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TestAmerica Laboratories, Inc.
TestAmerica Denver
4955 Yarrow Street
Arvada, CO 80002
Phone: 303-736-0100
Fax: 303-431-7171

Electronic Copy Only

SOP No. DV-LC-0012, Rev. 15
Effective Date: 5/7/2018
Page No.: 2 of 51
1.0

2.0

Scope and Application
1.1

This procedure describes the analysis of water and soil samples for
perfluorooctanoic acid (PFOA, CAS# 335-67-1), and other perfluorinatedhydrocarbons (PFCs) and perfluorinated hydrocarbon sulfonates (PFSs) or
perfluorinated hydrocarbon sulfonic acids using liquid chromatography / tandem
mass spectrometry (LC/MS/MS). See Table 6 for the list of compounds and theircommon names. Basic structure for the perfluorinated hydrocarbon carboxylic
acids is CnF(2n+1)COOH; basic structure for the perfluorinated hydrocarbon
sulfonates is CnF(2n+1)SO3- in the anion form or CnF(2n+1)SO3H in the acid form.

1.2

Throughout this procedure the term perfluorinated hydrocarbon sulfonates (PFSs)
is used to describe a sub-set of the compounds listed in Table 6. Namely, thesecompounds are PFOS, PFBS, PFHxS, and PFDS. The term PFC is used todescribe all of the compounds listed in Table 6, including the PFSs.

1.3

This procedure has also been adapted for the analysis of Perfluoro(2-propoxy
propanoic) acid (HFPO).

1.4

This procedure does not include the sample extraction. See SOP DV-OP-0019
Extraction of Perfluorooctanoic Acid (PFOA) and Perfluorooctanoic Sulfonate(PFOS) and other Perfluorinated Hydrocarbons (PFCs) in Water and Soil.

1.5

The method has been validated by TestAmerica Denver for reagent water, drinking
water, groundwater, and soil. Analytes and reporting limits in water and soil areprovided in Table 1.

Summary of Method
2.1

The extracts are analyzed by LC/MS/MS. The PFCs are separated from othercomponents on an HPLC C18 column with a mobile phase of 0.010 M ammonium
acetate in 90:10 water:methanol. The mass spectrometer detector is operated in
the electrospray (ESI) negative ion mode.
Ion transitions monitored forquantitation are listed in Tables 2 and 3. Sample results are quantitated using theisotope dilution (internal standard) method.

2.2

Labeled PFCs are used in this analysis as an isotope dilution standard. The
isotope dilution technique allows correction for analytical bias encountered whenanalyzing more chemically complex environmental samples. Known quantities ofthe labeled PFC compounds are added to every sample and to the batch quality
control samples prior to extraction. Because the isotopically labeled compounds
are chemically similar to the compounds of concern, they are affected by any
interfering substances in the sample to the same extent as the compound ofconcern. The recoveries of the labeled PFCs are then used to mathematicallycorrect the final results for the PFCs.

2.3

The perfluorinated hydrocarbon sulfonates are received from the vendor as
sodium or potassium salts. These compounds are treated as anions throughout
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the method, including the calibration concentrations, and the results are reported
as the anions or the acids depending upon client requests. The concentration of
the anions and the acids are the same to two significant figures, therefore, thesample concentration will be the same whether analyzes as the anion or the acidfor these compounds.
2.4

3.0

There are four separate methods that are described in this SOP:
2.4.1

PFC – for the analysis of the full spectrum of PFC compounds, except
FOSA in water samples

2.4.2

PFC_FOSA – for the analysis of FOSA in water samples

2.4.3

LCMS_PFOA – for the analysis of only PFOA, APFO and PFOS in bothwater and soil samples

2.4.4

LCMS_HFPO – for the analysis of HFPO in water samples

Definitions
Refer to the Glossary of the TestAmerica Denver Quality Assurance Manual (QAM) andpolicy DV-QA-003P, “Quality Assurance Program,” for definitions of general analytical and
QA/QC terms.

4.0Interferences
4.1

Perfluorinated compounds have been used in a wide variety of manufacturing
processes, and laboratory supplies should be considered potentially contaminateduntil they have been tested and shown to be otherwise. The materials andsupplies used during the method validation process have been tested and shown
to be clean. These items are listed below in Section 6.

4.2

To avoid contamination of samples, standards are prepared in a ventilation hood inan area separate from where samples are extracted.

4.3

PTFE products can be a source of PFOA contamination. The use of PTFE in theprocedure should be avoided.

4.4

Volumetric glassware and syringes are difficult to clean after being used forsolutions containing high levels of PFCs and PFSs. These items should belabeled for use only with similarly concentrated solutions or verified clean prior to
re-use. To the extent possible, disposable labware is used.

4.5

Aluminum foil and foilware has been identified as a possible source of PFOA
contamination. These materials should be avoided during the extraction process.

4.6

PFC compounds can adhere to glass surfaces to varying degrees. Glasscontainers should be avoided as much as possible during the extraction andCompany Confidential & Proprietary

SOP No. DV-LC-0012, Rev. 15
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analysis of the samples.
4.7

5.0

Solvents used on the HPLC instrumentation can also be a source ofcontamination. Highest purity solvents are used to minimize the presence ofcontaminants. Also, the solvents are passed through an analytical column(Section 6.1.1.3) prior to interaction with the sample, to chromatographicallyseparate any contaminants from the target compounds in the sample.

Safety5.1

Employees must abide by the policies and procedures in the Environmental Healthand Safety Manual, Radiation Safety Manual and this document.

5.2

This procedure may involve hazardous material, operations and equipment. ThisSOP does not purport to address all of the safety problems associated with its use.
It is the responsibility of the user of the method to follow appropriate safety, waste
disposal and health practices under the assumption that all samples and reagentsare potentially hazardous. Safety glasses, gloves, lab coats and closed-toe,nonabsorbent shoes are a minimum.

5.3

Specific Safety Concerns or Requirements

5.4

5.3.1

Preliminary toxicity studies indicate that PFC compounds could havesignificant toxic effects. In the interest of keeping exposure levels as low asreasonably achievable, PFCs must be handled in the laboratory as if they
are hazardous and toxic chemicals.

5.3.2

Eye protection that satisfies ANSI Z87.1 (as per the Environmental Healthand Safety Manual), laboratory coat, and nitrile gloves must be worn whilehandling samples, standards, solvents, and reagents. Disposable gloves
that have been contaminated will be removed and discarded; other gloves
will be cleaned immediately.

5.3.3

PFCs are acids and are not compatible with strong bases.

5.3.4

PFCs are not known to be highly flammable. However, methanol is highly
flammable and is used throughout this procedure. Methanol should be
stored away from any ignition sources and kept in closed containers withsecondary containment measures or within a fume hood.

Primary Materials Used
The following is a list of the materials used in this method, which have a serious orsignificant hazard rating. Note: This list does not include all materials used in
the method. The table contains a summary of the primary hazards listed in
the SDS for each of the materials listed in the table. A complete list of
materials used in the method can be found in the reagents and materials section.
Employees must review the information in the SDS for each material before using
it for the first time or when there are major changes to the SDS.
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SOP No. DV-LC-0012, Rev. 15
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Material (1)Exposure
Limit (2)Hazards

Hydrochloric
Acid

Corrosive

SodiumHydroxide

Corrosive

Formic Acid

Combustible

PoisonPoisonCorrosive

5 ppm(Ceiling)

2 mg/cm(Ceiling)

Signs and symptoms of exposure
Can cause pain and severe burns upon inhalation,
ingestion, eye or skin contact. Exposure to concentrated
solutions may cause deep ulcerations to skin, permanenteye damage, circulatory failure and swallowing may be fatal.

3

Severe irritant. Can cause severe burns upon inhalation,
ingestion, eye or skin contact. Exposure to concentrated
solutions may cause severe scarring of tissue, blindness,and swallowing may be fatal.

5 ppm
(TWA)

Causes severe burns, toxic by inhalation, harmful if
swallowed, possible sensitizer.

200 ppm(TWA)

A slight irritant to the mucous membranes. Toxic effects
exerted upon nervous system, particularly the optic nerve.
Symptoms of overexposure may include headache,drowsiness and dizziness. Methyl alcohol is a defatting
agent and may cause skin to become dry and cracked. Skin
absorption can occur; symptoms may parallel inhalation
exposure. Irritant to the eyes.

25 ppm(TWA)

Causes severe burns and possible irreversible eye damage.
May cause severe and permanent damage to the digestive
tract.

Flammable
MethanolFlammable
PoisonIrritant

Ammoniumhydroxide

CorrosiveToxic

1 – Always add acid to water to prevent violent reactions.
2 – Exposure limit refers to the OSHA regulatory exposure limit.

6.0

Equipment and Supplies
6.1

Instrumentation
6.1.1

HPLC system connected to a Waters Quattro Micro MS/MS detector(LCMS-4), Waters XEVO MS/MS detector (LCMS-7) or an Agilent 6460
MS/MS detector (LCMS-5).
6.1.1.1(LCMS_4 LCMS-7) Phenomenex Synergi 4u Hydro
75x3.0mm and security Guard cartridge or equivalent

6.1.1.2(LCMS_5) Phenomenex Gemini NX analytical column and
Security Guard cartridge, or equivalent.

6.1.1.3(LCMS_5):
ZORBAX Eclipse Plus C18 (Agilent) column,
4.6x30mm, 3.5-micron, installed after solvent mixer, prior to
injector.

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6.2

6.3

Supplies
6.2.1

Volumetric Flasks (Class A): 100 mL; 200 mL; 500 mL; 1000 mL.

6.2.2

2.0-mL autosampler vials, Waters PN 186000307, or equivalent.

6.2.3

Miscellaneous laboratory apparatus (beakers, test tubes, volumetric flasks,
pipettes etc.). These should be disposable where possible, or marked andsegregated for high-level versus low-level use.

Computer Software and Hardware
Please refer to the master list of documents and software located on
R:\QA\Read\Master List of Documents\Master List of Documents, Software andHardware.xls or current revision for the current software and hardware to be used
for data processing.

7.0

Reagents and Standards
7.1

The solvents used for this procedure must be HPLC grade or better. LCMS-grade
methanol is used during sample preparation, standards preparation and instrument
analysis to minimize potential contamination due to solvent impurities.

7.2

Mobile Phase
0.010 M Ammonium Acetate in 90:10 water : methanol – Weigh 0.72 - 0.88 g ofammonium acetate. Place in a 1 L class A volumetric flask and dilute to 1 L with a
90:10 solution of water: methanol.

