411

- 3 0.498 0.500 100 0.497 0.493 99.1

Average ± %RSD 101 % ± 1.5 % 98.8 % ± 0.42 %

LCS

- 120411

- 4 4.98 5.12 103 4.97 4.96 99.7

LCS

- 120411

- 5 4.98 5.19 104 4.97 4.79 96.4

LCS

- 120411

- 6 4.98 4.95 99.4 4.97 4.95 99.6

Average ± %RSD 102 % ± 2.4 % 98.6 % ± 1.9 %

LCS

- 120411

- 7 29.9 29.3 98.0 29.9 27.0 90.3

LCS

- 120 411

- 8 29.9 28.8 96.3 29.9 27.7 92.5

LCS

- 120411

- 9 29.9 29.0 97.0 29.9 28.3 94.8

Average ± %RSD 97.1 % ± 0.88 % 92.5 % ± 2.4 %

ETS -8- 044.1

Analyzed 4/16/12 **PFOS (Linear and Branched)** 

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

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

LCS

- 120411

- 1 0.462 0.348 75.3

LCS

- 120411

- 2 0.462 0.351 75.9

LCS

- 120411

- 3 0.462 0.344 74.5

Average ± %RSD 75.2 % ± 0.93 % (2)

LCS

- 120411

- 4 4.62 4.20 91.0

LCS

- 120411

- 5 4.62 4.17 90.3

LCS

- 120411

- 6 4.62 4.26 92.2

Average ± %RSD 91.2 % ± 1.1 %

LCS

- 120411

- 7 27.8 26.5 95.2

LCS

- 120411

- 8 27.8 25.8 92.8

LCS

- 120411

- 9 27.8 26.8 96.3

Average ± %RSD 94.8 % ± 1.9 %

NA = Not Applicable

(1) LCS spiked concentration exceeded the calibration range.

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

### Table 7 continued. Laboratory Control Spike Recovery

.

ETS -8- 044.1

Analyzed 4/ 27

/12 **PFBS PFHS** 

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

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

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %Recovery** 

LCS

- 120426

- 1 4.98 4.95 99.5 4.97 4.85 97.6

LCS

- 120426

- 2 4.98 5.09 102 4.97 5.00 101

LCS

- 120426

- 3 4.98 5.30 107 4.97 5.12 103

Average ± %RSD 103 % ± 3.7 % 101 % ± 2.7 %

LCS

- 120426

- 4 20.0 18.6 93.0 19.9 17.7 88.8

LCS

- 120426

- 5 20.0 18.5 92.6 19.9 18.5 93.1

LCS

- 120426

- 6 20.0 18.2 90.8 19.9 18.8 94.6

Average ± %RSD 92.1 % ± 1.3 % 92.2 % ± 3.3 %

LCS

- 120426

- 7 49.8 50.4 101 49.7 49.1 98.8

LCS

- 120 426

- 8 49.8 47.5 95.4 49.7 46.6 93.7

LCS

- 120426

- 9 49.8 47.1 94.6 49.7 47.0 94.6

Average ± %RSD 97.0 % ± 3.6 % 95.7 % ± 2.8 %

ETS -8- 044.1

Analyzed 4/27 /12 **PFOS (Linear and**  **Branched)**

**Lab ID Spiked** 

**Concentration**

**(ng/mL) Calculated** 

**Concentration**

**(ng/mL) %R**  **ecovery**

LCS

- 120426

- 1 4.62 4.26 92.1

LCS

- 120426

- 2 4.62 4.02 87.0

LCS

- 120426

- 3 4.62 4.21 91.1

Average ± %RSD 90.1 % ± 3.0 %

LCS

- 120426

- 4 18.5 15.7 85.0

LCS

- 120426

- 5 18.5 16.6 89.6

LCS

- 120426

- 6 18.5 16.8 90.6

Average ± %RSD 88.4 % ± 3.4 %

LCS

- 120426

- 7 46 .2 46.3 100

LCS

- 120426

- 8 46.2 43.8 94.9

LCS

- 120426

- 9 46.2 43.5 94.1

Average ± %RSD 96.3 % ± 3.3 %

NA = Not Applicable

(1) LCS spiked concentration exceeded the calibration range.

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

## 8.7 Analytical Me thod 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 5.91 ± 12 %

PFHS 5.88 ± 12 %

PFOS 17. 6 ± 35 %



## 8.8 Field Matrix Spikes (FMS)

Low, mid, and high field matrix spikes (FMS) were collected at each sampling point to verify that the

analytical method is applicable 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. Field matrix spikes are presented in section 9 of this report.

Table 9 . Field Matrix Spike Levels

.

Sampling Lo cation Spike Level PFBS,

ng/mL PFHS,

ng/mL PFOS,

ng/mL

All locations and Trip Blank Low 10 9.98 9.98

High 100 99.8 99.8

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 s of PFBS, PFHS, and/or PFOS detected in sampling locations TW2, TW3, TW4,

TW5, and TW6, the FMS spike levels were not appropriate for at least one of the analytes. Therefore,

laboratory matrix spikes (LMS) samples were prepared to verify that the analytical method is applicable

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

PFBS, PFHS, and PFO S (reference standard containing both linear and branched isomers) to an aliquot

of the primary sample. Since all samples required dilution prior to sample analysis, the laboratory matrix

spike added was based on the on

- column instrument concentration.

Sampling locationTW2; the primary sample (GLP10

- 01 -02-31- 005) was diluted 1:50, to which 20 ng/mL

(nominal) of target analytes w ere added for a LMS concentration of approximately 1000 ng/mL.

Sampling location TW3; the primary sample (GLP10

- 01

- 02 -31- 009) was diluted 1:100, to which the low

LMS had 20 ng/mL (nominal) of target analytes added for a LMS concentration of approximately 2000

ng/mL , and the high LMS had 50 ng/mL (nominal) of target analytes added for a LMS concentration of

approximately 5000 ng/mL

.

### Sampling location TW4; the primary sample (GLP10

- 01

- 02

- 31

- 013) was diluted 1:10, to which 20 ng/mL

(nominal) of target analytes were added for a LMS concentration of approximately 200 ng/mL.

Sampling location TW5; the primary sample (GLP10

- 01

- 02

- 31

- 017) was diluted 1: 5 0, to which 10 ng/mL

(nominal) of target analytes were added for a LMS concentration of approximately 500 ng/mL.

Sampling location TW6; the primary sample (GLP10

- 01

- 01

- 32

- 017) was diluted 1:10, to which 20 ng/mL

(nominal) of target analytes were added for a LMS concentration of approximately 200 ng/mL.

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 following 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 or lab 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 matrix spike recoveries meeting the method

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

their respective quantitative ranges

.

DAL GW TW1R 120405; The recovery of the high field matix spike for PFOS was 65.8%. Since the

anal ytical method uncertainty was calculated at ± 35% for PFOS, no further adjustment to the analytical

uncertainty is needed for DAL GW TW1R.

### Table 10

. DAL GW TW1R 120405

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

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

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

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

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 31

- 001 DAL

- GW

- TW1R

- 0

- 120405 33.8 NA 56.7 NA 27.7 NA

GLP10

- 01

- 02

- 31

- 002 DAL

- GW

- TW1R

- DB

- 120405 33.7 NA 57.9 NA 30.3 NA

GLP10

- 01

- 02

- 31

- 003 DAL

- GW

- TW1R

- LS

- 120405 42.8 NC 63.3 NC 35.4 NC

GLP10

- 01

- 02

- 31

- 004 DAL

- GW

- TW1R

- HS

- 120405 120 86.3 147 89.9 94.7 65.8 (1 )

**Average Concentration (ng/mL) ±**  **%RPD 33.8**  **ng/mL ±**  **0.30% 57.3**  **ng/mL ±**  **2.1 29.0**  **ng/mL ±**  **9 .0%** 

NA = Not Applicable

NC = Not Cal culated; the sample concentration is greater than 2x the spike level.

Samples analyzed using a 1:10 dilution factor

PFBS and PFHS reported from 4/13/12 analysis. PFOS results reported from 4/16/12 analysis.

(1 ) FMS recovery did not meet acceptance criter ia.



