TestAmerica Denver

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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-OP-0019, Rev. 9
Effective Date:12/08/2017
Page No.: 2 of 32
1.0

2.0

3.0

Scope and Application
1.1

This procedure describes the extraction of perfluorooctanoic acid (PFOA, CAS#
335-67-1), and perfluorooctanoic sulfonate (PFOS, no CAS number), and other
Perfluorinated Hydrocarbons (PFC’s) from water and soil samples. The method
has been validated by TestAmerica Denver for reagent water, drinking water,
groundwater, and soil. See Table 1 for analytes that can be extracted under thisprocedure.

1.2

This procedure does not describe the LC/MS/MS analysis of the extracts. For
those details, see DV-LC-0012 “Analysis of Perfluorooctanoic Acid (PFOA),
Perfluorooctanoic Sulfonate (PFOS), and other Perfluorinated Hydrocarbons(PFCs) by LC/MS/MS”

Summary of Method
2.1

For all compounds except perfluorooctane sulfonamide (FOSA), water samples
undergo solid phase extraction using a Strata X-AW 33u Polymeric Weak Anion
cartridge. The compounds are eluted from the cartridge with a 10% ammoniumhydroxide in methanol solution.

2.2

For perfluorooctane sulfonamide (FOSA), water samples undergo solid phaseextraction using a Waters tC18 cartridge. FOSA is eluted from the cartridge withmethanol.

2.3

Soil samples are mixed with a sodium hydroxide solution and allowed to sit for 1/2
hour. Methanol is then added and the soil/solvent mixture is tumbled for 24 hours.
The pH is adjusted to ≤7 then the mixture is centrifuged and the solvent filtered.
The extract is then brought up to final volume with methanol. This extraction
procedure is used for all PFCs.

2.4

Labeled PFC’s 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. A known quantity of
the labeled PFC compounds is added to every sample and to the batch quality
control samples prior to extraction. Because the isotopically labeled compound ischemically similar to the compound of concern, it is affected by any interfering
substances in the sample to the same extent as the compound of concern. The
recovery of the labeled PFC’s is then used to mathematically correct the final
results for the PFC’s.

Definitions
Refer to the Glossary of the TestAmerica Denver Quality Assurance Manual (QAM) andpolicy DV-QA-003P, Quality Control Program, for definitions of general analytical and
QA/QC terms.
3.1

PTFE: polytetrafluoroethylene (e.g., Teflon®)
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4.0

3.2

Preparation Batch: A group of up to 20 samples that are of the same matrix andare processed together in the same extraction event using the same procedureand lots of reagents and standards

3.3

Method Comments: The Method Comments are used to communicate to thebench level chemists special requirements and instructions from the client. Pleasereference WI-DV-0032 for details on Method Comments.

3.4

Quality Assurance Summary (QAS): Certain clients may require extensive specificproject instructions or program QC, which are too lengthy to fit conveniently in the
Method Comments field in LIMS. In these situations, laboratory Project Managers
describe the special requirements in a written QAS to address these requirements.
QASs are posted on a public drive for easy accessibility by all lab employees.
Normally, QASs are introduced to analysts in an initial project kick-off meeting tobe sure that the requirements are understood.

3.5

Aliquot: A part that is a definite fraction of a whole; as in “take an aliquot of asample for testing or analysis.” In the context of this SOP, “aliquot” is also used asa verb, meaning to take all or part of a sample for preparation, extraction, and/or
analysis.

Interferences
4.1

PFCs have been used in a wide variety of manufacturing processes, andlaboratory supplies should be considered potentially contaminated until they havebeen tested and shown to be otherwise. The materials and supplies 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 and PFC contamination. The use ofPTFE in the procedure should be avoided or at least thoroughly tested before use.

4.4

Volumetric glassware and syringes are difficult to clean after being used forsolutions containing high levels of PFOA and PFCs. 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 andPFC contamination. These materials should be avoided during the extraction
process.

4.6

PFOA and PFCs can readily adhere to glass surfaces, glass containers should be
avoided as much as possible during the extraction of the samples. Therefore water
samples will be aliquoted gravimetrically instead of using volumetric glassware.

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5.0

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 PFOA and PFOS could havesignificant toxic effects. In the interest of keeping exposure levels as low asreasonably achievable, PFOA and PFOS 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

Stock standards are purchased as high purity solids in a powder form or at a
certified concentration in solvent. Handling of the powder will be limited tomaking intermediate concentration standards in solution, which will be done
only in certified laboratory fume hoods. The pure powders are stored insealed bottles kept in cabinets with known or other presumed hazardous
chemicals. There are no steps in this method that would generate mists inthe open lab environment.