7.3

Stock Standard Materials
PFCs and labeled PFCs are purchased at certified concentrations in solution.
Standard materials are verified compared to a second source material at the timeof initial calibration. The Compounds Perfluorohexanesulfonate, Perfluorooctane
Sulfonate and Techinical Ammonium Perfluorooctanate are obtained as thebranched isomers from Wellington Laboratories (product code br-PFHxSK, brPFOSK and T-PFOA respectively)
7.3.1

The stock standard solutions are stored at ≤6°C. Stock standard
solutions should be brought to room temperature before using.
Standards are monitored for signs of degradation or evaporation.
Standard solutions must be replaced at least annually from the date of
opening or earlier if the vendor indicates an earlier expiration date.

7.3.2

All standards are stored in HDPE bottles and vials at ≤6°C and areassigned an expiration date of one year or earlier if the parent stock
materials have an earlier expiration date. Dilutions from stocks may not

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SOP No. DV-LC-0012, Rev. 15
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be assigned expiration dates that exceed the expiration of the stock
materials. Glass containers are avoided.
7.4

The perfluorinated hydrocarbon sulfonates are received from the vendor as
sodium or potassium salts. These compounds are treated as anions throughout
the method, including the calibration concentrations, and the results are reported
as the anions or the acids depending upon client requests. The concentration of
the anions and the acids are the same to two significant figures, therefore, thesample concentration will be the same whether analyzes as the anion or the acidfor these compounds.
7.4.1

The primary vendor lists both the concentration of the salt and the anion
on the certificate of analysis. The concentration of the anion isdetermined by multiplying the concentration of the salt by the ratio of themolecular weights of the anion to the salt, example:

MWanion [ Salt]
MWsaltEquation 1

MWacid [ Salt]
MWsaltEquation 2[Anion]=[Acid] =Example calculations:

MW Ratio(Anion / Na salt)

MW Ratio(Anion / K salt)

PFBS

0.929

0.885

PFHxS*

0.945

0.911

PFOS*

0.956

0.928

PFDS

0.963

0.939

*Both the branched and linear isomers are used to evaluate thesecompounds
7.5

LCS Standards (Spike Solutions)
The LCS stock solutions are prepared by diluting the appropriate amounts of PFC
stock solutions in LCMS-grade methanol. These standards are at a concentration of
0.5 µg/mL. These standards are stored in HDPE bottles at ≤6°C and given a 1 year
expiration date. See DV-OP-0019 for details. These standards are also used tospike the MS and MSD extracts.
7.5.1

PFC Method LCS:
A spike solution is prepared containing all PFC compounds at 0.5 g/mL byCompany Confidential & Proprietary

SOP No. DV-LC-0012, Rev. 15
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diluting 2.5 mL PFAC-MXB-Stk (2.0 g/mL) and 0.1 mL FOSA-Stock (50.0g/mL) to 10 mL in LCMS-grade methanol.
7.5.2

PFC_FOSA Method LCS:
A spike solution is prepared containing FOSA at 0.05 g/mL by diluting 0.1
mL FOSA-Stock (50.0 g/mL) to 100 mL in LCMS-grade methanol.

7.5.3

LCMS_PFOA Method LCS
A spike solution is prepared containing PFOA and PFOS at 0.5 g/mL by
diluting 1.0 mL PFOA-stock (50.0 g/mL) and 1.0 mL PFOS-Stock (50.0g/mL) to 100 mL in LCMS-grade methanol.

7.5.4

HFPO Method LCS
A spike solution is prepared containing HFPO at 0.5 μg/mL by diluting 1 mL
of HFPO-DA stock (50.0 μg/mL) to 100 mL in LCMS grade methanol.

7.5.5

Table of LCS spike volumes and final sample concentrations:

MethodPFC

PFC_FOSALCMS_PFOALCMS_HFPO

7.6

MatrixVolume of
Spike Added

Concentrationin Sample

Water

0.1 mL0.2 g/L

Soil

0.4 mL20 g/kg

Water

1.0 mL0.2 g/L

Soil

N/A

N/A

Water

0.1 mL0.2 g/L

Soil

0.4 mL20 g/kg

Water

0.1 mL0.2 μg/L

Soil

N/A

N/A

Surrogate Standard Solutions, 0.5 g/mL (Instrument)
A surrogate spike solution is prepared containing the labeled PFC isotopes, 13C8PFOA and 13C8 PFOS, at 0.5 g/mL by diluting 0.1 mL 13C8PFOA-stk (50.0 g/mL)
and 0.1 mL 13C8PFOS-stk (50.0 g/mL) to 10 mL in LCMS-grade methanol. Thissurrogate solution is utilized for the PFC method and LCMS_PFOA method, the
PFC_FOSA method does not utilize a surrogate. Standards are stored in HDPE
bottles at ≤6°C and given an expiration date of one year or earlier if the parent
stock materials have an earlier expiration date.

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7.7Internal Standard Solutions, 0.5 g/mL (Instrument)
All individual internal standards and surrogates are received from the vendor at aconcentration of 50.0 g/mL. Internal standard solutions are prepared for eachmethod by combining 0.1 mL of each compound listed below and diluting to 10 mL in
LCMS-grade methanol. Standards are stored in HDPE bottles at ≤6°C and given
an expiration date of one year or earlier if the parent stock materials have an
earlier expiration date.
PFC Method:

13

13

13

13

13

13

13

18

13

C4 PFBA
C4 PFNA
C4 PFDoAC2 PFHxAC2 PFDAO2 PFHxSC4 PFOAC2 PFUnA
C4 PFOS13

C8 FOSALCMS_PFOA Method: 13C4 PFOALCMS_HFPO Method
7.8

13

C4 PFOS13

C3 HFPOInternal Standards / Surrogates Spiking Solutions (Prep)
All individual internal standards and surrogates are received from the vendor at aconcentration of 50.0 g/mL. An internal standard / surrogate solution is prepared bydiluting 1.0 mL of each compound to 100 mL in LCMS-grade methanol. Standards
are stored in HDPE bottles at ≤6°C and given an expiration date of one year or
earlier if the parent stock materials have an earlier expiration date.
PFC Method:
Internal Standards:

13

13

13

13

13

13

13

18

13

13

13

13

C4 PFBA
C4 PFOAC2 PFUnA
C4 PFOSSurrogates:

C8 PFOAC2 PFHxA
C4 PFNA
C4 PFDoA
C8 FOSA
C8 PFOSPFC_FOSA Method:
Internal Standard:

13

C8 FOSALCMS_PFOA Method:
Internal Standards:

13

13

Surrogates:

13

13

C4 PFOAC8 PFOAC4 PFOSC8 PFOSLCMS_HFPO Method:
Internal Standard:

13

C3 HFPO

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13

Surrogate:
7.9

C3 HFPO

Calibration Stock Standard (Instrument)
The calibration stock solution is prepared by diluting the appropriate amounts of PFC
LCS solution and surrogate solution in 80% methanol in water. This standard is at aconcentration of 20 µg/L and is diluted with 80% methanol in water to produce otherinitial calibration standards. For the HFPO calibration a 50% methanol in water
solution is used for preparation of the calibration standards. For the PFC_FOSA
method, the PFC standards are used to calibrate the instrument. Standards are
stored in HDPE bottles at ≤6°C and given an expiration date of one year or earlier
if the parent stock materials have an earlier expiration date.

7.10

7.9.1

For the PFC method, take 0.4 mL of the PFC LCS Spike Solution and 0.4
mL of the Surrogate Standard Solution (Instrument) and dilute to 10 mL with
80% methanol in water.

7.9.2

For the LCMS_PFOA method, take 0.4 mL of the LCMS_PFOA LCS Spike
Solution and 0.4 mL of the Surrogate Standard Solution (Instrument) anddilute to 10 mL with 80% methanol in water.

Initial Calibration (ICAL) Levels (Instrument)
7.10.1

The calibration levels in the table below are provided as an example. Final
volume of each calibration level standard is 1 mL and standards arebrought to volume with 80% methanol in water. To each calibration
standard, 20 L of Internal Standard solution (Section 7.7) must be added.

7.10.2

Note that the PFS compounds are purchased in solutions at certified
concentrations as salts. They are reported as anions or acids depending
upon client request. The concentration is adjusted to be expressed as theanion or acid concentration. If a different salt is used (e.g. potassium
versus sodium), these concentrations would be different.

ICAL
Level

Volume Stk Soln
to Add (L)On
ColumnConc. ofPFCs
(g/L)

Equiv.
SampleConc
Water(ng/L)

Equiv.
SampleConc
Soil(g/kg)On
ColumnConc. ofPFBS asthe anionor acid(µg/L)On Column
Conc. ofPFHxS asthe anionor acid(µg/L)On
ColumnConc. ofPFOS asthe anionor acid(µg/L)On Column
Conc. ofPFDS asthe anion oracid (µg/L)

1

10L CALSTK

0.2

4

0.4

0.177

0.189

0.191

0.193

2

25L CALSTK

0.5

10

1.0

0.44250.47250.47750.4825

3

50L CALSTK

1.0

20

2.0

0.885

0.945

0.955

0.965

4

100L CALSTK

2.0

40

4.0

1.77

1.89

1.91

1.93

5

250L CALSTK

5.0

100

10.0

4.425

4.725

4.775

4.825

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Equiv.
SampleConc
Water(ng/L)

Equiv.
SampleConc
Soil(g/kg)On
ColumnConc. ofPFBS asthe anionor acid(µg/L)On Column
Conc. ofPFHxS asthe anionor acid(µg/L)On
ColumnConc. ofPFOS asthe anionor acid(µg/L)On Column
Conc. ofPFDS asthe anion oracid (µg/L)

ICAL
Level

Volume Stk Soln
to Add (L)On
ColumnConc. ofPFCs
(g/L)

6500 L CALSTK

10

200

20.0

8.85

9.45

9.55

9.65

7

40 L LCS & 40
L Surrogate

20

400

40.0

17.7

18.9

19.1

19.3

8

100 L LCS &100 L Surrogate

50

1000

100

44.25

47.25

47.75

48.25

9

250 L LCS &250 L Surrogate

125

2500

250

110.625118.125

119.375

120.625

The calibration range for HFPO varies slightly from the concentrations listed in thetable above, beginning with an on column concentration of 0.25 μg/L and ending withan on column concentration of 100 μg/L for a range of equivalent sample
concentration of 5 ng/L to 2000 ng/L. Actual HFPO on-column concentrations for theentire nine-point curve are: 0.25, 0.50, 1.0, 2.0, 5.0, 10, 25, 50 and 100 μg/L.
7.11

Initial Calibration Verification Standard (ICV), 2.0 g/L
7.11.1The second source standard must be obtained from a different source
than the standards used for initial calibration. This standard is used toverify the accuracy of the calibration standards.