### Table 11

. DAL GW TW2R 120405

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

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

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

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

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 31

- 005 DAL

- GW

- TW2R

- 0

- 120405 120 NA 563 NA 995 NA

GLP10

- 01

- 02

- 31

- 0 06 DAL

- GW

- TW2R

- DB

- 120405 114 NA 595 NA 1180 NA

GLP10

- 01

- 02

- 31

- 007 DAL

- GW

- TW2R

- LS

- 120405 125 NC NA (1 ) NA (1 ) NA (1 ) NA (1 )

GLP10

- 01

- 02

- 31

- 008 DAL

- GW

- TW2R

- HS

- 120405 217 100 NA (1 ) NA (1 ) NA (1 ) NA (1 )

GLP10

- 01

- 02

- 31

- 005 ; 1000ppb LMS DAL

- GW

- TW2R

- 0

- LMS NA (1 ) NA (1 ) 1520 94.5 2020 101

**Average Concentration (ng/mL) ±**  **%RPD 117**  **ng/mL ±**  **5.1% 579**  **ng/mL ±**  **5.5 %**  **1090 ng/mL**  **± 17**  **%**

NA = Not Applicable

NC = Not Calculated; the sample concentration is greater than 2x the spike level.

Samples analyzed on 4/13/12 for PFBS using a 1:10 dilution factor), and on 4/27/12 for PFHS and PFOS using a 1:50 dilution factor.

(1 ) Sample not analyzed for this analyte due to inappropriate spike level.

### Table 1 2

. DAL GW TW3R 120405

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

**3M LIMS ID Description**  **Concentra tion** 

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

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

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 31

- 009 DAL

- GW

- TW3R

- 0

- 120405 292 NA 1210 NA 3340 NA

GLP10

- 01

- 02

- 31

- 010 DAL

- GW

- TW3R

- DB

- 120405 315 NA 1410 NA 3970 NA

GLP10

- 01

- 02

- 31

- 011 DAL

- GW

- TW3R

- LS

- 12 0405 NA (1) NA (1) NA (1) NA (1) NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 012 DAL

- GW

- TW3R

- HS

- 120405 NA (1) NA (1) NA (1) NA (1) NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 00 9; 2000ppb LMS DAL

- GW

- TW3 R

- 0

- LMS 2280 98.8 3160 93.0 5240 85.7

GLP10

- 01

- 02

- 31

- 00 9; 5000ppb LMS DAL

- GW

- TW3 R

- 0

- LMS 4960 93.5 5940 93.2 8480 104

**Average Concentration (ng/mL) ±**  **%RPD 304**  **ng/mL ±**  **7.6% 1310**  **ng/mL ±**  **15% 3660**  **ng/mL ±**  **17%**

NA = Not Applicable

Samples analyzed on 4/27/12 using a 1:100 dilution factor ; 5000ppb LMS analyzed using a 1:250 dilution facto r.

(1 ) Sample not analyzed for this analyte due to inappropriate spike level.



### Table 1 3

. DAL GW TW4R 120405

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

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

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

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

**(ng/mL) %Recovery** 

GLP1 0

- 01

- 02

- 31

- 013 DAL

- GW

- TW4R

- 0

- 120405 179 NA 19.0 NA 244 NA

GLP10

- 01

- 02

- 31

- 014 DAL

- GW

- TW4R

- DB

- 120405 188 NA 19.2 NA 250 NA

GLP10

- 01

- 02

- 31

- 015 DAL

- GW

- TW4R

- LS

- 120405 NA (1) NA (1) 28.2 91.2 NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 016 DAL

- GW

- TW4R

- HS

- 120405 278 94.5 102 83.1 NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 013; 200ppb LMS DAL

- GW

- TW4 R

- 0

- LMS NA (1 ) NA (1) NA (1) NA (1) 450 110

**Average Concentration (ng/mL) ±**  **%RPD 184**  **ng/mL ±**  **4.9% 19.1**  **ng/mL ±**  **1.0% 247**  **ng/mL ±**  **2.4%**

NA = Not Applicable

Samples analyzed on 4/16/12 for PFBS u sing a 1:5 0 dilution factor and on 4/27/12 for PFHS and PFOS using a 1:10 dilution factor.

(1 ) Sample not analyzed for this analyte due to inappropriate spike level.

### Table 14

. DAL GW TW5R 120405

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

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

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

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

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 31

- 017 DAL

- GW

- TW5R

- 0

- 120405 533 NA 43.3 NA 704 NA

GLP10

- 01

- 02

- 31

- 018 DAL

- GW

- TW5R

- DB

- 120405 510 NA 42.6 NA 688 NA

GLP10

- 01

- 02

- 31

- 019 DAL

- GW

- TW5R

- LS

- 120405 NA (1) NA (1) 51.1 NC NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 020 DAL

- GW

- TW5R

- HS

- 120405 NA (1) NA (1) 136 93.2 NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 017; 500ppb LMS DAL

- GW

- TW5 R

- 0

- LMS 988 93.3 NA (1) NA (1) 696 91.4

**Average Concentration (ng/mL) ±**  **%RPD 522**  **ng/mL ±**  **4.4% 43.0 ng**  **/mL ±**  **1.6% 696**  **ng/mL ±**  **2.3%**

NA = Not Applicable

NC = Not Calculated; the sample concentration is greater than 2x the spike level.

Samples analyzed on 4/27/12 for PFBS and PFOS using a 1:5 0 dilution factor and on 4/13/12 for PFHS using a 1:10 dilution fa ctor

. (1 ) Sample not analyzed for this analyte due to inappropriate spike level.



### Table 15. DAL GW TW6R 120405

**PF BS**  **(1) PF**  **HS (2)**  **PFOS (1)** 

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

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

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

**(ng/mL) %Re**  **covery**

GLP10

- 01

- 02

- 31

- 021 DAL

- GW

- TW6R

- 0

- 120405 156 NA 48.0 NA 287 NA

GLP10

- 01

- 02

- 31

- 022 DAL

- GW

- TW6R

- DB

- 120405 155 NA 45.4 NA 274 NA

GLP10

- 01

- 02

- 31

- 023 DAL

- GW

- TW6R

- LS

- 120405 NA (1) NA (1) 54.6 NC NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 024 DAL

- GW

- TW6R

- HS

- 120405 23 8 82.5 131 84.5 NA (1) NA (1)

GLP10

- 01

- 02

- 31

- 021; 200ppb LMS DAL

- GW

- TW6 R

- 0

- LMS NA (1) NA (1) NA (1) NA (1) 456 94.9

**Average Concentration (ng/mL) ±**  **%RPD 156**  **ng/mL ±**  **0.64% 46.7**  **ng/mL ±**  **5.6% 281**  **ng/mL ±**  **4.6%**

NA = Not Applicable

NC = Not Calculated; the sample concentration is greater than 2x the spike level.

Samples analyzed on 4/27/12 for PFBS and PFOS using a 1:50 dilution factor and on 4/13/12 for PFHS using a 1:10 dilution factor.

(1 ) Sample not analyzed for this analyte due to inappropriate spike level.

### Table 16. DAL GW TRIP 01 120405

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

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

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

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

**(ng/mL) %Recovery** 

GLP10

- 01

- 02

- 31

- 026 DAL

- GW

- TRIP01

- 0

- 120405 <0.250 NA <0.250 NA <0.232 NA

GLP10

- 01

- 02

- 31

- 027 DA L

- GW

- TRIP01

- LS

- 120405 8.51 85.1 8.60 86.2 6.96 69.7

GLP10

- 01

- 02

- 31

- 028 DAL

- GW

- TRIP01

- HS

- 120405 92.0 92.0 88.4 88.6 70.1 70.2

NA = Not Applicable

Samples analyzed for PFBS and PFHS on 4/13/12 and on 4/16/12 for PFOS using a 1:10 dilution factor.



### Table 17. Rinseate Blank

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

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

**(ng/mL) Concentration** 

**(ng/mL) Concentration** 

**(ng/mL)**

GLP10

- 01

- 02

- 31

- 028 DAL

- GW

- TW3R

- RB

- 120405 <0.250 <0.250 <0.232

Samples analyzed for PFBS and PFHS on 4/13/12 and on 4/16/12 for PFOS using a 1:10 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 Retention**

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 Amendmen t 31 (General Project Outline)

Attachment B: Representative Chromatograms and Calibration Curves

Attachment C: Analytical Method

- ETS -8- 044. 1

Attachment D: Method DeviationGLP10-01-02 ; Interi m Repor t 3 1

Analysi s o f PFBS

, PFHS

, an d PFO S I n Groundwate r

Ne w On-Sit e WWT P Well s - Apri l 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 Departmen t Manage r Dat e



# Hi

/ 2

Gar y Hohenstein

, Sponso r Representativ e Dat e

Jaisimh a Keg^ri

, Stud y Directo r

Pag e 2 4 o f 8 9

# A TTACHMENT 

# A: P ROTOCOL 

# A MENDMENT

# A TTACHMENT 

# B: R EPRESENTATIVE 

# S AMPLE 

# C HROMATOGRAMS AND



# C ALIBRATION 

# C URVE 

# ( S 

# )g120413a.rdb (PFHS): "Quadratic" Regression ("1 / x" weighting): y = -1.61e+004 x^2 + 1.24e+006 x + 6.41e+003 (r = 0.9999)0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100Concentration, ng/mL0.02.0e64.0e66.0e68.0e61.0e71.2e71.4e71.6e71.8e72.0e72.2e72.4e72.6e72.8e73.0e73.2e73.4e73.6e73.8e74.0e74.2e74.4e74.6e7