5.3.4

The melting point for PFOA is 53C. Therefore, the vapor pressure is
extremely low at ambient lab temperatures. As a result, inhalation of vapors
during this procedure is not considered to be a significant health risk.

5.3.5

PFOA is an acid and is not compatible with strong bases.

5.3.6

PFOA and PFOS 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 inclosed containers with secondary containment measures or within a fumehood.

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
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SOP No. DV-OP-0019, Rev. 9
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method. The table contains a summary of the primary hazards listed in the SDS foreach 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 aremajor changes to the SDS.

Material (1)Hazards

Exposure
Limit (2)Signs and symptoms of exposure

Hydrochloric
Acid

Corrosive
Poison5 ppm(Ceiling)

SodiumHydroxide

Corrosive
Poison2 mg/cm(Ceiling)

Formic Acid

Combustible
Corrosive
Flammable

5 ppm
(TWA)

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

MethanolFlammable

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.

PoisonIrritant

Ammoniumhydroxide

Corrosive
Toxic

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 may be fatal if swallowed.

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
All equipment IDs for any support equipment (pipettes, thermometers, etc.) must be recordedin the batch record.
6.1

Equipment
6.1.1

Vacuum manifold for SPE cartridges. Capable of maintaining approximately
66 cm (26”) of Hg. After each use, the valves are removed from the
manifold and placed in a jar with acetonitrile. The jar is placed into asonication bath overnight.

6.1.2

Bottle rotating apparatus for the extraction of soil samples.

6.1.3

Nitrogen evaporation apparatus (N-EVAP) for the concentration of somewater extracts.

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6.2

6.1.4

Balance capable of measuring ± 0.1 g. Calibration checked per SOP DVQA-0014. Used to measure the initial sample mass and volume.

6.1.5

Pipettor with disposable 1.0 mL tips. Calibration checked per SOP DV-QA0008. Used to add internal standard and spike solution to samples.

6.1.6

Pipettor with disposable 0.1 mL tips. Calibration checked per SOP DV-QA0008. Used to add internal standard and spike solution to samples.

6.1.7

Pipettor capable of dispensing 5 to 50 mL. Calibration checked per SOP
DV-QA-0008. Used to calibrate vials to hold 5 mL for the final volume
determination for water extracts.

6.1.8

Pipettor capable of dispensing 0.6 to 5.6 mL. Calibration checked per SOP
DV-QA-0008. Used to bring soil extracts to a 20 mL final volume.

6.1.9

Centrifuge capable of clarifying water samples and soil extracts that containsediment.

Supplies
6.2.1

10-mL HDPE vials, screw thread – used to store final extracts.
NOTE:

Vials used to store the final extracts of water samples are calibrated
to hold a volume of 5 mL by using a calibrated pipette to deliver 5
mL of HPLC grade methanol into the vial and marking the meniscus
with a fine tipped permanent marker. This volume is accurate to ± 2
%.

6.2.2

125 mL HDPE wide-mouth bottles – used in the extraction of soil samples.

6.2.3

Syringe filter, 0.45 μm, PVDF, Millipore Millex-HV, or equivalent – used tofilter soil extracts.

6.2.4

Strata X-AW 33u Polymeric Weak Anion 200 mg packed in 6 mL tubes,
Phenomenex part no. 8B-S038-FCH (or equivalent) – used in theextraction of all PFCs described in Section 10.5, except FOSA. See
Sections 6.2.5 and 10.6 for FOSA cartridge and extraction procedure.

6.2.5

Waters SPE cartridge “tC18” part number WAT036795 (or equivalent) –used in the extraction of FOSA that is described in Section 10.6.

6.2.6

SPE tubing, non-PTFE in composition, Tubing is cleaned before and after
use with HPLC grade methanol and HPLC grade water.

6.2.7

Miscellaneous laboratory apparatus (beakers, test tubes, volumetric flasks,
pipettes etc.). These should be disposable or HDPE where possible, glass
should be avoided.
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6.2.8
6.3

pH paper – wide range.