7.11.2In the case of the PFC and PFC_FOSA methods, a custom standard ispurchased from Supelco with most PFC compounds at 1 g/mL; asnecessary, the concentrations of the sulfonate salts are adjusted to theanion concentration. Second source standards for PFDS and FOSA are
not currently available, so these compounds are added from separate
vials purchased from the primary vendor. If a second lot is not available,
the standard is prepared by a different analyst. These standards are
combined and diluted to a final concentration of 2.0 g/L for analysis at
the instrument.

7.11.3In the case of the LCMS_PFOA method, the second sources of PFOA
and PFOS are purchased from Accustandard and diluted to a final
concentration of 2.0 g/L for analysis at the instrument.

7.11.4In the case of HFPO, the second source standards are not currently
available. These compounds are added from separate vials purchasedfrom the primary vendor. The standard is diluted to a final concentrationof 2.0 μg/L for analysis at the instrument.

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7.12

Continuing Calibration Verification Standards (CCV), 5.0 g/L and 10.0 g/L
The continuing calibration verification (CCV) standards alternate between the 5.0
g/L solution (L5) and the 10.0 g/L solution (L6) of PFCs in 80% methanol in water
made from the same source as the initial calibration standards.

8.0

Sample Collection, Preservation, Shipment and Storage
Sample container, preservation techniques and holding times may vary and are dependent
on sample matrix, method of choice, regulatory compliance, and/or specific contract or client
requests. Listed below are the holding times and the references that include preservation
requirements.

9.0

Min.
SampleSize

Preservation

HDPE

250 mL3(x2)

Cool, < 6 C

HDPE

10 g

Cool, < 6 C

MatrixSampleContainer

Water
Soil

2

Extraction
Holding1Time

Analytical
HoldingTime

Referenceo

14 Days

40 DaysN/Ao

14 Days

40 Days

N/A

4

4

1

Extraction holding times were determined based on stability studies performed during thedevelopment of this method. TestAmerica Denver has conducted stability studies indicating thatmedium- and low-level standard solutions of PFOA are stable for at least two months in glass,polystyrene, and polypropylene plastics at 0-6 °C.

2PFCs have been shown to adhere to glass. Glass containers are not an appropriate sample
collection vessel.

3

If FOSA is a target analyte for a specific client’s samples in addition to other PFCs, duplicate water
samples are required because FOSA is extracted separately and therefore 2-250 mL bottles are
required.

4This method was developed and validated by TestAmerica Denver.

Quality Control
9.1

The minimum quality controls (QC), acceptance criteria, and corrective actions aredescribed in this section. When processing samples in the laboratory, use the
LIMS Method Comments to determine specific QC requirements that apply.
9.1.1

The laboratory’s standard QC requirements, the process of establishing
control limits, and the use of control charts are described more
completely in TestAmerica Denver policy DV-QA-003P, Quality
Assurance Program.

9.1.2

Specific QC requirements for Federal programs, e.g., Department of
Defense (DoD) and Department of Energy (DOE), are described inCompany Confidential & Proprietary

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TestAmerica Denver policy DV-QA-024P, Requirements for Federal
Programs. This procedure meets all criteria for DoD QSM 5.0 unlessotherwise stated.
Any deviation or exceptions from QSM 5.0
requirements must have prior approval of the client in the projectrequirements. See Table 5 for a summary of the DoD QSM 5.0
requirements for this method and possible client approved technicalspecifications.

9.2

9.3

9.1.3

Project-specific requirements can override the requirements presented inthis section when there is a written agreement between the laboratory
and the client, and the source of those requirements should be describedin the project documents.
Project-specific requirements arecommunicated to the analyst via Method Comments in the LIMS and the
Quality Assurance Summaries (QAS) in the public folders.

9.1.4

Any QC result that fails to meet control criteria must be documented in a
Nonconformance Memo (NCM). The NCM is automatically sent to thelaboratory Project Manager by e-mail so that the client can be notified as
appropriate. The QA group periodically reviews NCMs for potential
trends. The NCM process is described in more detail in SOP DV-QA0031. This is in addition to the corrective actions described in thefollowing sections.

Batch Definition
9.2.1

Batches are defined at the sample preparation stage. The batch is a set of
up to 20 samples of the same matrix, plus required QC samples, processedusing the same procedures and reagents within the same time period.
Batches should be kept together through the whole analytical process as far
as possible. See Policy DV-QA-003P for further details.

9.2.2

Since the extraction method for waters has a separate FOSA method, eachjob of water samples submitted for PFC testing will be separated into atleast two batches, one for FOSA and one for all other PFCs. There will also
be two separate runs on the instrument.

QC Samples
9.3.1

Method Blank
9.3.1.1

A method blank is prepared with each batch of samples. The
method blank demonstrates freedom from contamination in thelaboratory. The standard matrix used for method blanks
consists of reagent water (for aqueous sample batches) andreagent sand (for soil sample batches) plus the isotopicallylabeled internal standard / surrogate spike (Section 7.8).

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9.3.1.2

9.3.2

For the FOSA extraction from water samples, surrogates are notadded to any sample or QC, recovery of the internal standard isused to evaluate extraction performance.
Acceptance Criteria:

The method blank result must be lessthan ½ the reporting limit (RL) for all
target compounds.

Corrective Action:

If the method blank exceeds ½ the RL,
the associated samples must be
evaluated and any samples with hits
less than 10x the concentration in the
MB must be re-extracted andreanalyzed. If the associated samples
are reported as ND for the compounds
that are found in the MB the data may
be reported if the program or client
requirements allow.
If results arereported, they must be qualified and an
NCM generated.

Laboratory Control Standard (LCS)
Each batch of samples is prepared with an LCS, spiked as described in
Section 0. This QC sample serves to verify within-batch accuracy and longterm precision at mid-level concentrations. The LCS matrix for aqueous
samples consists of reagent water and the LCS matrix for soil samples
consists of reagent sand (Ottawa sand).

9.3.3

Acceptance Criteria:

The percent recovery must fall within control limits,which are set at  3 standard deviations aroundthe historical mean. Historical limits are stored in
the LIMS.

Corrective Action:

If the recovery falls outside of the control limits,corrective action must be taken. All associatedsamples must be re-extracted and reanalyzed. If
the recovery is biased high and the analyte ofinterest is not detected in the samples, results canbe reported with a comment in the final report casenarrative.
Flag the data and document thedecision in an NCM. This is an exception to DoD
QSM 5.0 criteria and must be approved by theclient as documented in the project records.

Matrix Spike Samples
The matrix spike (MS) is an aliquot of a selected field sample that is spiked
with the analytes of interest. The matrix spike duplicate (MSD) is a second

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aliquot of the selected sample that is spiked in the same manner as the MS.
At a minimum, the laboratory must spike one sample in every batch. The
MS/MSD must be analyzed at the same dilution level as the un-spiked
sample, unless the matrix spike components would then be above thecalibration range. The matrix spike sample is prepared as described in
Section 0. The matrix spike demonstrates acceptable accuracy for
individual samples.
Acceptance Criteria: The MS and MSD recoveries and the relative
percent difference (RPD) between the MS and
MSD results must be within the establishedcontrol limits. Percent recovery control limits are
set at  3 standard deviations around thehistorical mean of the LCS recovery data, unlessotherwise dictated by the client or project. The
RPD control limit is set at 3 standard deviations
above the mean of the historical data.
NOTE:

Corrective Actions:

DOD QSM 5 limits apply to projects
performed under this program.

The information obtained from MS data aresample/matrix specific and are not normally used
to determine the validity of the entire batch. If the
MS and/or MSD recovery falls outside of theestablished control limits, the bracketing CCV
and batch LCS recoveries must be within control
limits in order to accept results for the associatedsamples. The following corrective actions are
required for MS/MSD recovery failures to rule out
lab error:Checkcalculation
performance;Verify, if possible, that the MS and MSD werespiked correctly (e.g., very low or very highrecoveries);Consider objective evidence of matrix
interference (e.g., heterogeneous sample,
interfering peaks seen on chromatograms, orinterference demonstrated by prior analyses);

Flag the data for any results outside of
acceptance limits.

For any single RPD failure, checkcalculations; verify, if possible, that the MS
and MSD were spiked correctly; check
instrument performance; consider objective

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evidence of matrix interference or sample
inhomogeneity; and flag the data.


If both the parent sample and associatedmatrix spike results are over range the parent
and the spikes shall be diluted by the sameamount and the results from the reanalysis
reported for both. If the analyte concentrationin the parent sample is greater than fourtimes the concentration of spike added, thenspike recovery results are not compared tocontrol limits, and the recovery is either
reported as “NC” (not calculated) or with aqualifier flag to indicate that the spike wasless than four times the analyte concentrationin the sample. If the dilution will cause thespike to be less than two times the reporting
limit, the MS/MSD do not need to be diluted
and the recovery reported as “NC” (notcalculated).



For MS/MSD that serve as batch QC, if theparent sample result is within the calibration
range and the MS/MSD results are above thecalibration range, the results are reportedwith the MS/MSD result being flagged as anover-range measurement (e.g., the E-flag
qualifier).



If the MS/MSD are client requested, theparent sample result is within calibration
range and the MS/MSD results are above thecalibration range, the sample and spikeshould be diluted, keeping in mind that weneed to assess whether or not the dilution will
best serve the client’s needs. Consult with
the PM as needed. Both the parent sample
and MS/MSD samples must have the same
dilution factor. Some EDDs do not accept
data that are at different dilution factors.



If the native analyte concentration in the
MS/MSD sample exceeds 4 times the spikelevel for that analyte, the recovery data arereported as NC (i.e., not calculated) and theappropriate qualifier flags are added.

NOTE: See Denver Policy Memorandum P16-001 andCorporate Policy Memorandum CA-Q-QM-013
for more detail.