Area, counts*** Ginger AG01330509Results Name: g120413a.rdbData printed by KJUPrinting Time: 4:46:58 PMPrinting Date: Monday, April 16, 2012Page 1 of 1g120413a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -1.84e+004 x^2 + 1.27e+006 x + 4.06e+003 (r = 0.9998)0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100Concentration, ng/mL0.02.0e64.0e66.0e68.0e61.0e71.2e71.4e71.6e71.8e72.0e72.2e72.4e72.6e72.8e73.0e73.2e73.4e73.6e73.8e74.0e74.2e74.4e7

Area, counts*** Ginger AG01330509Results Name: g120413a.rdbData printed by KJUPrinting Time: 4:46:30 PMPrinting Date: Monday, April 16, 2012Page 1 of 1Sample Name: "g120413a017" Sample ID: "11012-10-11" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q water" Annotation: ""Sample Index: 17Sample Type: StandardConcentration: 10.0 ng/mLCalculated Conc: 9.83 ng/mLAcq. Date: 4/14/2012Acq. Time: 12:18:40 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.3 minArea: 10731882 countsHeight: 2970000. cpsStart Time: 13.2 minEnd Time: 13.7 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e6

Intensity, cps13.2713.12Sample Name: "g120413a024" Sample ID: "TN12-0139" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "Method Blank" Annotation: ""Sample Index: 24Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 2:46:48 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.0 minArea: 2284 countsHeight: 566. cpsStart Time: 12.9 minEnd Time: 13.1 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0200400600800100012001400160018002000220024002600280030003200

Intensity, cpsSample Name: "g120413a025" Sample ID: "LCS-120411-1" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "0.5ppb LCS" Annotation: ""Sample Index: 25Sample Type: QCConcentration: 0.498 ng/mLCalculated Conc: 0.479 ng/mLAcq. Date: 4/14/2012Acq. Time: 3:08:04 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.1 minArea: 609352 countsHeight: 187000. cpsStart Time: 13.0 minEnd Time: 13.3 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e5

Intensity, cps13.1113.7512.9515.22Sample Name: "g120413a028" Sample ID: "LCS-120411-4" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 28Sample Type: QCConcentration: 4.98 ng/mLCalculated Conc: 4.69 ng/mLAcq. Date: 4/14/2012Acq. Time: 4:11:21 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.1 minArea: 5563369 countsHeight: 1630000. cpsStart Time: 13.0 minEnd Time: 13.6 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e6

Intensity, cps13.07*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:42 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 9Sample Name: "g120413a031" Sample ID: "LCS-120411-7" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "30ppb LCS" Annotation: ""Sample Index: 31Sample Type: QCConcentration: 29.9 ng/mLCalculated Conc: No InterceptAcq. Date: 4/14/2012Acq. Time: 5:14:51 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.0 minArea: 23742500 countsHeight: 5920000. cpsStart Time: 12.9 minEnd Time: 13.5 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.05.0e51.0e61.5e62.0e62.5e63.0e63.5e64.0e64.5e65.0e65.5e6

Intensity, cps13.01Sample Name: "g120413a043" Sample ID: "GLP10-01-02-31-001" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW1R-0 (dil 1:10)" Annotation: ""Sample Index: 43Sample Type: UnknownConcentration: N/ACalculated Conc: 33.8 ng/mLAcq. Date: 4/14/2012Acq. Time: 9:28:50 AMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.0 minArea: 4094319 countsHeight: 1180000. cpsStart Time: 12.8 minEnd Time: 13.4 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e61.10e61.15e6

Intensity, cps13.04Sample Name: "g120413a046" Sample ID: "GLP10-01-02-31-004" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW1R-HS (dil 1:10)" Annotation: ""Sample Index: 46Sample Type: UnknownConcentration: N/ACalculated Conc: 120. ng/mLAcq. Date: 4/14/2012Acq. Time: 10:32:24 AMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.0 minArea: 12616952 countsHeight: 3430000. cpsStart Time: 12.8 minEnd Time: 13.4 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e6

Intensity, cps13.01Sample Name: "g120413a050" Sample ID: "GLP10-01-02-31-005" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:10)" Annotation: ""Sample Index: 50Sample Type: UnknownConcentration: N/ACalculated Conc: 116. ng/mLAcq. Date: 4/14/2012Acq. Time: 11:57:29 AMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.3 minArea: 2851509 countsHeight: 801000. cpsStart Time: 13.0 minEnd Time: 13.6 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e5

Intensity, cps13.27*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:42 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 9Sample Name: "g120413a081" Sample ID: "GLP10-01-02-31-005" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:10)" Annotation: ""Sample Index: 81Sample Type: UnknownConcentration: N/ACalculated Conc: 120. ng/mLAcq. Date: 4/14/2012Acq. Time: 10:53:53 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.0 minArea: 12579452 countsHeight: 3350000. cpsStart Time: 12.8 minEnd Time: 13.4 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e6

Intensity, cps13.0112.86Sample Name: "g120413a055" Sample ID: "GLP10-01-02-31-009" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW3R-0 (dil 1:10)" Annotation: ""Sample Index: 55Sample Type: UnknownConcentration: N/ACalculated Conc: 309. ng/mLAcq. Date: 4/14/2012Acq. Time: 1:42:48 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.3 minArea: 7168230 countsHeight: 2040000. cpsStart Time: 13.0 minEnd Time: 13.6 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e6

Intensity, cps13.28Sample Name: "g120413a062" Sample ID: "GLP10-01-02-31-013" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW4R-0 (dil 1:10)" Annotation: ""Sample Index: 62Sample Type: UnknownConcentration: N/ACalculated Conc: 185. ng/mLAcq. Date: 4/14/2012Acq. Time: 4:10:50 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.0 minArea: 17265648 countsHeight: 4550000. cpsStart Time: 12.8 minEnd Time: 13.5 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e63.6e63.8e64.0e64.2e64.4e6

Intensity, cps13.04Sample Name: "g120413a064" Sample ID: "GLP10-01-02-31-015" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW4R-LS (dil 1:10)" Annotation: ""Sample Index: 64Sample Type: UnknownConcentration: N/ACalculated Conc: 195. ng/mLAcq. Date: 4/14/2012Acq. Time: 4:53:12 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.1 minArea: 17807061 countsHeight: 4820000. cpsStart Time: 12.8 minEnd Time: 13.6 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e63.6e63.8e64.0e64.2e64.4e64.6e6

Intensity, cps13.06*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:43 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 9Sample Name: "g120413a067" Sample ID: "GLP10-01-02-31-017" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW5R-0 (dil 1:10)" Annotation: ""Sample Index: 67Sample Type: UnknownConcentration: N/ACalculated Conc: No InterceptAcq. Date: 4/14/2012Acq. Time: 5:57:06 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.1 minArea: 33576423 countsHeight: 7580000. cpsStart Time: 12.9 minEnd Time: 13.5 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.05.0e51.0e61.5e62.0e62.5e63.0e63.5e64.0e64.5e65.0e65.5e66.0e66.5e67.0e67.5e6

Intensity, cps13.08Sample Name: "g120413a074" Sample ID: "GLP10-01-02-31-021" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW6R-0 (dil 1:10)" Annotation: ""Sample Index: 74Sample Type: UnknownConcentration: N/ACalculated Conc: 156. ng/mLAcq. Date: 4/14/2012Acq. Time: 8:25:30 PMModified: YesRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.3 minArea: 15369048 countsHeight: 4190000. cpsStart Time: 13.0 minEnd Time: 13.7 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e63.6e63.8e64.0e6

Intensity, cps13.29Sample Name: "g120413a038" Sample ID: "GLP10-01-02-31-025" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-RW3R-RB (dil 1:10)" Annotation: ""Sample Index: 38Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 7:43:05 AMModified: YesProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.3 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 13.3 minArea: 1643 countsHeight: 558. cpsStart Time: 13.2 minEnd Time: 13.4 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0200400600800100012001400160018002000220024002600280030003200

Intensity, cpsSample Name: "g120413a039" Sample ID: "GLP10-01-02-31-026" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TRIP01-0 (dil 1:10)" Annotation: ""Sample Index: 39Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 8:04:09 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.1 minArea: 1065 countsHeight: 281. cpsStart Time: 13.0 minEnd Time: 13.1 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0200400600800100012001400160018002000220024002600280030003200

