3M Environmental Laboratory

Report No. ISO12-01-01-02

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Final Report

Analysis of Water Samples from the Tennessee River – April 2012

Laboratory Request Number: ISO12-01-01-02

Method Requirement: 3M Method ETS-8-044.1

Report Date – April 26, 2012

Testing Laboratory

3M Environmental Health and Safety Operations

Environmental Laboratory

3M Center, Bldg 260-05-N-17

St. Paul, MN 55144

Requester 

Gary Hohenstein

3M EHS Operations

3M Building 224-5W-03

Saint Paul, MN 55144-1000

Phone: (651) 737-3570

Testing Cert #2052.01

The testing reported herein meet the requirements of ANSI/ ISO/IEC 17025:2005 “General 

Requirements for the Competence of Testing and Calibration Laboratories”, in accordance with 

the A2LA Testing Certificate # 2052.01. Testing that complies with this International Standard 

also meets principles of ISO 9001:2000. 
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Report No. ISO12-01-01-02

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3M Environmental Laboratory

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

3M Principal Analytical Investigator and Report Author: Susan Wolf

Analytical Report ISO12-01-01-02

Analysis of Water Samples from the Tennessee River –

April 2012

Report Date: April 26, 2012

1 Summary/Introduction

The 3M Environmental Laboratory prepared and analyzed samples collected by Weston personnel 

from the Tennessee River, conducted per the study work plan titled "Work Plan for Fish Sampling and 

Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M Decatur 

Facility". Samples were collected April 5 and 6, 2012. Samples were returned to the 3M 

Environmental Laboratory on April 13, 2012 at ambient conditions and analyzed for perfluorooctane 

sulfonate (PFOS) under 3M Environmental Laboratory project number ISO12-01-01-02.

The 3M Environmental Laboratory prepared sample containers for nine sampling locations, however 

only three sampling locations were collected. Each empty container was marked with a “fill to here” line 

that corresponded to a final volume of 200 mL. Each sample set consisted of a field sample, field 

sample duplicate, and a field matrix spike. All samples bottles included the addition of internal standard 

[

13C8]-PFOS and surrogate recovery standard [13C4]-PFOS, which were added to the sample containers 

prior to being sent to the field for sample collection. Sample bottles reserved for field matrix spikes 

were fortified with appropriate matrix spike solutions containing PFOS, internal standard [13C8]-PFOS 

and surrogate recovery standard [13C4]-PFOS prior to being sent to the field for sample collection. 

Samples were prepared and analyzed using either direct injection or solvent dilution according to 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”. The internal standard

[

13C8]-PFOS was used to aid in the quantitation of the samples. 

Table 1 summarizes the sample results using the analytical method identified above. All results for 

quality control samples prepared and analyzed with the samples will be reported and discussed 

elsewhere in this report. 
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Table 1. Sample Results Summary.

3M LIMS ID Sample Description

PFOS

Concentration 

(ng/mL)

ISO12-01-01-02-001 Location 1 - Sample 0.516

ISO12-01-01-02-002 Location 1 – Sample Duplicate 0.450

Average 0.483

%RPD Sample/Sample Dup 14

ISO12-01-01-02-004 Location 2 - Sample 0.720

ISO12-01-01-02-005 Location 2 – Sample Duplicate 0.661

Average 0.691

%RPD Sample/Sample Dup 8.5

ISO12-01-01-02-007 Location 3 - Sample 0.0210

ISO12-01-01-02-008 Location 3 – Sample Duplicate 0.0263

Average 0.0237

%RPD Sample/Sample Dup 22 (2)

ISO12-01-01-02-010 Equipment Blank 1 <0.00928

ISO12-01-01-02-011 Equipment Blank 2 <0.00928

ISO12-01-01-02-012 Equipment Blank 3 <0.00928

ISO12-01-01-02-013 Trip Blank <0.00928

NA = Not Applicable

(1) The analytical method uncertainty for the reported results using method ETS-8-044.1 by direct inject for PFOS is ± 18%.

(2) The sample / sample duplicate RPD value did not meet method acceptance criteria of ≤20%.

2 Method Summary

2.1 Methods

Analysis for PFOS was completed following 3M Environmental Laboratory method ETS-8-044.1

“Method of Analysis for the Determination of Perfluorinated Compounds in Water by LC/MS/MS”. 

Table 2. Target Analytes.