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NOTE: Some client programs require reanalysis toconfirm matrix interferences. Check special
project requirements for this corrective action.
NOTE:

9.3.4

If the recovery is outside the control limits forboth the matrix spikes and the LCS, then theentire batch should be re-prepared andreanalyzed.

Sample Duplicate and/or Matrix Spike Duplicate (MSD)
Depending on client requirements, a sample duplicate and/or a matrix spike
duplicate (MSD) may be prepared and analyzed. Comparison of duplicate
results provides an assessment of the precision of the method in actual
sample matrices.

9.3.5

Acceptance Criteria:

Acceptance criteria for the relative percent
difference (RPD) between the duplicate sample
results are specified by client or projectrequirements. Control limits for the RPD may bebased on historical precision of the method or may
be project-specific limits. If the duplicate is an
MSD, then the recovery for the analyte(s) ofinterest must also meet the established control
limits as described in Section 9.3.3 above. For
DoD QSM 5.0, the acceptance limits for the RPD
for all analytes is < 30%.

Corrective Action:

Corrective actions or data reporting based on
duplicate results are specified in client or projectrequirements, which are provided outside of thescope of this SOP. The analyst must ensure that
MS/MSD recoveries or RPD limits outside of
acceptance criteria are not due to lab error.

Isotopically Labeled Surrogate Standard
The surrogate spike solution is added to each field and QC sample as
described in Sections 7.6 and 7.8. For the FOSA extraction from water
samples, surrogates are not added to any sample or QC, recovery of theinternal standard is used to evaluate extraction performance.
Acceptance Criteria:

The recovery of the labeled surrogate standards
are based on historical data, and are 3 standard
deviations around the mean. Historical limits are
stored in the LIMS.

Corrective Action:

If the recoveries are not within these ranges, thesample should be re-extracted to determine if thisCompany Confidential & Proprietary

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is an extreme sample matrix effect.
9.3.6

Isotopically Labeled (Internal) Standard (IS)The isotope dilution spike solution is added to each field and QC sample as
described in Section 7.7. This is also the internal standard.
Acceptance Criteria:

The recoveries of the labeled internal standards
must be within 50% -150% of the true value or of
the expected concentration for a dilution up to x50.

Corrective Action:

If the recoveries are not within these ranges insamples, the recovery for the batch QC (MB, LCS)
must be checked. If these recoveries are within
limits and the labeled standard(s) are outside ofcontrol limits in the sample, then the sample isconsidered to have an extreme matrix effect andan NCM must be generated.

If the batch QC (MB, LCS) recoveries are also out of control, the samples
must be re-extracted and reanalyzed.
9.4

Instrument QC
9.4.1

Initial Calibration Blank (ICB)
Immediately following the ICAL, a calibration blank is analyzed that consistsof an injection of 80% methanol in water containing the PFC internal
standards.

9.4.2

Acceptance Criteria:

The result for the calibration blank must be lessthan two times the method detection limit, or lessthan ½ the RL, which ever is smaller.

Corrective Action:

If the ICB does not meet the acceptance criteria,then the source of contamination must be
identified and any necessary cleaning completed,and then the instrument should be recalibrated.

Initial Calibration Verification (ICV)
Following the ICAL and the ICB, the ICV standard described in Section 7.11
is analyzed.

Acceptance Criteria:

The recovery for the ICV must be within 70-130%. DoD
QSM 5.0 requires ICV limits of 75-125%.

Corrective Action:

If results are outside these limits, the standards should be

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verified and/or remade, and then the instrument should berecalibrated.
9.4.3

Detection Limit Calibration Verification (DLCK)
Following the ICAL, ICB and ICV, the 0.5 g/L calibration standard (L2) is
re-analyzed to verify that detections at the reporting limit are accurately
quantified by the calibration curve.
Acceptance Criteria: The recovery for the DLCK must be within 70-130%.
Some programs (like West Sacramento) require a L1standard to be analyzed at the start of the sequence.
The recovery for the L1 standard must be within 50150%.
Corrective Action:

9.4.4

If results are outside these limits, the standards shouldbe verified and/or remade, and then the instrument
should be recalibrated.

Continuing Calibration Verification (CCV)The 5.0 g/L (L5) and 10.0 g/L (L6) CCV standards described in Section
7.12 are alternately analyzed after every 10 injections and at the end of theanalytical sequence.

10.0

Acceptance Criteria:

The recovery for the CCV standards must be 70-130%.
DoD QSM 5.0 requires CCV limits of 75-125%.

Corrective Action:

If this is not achieved, the instrument has drifted outside thecalibration limits. The instrument must be recalibrated or
two CCVs must be analyzed consecutively and pass. All
samples analyzed since the last passing CCV must bereanalyzed. If CCV recovery is >130% (125% for DoD QSM
5.0) and the samples are ND, the samples can be reported
without reanalysis. Flag the data and document thedecision in an NCM. This is an exception to DoD QSM 5.0
criteria and must be approved by the client as documented
in the project records.

Procedure
10.1

One-time procedural variations are allowed only if deemed necessary in theprofessional judgment of supervision to accommodate variation in sample matrix,
radioactivity, chemistry, sample size, or other parameters. Any variation inprocedure shall be completely documented using an NCM. The NCM is
automatically sent to the laboratory Project Manager by e-mail so that the client
can be notified as appropriate. The QA group periodically reviews NCMs forpotential trends. The NCM process is described in more detail in SOP DV-QA-

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0031. The NCM shall be filed in the project file and addressed in the casenarrative.
10.2

Any deviations from this procedure identified after the work has been completed
must be documented in an NCM, with a cause and corrective action described.

10.3

Sample Extraction
Refer to SOP DV-OP-0019 “Extraction of Perfluorooctanoic Acid (PFOA) and
Perfluorooctanoic Sulfonate (PFOS) and other Perfluorinated Hydrocarbons (PFCs)
in Water and Soil”.

10.4

Run Sequence
Analyze calibration standards, samples and quality control samples following theanalysis sequence shown in Table 4.

10.5

Sample Analysis
10.5.1

If results are to be reported as ammonium perfluorooctanoate (APFO),
instead of PFOA, a multiplier of 1.0406 is applied to the sample results tocorrect for the molecular weight differences between PFOA and APFO.
This adjustment is made during the preparation of the standards used forcalibration. APFO is calibrated using the same peak as PFOA.

10.5.2

Dilutions10.5.310.5.2.1

Samples that exceed the working range must be diluted with
80:20 methanol:water (for HFPO use 50:50 methanol:water) toconcentrations within the working range and reanalyzed.
Dilutions are done without fortifying the internal standard
concentrations as this is an isotope dilution method. Therefore,
maximum dilution factors will be dependent on the recovery of
the internal standards for each sample matrix. The maximum
dilution performed on an extract will be 50x. Beyond the
maximum dilution, the data will be E-flagged and reported as agreater than concentration.

10.5.2.2

If samples are known to be of high concentration the dilution
may be performed at the preparation step. Reporting limits will
be adjusted based on the dilution and further dilution of thesample extract may be performed if needed as described in
Section 10.5.2.1. This prep dilution allows the laboratory toextend the calibration range beyond that provided by extract
dilution alone.

Raw data, calibration summaries, QC data, and sample results arereviewed by the analyst. These must also be thoroughly reviewed by asecond qualified person. These reviews are documented on the LC/MS
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Data Review Checklist. The data review process is described in DV-QA0020.
10.6

Troubleshooting and Maintenance
For instrument maintenance guidelines see the Denver Quality Assurance Manual.

11.0

Calibration
11.1

Instrument Tuning
11.1.1

Prior to analyzing samples, evaluate the lowest standard for the ability to
see compounds. If sensitivity and/or resolution are compromised, infuse asolution of each target compound into the mobile phase at a point justbefore the entrance to the electrospray probe. The responses for the
precursor ions and product ions are observed and optimized for sensitivity
and resolution.

11.1.2

Instrument mass calibration is performed if the analyst notices mass
assignment errors, see Table 7 for solutions and masses to use.

11.1.3DoD QSM 5.0 requires a tune check containing all analytes of interest be
analyzed prior to an initial calibration and after any mass calibration or
maintenance is performed. The ICV may be used for this tune.
Acceptance Criteria: Mass assignments of tuning standard must bewithin 0.5 amu of true value.
Corrective Action:

11.2

Retune instrument. If the tuning does not meet
acceptance criteria, an instrument mass
calibration must be performed and the tuning
redone. No samples shall be analyzed without
acceptable tuning.

Initial Calibration (ICAL)
11.2.1

Detailed information regarding calibration models and calculations can befound in Section 11 and Corporate SOP, CA-Q-P-003, Calibration Curves
and the Selection of Calibration Points.

11.2.2

Routine instrument operating conditions are listed in Tables 2 & 3.

11.2.3

Target compound recommended internal standard associations are listed inTable 6.

11.2.4The instrument is calibrated using nine concentration levels (see Section
7.9 and 7.10 for details). These concentrations define the working range foranalysis.
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11.3

11.4

Rejection of Calibration Points
11.3.1

If the analyst believes that a point on the curve is inaccurate, the preferredpractice is to rerun the ICAL. Generally it is NOT acceptable to remove
points from a calibration for the purposes of meeting calibration criteria,
unless the points are the highest or lowest on the curve AND the reporting
limit and/or linear range is adjusted accordingly. A “Huge Error Test” orother statistical test should be used to determine that a point does not fit thecalibration model.

11.3.2The only exception is that a level may be removed from the calibration if thereason is clearly documented, for example a broken vial. A minimum of fivelevels must remain for linear fits, and there must be a minimum of six levels
for a quadratic fit. All raw data, including data from rejected calibration
points, will be included in the study package.

Linear Calibration Using Average Response Factors
For each target analyte, calculate the response factor of each calibration level asfollows:RFi 

Rx Cis
Ris

RFi
Rx
Cis
Ris

=
=
=
=

Where:

Equation 3
Response factor for the ith level
Response for analyte
Concentration for internal standard
Response for internal standard

For each target analyte, calculate the average response factor as follows:n RFi
Average Response Factor  RF  i 1nWhere:nRFi==

Equation 4

Number of calibration levels
Response factor for the ith level

The relative standard deviation (RSD) is calculated as follows:% RSD SD
 100 %
RF

Equation 5

Where SD is the standard deviation of the average RF, which is
calculated as follows: RF  RF 
N

SD 

i 1

2

i

N 1

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11.5

Average Calibration Factor Evaluation
The calibration relationship can be graphically represented as a line through theorigin with a slope equal to the average calibration factor.
Acceptance Criteria: The RSD of the average response factor must be <20%.
Also examine the residuals, especially for the high points
versus the fitted function. If the residual values are large,
a linear regression should be considered.
Corrective Action:

11.6

If the RSD is > 20%, average response factor cannot be
used and least-squares linear regression should beattempted.