Intensity, cps13.6913.8915.0015.3514.0114.7213.0613.5811.1712.93*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:43 AMPrinting Date: Wednesday, May 09, 2012Page 4 of 9Sample Name: "g120413a040" Sample ID: "GLP10-01-02-31-027" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TRIP01-LS (dil 1:10)" Annotation: ""Sample Index: 40Sample Type: UnknownConcentration: N/ACalculated Conc: 8.51 ng/mLAcq. Date: 4/14/2012Acq. Time: 8:25:25 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.0 minArea: 1073895 countsHeight: 318000. cpsStart Time: 12.9 minEnd Time: 13.3 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e5

Intensity, cps13.0513.7012.90Sample Name: "g120413a041" Sample ID: "GLP10-01-02-31-028" File: "g120413a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TRIP01-HS (dil 1:10)" Annotation: ""Sample Index: 41Sample Type: UnknownConcentration: N/ACalculated Conc: 92.0 ng/mLAcq. Date: 4/14/2012Acq. Time: 8:46:37 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 2Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.0 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.1 minArea: 10148792 countsHeight: 2920000. cpsStart Time: 13.0 minEnd Time: 13.4 min11.0 11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.51714 1792 1870 1947 2025 2103 2181 2259 2337 2415Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e6

Intensity, cps13.09Sample Name: "g120413a017" Sample ID: "11012-10-11" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q water" Annotation: ""Sample Index: 17Sample Type: StandardConcentration: 9.98 ng/mLCalculated Conc: 9.91 ng/mLAcq. Date: 4/14/2012Acq. Time: 12:18:40 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.8 minArea: 10664568 countsHeight: 3180000. cpsStart Time: 14.7 minEnd Time: 15.1 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e6

Intensity, cps14.81Sample Name: "g120413a024" Sample ID: "TN12-0139" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "Method Blank" Annotation: ""Sample Index: 24Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 2:46:48 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 4389 countsHeight: 1260. cpsStart Time: 14.5 minEnd Time: 14.6 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0500100015002000250030003500400045005000550060006500

Intensity, cps*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:43 AMPrinting Date: Wednesday, May 09, 2012Page 5 of 9Sample Name: "g120413a025" Sample ID: "LCS-120411-1" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "0.5ppb LCS" Annotation: ""Sample Index: 25Sample Type: QCConcentration: 0.497 ng/mLCalculated Conc: 0.472 ng/mLAcq. Date: 4/14/2012Acq. Time: 3:08:04 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 586064 countsHeight: 185000. cpsStart Time: 14.5 minEnd Time: 14.8 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e5

Intensity, cps14.6515.18Sample Name: "g120413a028" Sample ID: "LCS-120411-4" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 28Sample Type: QCConcentration: 4.97 ng/mLCalculated Conc: 4.73 ng/mLAcq. Date: 4/14/2012Acq. Time: 4:11:21 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 5490381 countsHeight: 1710000. cpsStart Time: 14.5 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e6

Intensity, cps14.61Sample Name: "g120413a031" Sample ID: "LCS-120411-7" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "30ppb LCS" Annotation: ""Sample Index: 31Sample Type: QCConcentration: 29.9 ng/mLCalculated Conc: 37.1 ng/mLAcq. Date: 4/14/2012Acq. Time: 5:14:51 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.5 minArea: 23673721 countsHeight: 6390000. cpsStart Time: 14.4 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.05.0e51.0e61.5e62.0e62.5e63.0e63.5e64.0e64.5e65.0e65.5e66.0e6

Intensity, cps14.55Sample Name: "g120413a043" Sample ID: "GLP10-01-02-31-001" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW1R-0 (dil 1:10)" Annotation: ""Sample Index: 43Sample Type: UnknownConcentration: N/ACalculated Conc: 56.7 ng/mLAcq. Date: 4/14/2012Acq. Time: 9:28:50 AMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.6 minArea: 6486895 countsHeight: 1730000. cpsStart Time: 14.3 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e6

Intensity, cps14.57*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:43 AMPrinting Date: Wednesday, May 09, 2012Page 6 of 9Sample Name: "g120413a046" Sample ID: "GLP10-01-02-31-004" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW1R-HS (dil 1:10)" Annotation: ""Sample Index: 46Sample Type: UnknownConcentration: N/ACalculated Conc: 147. ng/mLAcq. Date: 4/14/2012Acq. Time: 10:32:24 AMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.5 minArea: 14709560 countsHeight: 3990000. cpsStart Time: 14.3 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e63.2e63.4e63.6e63.8e6

Intensity, cps14.5414.40Sample Name: "g120413a050" Sample ID: "GLP10-01-02-31-005" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:10)" Annotation: ""Sample Index: 50Sample Type: UnknownConcentration: N/ACalculated Conc: 580. ng/mLAcq. Date: 4/14/2012Acq. Time: 11:57:29 AMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.8 minArea: 12168421 countsHeight: 3010000. cpsStart Time: 14.5 minEnd Time: 15.1 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e6

Intensity, cps14.8114.66Sample Name: "g120413a081" Sample ID: "GLP10-01-02-31-005" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:10)" Annotation: ""Sample Index: 81Sample Type: UnknownConcentration: N/ACalculated Conc: No InterceptAcq. Date: 4/14/2012Acq. Time: 10:53:53 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.6 minArea: 43613173 countsHeight: 8020000. cpsStart Time: 14.2 minEnd Time: 15.0 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.05.0e51.0e61.5e62.0e62.5e63.0e63.5e64.0e64.5e65.0e65.5e66.0e66.5e67.0e67.5e68.0e6

Intensity, cps14.5614.42Sample Name: "g120413a055" Sample ID: "GLP10-01-02-31-009" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW3R-0 (dil 1:10)" Annotation: ""Sample Index: 55Sample Type: UnknownConcentration: N/ACalculated Conc: No InterceptAcq. Date: 4/14/2012Acq. Time: 1:42:48 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.8 minArea: 24528502 countsHeight: 5490000. cpsStart Time: 14.5 minEnd Time: 15.1 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.05.0e51.0e61.5e62.0e62.5e63.0e63.5e64.0e64.5e65.0e6

Intensity, cps14.8114.67*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:44 AMPrinting Date: Wednesday, May 09, 2012Page 7 of 9Sample Name: "g120413a062" Sample ID: "GLP10-01-02-31-013" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW4R-0 (dil 1:10)" Annotation: ""Sample Index: 62Sample Type: UnknownConcentration: N/ACalculated Conc: 19.0 ng/mLAcq. Date: 4/14/2012Acq. Time: 4:10:50 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.6 minArea: 2294370 countsHeight: 618000. cpsStart Time: 14.3 minEnd Time: 14.8 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps14.58Sample Name: "g120413a064" Sample ID: "GLP10-01-02-31-015" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW4R-LS (dil 1:10)" Annotation: ""Sample Index: 64Sample Type: UnknownConcentration: N/ACalculated Conc: 28.2 ng/mLAcq. Date: 4/14/2012Acq. Time: 4:53:12 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.6 minArea: 3365504 countsHeight: 935000. cpsStart Time: 14.3 minEnd Time: 14.8 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e58.5e59.0e5

Intensity, cps14.6014.46Sample Name: "g120413a067" Sample ID: "GLP10-01-02-31-017" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW5R-0 (dil 1:10)" Annotation: ""Sample Index: 67Sample Type: UnknownConcentration: N/ACalculated Conc: 43.3 ng/mLAcq. Date: 4/14/2012Acq. Time: 5:57:06 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.6 minArea: 5057137 countsHeight: 1400000. cpsStart Time: 14.3 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e6

Intensity, cps14.6214.48Sample Name: "g120413a074" Sample ID: "GLP10-01-02-31-021" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW6R-0 (dil 1:10)" Annotation: ""Sample Index: 74Sample Type: UnknownConcentration: N/ACalculated Conc: 48.0 ng/mLAcq. Date: 4/14/2012Acq. Time: 8:25:30 PMModified: YesRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 14.8 minArea: 5564441 countsHeight: 1470000. cpsStart Time: 14.5 minEnd Time: 15.1 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e6

Intensity, cps14.8314.68*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:44 AMPrinting Date: Wednesday, May 09, 2012Page 8 of 9Sample Name: "g120413a038" Sample ID: "GLP10-01-02-31-025" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-RW3R-RB (dil 1:10)" Annotation: ""Sample Index: 38Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 7:43:05 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.8 minArea: 2254 countsHeight: 697. cpsStart Time: 14.8 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min050010001500200025003000350040004500500055006000

Intensity, cpsSample Name: "g120413a039" Sample ID: "GLP10-01-02-31-026" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TRIP01-0 (dil 1:10)" Annotation: ""Sample Index: 39Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/14/2012Acq. Time: 8:04:09 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 1832 countsHeight: 542. cpsStart Time: 14.5 minEnd Time: 14.7 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min05001000150020002500300035004000450050005500