Computer Software and Hardware
Please refer to the master list of documents, software and hardware 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
Reagent grade chemicals shall be used in all tests. Unless otherwise indicated, it isintended that all reagents shall conform to the specifications of the Committee on
Analytical Reagents of the American Chemical Society, where such specifications areavailable. Other grades may be used, provided it is first ascertained that the reagent is ofsufficiently high purity to permit its use without lessening the accuracy of thedetermination.
7.1

HPLC Grade Methanol – Used in rinsing SPE lines and ports, in the making ofstandards, conditioning of SPE cartridges, the extraction of water samples for FOSA,
and the extraction of soil samples.

7.2

Ottawa Sand – Used to prepare method blanks and LCSs for the extraction of soil
samples. Ottawa Sand is baked at 400C for at least 4 hours.

7.3

HPLC Grade Water – Used to prepare method blanks and LCSs for the extraction ofwater samples.

7.4

0.67 N NaOH solution – Used in the extraction of soil samples.
Using a Class A graduated cylinder, transfer 67 mL of 10 N NaOH to a 1000 mL
Class A volumetric flask and bring to volume with HPLC grade water.

7.5

2.4 M HCl solution – Used in the extraction of soil samples.
Fill a 500 mL Class A volumetric flask approximately half full with HPLC grade water.
Using a Class A graduated cylinder, transfer 100 mL of reagent grade concentrated
HCL to the flask and then bring to volume (500 mL) with HPLC grade water.

7.6

10% Ammonium Hydroxide in Methanol – Used in the elution of PFOA, PFOS, andPFCs except FOSA from the Strata X-AW 33u Polymeric Weak Anion (orequivalent).
Fill a 1000 mL Class A volumetric flask approximately half full with HPLC grade
methanol. Using a Class A graduated cylinder, transfer 100 mL of ammoniumhydroxide to the flask and then bring to volume (1000 mL) with HPLC grade
methanol.

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7.7

5% Ammonium Hydroxide in 50/50 Water/Methanol – Used in the extraction of
HFPO-DA from soils.
Transfer 50 mL of ammonium hydroxide into a 1000 mL Class A graduated cylinder.
Bring to volume (1000 mL) with equal parts HPLC water/HPLC grade methanol.

7.8

2% Formic Acid – Used to rinse the Strata X-AW 33u Polymeric Weak Anion (orequivalent) in the extraction of PFOA, PFOS, and PFCs except FOSA. Also used toneutralize the pH in the extraction of HFPO-DA from soils.
Fill a 1000 mL Class A volumetric flask approximately half full with HPLC grade water.
Using a Class A graduated cylinder, transfer 23 mL of 88% formic acid to the flask
and bring to volume (1000 mL) with HPLC grade water.

7.9

7.10

7.11

Stock Standard Materials
7.9.1

The PFCs are purchased as a certified concentration in solution.
Standard materials are verified compared to a second source material at
the time of initial calibration.

7.9.2

Stock standard solutions are stored at 0-6°C. Stock standard solutions
should be brought to room temperature before using. Standards aremonitored for signs of degradation or evaporation. Standard solutions
must be replaced at least annually from the date of preparation.

7.9.3

All standards are stored in HDPE bottles and vials. Glass containers are
avoided.

“PFOA/S_Spike”, 0.5 g/mL
7.10.1

A spike solution is prepared containing PFOA and PFOS at 0.5 g/mL eachin HPLC grade methanol. This standard is stored in HDPE bottles at 0-6C
and given a 1 year expiration date.

7.10.2For soil samples, when PFOA and PFOS are the only analytes of concern,
0.4 mL of this solution is spiked into the LCS and matrix spike samples,
which results in a 20 g/Kg sample concentration.

7.10.3For water samples, when PFOA and PFOS are the only analytes ofconcern, 0.1 mL of this solution is spiked into the LCS and matrix spike
samples, which results in a 0.2 µg/L sample concentration.

“PFC-LCS-BR”, 0.5 g/mL

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7.12

7.13

7.14

7.11.1

A spike solution is prepared containing all PFCs listed in Table 1 at 0.5
g/mL each in LCMS grade methanol. This standard is stored in HDPE
bottles at 0-6C and given a 1 year expiration date.

7.11.2For soils samples, when more PFCs than just PFOA and PFOS are theanalytes of concern, 0.4 mL of this solution is spiked into the LCS andmatrix spike samples, which results in a 20 g/Kg sample concentration.

7.11.3For water samples, when more PFCs than just PFOA and PFOS are theanalytes of concern, 0.1 mL of this solution is spiked into the LCS andmatrix spike samples, which results in a 0.2 µg/L sample concentration.