Linear Fit
Calibration using least-squares linear regression produces a straight line that does
not pass through the origin. The calibration relationship is constructed by
performing a linear regression of the instrument response (peak area) versus theconcentration of the standards. The instrument response is treated as thedependent variable (y) and the concentration as the independent variable (x). The
weighting used is the reciprocal of the concentration or the reciprocal of the squareof the concentration. The regression produces the slope and intercept terms for alinear equation in the following form for an internal standard calibration:

Rx Cis m1 Cs   b
Ris
Where:

Cs

=

Rx
Ris
Cis
b
m1

=
=
=
=
=

Equation 7

Analyte concentration in calibration
standard, µg/L
Response for analyte
Response for internal standard
Concentration of internal standard
y - Intercept
Slope

To calculate the concentration in an unknown sample extract, the regression
equation is solved for concentration, resulting in the following equation:

 Rx Cis R  bCex   ism1
Where:

Cex
Rx
Ris
Cis
b

=
=
=
=
=

Equation 8
Extract analyte concentration, µg/L
Response for analyte
Response for internal standard
Concentration of internal standard
y - Intercept

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11.7

Evaluation of the Linear Least-Squares Regression Calibration Function:
11.7.1

With an unweighted linear regression, points at the lower end of thecalibration curve have less weight in determining the curve than points at
the high concentration end of the curve. For this reason, inverse
weighting of the linear function is recommended to optimize the accuracy
at low concentrations.

11.7.2

Note that the August 7, 1998 EPA memorandum “Clarification Regarding
Use of SW-846 Methods”, Attachment 2, Page 9, includes the statement“The Agency further recommends the use of this for weighted regression
over the use of an unweighted regression.”
Acceptance Criteria: The linear regression must have a correlation
coefficient (r) ≥ 0.99 (r2 ≥ 0.98). In order to meet
support work for the West Sacramento laboratory
the linear regression must have a correlation
coefficient (r) ≥ 0.995 (r2 ≥ 0.990).

11.8

Quadratic Fit
When the instrument response does not follow a linear model over a sufficiently
wide working range, or when the previously described calibration approaches failacceptance criteria, a non-linear, second-order calibration model may be
employed. The second-order calibration uses the following equation for an internal
standard calibration:

RxCis2
 m2 Cs   m1 Cs   b
Ris
Where:

Cs

=

Rx
Ris
Cis
m2
m1
b

=
=
=
=
=
=

Equation 9

Analyte concentration in calibration
standard, µg/L
Response for analyte
Response for internal standard
Concentration of internal standard
Curvature
Slopey - Intercept

To calculate the concentration in an unknown sample extract, the roots of thequadratic equation are solved for:

 m1 Cex 
Where:







Ris 

m1 2  4m2  b  RxCis 
2m2

Cex
Rx

=
=

Equation 10

Extract analyte concentration, µg/L
Response for analyte

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Ris
Cis
m2
m1
b
11.9

=
=
=
=
=

Response for internal standard
Concentration of internal standard
Curvature
Slopey – Intercept

Evaluation of Second-Order Regression Calibration:
A minimum of six points must be used for a second-order regression fit.
Acceptance Criteria:
11.9.1

Second-order regressions should be the last option, and note that some
programs (e.g., South Carolina) do not allow the use of second-order
regressions.

11.9.2The coefficient of determination (COD, r2) must be ≥ 0.99.

11.9.3

The response increases significantly with increasing standard
concentration (i.e., the instrument response does not plateau at highconcentrations).

11.9.4The distribution of concentrations is adequate to characterize thecurvature.

11.9.5

Calibrations are modeled as calibration curves.
The following requirements must be met for any calibration to be used. Ifthese criteria are not met, instrument conditions and standards will bechecked, and the ICAL successfully repeated before continuing.

12.0

11.9.6

Response must increase with increasing concentration.

11.9.7

The absolute value of the intercept of the curve at zero response must be
less than the concentration that corresponds to ½ the Reporting Limit.

11.9.8Per Minnesota QAS for sample results from sites in Minnesota, therecovery (accuracy) for each point in the calibration curve must be 75-125%
except for the lowest point in the curve which must be 70-130%.

Calculations/ Data Reduction
12.1

Qualitative Identification
12.1.1An analyte is identified by retention time and ion mass (see Tables 2 and
3). The sample component retention time must compare to within ± 0.5
minute of the average retention time of the standard component in theinitial calibration. If a compound cannot be verified by all of the above

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criteria, but in the technical judgment of the analyst the identification iscorrect, the analyst shall report that identification and proceed withquantitation. This decision is documented in an NCM.

12.2

12.1.2The perfluorinated hydrocarbon acids and sulfonates may have bothlinear and branched isomers due to the manufacturing process. These
isomers have been studied and identified for PFOA and PFOS, however,
the other compounds are also likely to have isomers. The linear andbranched isomers have the same MS/MS transitions; however theretention time of the branched isomers is generally earlier and the ratio of
the quant and qual ions may be different. The calibration standards arecomposed primarily of the linear isomers. For samples, all isomers
should be integrated and quantified; if there is a double peak due to thedifference in retention time, the entire area of both peaks should be
integrated.

12.1.3For all compounds except PFBA and PFPA, two product ions aremeasured, a quant ion and a qual ion. The ratio of the peak areas of thetwo PFC ions is calculated and generally within  30% of the ion ratios forthe mid-point ICAL.

12.1.4

While the linear and branched isomers described in Section 12.1.2 produce
the same product ions, the ratio is not necessarily the same for both types
of isomers. If the ion ratio is outside of this 30% window, but in theanalyst’s judgment, there are still PFCs present due to retention time andpeak presence, the results will still be reported as valid detections.

Manual Integrations
Upon completion of the analytical sequence, transfer the raw instrument data to
Chrom for further processing. Review the chromatograms to ensure correct
assigning of peaks and correct integration of each peak. If manual data
manipulations are necessary, they must be justified and documented. See DVQA-011P requirements for manual integration.

12.3

Calculations for Calibration Standards and Sample Extracts
The concentration of each identified analyte and surrogate in the extract is
calculated from the linear or quadratic curve fitted to the initial calibration points, orfrom the average response factor (RF) of the initial calibration.

12.4

Calculations for reporting original samples
12.4.1

Calculating concentration in an aqueous / soil samples:
Note: Perfluorinated hydrocarbon sulfonate compounds are calibrated
as the anion and reported as either the anion concentration oracid concentration based on client request. (See Section 7.4)

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The concentration in a water sample is calculated as follows:

Concentration, g/L =

CexVt
xDF
Vo

Where:

=
=
=
=

Cex
Vt
Vo
DF

Equation 11

Extract analyte concentration, µg/L
Volume of total extract in liters
Volume of water extracted in liters
Dilution factor

The concentration in a sediment/soil, sludge (on a dry-weight basis) andwaste (normally on a wet-weight basis) is calculated as follows:

Concentration, g/kg =Where:

12.4.2

CexVt
 DF
WsD

Cex
Vt
Ws

=
=
=

D

=

DF

=

Extract analyte concentration, µg/L
Volume of total extract in liters
Weight of sample extracted or
diluted in kilograms(100 - % moisture in sample)/100,
for dry weight basis or 1 forwet-weight basis
Dilution factor

LCS and CCV Percent Recovery

ControlSpike Recovery =12.4.3Equation 12

Where (in g/L):
SSR

=

SA

=

S SR
 100 %
SA

Equation 13

Calculated analyte concentration of
spiked sample
Concentration of standard added

MS / MSD Percent Recovery Calculation

Matrix Spike Recovery 

SSR  SR
100 %SA

Where (in g/L):
SSR

=

SR

=

SA

=

Equation 14

Calculated analyte concentration of
spiked sample
Calculated analyte concentration ofparent sample
Concentration of standard added

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12.4.4

Relative Percent Difference Calculation for the MS/MSD

RPD 

MSR  MSDR
1 / 2( MSR  MSDR ) 100

Equation 15

Where:RPD =MSR =MSDR =
13.0

Relative percent difference
Matrix spike result of analyte
Matrix spike duplicate result of analyte

Method Performance
13.1

Method Detection Limits (MDL)
An initial method detection limit study is performed for each analyte and eachsample matrix type in accordance with the method validation protocol. MDLs are
stored in the LIMS. The procedure for determining detection limits is defined in
Policy CA-Q-S-006.

13.2

13.3

Method Detection Limit Verification (MDLV)
13.2.1

Calculated MDLs from the annual studies are subject to verification by
analyzing an MDLV standard prepared at 1-4 times the calculated MDL
concentration. An MDLV standard is analyzed immediately after each
MDL study and annually thereafter to satisfy the NELAC requirements.
For DoD and DOE projects, the MDL is verified quarterly. DoD QSM 5.0
requires that the MDLV (LODV) be spiked at 2-4 times the MDL
concentration. This standard is subject to the entire preparation andanalysis process. The calculated MDL is verified if the MDLV standard is
detected under routine instrument conditions.

13.2.2

If the first MDLV is not detected, the MDLV standard will be re-prepared
and analyzed at twice the original concentration.
The lowest
concentration that produces a detectable signal will then be reported as
the MDL.

13.2.3

DoD also requires a quarterly LOQV sample that is spiked at 1-2x theLOQ (RL) and must meet laboratory determined control limits for
precision and accuracy at the spike level.

Demonstration of Capabilities
All personnel are required to perform an initial demonstration of proficiency (IDOC)
on an instrument they will be using for analysis prior to testing samples. On-goingproficiency must be demonstrated annually. IDOCs and on-going proficiency
demonstrations are conducted as follows.

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13.4

13.3.1

Four aliquots of the QC check sample are analyzed using the sameprocedures used to analyze samples, including sample preparation. The
concentration of the QC check sample should be equivalent to a mid- level
calibration.

13.3.2

Calculate the average recovery and standard deviation of the recovery foreach analyte of interest.

13.3.3

If any analyte does not meet the acceptance criteria, the test must be
repeated. Only those analytes that did not meet criteria in the first test need
to be evaluated. TNI 2009 requires consecutive passing results. Repeated
failure for any analyte indicates the need for the laboratory to evaluate theanalytical procedure and take corrective action.