Intensity, cps15.1115.3614.6016.7116.6115.78Sample Name: "g120413a040" Sample ID: "GLP10-01-02-31-027" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TRIP01-LS (dil 1:10)" Annotation: ""Sample Index: 40Sample Type: UnknownConcentration: N/ACalculated Conc: 8.60 ng/mLAcq. Date: 4/14/2012Acq. Time: 8:25:25 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 1057216 countsHeight: 333000. cpsStart Time: 14.5 minEnd Time: 14.8 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e53.2e5

Intensity, cps14.5815.11Sample Name: "g120413a041" Sample ID: "GLP10-01-02-31-028" File: "g120413a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TRIP01-HS (dil 1:10)" Annotation: ""Sample Index: 41Sample Type: UnknownConcentration: N/ACalculated Conc: 88.4 ng/mLAcq. Date: 4/14/2012Acq. Time: 8:46:37 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 14.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 14.6 minArea: 9665200 countsHeight: 2890000. cpsStart Time: 14.5 minEnd Time: 14.9 min12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.01947 2025 2103 2181 2259 2337 2415 2492 2570 2648Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e6

Intensity, cps14.62*** Ginger AG01330509Results Name: g120413a.rdbData printed by STWPrinting Time: 11:15:44 AMPrinting Date: Wednesday, May 09, 2012Page 9 of 9b120416a.rdb (PFOS): "Quadratic" Regression ("1 / x" weighting): y = -3.78e+003 x^2 + 6.38e+005 x + 1.29e+004 (r = 0.9995)0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46Concentration, ng/mL0.01.0e62.0e63.0e64.0e65.0e66.0e67.0e68.0e69.0e61.0e71.1e71.2e71.3e71.4e71.5e71.6e71.7e71.8e71.9e72.0e72.1e72.2e7

Area, countsWorkstation: ETSBUSTERBatch Name: b120416a.dab*Data worked up by KJUPage 1 of 1Printing Time: 2:17:59 PMPrinting Date: Wednesday, April 18, 2012b120416a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -3.75e+003 x^2 + 5.22e+005 x + 5.55e+003 (r = 0.9993)0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50Concentration, ng/mL0.01.0e62.0e63.0e64.0e65.0e66.0e67.0e68.0e69.0e61.0e71.1e71.2e71.3e71.4e71.5e71.6e71.7e7

Area, countsWorkstation: ETSBUSTERBatch Name: b120416a.dab*Data worked up by KJUPage 1 of 1Printing Time: 2:17:43 PMPrinting Date: Wednesday, April 18, 2012Sample Name: "b120416a020" Sample ID: "11012-10-11" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q Water" Annotation: ""Sample Index: 20Sample Type: StandardConcentration: 9.28 ng/mLCalculated Conc: 9.80 ng/mLAcq. Date: 4/16/2012Acq. Time: 10:15:48 PMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 5899151 countsHeight: 1.34e+006 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e58.5e59.0e59.5e51.0e61.1e61.1e61.2e61.2e61.3e61.3e6

Intensity, cps16.0315.8915.71Sample Name: "b120416a024" Sample ID: "Solvent Blank" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "Milli Q Water TN12-0142" Annotation: ""Sample Index: 24Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/16/2012Acq. Time: 11:40:12 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 16.0 minArea: 1091 countsHeight: 4.17e+002 cpsStart Time: 16.0 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min02004006008001000120014001600180020002200240026002800300032003400360038004000

Intensity, cpsSample Name: "b120416a027" Sample ID: "LCS-120411-1" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "0.5ppb LCS" Annotation: ""Sample Index: 27Sample Type: QCConcentration: 0.462 ng/mLCalculated Conc: 0.348 ng/mLAcq. Date: 4/17/2012Acq. Time: 12:43:31 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 16.0 minArea: 234201 countsHeight: 5.09e+004 cpsStart Time: 15.8 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.02000.04000.06000.08000.01.0e41.2e41.4e41.6e41.8e42.0e42.2e42.4e42.6e42.8e43.0e43.2e43.4e43.6e43.8e44.0e44.2e44.4e44.6e44.8e45.0e4

Intensity, cps16.0215.8816.4217.7215.7117.3314.36 16.5917.1714.5915.44Sample Name: "b120416a030" Sample ID: "LCS-120411-4" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 30Sample Type: QCConcentration: 4.62 ng/mLCalculated Conc: 4.20 ng/mLAcq. Date: 4/17/2012Acq. Time: 1:46:54 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 2626788 countsHeight: 6.19e+005 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps16.0215.8815.70*** Buster J2930203Printing Time: 11:27:17 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 3Sample Name: "b120416a033" Sample ID: "LCS-120411-7" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "30ppb LCS" Annotation: ""Sample Index: 33Sample Type: QCConcentration: 27.8 ng/mLCalculated Conc: 26.5 ng/mLAcq. Date: 4/17/2012Acq. Time: 2:50:10 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 16.0 minArea: 14250414 countsHeight: 3.03e+006 cpsStart Time: 15.6 minEnd Time: 16.3 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e6

Intensity, cps16.0215.8715.70Sample Name: "b120416a044" Sample ID: "GLP10-01-02-31-001" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW1R-0" Annotation: ""Sample Index: 44Sample Type: UnknownConcentration: N/ACalculated Conc: 27.7 ng/mLAcq. Date: 4/17/2012Acq. Time: 6:42:17 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 15.9 minArea: 1747791 countsHeight: 1.96e+005 cpsStart Time: 15.5 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e51.9e5

Intensity, cps15.8616.01Sample Name: "b120416a047" Sample ID: "GLP10-01-02-31-004" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW1R-HS" Annotation: ""Sample Index: 47Sample Type: UnknownConcentration: N/ACalculated Conc: 94.7 ng/mLAcq. Date: 4/17/2012Acq. Time: 7:45:24 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 5713870 countsHeight: 9.27e+005 cpsStart Time: 15.5 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e58.5e59.0e5

Intensity, cps16.0115.8615.70Sample Name: "b120416a040" Sample ID: "GLP10-01-02-31-025" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-RW3R-RB" Annotation: ""Sample Index: 40Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/17/2012Acq. Time: 5:17:54 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 16.0 minArea: 1220 countsHeight: 3.94e+002 cpsStart Time: 15.9 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min020040060080010001200140016001800200022002400260028003000320034003600

Intensity, cps*** Buster J2930203Printing Time: 11:27:17 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 3Sample Name: "b120416a041" Sample ID: "GLP10-01-02-31-026" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TRIP01-0" Annotation: ""Sample Index: 41Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/17/2012Acq. Time: 5:39:01 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 15.9 minArea: 271 countsHeight: 2.02e+002 cpsStart Time: 15.9 minEnd Time: 15.9 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min01002003004005006007008009001000110012001300140015001600170018001900200021002200230024002500

Intensity, cps16.4117.7216.2917.3917.9015.5416.5917.1714.3218.2615.4715.6215.9114.4114.7914.2814.03Sample Name: "b120416a042" Sample ID: "GLP10-01-02-31-027" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TRIP01-LS" Annotation: ""Sample Index: 42Sample Type: UnknownConcentration: N/ACalculated Conc: 6.96 ng/mLAcq. Date: 4/17/2012Acq. Time: 6:00:04 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: Base To BaseRetention Time: 16.0 minArea: 454899 countsHeight: 8.94e+004 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e48.0e48.5e4

Intensity, cps16.0115.8716.4115.7017.7014.3416.6917.2815.4616.99Sample Name: "b120416a043" Sample ID: "GLP10-01-02-31-028" File: "b120416a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TRIP01-HS" Annotation: ""Sample Index: 43Sample Type: UnknownConcentration: N/ACalculated Conc: 70.1 ng/mLAcq. Date: 4/17/2012Acq. Time: 6:21:07 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 4299439 countsHeight: 8.42e+005 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e5

Intensity, cps16.0215.8715.70*** Buster J2930203Printing Time: 11:27:17 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 3Sample Name: "b120416a020" Sample ID: "11012-10-11" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q Water" Annotation: ""Sample Index: 20Sample Type: StandardConcentration: 10.0 ng/mLCalculated Conc: 10.2 ng/mLAcq. Date: 4/16/2012Acq. Time: 10:15:48 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 4930110 countsHeight: 1.34e+006 cpsStart Time: 13.5 minEnd Time: 13.8 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e58.5e59.0e59.5e51.0e61.1e61.1e61.2e61.2e61.3e61.3e6

Intensity, cps13.56Sample Name: "b120416a024" Sample ID: "Solvent Blank" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "Milli Q Water TN12-0142" Annotation: ""Sample Index: 24Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/16/2012Acq. Time: 11:40:12 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.5 minArea: 1362 countsHeight: 3.91e+002 cpsStart Time: 13.5 minEnd Time: 13.6 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min010020030040050060070080090010001100120013001400150016001700180019002000