Target Analytes Acronym

Reference 

Material Structure

Perfluorooctanesulfonate (C8 Sulfonate) PFOS Linear + Branched

2.2 Sample Collection

Water samples were collected in 250 mL NalgeneTM (high-density polyethylene) bottles prepared at the 

3M Environmental Laboratory. A laboratory prepared Trip Blank and Trip Blank field matrix spike were 

sent with the set of collection bottles. Sample bottles were received by the laboratory at ambient 

conditions on April 13, 2012. Samples were stored refrigerated at the laboratory after receipt. 

2.3 Sample Preparation

An aliquot of sample was removed for analysis and placed in an autovial. 

During the preparation of the laboratory control samples, an aliquot of a separate internal standard 

spiking solution and surrogate recovery standard solution was added to the laboratory control samples 

(nominal concentration of 0.1 ng/mL). The sample bottles were spiked with an internal standard at a 
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Report No. ISO12-01-01-02

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nominal concentration of 0.1 ng/mL and a surrogate recovery standard at a nominal concentration of 

0.025 ng/ml prior to being sent to the field for sample collection.

2.4 Analysis

All samples and quality control samples were analyzed for PFOS and the surrogate recovery standard 

[

13C4]-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 Kirk

Analysis Dates 4/24/12

Analytical Method ETS-8-044.1 

Liquid Chromatograph Agilent 1290

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

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

Injection Volume 50 μL

Mass Spectrometer Applied Biosystems API 5500

Ion Source Turbo Spray

Electrode Turbo ion electrode

Polarity Negative

Software Analyst 1.5.2

Table 4. Liquid Chromatography Conditions.

Step 

Number

Total Time 

(min)

Flow Rate 

(μL/min)

Percent A

(5mM ammonium 

acetate:0.1% acetic acid)

Percent B

(Methanol)

0 0.00 750 90.0 10.0

1 0.50 750 90.0 10.0

2 4.00 750 70.0 30.0

3 6.00 750 70.0 30.0

4 11.0 750 20.0 80.0

5 13.0 750 20.0 80.0

6 13.5 750 10.0 90.0

7 16.0 750 10.0 90.0

8 16.5 750 90.0 10.0

9 19.0 750 90.0 10.0
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Report No. ISO12-01-01-02

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Table 5. Mass Transitions. 

Analyte

Mass Transition

Q1/Q3

Internal Standard Mass Transition

Q1/Q3

PFOS

499/99

[

13 499/80 C8]-PFOS 507/80

499/130

[

13C4]-PFOS 503/80 [13C8]-PFOS 507/80

Dwell time was 30 msec for each transition. The individual transitions were summed to produce a 

“total ion chromatogram” (TIC), which was used for quantitation. 

3 Analytical Results

3.1 Calibration

Samples were analyzed for PFOS and [

13C4]-PFOS against a stable isotope internal standard 

calibration curve. Calibration standards were prepared by spiking known amounts of the stock solution 

containing the target analyte and surrogate recovery standard into a laboratory-prepared synthetic 

groundwater containing calcium and magnesium. A separate internal standard spiking solution was 

prepared and an aliquot was added at the same level to all calibration standards and laboratory control 

samples at a nominal concentration of 1 ng/mL. A calibration curve ranging from approximately 0.005 

ng/mL to 15 ng/mL was analyzed (0.005 ng/mL to 5 ng/mL for [

13C4]-PFOS). The low and high curve 

points were disabled for PFOS to meet accuracy or method blank criteria. A quadratic, 1/x weighted, 

calibration curve of the standard peak area ratios 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 ratios and the resultant calibration curve 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 PFOS and [13C4]-PFOS.

It should be noted that the calibration curve that was used to analyze the samples on 4/24/12, expired 

on 4/17/12. The expiration date assigned to the curve was based on the expiration date of the solutions 

containing the surrogate recovery standard [13C4]-PFOS, which were used to prepared the calibration 

standards. The solutions containing PFOS that were used to prepare the calibation curve were not 

expired. The solutions containing [13C4]-PFOS that were used to prepared the high set of LCS samples 

and the sample collection bottles, were not expired. The high set of LCSs for [

13C4]-PFOS surrogate met 

method acceptance criteria, as did the [

13C4]-PFOS surrogate in the Tennessee River samples. A 

method deviation is included in the raw data.