13.3.4

Until the IDOC is approved by the QA Manager (or designee); the trainer
and trainee must be identified in the batch record.

13.3.5Further details concerning demonstrations of proficiency are described in
SOP DV-QA-0024.

Training Requirements
The Group Leader is responsible for ensuring that this procedure is performed byan associate who has been properly trained in its use and has the required
experience. A new analyst must be working under documented supervision prior
to approval of the IDOC. Documentation that a new analyst is performing under
supervision must be entered into the batch record (View Batch Information) untilthat analyst’s IDOC has been approved by the QA Manager (or designee). See
requirements for demonstration of analyst proficiency in SOP DV-QA-0024.

14.0

Pollution Control
Standards and reagents are prepared in volumes consistent with laboratory use tominimize the volume of expired standards and reagents requiring disposal.

15.0

Waste Management
15.1

All waste will be disposed of in accordance with Federal, State, and local
regulations. Where reasonably feasible, technological changes have been
implemented to minimize the potential for pollution of the environment. Employees
will abide by this procedure, the policies in section 13, “Waste Management and
Pollution Prevention”, of the Corporate Safety Manual, and HS-001, “Waste
Management Program.”

15.2

The following waste streams are produced when this method is carried out:
15.2.1

Vial waste – Collect in PFC waste containers

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16.0

15.2.2

Instrument process waste – Flammable Solvent (C)15.2.3

Expired Chemicals/Reagents – Contact Waste Coordinator15.2.4

Soil Samples, post extraction, standards, and any solid waste contacting
solutions which contained PFCs, and all other solid waste generated by thisprocedure, such as disposable pipette tips and extraction bottles arecollected in special PFC waste containers and turned into the Waste
Coordinator for incineration.

NOTE:Radioactive and potentially radioactive or mixed waste must be segregatedfrom non-radioactive waste as appropriate. Contact the Waste Coordinatorfor proper management of radioactive or potentially radioactive waste
generated by this procedure.

References / Cross-References
16.1

Cheryl Moody, Wai Chi Kwan, Johnathan W. Martin, Derek C. G. Muir, Scott A.
Mabury, “Determination of Perfluorinated Surfactants in Surface Water Samples by
Two Independent Analytical Techniques: Liquid Chromatography/Tandem Mass
Spectrometry and 19FNMR,” Analytical Chemistry, 2001, 73, 2200-2206.

16.2John Giesy et al., “Accumulation of Perfluorooctane Sulfonate in Marine
Mammals”, Environmental Science & Technology, 2001 Vol. 35, No. 8, pages
1593-1598.

16.3

U.S. EPA, “Residue Chemistry Test Guidelines, OPPTS 860.1340, Residue
Analytical Method”, EPA 712-C-95-174, August 1995.

16.4

STL Denver White Paper DEN-W-LC-002, “Method Validation Study for Analysisof Ammonium Perfluorooctanoate in Soil Matrices by High Performance Liquid
Chromatography/Mass Spectrometry (HPLC/MS/MS)”, Mark Dymerski, September
5, 2003.

16.5

STL Denver White Paper DEN-W-LC-003, “Addendum A to Method Validation
Study for Analysis of Ammonium Perfluorooctanoate in Soil Matrices by High
Performance Liquid Chromatography/Mass Spectrometry (HPLC/MS/MS)”, Mark
Dymerski, August 6, 2003.

16.6

STL Denver White Paper DEN-W-LC-004, “Method Validation Study for Analysis
of Perfluorooctanoic Acid in Waters by High Performance Liquid
Chromatography/Tandem Mass Spectrometry (HPLC/MS/MS)”, Mark Dymerski,
January 26, 2005.

16.7

SOP No. DV-OP-0019 “Extraction of Perfluorooctanoic Acid (PFOA) and
Perfluorooctanoic Sulfonate (PFOS) and other Perfluorinated Hydrocarbons(PFCs) in Water and Soil”.

16.8

SW-846, Test Methods for Evaluating Solid Waste, Physical/Chemical Methods,
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SOP No. DV-LC-0012, Rev. 15
Effective Date: 5/7/2018
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Third Edition and all promulgated updates, EPA Office of Solid Waste, January
2005.

16.9

17.0

18.0

16.8.1

Method 8321A, Solvent Extractable Nonvolatile Compounds by High
Performance Liquid Chromatography/Thermospray/Mass Spectrometry(HPLC/TS/MS) or Ultraviolet (UV) Detection, Revision 1, December,
1996,

16.8.2

Method 8321B, Solvent Extractable Nonvolatile Compounds by High
Performance Liquid Chromatography/Thermospray/Mass Spectrometry(HPLC/TS/MS) or Ultraviolet (UV) Detection, Revision 2, February, 2007.

16.8.3

Method 8000B, Determinative Chromatographic Separations, Revision 2,
December 1996.

16.8.4

Method 8000C, Determinative Chromatographic Separations, Revision 3,
March 2003.

16.8.5

Method 3535A, Solid-Phase Extraction (SPE), Revision 1, February 2007

J. A. Shoemaker, P.E. Grimmett, B. K. Boutin, “Method 537: Determination of
Selected Perfluorinated Alkyl Acids in Drinking Water by Solid Phase Extraction
and Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS),”
EPA/600/R-08/092, Version 1.1, September 2009.

Method Modifications:17.1

This SOP was developed during an extensive method development and evaluation
process. This SOP presents the best practices that were demonstrated during thedevelopment process for the full suite of target compounds.

17.2

As of this writing, Method 537 provides for 14 day holding time for water samples
preserved with Trizma buffer. The scientific literature indicates the perfluorinatedsubstances are highly persistent in the environment. TestAmerica Sacramento has
conducted holding time studies that support 14 day holding time for aqueous
samples with and without Trizma preservation. TestAmerica Denver has
conducted stability studies indicating that medium and lower-level solutions of
PFOA are stable for at least three months in polystyrene and polypropylene
plastics at 0-6 degrees C.

Tables / Figures / Attachments
Table 1
Table 2
Table 3
Table 4
Table 5

Standard Reporting Limits for Waters and Soils
Recommended Agilent (LCMS_5) Instrument Operating Conditions
Recommended Waters (LCMS_4) Instrument Operating Conditions foranalysis of HFPO
Analytical Sequence and Summary of QC Criteria
DoD QSM 5.0 Requirements

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SOP No. DV-LC-0012, Rev. 15
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Table 6
Table 7
Figure 1
Figure 2
Figure 3
Figure 4
19.0

PFC names and abbreviations and Recommended Internal Standard andSurrogate Associations
Mass Calibrations
Example Chromatograms for the PFC Method
Example Chromatograms for the PFC_FOSA Method
Example Chromatograms for the LCMS_PFOA Method
Example Chromatograms for the HFPO Method

Revision History
Revision 15, dated 7 May 2018

Annual Review

Revision 14, dated 31 January 2017


Added language to Section 7 regarding standard expiration dates
Revised Section 9.3.3 to reflect current MS/MSD policy
Moved instrument tuning, initial calibration, ICV and CCV criteria to Section 11 andrenumbered remainder of document accordingly.
Updated gradient in Table 3 to reflect current practice

Revision 13, dated 6 January 2016











Removed Confidentiality Restriction Note boxes
Added procedure and requirements for analysis of HFPO in water throughout
Added description of quantitation of perfluorinated sulfonates as either the anion or
the acid (throughout)
Revised Sections 3, 9, and 12 to reflect current practiceMoved Instrument maintenance statement from Section 9.4.1 to Section 10.6
Revised Section 10.5.2.2 to describe use of dilution at the preparation step if
expected concentration of sample is known and removed Section 10.5.3.
Revised Equations 11 & 12 in Section 11.4.1 to account for dilution factor of extract.
Removed all references to WI-DV-0049. Work instruction is being permanently
archived as the process for calculation of dilution factors is now handled by Chrom.
Added surrogate associations to Table 6
Formatting and editorial changes throughout

Revision 12, dated 13 October 2014








Revised holding time for water samples to 14 days, consistent with stability studies
performed during method development.
Added statement to Section 9.1.2 regarding DoD QSM 5.0 compliance.
Added new Section 11.4.2 for reference to WI-DV-0049, PFC Method Data Review
which addresses calculations of dilutions.
Added requirements for compliance with DoD QSM 5.0 in text when different fromlaboratory requirements and as new Table 5
Renumbered existing Tables 5 and 6 to accommodate addition of new table.
Removed Appendix I (client-specific information)
Formatting and editorial changes throughout

Revision 11, dated Dec 04, 2013
Confidential information included

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SOP No. DV-LC-0012, Rev. 15
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Revision 10, dated July 12, 2013


Annual review
Added Figure 1, Figure 2 and Figure 3 of chromatograms
Removed Appendix 2
Updated sections 9.1 and 10.1 and inserted new section 10.2

Revision 9.2 dated July 13, 2012
Updated solvent grade requirements (Section 7.1)

Revision 9.1 dated April 4, 2012Annual review

Revision 9 dated February 28, 2011












Added Table 1 for the standard reporting limits for waters and soils.
Moved Table 1 to Table 6 for the mass calibrations.
Added 13C8 PFOA and 13C8 PFOS as surrogates.
Expanded dilution section to reflect isotope dilution method.
Updated standards section to explicitly list all standards used in this method.
Updated Table 2 with Waters Instrument method conditions.
Added Table 3 to include Agilent Instrument (LCMS_5) method conditions.
Changed numbering of Attachments to incorporate the addition of Table 3 for the
Agilent Instrument.
Removed references to client-specific requirements.
Added Appendices 1 and 2 for specific client requirements.
Updated references
Earlier revision histories have been archived and are available upon request.