Intensity, cpsSample Name: "b120416a027" Sample ID: "LCS-120411-1" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "0.5ppb LCS" Annotation: ""Sample Index: 27Sample Type: QCConcentration: 0.498 ng/mLCalculated Conc: 0.505 ng/mLAcq. Date: 4/17/2012Acq. Time: 12:43:31 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 268240 countsHeight: 7.52e+004 cpsStart Time: 13.5 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05000.01.0e41.5e42.0e42.5e43.0e43.5e44.0e44.5e45.0e45.5e46.0e46.5e47.0e47.5e4

Intensity, cps13.5511.7114.2115.7815.1214.7711.8414.6313.43Sample Name: "b120416a030" Sample ID: "LCS-120411-4" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 30Sample Type: QCConcentration: 4.98 ng/mLCalculated Conc: 5.12 ng/mLAcq. Date: 4/17/2012Acq. Time: 1:46:54 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.5 minArea: 2576292 countsHeight: 7.20e+005 cpsStart Time: 13.4 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e5

Intensity, cps13.55*** Buster J2930203Printing Time: 11:28:38 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 3Sample Name: "b120416a033" Sample ID: "LCS-120411-7" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "30ppb LCS" Annotation: ""Sample Index: 33Sample Type: QCConcentration: 29.9 ng/mLCalculated Conc: 29.3 ng/mLAcq. Date: 4/17/2012Acq. Time: 2:50:10 AMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 12068689 countsHeight: 3.04e+006 cpsStart Time: 13.4 minEnd Time: 13.8 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.02.0e54.0e56.0e58.0e51.0e61.2e61.4e61.6e61.8e62.0e62.2e62.4e62.6e62.8e63.0e6

Intensity, cps13.56Sample Name: "b120416a057" Sample ID: "GLP10-01-02-31-013" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW4R-0" Annotation: ""Sample Index: 57Sample Type: UnknownConcentration: N/ACalculated Conc: 179. ng/mLAcq. Date: 4/17/2012Acq. Time: 11:15:54 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 1824040 countsHeight: 4.95e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e53.2e53.4e53.6e53.8e54.0e54.2e54.4e54.6e54.8e5

Intensity, cps13.52Sample Name: "b120416a059" Sample ID: "GLP10-01-02-31-016" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW4R-HS" Annotation: ""Sample Index: 59Sample Type: UnknownConcentration: N/ACalculated Conc: 278. ng/mLAcq. Date: 4/17/2012Acq. Time: 11:58:03 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 2793103 countsHeight: 7.74e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e5

Intensity, cps13.53Sample Name: "b120416a066" Sample ID: "GLP10-01-02-31-021" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW6R-0" Annotation: ""Sample Index: 66Sample Type: UnknownConcentration: N/ACalculated Conc: 156. ng/mLAcq. Date: 4/17/2012Acq. Time: 2:25:37 PMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 1599713 countsHeight: 4.53e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e53.2e53.4e53.6e53.8e54.0e54.2e54.4e5

Intensity, cps13.51*** Buster J2930203Printing Time: 11:28:38 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 3Sample Name: "b120416a068" Sample ID: "GLP10-01-02-31-024" File: "b120416a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW6R-HS" Annotation: ""Sample Index: 68Sample Type: UnknownConcentration: N/ACalculated Conc: 238. ng/mLAcq. Date: 4/17/2012Acq. Time: 3:07:39 PMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 2399904 countsHeight: 6.76e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e5

Intensity, cps13.51*** Buster J2930203Printing Time: 11:28:38 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 3b120427a.rdb (PFOS): "Quadratic" Regression ("1 / x" weighting): y = -348 x^2 + 2.79e+005 x + 9.24e+003 (r = 0.9998)5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90Concentration, ng/mL0.01.0e62.0e63.0e64.0e65.0e66.0e67.0e68.0e69.0e61.0e71.1e71.2e71.3e71.4e71.5e71.6e71.7e71.8e71.9e72.0e72.1e72.2e72.3e7

Area, countsWorkstation: ETSBUSTERBatch Name: b120427a.dab*Data worked up by KJUPage 1 of 1Printing Time: 2:04:20 PMPrinting Date: Monday, April 30, 2012b120427a.rdb (PFHS): "Quadratic" Regression ("1 / x" weighting): y = -642 x^2 + 2.39e+005 x + 6.09e+004 (r = 0.9996)5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100Concentration, ng/mL0.01.0e62.0e63.0e64.0e65.0e66.0e67.0e68.0e69.0e61.0e71.1e71.2e71.3e71.4e71.5e71.6e71.7e71.8e7

Area, countsWorkstation: ETSBUSTERBatch Name: b120427a.dab*Data worked up by KJUPage 1 of 1Printing Time: 2:03:53 PMPrinting Date: Monday, April 30, 2012b120427a.rdb (PFBS): "Quadratic" Regression ("1 / x" weighting): y = -525 x^2 + 2.3e+005 x + 5.54e+004 (r = 0.9996)5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100Concentration, ng/mL0.01.0e62.0e63.0e64.0e65.0e66.0e67.0e68.0e69.0e61.0e71.1e71.2e71.3e71.4e71.5e71.6e71.7e71.8e7

Area, countsWorkstation: ETSBUSTERBatch Name: b120427a.dab*Data worked up by KJUPage 1 of 1Printing Time: 2:04:06 PMPrinting Date: Monday, April 30, 2012Sample Name: "b120427a012" Sample ID: "11012-10-11" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q Water" Annotation: ""Sample Index: 12Sample Type: StandardConcentration: 10.0 ng/mLCalculated Conc: 10.6 ng/mLAcq. Date: 4/27/2012Acq. Time: 5:30:22 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.5 minArea: 2433472 countsHeight: 5.89e+005 cpsStart Time: 13.4 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e5

Intensity, cps13.5313.70Sample Name: "b120427a020" Sample ID: "Solvent Blank" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "Milli Q Water TN12-0158" Annotation: ""Sample Index: 20Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/27/2012Acq. Time: 8:18:29 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.4 minArea: 611 countsHeight: 1.77e+002 cpsStart Time: 13.3 minEnd Time: 13.4 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min010020030040050060070080090010001100120013001400

Intensity, cpsSample Name: "b120427a021" Sample ID: "LCS-120426-1" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 21Sample Type: QCConcentration: 4.98 ng/mLCalculated Conc: 4.95 ng/mLAcq. Date: 4/27/2012Acq. Time: 8:39:30 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 1183526 countsHeight: 2.96e+005 cpsStart Time: 13.4 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e5

Intensity, cps13.5614.21Sample Name: "b120427a024" Sample ID: "LCS-120426-4" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "20ppb LCS" Annotation: ""Sample Index: 24Sample Type: QCConcentration: 20.0 ng/mLCalculated Conc: 18.6 ng/mLAcq. Date: 4/27/2012Acq. Time: 9:42:39 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 4157125 countsHeight: 1.04e+006 cpsStart Time: 13.4 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e6

Intensity, cps13.57*** Buster J2930203Printing Time: 11:31:04 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 3Sample Name: "b120427a027" Sample ID: "LCS-120426-7" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "50ppb LCS" Annotation: ""Sample Index: 27Sample Type: QCConcentration: 49.8 ng/mLCalculated Conc: 50.4 ng/mLAcq. Date: 4/27/2012Acq. Time: 10:45:36 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 50Base. Sub. Window: 1.00 minPeak-Split. Factor: 1Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ValleyRetention Time: 13.6 minArea: 10333827 countsHeight: 2.40e+006 cpsStart Time: 13.5 minEnd Time: 13.8 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e62.1e62.2e62.3e6

Intensity, cps13.57Sample Name: "b120427a038" Sample ID: "GLP10-01-02-31-009" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW3R-0 (dil 1:100)" Annotation: ""Sample Index: 38Sample Type: UnknownConcentration: N/ACalculated Conc: 292. ng/mLAcq. Date: 4/28/2012Acq. Time: 2:36:55 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.6 minArea: 723278 countsHeight: 1.81e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.01.0e42.0e43.0e44.0e45.0e46.0e47.0e48.0e49.0e41.0e51.1e51.2e51.3e51.4e51.5e51.6e51.7e51.8e5

Intensity, cps13.56Sample Name: "b120427a040" Sample ID: "GLP10-01-02-31-009; LMS" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW3R-0 2000ppb LMS (dil 1:100)" Annotation: ""Sample Index: 40Sample Type: UnknownConcentration: N/ACalculated Conc: 2280. ng/mLAcq. Date: 4/28/2012Acq. Time: 3:18:57 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 5024434 countsHeight: 1.23e+006 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e61.10e61.15e61.20e6