 

3.2 System Suitability

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

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

standard deviation (RSD) for peak area ratio and retention time criteria of less than or equal to 2% RSD

were met for PFOS and [13C4]-PFOS.

3.3 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 are at least twice those of the appropriate 

blanks. The LOQ for PFOS was 0.00928 ng/mL.
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Report No. ISO12-01-01-02

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3.4 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 

reported sample results were bracketed by CCVs that met method criteria of 100% ± 25%.

3.5 Blanks

Three types of blanks were prepared and analyzed with the samples: solvent blanks (procedural 

blanks), field/trip blanks, and equipment rinseate blanks for the groundwater samples. Each blank result 

was reviewed and used to evaluate method performance. Procedural blank results were reviewed 

according to the method and used to evaluate method performance to determine the LOQ for each 

analyte.

3.6 Lab Control Spikes (LCSs) 

Low, mid, and high lab control spikes were prepared and analyzed in triplicate for PFOS, while only low 

and high lab control spikes were prepared for the [13C4]-PFOS surrogate. The LCS samples were

prepared by spiking known amounts of the analyte into 10 mL of synthetic groundwater to produce the 

desired concentration. The spiked water samples (LCSs) were then analyzed in the same manner as 

the samples. 

The method acceptance criteria states that the average recovery of LCSs be 100% ± 20% with a RSD 

≤20%, when evaluated independently at each concentration level. All LCSs met these criteria. The LCS 

results were used to determine overall method uncertainty in Section 3.7. 

The following calculations were used to generate data in Table 6 for laboratory control spikes.

* 100%

Spike Concentration

Calculated Concentration LCSPercent Recovery =

* 100%

average LCS recovery

standard deviation LCS replicates LCS% RSD =
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Report No. ISO12-01-01-02

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Table 6. Laboratory Control Spike Recovery.

ETS-8-044.1 

Analyzed 4/24/12 PFOS (Linear + Branched)

Lab ID

Spiked 

Concentration 

(ng/mL)

Calculated 

Concentration 

(ng/mL) %Recovery

LCS-120424-1 0.0231 0.0273 118

LCS-120424-2 0.0231 0.0257 111

LCS-120424-3 0.0231 0.0253 110

Average ± %RSD 113% ± 3.9%

LCS-120424-4 0.185 0.174 93.8

LCS-120424-5 0.185 0.186 100

LCS-120424-6 0.185 0.175 94.4

Average ± %RSD 96.1% ± 3.6%

LCS-120424-7 4.64 3.75 80.7

LCS-120424-8 4.64 4.01 86.4

LCS-120424-9 4.64 4.19 90.4

Average ± %RSD 85.8% ± 5.7%

ETS-8-044.1 

Analyzed 4/24/12 [

13C4]-PFOS

Lab ID

Spiked 

Concentration 

(ng/mL)

Calculated 

Concentration 

(ng/mL) %Recovery

LCS-120424-1 (1) 0.0238 0.0246 103

LCS-120424-2 (1) 0.0238 0.0249 105

LCS-120424-3 (1) 0.0238 0.0240 101

Average ± %RSD 103% ± 1.9%

LCS-120424-4 0.190 0.204 107

LCS-120424-5 0.190 0.200 105

LCS-120424-6 0.190 0.202 106

Average ± %RSD 106% ± 0.94%

(1) The solution used to prepare this set of LCS samples expired 4/17/12.
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3.7 Analytical Method Uncertainty

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

method accuracy and precision. The method uncertainty is calculated following ETS-12-012.2. The 

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

method ETS-8-044.1, the most recent fifty QC samples were used. The analytical method uncertainty 

is calculated by multiplying the standard deviation by a factor of 2, which corresponds to a confidence 

level of 95%. 

Table 7. Analytical Method Uncertainty 

3.8 Field Matrix Spikes (FMS)

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

applicable to the collected matrix. Field matrix spikes are generated by adding a measured volume of 

field sample to a container spiked by the laboratory with the target analytes 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. The reference standards for PFOS in the field matrix 

spiking solution consisted of linear and branched isomers. 

In addition, field matrix spikes for this project also included the use of stable isotope surrogate spikes of 

[

13C4]-PFOS, which was added at a nominal concentration of 0.025 ng/mL to all sample bottles prior to 

sample collection. Surrogate matrix spike recoveries within method acceptance criteria of 100±30% 

confirm that “unknown” components in the sample matrix do not significantly interfere with the 

preparation and analysis of the analytes of interest.