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SOP No. DV-LC-0012, Rev. 15
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Table 1: Standard Reporting Limits
CompoundPFC Method
Water
5 mL
FV
(µg/L)

Perfluorobutyric Acid
Perfluoropentanoic Acid
Perfluorohexanoic Acid
Perfluoroheptanoic Acid
Brch-Perfluorooctanoic Acid
Perfluorononanoic Acid
Perfluorodecanoic Acid
Perfluoroundecanoic Acid
Perfluorododecanoic Acid
Perfluorotridecanoic Acid
Perfluorotetradecanoic Acid
Perfluorobutane Sulfonate
Perfluorobutanesulfonic Acid
Brch-Perfluorohexane Sulfononic
Acid
Perfluorohexanesulfonic Acid
Brch-Perfluorooctane Sulfonic Acid
Perfluorooctanesulfonic Acid
Perfluorodecane Sulfonate
Perfluorodecanesulfonic Acid
Perfluorooctanesulfonamide

Water
1 mL
FV
(µg/L)

Soil(µg/kg)

PFC_FOSA
Method
Water
Water
1 mL
5 mL
FV
FV
(µg/L)(µg/L)--------------LCMS_PFOA
MethodWater(µg/L)

Soil(µg/kg)----0.02-------------0.8---------

0.02
0.03
0.02
0.03
0.02
0.04
0.02
0.02
0.03
0.04
0.03
0.02
0.02

0.8
0.8
0.8
0.8
0.8
0.8
0.8
0.8
2.0
0.8
2.0
0.8
0.8

0.03

0.8

--

--

--

0.03
0.03
0.03
0.02
0.02
--

0.8
0.8
0.8
0.8
0.8
0.8

------

-0.03
-----

-0.8
-----

0.05

Brch= quantitation using standards containing branched isomers

Compound

Perfluoro(2-propoxypropanoic) acid

HFPO Method
Water (g/L)

Soil (μg/kg)

0.01

--

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Table 2: Agilent (LCMS_5) Recommended Instrument Operating Conditions
NOTE:

The conditions listed below for HPLC conditions and MS interface gas flows,
temperatures, probe voltage, and collision energy are subject to final fineadjustments to maximize sensitivity. Changes to the below conditions will be
documented in the project data file. Argon or nitrogen collision gas pressure is set
according to the instrument manufacturer.

Mass Spectrometer Interface Settings (QQQ)
MS Interface ModeoGas Temp ( C)
Gas Flow (L/min)
Nebulizer (psi)oSheath Gas Temp ( C)
Sheath Gas Flow (L/min)
Negative Capillary (V)Nozzle Voltage (V)ESI + Agilent Jet Stream Negative Ion
320
4
45
350
12
2500
500

Water PFC_FOSA Method:
HPLC Conditions (Agilent 1200)
Column (equivalent may be used)oColumn Temp ( C)Flow Rate
Mobile Phase Composition
Gradient for FOSA only (waters)

Injection SizeFOSA run time

Phenomenex Gemini NX plus Gemini guard cartridge
25
0.5 mL/min
A=0.005 M NH4OAc (aq, 10% MeOH),
B=LCMS grade methanol
Time
Flow
%B
0
0.4
10
0.5
0.4
10
2.5
0.4
50
6
0.4
95
8.5
0.4
95
9
0.8
10
9.5
0.4
10
Hold for 2.5 min. before next injection.
25 L, with Needle Wash
12 minutes

QQQ Mass Spectrometer Scan Settings
AnalyteParent Ion(Da)

Product Ions
(Da)

Dwell (s)Fragment (V)Collision
Energy (V)

13C8 FOSA
FOSA

505.9
497.9

78
78

0.100
0.100

127
127

36
36

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Table 2 (continued)
PFOA / PFOS Only Method (LCMS_PFOA):
HPLC Conditions (Agilent 1200)
Column (equivalent may be used)oColumn Temp ( C)Flow Rate
Mobile Phase Composition
Gradient for FOSA only (waters)

Injection Size
PFOA/PFOS run time

Phenomenex Gemini NX plus Gemini guard cartridge
40
0.4 mL/min
A=0.005 M NH4OAc (aq, 10% MeOH),
B=LCMS grade methanol
30% B → 100% B over 3 minutes. Hold for 1 minute.
Back to initial conditions over 0.5 minute. Hold for 1 min.
before next injection.
25 L, with Needle Wash
5 minutes

QQQ Mass Spectrometer Scan Settings
AnalyteParent Ion(Da)

Product Ions
(Da)

Dwell (s)Fragment (V)Collision
Energy (V)

PFOA
PFOA
PFOA
13
C4 PFOA
13
C8 PFOA
PFOS
PFOS
PFOS
13
C4 PFOS13
C8 PFOS

412.9
412.9
412.9
417.0
421.0
498.8
498.8
502.9
502.9
502.9

169
219
368.9
371.9
375.9
79.9
98.9
130
80
80

0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05

70
70
70
70
70
159
159
159
159
159

12
6
0
0
0
72
48
96
72
72

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Table 2 Continued.
PFC Method:
HPLC Conditions (Agilent 1200)
Column (equivalent may be used)oColumn Temp ( C)Flow Rate
Mobile Phase Composition
Gradient for PFCs

Injection Size
Time Segments

PFC run time

Phenomenex Gemini NX plus Gemini guard cartridge
25
0.5 mL/min
A=0.005 M NH4OAc (aq, 10% MeOH),
B=LCMS grade methanol
Time
Flow
%B
0
0.4
10
0.5
0.4
10
2.5
0.4
50
6
0.4
95
8.5
0.4
95
9
0.8
10
9.5
0.4
10
Hold for 2.5 min. before next injection.
25 L, with Needle Wash
0 – 1 min. – MS2 Scan – to Waste
1 – 9 min. – MRM – to MS
9 – 9.5 min. – MS2 Scan – to Waste
12 minutes

QQQ Mass Spectrometer Scan Settings
AnalyteParent Ion(Da)

Product Ions
(Da)

Dwell (s)Fragment (V)Collision
Energy (V)PFBA
C4 PFBA
PFPA
PFBS
PFBS
18
O2 PFBS
PFHxA
PFHxA
PFHxA
13
C2 PFHxA
PFHpA
PFHpA
Br-PFHxSBr-PFHxS18
O2 PFHxS18
O2 PFHxSBr-PFOABr-PFOA

213
216.7
263
298.9
298.9
303
313
313
313
314.9
362.9
362.9
398.9
398.9
402.9
402.9
412.9
412.9

168.9
172
218.9
79,9
98.9
84
118.6
169
268.9
269.9
318.9
168.9
80
98.9
84
103
169
219

0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008

42
42
42
121
121
121
62
62
62
62
68
68
130
130
130
130
70
70

4
4
0
32
24
32
8
8
0
0
0
8
48
32
32
32
12
6

13

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Table 2 continued
QQQ Mass Spectrometer Scan Settings
AnalyteParent Ion(Da)

Product Ions
(Da)

Dwell (s)Fragment (V)Collision
Energy (V)Br-PFOAC4 PFOA
13
C8 PFOA
PFHpS
PFHpS
PFNA
PFNA
13
C5 PFNAFOSA
Br-PFOSBr-PFOSBr-PFOS13
C4 PFOS13
C8 FOSA
13
C8 PFOS
PFDA
PFDA
13
C2 PFDA
PFUnA
PFUnA
PFUnA
13
C2 PFUnA
PFDS
PFDS
PFDoA
PFDoA
13
C2 PFDoAPFTriA
PFTeA

412.9
417.0
421
449.9
449.9
463
463
467.9
497.9
498.8
498.8
498.8
502.9
505.9
506.9
513
513
514.9
563
563
563
564.9
598.9
598.9
613
613
614.9
663
712.9

368.9
371.9
375.9
80
98.9
218.9
418.9
423.0
78
79.9
98.9
130
80
78
80
219
269
470
219
268.9
518.9
520
79.9
99
168.9
568.9
569.9
618.9
169

0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008
0.008

70
70
70
153
153
71
71
71
127
159
159
159
159
127
159
77
77
77
83
83
83
83
174
174
77
77
77
86
86

0
0
0
44
48
8
4
4
36
72
48
96
72
36
72
8
12
4
8
12
0
0
84
56
24
4
4
8
28

13

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Table 3: Waters (LCMS_4, LCMS-7) Recommended Instrument Operating
Conditions
for Analysis of HFPONOTE:The conditions listed below for HPLC conditions and MS interface gas flows,
temperatures, probe voltage, and cone voltage are subject to final fine adjustments
to maximize sensitivity. Changes to the below conditions will be documented in the
project data file. Argon or nitrogen collision gas pressure is set according to the
instrument manufacturer.

HPLC Conditions (Waters Alliance/Acquity)
Column (equivalent may be used)
Flow Rate
Mobile Phase Composition
Gradient for HFPOInjection Size
HFPO run timeoPhenomenex Synergi Hydro RP plus C18 guard cartridge 35 C
0.5mL/min
A=LCMS grade methanol B=0.01 M NH4OAc (aq),
70% A and 30% B isocratic.

40uL (Alliance) 20 L (Acquity)
2 minutes

Mass Spectrometer Interface Settings (Quattro Premier/Xevo)
MS Interface Mode
Capillary (kV)
Source Temp
Desolvation Temp
Cone Gas (nitrogen) Flow
Desolvation Gas (nitrogen) FlowESI Negative Ion
1.0
o
100 C
o
300 C
100 L/Hr
500 L/Hr

Waters Acquity Mass Spectrometer Scan Settings
Function

Channel112 (IS)

AnalyteParentIons (Da)

Product
Ions (Da)

Dwell
(s)

Cone
(V)

Collision
(eV)

Delay
(s)

HFPO
C3 HFPO

328.8
331.8

284.8
286.8

0.6
0.6

9
9

6
6

0.1
0.1

13

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Table 4: Analytical Sequence and Summary of QC Criteria for Commercial Workand DoD QSM 4.2
Parameter

Description

Acceptance Criteria

Corrective Action
Perform instrument
maintenance and/or
clean system.
Recalibrate.

1

Tune solution

Injection of the Level 1
calibration standard.

Acceptable peak shapeand recovery.

2

ICAL

r > 0.990 or %RSD≤20%

3

ICB

4

ICV

Calibration standards 0.2 –125 g/L.
Analyze HPLC-grade
methanol spiked with IS.
Second-source, 2.0 g/L
standard.

70-130% recovery for
PFCs

Clean system &recalibrate.
Check standards &recalibrate.

5

DLCK

Detection Limit Check, 0.5
g/L standard.

70-130% recovery for
PFCs

Check standards &recalibrate.

6

Method Blank

Extracted reagent water.

< ½RL

7

LCS

Extracted 0.2 g/L or 20.0
g/kg spiked blank.

±3 standard deviations
from the historical mean

8

Sample---9Matrix Spike andMatrix Spike
Duplicate

Spiked second & third
portions of sample.