Intensity, cps13.55Sample Name: "b120427a046" Sample ID: "GLP10-01-02-31-017" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW5R-0 (dil 1:50)" Annotation: ""Sample Index: 46Sample Type: UnknownConcentration: N/ACalculated Conc: 533. ng/mLAcq. Date: 4/28/2012Acq. Time: 5:25:16 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 2453229 countsHeight: 6.10e+005 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps13.53*** Buster J2930203Printing Time: 11:31:04 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 3Sample Name: "b120427a048" Sample ID: "GLP10-01-02-31-017; LMS" File: "b120427a.wiff"Peak Name: "PFBS" Mass(es): "299.0/99.0 amu,299.0/80.0 amu"Comment: "DAL-GW-TW5R-0 LMS (dil 1:50)" Annotation: ""Sample Index: 48Sample Type: UnknownConcentration: N/ACalculated Conc: 988. ng/mLAcq. Date: 4/28/2012Acq. Time: 6:07:16 AMModified: YesRT Window: 30.0 secExpected RT: 13.6 minUse Relative RT: NoInt. Type: ManualRetention Time: 13.5 minArea: 4403931 countsHeight: 1.06e+006 cpsStart Time: 13.3 minEnd Time: 13.7 min11.5 12.0 12.5 13.0 13.5 14.0 14.5 15.0 15.5 16.0Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e6

Intensity, cps13.52*** Buster J2930203Printing Time: 11:31:04 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 3Sample Name: "b120427a012" Sample ID: "11012-10-11" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q Water" Annotation: ""Sample Index: 12Sample Type: StandardConcentration: 9.28 ng/mLCalculated Conc: 9.65 ng/mLAcq. Date: 4/27/2012Acq. Time: 5:30:22 PMModified: NoProc. Algorithm: IntelliQuan - MQIINoise Percentage: 95Base. Sub. Window: 1.00 minPeak-Split. Factor: 8Report Largest Peak: YesMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 16.0 minArea: 2668335 countsHeight: 5.03e+005 cpsStart Time: 15.6 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e53.2e53.4e53.6e53.8e54.0e54.2e54.4e54.6e54.8e55.0e5

Intensity, cps15.9915.83Sample Name: "b120427a020" Sample ID: "Solvent Blank" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "Milli Q Water TN12-0158" Annotation: ""Sample Index: 20Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/27/2012Acq. Time: 8:18:29 PMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 4791 countsHeight: 7.78e+002 cpsStart Time: 15.8 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min020040060080010001200140016001800200022002400260028003000320034003600380040004200440046004800500052005400

Intensity, cpsSample Name: "b120427a021" Sample ID: "LCS-120426-1" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 21Sample Type: QCConcentration: 4.62 ng/mLCalculated Conc: 4.26 ng/mLAcq. Date: 4/27/2012Acq. Time: 8:39:30 PMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 1190720 countsHeight: 2.30e+005 cpsStart Time: 15.6 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, 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, cps16.0015.8616.3715.69Sample Name: "b120427a024" Sample ID: "LCS-120426-4" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "20ppb LCS" Annotation: ""Sample Index: 24Sample Type: QCConcentration: 18.5 ng/mLCalculated Conc: 15.7 ng/mLAcq. Date: 4/27/2012Acq. Time: 9:42:39 PMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 4313013 countsHeight: 8.07e+005 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e5

Intensity, cps16.0015.86*** Buster J2930203Printing Time: 11:33:49 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 4Sample Name: "b120427a027" Sample ID: "LCS-120426-7" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "50ppb LCS" Annotation: ""Sample Index: 27Sample Type: QCConcentration: 46.2 ng/mLCalculated Conc: 46.3 ng/mLAcq. Date: 4/27/2012Acq. Time: 10:45:36 PMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 12177430 countsHeight: 2.20e+006 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e62.1e6

Intensity, cps16.0015.86Sample Name: "b120427a034" Sample ID: "GLP10-01-02-31-005" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:50)" Annotation: ""Sample Index: 34Sample Type: UnknownConcentration: N/ACalculated Conc: 995. ng/mLAcq. Date: 4/28/2012Acq. Time: 1:12:43 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 5421689 countsHeight: 6.29e+005 cpsStart Time: 15.5 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps16.0015.84Sample Name: "b120427a036" Sample ID: "GLP10-01-02-31-005; LMS" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW2R-0 LMS (dil 1:50)" Annotation: ""Sample Index: 36Sample Type: UnknownConcentration: N/ACalculated Conc: 2020. ng/mLAcq. Date: 4/28/2012Acq. Time: 1:54:50 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 10706006 countsHeight: 1.53e+006 cpsStart Time: 15.5 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e6

Intensity, cps15.9915.8415.69Sample Name: "b120427a038" Sample ID: "GLP10-01-02-31-009" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW3R-0 (dil 1:100)" Annotation: ""Sample Index: 38Sample Type: UnknownConcentration: N/ACalculated Conc: 3340. ng/mLAcq. Date: 4/28/2012Acq. Time: 2:36:55 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 8941101 countsHeight: 1.12e+006 cpsStart Time: 15.5 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, 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, cps15.9915.8315.69*** Buster J2930203Printing Time: 11:33:49 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 4Sample Name: "b120427a041" Sample ID: "GLP10-01-02-31-009; LMS" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW3R-0 5000ppb LMS (dil 1:100)" Annotation: ""Sample Index: 41Sample Type: UnknownConcentration: N/ACalculated Conc: 8480. ng/mLAcq. Date: 4/28/2012Acq. Time: 3:39:57 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 9070510 countsHeight: 1.45e+006 cpsStart Time: 15.5 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e6

Intensity, cps15.9715.82Sample Name: "b120427a056" Sample ID: "GLP10-01-02-31-013" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW4R-0 (dil 1:10)" Annotation: ""Sample Index: 56Sample Type: UnknownConcentration: N/ACalculated Conc: 244. ng/mLAcq. Date: 4/28/2012Acq. Time: 8:55:36 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 6604817 countsHeight: 1.09e+006 cpsStart Time: 15.6 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e6

Intensity, cps15.9715.82Sample Name: "b120427a058" Sample ID: "GLP10-01-02-31-013;LMS" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW4R-0 LMS (dil 1:10)" Annotation: ""Sample Index: 58Sample Type: UnknownConcentration: N/ACalculated Conc: 450. ng/mLAcq. Date: 4/28/2012Acq. Time: 9:37:38 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 11864095 countsHeight: 1.98e+006 cpsStart Time: 15.6 minEnd Time: 16.2 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e6

Intensity, cps15.9615.81Sample Name: "b120427a046" Sample ID: "GLP10-01-02-31-017" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW5R-0 (dil 1:50)" Annotation: ""Sample Index: 46Sample Type: UnknownConcentration: N/ACalculated Conc: 704. ng/mLAcq. Date: 4/28/2012Acq. Time: 5:25:16 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 3870595 countsHeight: 5.90e+005 cpsStart Time: 15.6 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e5

Intensity, cps15.9715.82*** Buster J2930203Printing Time: 11:33:49 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 4Sample Name: "b120427a048" Sample ID: "GLP10-01-02-31-017; LMS" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW5R-0 LMS (dil 1:50)" Annotation: ""Sample Index: 48Sample Type: UnknownConcentration: N/ACalculated Conc: 1120. ng/mLAcq. Date: 4/28/2012Acq. Time: 6:07:16 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 6078162 countsHeight: 9.88e+005 cpsStart Time: 15.5 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e57.0e57.5e58.0e58.5e59.0e59.5e5

Intensity, cps15.9715.82Sample Name: "b120427a050" Sample ID: "GLP10-01-02-31-021" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW6R-0 (dil 1:10)" Annotation: ""Sample Index: 50Sample Type: UnknownConcentration: N/ACalculated Conc: 287. ng/mLAcq. Date: 4/28/2012Acq. Time: 6:49:24 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 7742598 countsHeight: 1.25e+006 cpsStart Time: 15.6 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e61.05e61.10e61.15e61.20e6

Intensity, cps15.9715.82Sample Name: "b120427a052" Sample ID: "GLP10-01-02-31-021; LMS" File: "b120427a.wiff"Peak Name: "PFOS" Mass(es): "499.0/130.0 amu,499.0/99.0 amu,499.0/80.0 amu"Comment: "DAL-GW-TW6R-0 LMS (dil 1:10)" Annotation: ""Sample Index: 52Sample Type: UnknownConcentration: N/ACalculated Conc: 456. ng/mLAcq. Date: 4/28/2012Acq. Time: 7:31:25 AMModified: YesRT Window: 30.0 secExpected RT: 16.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 16.0 minArea: 12003443 countsHeight: 1.93e+006 cpsStart Time: 15.5 minEnd Time: 16.1 min14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5 18.0 18.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e6