Table 8. Field Matrix Spike Level.

Sampling location PFOS ng/mL

All locations and Trip Blank 0.0232

*100%

Spike Concentration

(SampleConcentration of FMS Average Concentration : FieldSample& FieldSample Dup.) FMSRecovery − =

4 Data Summary and Discussion

The tables below summarize the sample results and field matrix spike and surrogate recoveries for the 

sampling locations as well as the Trip Blanks. 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 surrogate recoveries meeting the method 

acceptance criteria of ± 30%, demonstrate that the method(s) were appropriate for the given matrix and 

their respective quantitative ranges. 

Analyte Standard Deviation Method Uncertainty (%)

PFOS 8.75 ± 18

[

13C4]-PFOS 6.74 ± 13
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Report No. ISO12-01-01-02

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Table 9. Tennessee River; Location 1

PFOS [

13C4]-PFOS

3M LIMS ID Description

Concentration 

(ng/mL) %Recovery %Recovery

ISO12-01-01-02-001 Location 1 0.516 NA 108

ISO12-01-01-02-002 Location 1 Sample Duplicate 0.450 NA 105

ISO12-01-01-02-003 Location 1 FMS 0.537 NC 106

Average Concentration (ng/mL) ± %RPD / %RSD 0.483 ng/mL± 14% 106% ± 0.99%

NA = Not Applicable

NC = Not Calculated; Endogenous sample concentration is greater than 2x spiking level.

Table 10. Tennessee River; Location 2

PFOS [

13C4]-PFOS

3M LIMS ID Description

Concentration 

(ng/mL) %Recovery %Recovery

ISO12-01-01-02-004 Location 2 0.720 NA 108

ISO12-01-01-02-005 Location 2 Sample Duplicate 0.661 NA 106

ISO12-01-01-02-006 Location 2 FMS 0.654 NC 109

Average Concentration (ng/mL) ± %RPD / %RSD 0.691 ng/mL± 8.5% 108% ± 1.5%

NA = Not Applicable

NC = Not Calculated; Endogenous sample concentration is greater than 2x spiking level.

Table 11. Tennessee River; Location 3

PFOS [

13C4]-PFOS

3M LIMS ID Description

Concentration 

(ng/mL) %Recovery %Recovery

ISO12-01-01-02-007 Location 3 0.0210 NA 105

ISO12-01-01-02-008 Location 3 Sample Duplicate 0.0263 NA 108

ISO12-01-01-02-009 Location 3 FMS 0.0430 83.4 106

Average Concentration (ng/mL) ± %RPD / %RSD 0.0237 ng/mL± 22% (1) 106% ± 1.6%

NA = Not Applicable

(1) The sample / sample duplicate RPD did not meet method acceptance criteria of ≤20%.
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Report No. ISO12-01-01-02

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Table 12. Rinseate Blanks

PFOS [

13C4]-PFOS

3M LIMS ID Description

Concentration 

(ng/mL) %Recovery

ISO12-01-01-02-010 Equipment Blank 1 <0.00928 108

ISO12-01-01-02-011 Equipment Blank 2 <0.00928 105

ISO12-01-01-02-012 Equipment Blank 3 <0.00928 108

Table 13. Trip Blanks

PFOS [

13C4]-PFOS

3M LIMS ID Description

Concentration 

(ng/mL) %Recovery %Recovery

ISO12-01-01-02-013 Trip Blank <0.00928 NA 105

ISO12-01-01-02-014 Trip Blank FMS 0.0209 90.1 103
3M Envimnmental Laboratory 

Report No. ISO12-01-01-02 

5 Conclusion 

Laboratory control spikes were used to determine the analytical method accuracy and precision for 

PFOS. The accuracy and precision were then used to estimate the method uncertainty for the results. 

Field matrix spike recoveries demonstrated that the analytical method was appropriate for the given 

sample matrix. Analysis was completed using 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". Analytical results are reported in Tables 1 and 9-13 of this report. 

6 Data/Sample Retention 

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

to 3M Environmental Laboratory standard operating procedures. 

7 Signatures 

Susan Wolf, 3M Princi^Analytical Investigator Date 

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

The 3M Environmental Laboratory's Quality Assurance Unit has audited the data and report for this 

project. 

Quality Assurance Representative Date 

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