Positive results must bewithin working range±3 standard deviations
from the historical mean

10

Samples

---

11

CCV

12

Samples

Alternate 5.0 g/L and 10.0
g/L standards, analyze
after every 10 injections.
---

Identify & fix sourceof contamination, reextract batch.
Re-extract &reanalyze batch,
unless biased highand samples ND.
Dilute to within
working range.
Verify results byreanalysis. Reextract & rerun if LCS
also outside limits.
Dilute to within
working range
Recalibrate &reanalyze samples.

13

Closing CCV

14Internal
Standards

5.0 g/L or 10.0 g/L
standard< 2x MDL or < ½RL
whichever is smaller

Positive results must bewithin working range
70-130% recovery

Positive results must bewithin working range
70 - 130% Recovery
50-150% Recovery

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Dilute to within
working range
Recalibrate &reanalyze samples.
If out in QC, reextract, reanalyze
If out in samples and
QC is acceptable,
sample considered tohave matrix effect(NCM)SOP No. DV-LC-0012, Rev. 15
Effective Date: 5/7/2018
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Table 5: Summary of QC Criteria for DoD QSM 5.0
QSM 5.0 Table 15. Perfluorinated Compounds by Liquid Chromatography/Mass Spectrometry
Requirement
Tune Check

DoD QSM 5.0 and DoE QSAS 3.0
Prior to ICAL and after any mass calibration or maintenance is performed. Tuning
standards must contain the analytes of interest or appropriate substitute.
Mass assignments of tuning standard must be within 0.5 amu of true value. Retune
instrument and verify. If the tune check will not meet acceptance criteria, an instrument
mass calibration must be performed and the tuning redone.
No samples shall be analyzed without an acceptable tune check.

Initial Calibration (ICAL)

At instrument setup and after major maintenance. Minimum 5 points for linear or 6 points
for quadratic; weighting is allowed.
Each calibration point for each analyte must calculate to be within 75-125%, except thelowest cal point which must calculate to within 70-130%.
Any problems must be corrected and ICAL repeated. No sample shall be run until ICAL is
successful.

Initial Calibration
Verification (ICV)

Measure a second-source standard with concentration at the midpoint of the calibration
once after each ICAL, prior to sample analysis.
All reported analytes and surrogates must be within ± 25% of the true value. Any
problems must be corrected. No samples shall be analyzed until the second-source
calibration verification is successful.

Continuing calibration
Verification (CCV)

Analysis of mid-level standard after every 10 field samples and the end of the analyticalsequence. On days an ICAL is not performed, at the beginning of the sequence, after
every 10 field samples and at the end of the analytical sequence.
All reported analytes must be within + 25% of the true value.
If the CCV is above the project acceptance limits and there are no detections in thesamples, TestAmerica will report the non-detect results with a case narrative comment inaddition to applying any data qualifier flags required by the project (3HR).
Correct any problems then rerun CCV. If that fails, then repeat ICAL. Reanalyze all
samples since last successful CCV. Results cannot be reported without a valid CCV.
Or
Immediately (within one hour) analyze two additional consecutive CCVs. If both pass,
samples may be reported without reanalysis. If either fails, take corrective action(s) andre-calibrate; then reanalyze all affected samples since the last acceptable CCV.
If reanalysis cannot be performed, data must be qualified and explained in the casenarrative. Apply Q-flag to all results for the specific analytes(s) in all samples since thelast acceptable CCV. Flagging is only appropriate in cases where the samples cannot bereanalyzed.

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Effective Date: 5/7/2018
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Table 5: Summary of QC Criteria for DoD QSM 5.0 (continued)
QSM 5.0 Table 15. Perfluorinated Compounds by Liquid Chromatography/Mass Spectrometry
Requirement

DoD QSM 5.0 and DoE QSAS 3.0

Internal Standards (IS)

Isotopically-labeled analytes must be added to all field samples, QC samples (batch andinstrument) and standards.
Absolute areas of the quantitation ions of the IS(s) must be within 50-150% from theaverage areas measured during ICAL.
If recoveries are acceptable for QC samples but not field samples, the field samples may
be considered to suffer from a matrix effect. Apply Q-flag and explain in the casenarrative.
For failed QC samples, correct problem and rerun all associated failed field samples.
Failing internal standard should be thoroughly documented in the case narrative.

Method Blank (MB)One per prep batch. No analytes detected > ½ LOQ (RL) or >1/10 the amount measured
in any sample or 1/10 the regulatory limit, whichever is greater.
If criteria not met, correct problem. If required, reprep and reanalyze MB and all samples
processed with the contaminated blank.
If reanalysis is not possible, apply B-flag to all results for the specific analyte(s) in all
samples processed with the contaminated blank. Must be explained in the case narrative.
Flagging is only appropriate when samples cannot be reanalyzed.

LCS

One per prep batch.
As DoD limits are not available, use in-house LCS limits if project limits are not specified.
If in-house limits do not exist, use 70-130% until limits are established.
If the LCS recovery is above the project acceptance limits and there are no detections inthe samples, TestAmerica will report the non-detect results with a case narrative
comment in addition to applying any data qualifier flags required by the project (3HR).
Correct any problems then re-prep and reanalyze the LCS and all associated samples for
failed analytes. If insufficient sample or corrective action fails, then apply Q-flag to
specific analyte(s) in all samples in the associated prep batch. Must be explained in thecase narrative.

Matrix SpikeOne per prep batch. As DoD limits are not available, use in-house LCS limits if project
limits are not specified. For failures, consult project-specific DQOs and contact client for
additional measures to be taken.
For specific analyte(s) in parent sample, apply J-flag if acceptance criteria are not met.
If MS falls outside LCS limits, evaluate data to determine the source of the difference, i.e.,
matrix effect or analytical error.

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Effective Date: 5/7/2018
Page No.: 43 of 51

Table 5: Summary of QC Criteria for DoD QSM 5.0 (continued)
QSM 5.0 Table 15. Perfluorinated Compounds by Liquid Chromatography/Mass Spectrometry
Requirement
MSD or Sample
Duplicate

DoD QSM 5.0 and DoE QSAS 3.0
One per prep batch. Analyze MS/MSD for low concentration samples and Sample/MD forhigh concentration samples. Use DoD-specific criteria for LCS. If DoD limits are notavailable, use in-house LCS limits if project limits are not specified. RPD for duplicates ≤30%. For failures, consult project-specific DQOs and contact client for additional
measures to be taken.
For specific analyte(s) in parent sample, apply J-flag if acceptance criteria are not met.
If MS falls outside LCS limits, evaluate data to determine the source of the difference, i.e.,
matrix effect or analytical error.

Surrogate SpikeAll field and QC samples. As DoD limits are not available, use in-house LCS limits if
project limits are not specified. If in-house limits do not exist, use 70-130% until limits are
established.
If recoveries are acceptable for QC samples, but not field samples, the field samples may
be considered to suffer from a matrix effect. For failed QC samples, correct problem andrerun all failed samples.
If samples cannot be reanalyzed, apply Q-flag and discuss in the case narrative.

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Effective Date: 5/7/2018
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Table 6: PFC Names, Abbreviations andRecommended Internal Standard and Surrogate Associations
PFC
Abbreviation

PFC Name

Internal Standard

Surrogate

Perfluorobutyric Acid

PFBA

13

C4 PFBA

13

Perfluoropentanoic Acid

PFPA

13

C2 PFHxA

13

Perfluorohexanoic Acid

PFHxA

13

13

Perfluoroheptanoic Acid

PFHpA

13

13

Perfluorooctanoic Acid

PFOA

13

13

Perfluorononanoic Acid

PFNA

13

13

Perfluorodecanoic Acid

PFDA

13

C2 PFDA

13

Perfluoroundecanoic Acid

PFUnA

13

C2 PFUnA

13

Perfluorododecanoic Acid

PFDoA

13

13

Perfluorotridecanoic Acid

PFTriA13

13

Perfluorotetradecanoic Acid

PFTeA

13

13

Perfluorobutane Sulfonate

PFBS

18

13

Perfluorobutanesulfonic Acid

PFBS

18

O2 PFHxS

13

Perfluorohexane Sulfonate

PFHxS

18

O2 PFHxS

13

Perfluorohexanesulfonic Acid

PFHxS

18

Perfluorooctane Sulfonate

PFOS

13

Perfluorooctanesulfonic Acid

PFOS

13

C4 PFOS13

Perfluorodecane Sulfonate

PFDS

13

C4 PFOS13

Perfluorodecanesulfonic Acid

PFDS

13

C4 PFOS13

Perfluorooctanesulfonamide

FOSA

13

C8 FOSA

13

13

C8 PFOA

13

--

13

C8 PFOS

13

--

HFPO

13

C2 PFHxA
C4 PFOAC4 PFOAC5 PFNAC2 PFDoAC2 PFDoAC2 PFDoAO2 PFHxSO2 PFHxSC4 PFOSC4 PFBA

C8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOAC8 PFOSC8 PFOSC8 PFOS13

C8 PFOS13

C8 PFOSC8 PFOSC8 PFOSC8 PFOSC8 PFOA

13

C8 Perfluorooctanoic Acid
(Surrogate)
13
C8 Perfluorooctane Sulfonate(Surrogate)
Perfluoro(2-propoxypropanoic) Acid

C4 PFOAC4 PFOSC8 HFPO

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13

C8 HFPOSOP No. DV-LC-0012, Rev. 15
Effective Date: 5/7/2018
Page No.: 45 of 51

Table 7: Mass Calibrations
PEG

NaCsI

Na Formate

89.06
133.09177.11221.14239.15283.18327.20371.23415.25459.28503.31564.36608.39652.41
696.44740.46784.49828.52872.54916.57960.601004.6222.99
132.91172.88322.78472.67622.57772.46922.361072.25

22.99
90.98
105.04158.96226.95294.94362.93430.91498.90566.89634.88702.86770.85838.84906.83974.811042.80

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Figure 1: Example Chromatograms for the PFC Method

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Effective Date: 5/7/2018
Page No.: 47 of 51

Figure 1 (cont.): Example Chromatograms for the PFC Method

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Effective Date: 5/7/2018
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Figure 1 (cont.): Example Chromatograms for the PFC Method

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Effective Date: 5/7/2018
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Figure 1 (cont.): Example Chromatograms for the PFC Method

Figure 2: Example Chromatograms for the PFC_FOSA Method

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Effective Date: 5/7/2018
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Figure 3: Example Chromatograms for the LCMS_PFOA Method

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Figure 4: Example Chromatograms for LCMS - HPFO

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