Intensity, cps15.9715.82*** Buster J2930203Printing Time: 11:33:49 AMPrinting Date: Wednesday, May 09, 2012Page 4 of 4Sample Name: "b120427a012" Sample ID: "11012-10-11" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "10 ng/mL FC std in Milli Q Water" Annotation: ""Sample Index: 12Sample Type: StandardConcentration: 9.98 ng/mLCalculated Conc: 10.7 ng/mLAcq. Date: 4/27/2012Acq. Time: 5:30:22 PMModified: NoProc. Algorithm: IntelliQuan - IQAMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 15.0 minArea: 2553392 countsHeight: 6.63e+005 cpsStart Time: 14.9 minEnd Time: 15.3 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e56.5e5

Intensity, cps15.04Sample Name: "b120427a020" Sample ID: "Solvent Blank" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "Milli Q Water TN12-0158" Annotation: ""Sample Index: 20Sample Type: UnknownConcentration: N/ACalculated Conc: < 0Acq. Date: 4/27/2012Acq. Time: 8:18:29 PMModified: NoProc. Algorithm: IntelliQuan - IQAMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 15.1 minArea: 1272 countsHeight: 2.87e+002 cpsStart Time: 15.0 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min020040060080010001200140016001800200022002400260028003000320034003600380040004200440046004800

Intensity, cpsSample Name: "b120427a021" Sample ID: "LCS-120426-1" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "5ppb LCS" Annotation: ""Sample Index: 21Sample Type: QCConcentration: 4.97 ng/mLCalculated Conc: 4.85 ng/mLAcq. Date: 4/27/2012Acq. Time: 8:39:30 PMModified: NoProc. Algorithm: IntelliQuan - IQAMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 15.1 minArea: 1207355 countsHeight: 3.12e+005 cpsStart Time: 14.9 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.02.0e44.0e46.0e48.0e41.0e51.2e51.4e51.6e51.8e52.0e52.2e52.4e52.6e52.8e53.0e5

Intensity, cps15.0617.4615.5217.10Sample Name: "b120427a024" Sample ID: "LCS-120426-4" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "20ppb LCS" Annotation: ""Sample Index: 24Sample Type: QCConcentration: 19.9 ng/mLCalculated Conc: 17.7 ng/mLAcq. Date: 4/27/2012Acq. Time: 9:42:39 PMModified: NoProc. Algorithm: IntelliQuan - IQAMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 15.1 minArea: 4091996 countsHeight: 1.04e+006 cpsStart Time: 14.9 minEnd Time: 15.3 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.005.00e41.00e51.50e52.00e52.50e53.00e53.50e54.00e54.50e55.00e55.50e56.00e56.50e57.00e57.50e58.00e58.50e59.00e59.50e51.00e6

Intensity, cps15.07*** Buster J2930203Printing Time: 11:32:19 AMPrinting Date: Wednesday, May 09, 2012Page 1 of 3Sample Name: "b120427a027" Sample ID: "LCS-120426-7" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "50ppb LCS" Annotation: ""Sample Index: 27Sample Type: QCConcentration: 49.7 ng/mLCalculated Conc: 49.1 ng/mLAcq. Date: 4/27/2012Acq. Time: 10:45:36 PMModified: NoProc. Algorithm: IntelliQuan - IQAMin. Peak Height: 0.00 cpsMin. Peak Width: 0.00 secSmoothing Width: 3 pointsRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ValleyRetention Time: 15.1 minArea: 10271750 countsHeight: 2.53e+006 cpsStart Time: 14.9 minEnd Time: 15.3 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e61.7e61.8e61.9e62.0e62.1e62.2e62.3e62.4e62.5e6

Intensity, cps15.07Sample Name: "b120427a034" Sample ID: "GLP10-01-02-31-005" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW2R-0 (dil 1:50)" Annotation: ""Sample Index: 34Sample Type: UnknownConcentration: N/ACalculated Conc: 563. ng/mLAcq. Date: 4/28/2012Acq. Time: 1:12:43 AMModified: YesRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 15.1 minArea: 2678296 countsHeight: 5.69e+005 cpsStart Time: 14.8 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e5

Intensity, cps15.06Sample Name: "b120427a036" Sample ID: "GLP10-01-02-31-005; LMS" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW2R-0 LMS (dil 1:50)" Annotation: ""Sample Index: 36Sample Type: UnknownConcentration: N/ACalculated Conc: 1520. ng/mLAcq. Date: 4/28/2012Acq. Time: 1:54:50 AMModified: YesRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 15.0 minArea: 6734170 countsHeight: 1.60e+006 cpsStart Time: 14.8 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e6

Intensity, cps15.0514.91Sample Name: "b120427a038" Sample ID: "GLP10-01-02-31-009" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW3R-0 (dil 1:100)" Annotation: ""Sample Index: 38Sample Type: UnknownConcentration: N/ACalculated Conc: 1210. ng/mLAcq. Date: 4/28/2012Acq. Time: 2:36:55 AMModified: YesRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 15.1 minArea: 2858749 countsHeight: 6.16e+005 cpsStart Time: 14.8 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.05.0e41.0e51.5e52.0e52.5e53.0e53.5e54.0e54.5e55.0e55.5e56.0e5

Intensity, cps15.0514.92*** Buster J2930203Printing Time: 11:32:19 AMPrinting Date: Wednesday, May 09, 2012Page 2 of 3Sample Name: "b120427a040" Sample ID: "GLP10-01-02-31-009; LMS" File: "b120427a.wiff"Peak Name: "PFHS" Mass(es): "399.0/99.0 amu,399.0/80.0 amu"Comment: "DAL-GW-TW3R-0 2000ppb LMS (dil 1:100)" Annotation: ""Sample Index: 40Sample Type: UnknownConcentration: N/ACalculated Conc: 3160. ng/mLAcq. Date: 4/28/2012Acq. Time: 3:18:57 AMModified: YesRT Window: 30.0 secExpected RT: 15.1 minUse Relative RT: NoInt. Type: ManualRetention Time: 15.0 minArea: 6984904 countsHeight: 1.66e+006 cpsStart Time: 14.7 minEnd Time: 15.2 min13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0 17.5Time, min0.01.0e52.0e53.0e54.0e55.0e56.0e57.0e58.0e59.0e51.0e61.1e61.2e61.3e61.4e61.5e61.6e6

Intensity, cps15.0314.90*** Buster J2930203Printing Time: 11:32:19 AMPrinting Date: Wednesday, May 09, 2012Page 3 of 3

# 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 

# )

# R ECORD OF 

# D EVIATION 

# /N ONCONFORMANCE

ETS -4- 008.7

Documentation of Deviations and Control of Nonconforming Testing Page 1 of 1 

## I. Identification

Study / Project No.

GLP10

- 01

- 0 2

- 31 Date(s) of Occurrence:

4/ 16 /12 Document Number:

ETS

- 8

- 044. 1

Deviation type  SOP  Equipment Procedure  Method

(Check one)  Protocol  GPO  Other:



## II. Description (attach extra pages as needed)



## Method Requirements:

1. LCS recovery within ±20% (section 13.4).

2. FMS re covery within ±30% (section 13.4 ).



## Actual procedure/process:

1. The low set of LCSs had an average recovery of 75.2% for PFOS.

2. The sampling location DAL GW TW1R HS had a PFOS FMS recovery of 65.8 %.



## III. Actions Taken

(such as amendment issued, SOP revision, etc.)



## Corrective Action (  Yes  No) Reference :

Acceptability of the nonconforming work:

1. The non

- compliant LCSs will be flagged in the final report. Al l LCSs were used in the determination of the

analytical method uncertainty. Since the %bias wa s less than the c alculated method uncertainty for

PFOS of ±35% , the method uncertainty does not need to be expanded further.

2. The sampling location with a FMS recovery outside method acceptance criteria will be flagged in the

report . Since the analytical method uncertainty was calculated at 3 5 %, the uncertainty for DAL GW

TW1R does not need to be expanded further

.



### Actions:  Halting of Work  Client Notification  Work Recall  Withholding of Report

 Other: Deviations will be noted in final report.



### Project Lead /P AI Approval: Date:

Study Director (if GLP): Date:

Sponsor Approval (for GLP protocol deviations): NA Date: NA

T echnical Reviewer (optional): NA Date: NA

Laboratory Department Manager Approval: Date:



## IV. Authorization to Resume Work

Where haltin g of work occurred, resumption of work must first be approved by Laboratory Management



### Laboratory Department Manager Approval: NA Date: NA



## Deviation No. ______________

(assigned by Study Director or Team Leader at the end of study or project)