“FOSA-H2O-LCS”, 0.05 g/mL
7.12.1

A spike solution is prepared containing only perfluorooctane sulfamide(FOSA) at 0.05 µg/mL in HPLC grade methanol. This standard is stored in
HDPE bottles at 0-6C and given a 1 year expiration date.

7.12.2

For water samples, when FOSA is the analyte of concern, 1 mL of thissolution is spiked into the LCS and matrix spike samples, which results in a
0.2 µg/L sample concentration.

“PFOA/S_Sur/IS”, 0.5 g/mL
7.13.1

An isotope dilution spike solution is prepared containing the internal
standards 13C4 PFOA and 13C4 PFOS at 0.5 g/mL in HPLC grade
methanol. This standard also contains the surrogate compounds 13C8PFOA and 13C8 PFOS at 0.5 g/mL. This standard is stored in HDPE
bottles at 0-6C and given a 1 year expiration date.

7.13.2For soil samples, when PFOA and PFOS are the only analytes of concern,
0.4 mL of this standard is added into each sample (field and laboratory QC
samples) at the beginning of the preparation. This produces a 20 g/kg
spike concentration.

7.13.3For water samples, when PFOA and PFOS are the only analytes ofconcern, 0.1 mL of this standard is added into each sample (field andlaboratory QC samples) at the beginning of the preparation. This produces
a 0.2 µg/L spike concentration.

“PFC_Surr&IS”, 0.5 g/mL
7.14.1

An isotope dilution spike solution is prepared containing all of the labeled
PFC isotopes listed in Table 2 and the surrogate compounds listed in Table
3 at 0.5 g/mL in HPLC grade methanol. This standard is stored in HDPE
bottles at 0-6C and given a 1 year expiration date.

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7.14.2For soil samples, when more PFCs than just PFOA and PFOS are theanalytes of concern, 0.4 mL of this standard is added into each sample (field
and laboratory QC samples) at the beginning of the preparation. This
produces a 20 g/kg spike concentration.

7.14.3For water samples, when more PFCs than just PFOA and PFOS are theanalytes of concern, 0.1 mL of this standard is added into each sample (field
and laboratory QC samples) at the beginning of the preparation. This
produces a 0.2 µg/L spike concentration.

7.15 “FOSA8-IS-SPK”, 0.05µg/mL
7.15.1

A spike solution is prepared containing only 13C8 FOSA at 0.05 µg/mL in
HPLC grade methanol. This standard is stored in HDPE bottles at 0-6C
and given a 1 year expiration date.

7.15.2

For water samples, when FOSA is the analyte of concern, 1 mL of thissolution is added into each sample (field and laboratory QC samples) at the
beginning of the preparation.
This produces a 0.2 µg/L sample
concentration.

7.16 “HFPO I.S”, 0.5 µg/mL
7.16.1

A spike solution is prepared containing only 13C3 HFPO-DA at 0.5 µg/mL in
LCMS grade methanol. This standard is stored in HDPE bottles at 0-6C
and given a 1 year expiration date.

7.16.2

For water samples, when HFPO is the analyte of concern, 0.1 mL of thissolution is added into each sample (field and laboratory QC samples) at the
beginning of the preparation.
This produces a 0.2 µg/L sample
concentration.

7.17 “HFPO Spike”, 0.5 µg/mL

8.0

7.17.1

A spike solution is prepared containing only HFPO-DA (Perfluoro(2propoxypropanoic acid) at 0.5 µg/mL in HPLC grade methanol. This
standard is stored in HDPE bottles at 0-6C and given a 1 year expirationdate.

7.17.2

For water samples, when HFPO is the analyte of concern, 0.1 mL of thissolution is added into each sample (field and laboratory QC samples) at the
beginning of the preparation.
This produces a 0.2 µg/L sample
concentration.

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

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requests. Listed below are the holding times and the references that include preservation
requirements.

9.0

MatrixSampleContainer

Min.
SampleSize

Preservation

Extraction
Holding
Time1

Analytical
HoldingTime

Reference

Waters

HDPE2

250 mL(x2)3Cool, < 6oC

14 Days

40 Days

N/A4

Soils

HDPE

10 g

Cool, < 6oC

14 Days

40 Days

N/A4

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. For SOPs that
address only preparation, QC acceptance limits on the analytical results are notincluded. Refer to the appropriate SOP that describes the determinative method.
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 Control Program.

9.1.2

Specific QC requirements for Federal programs, e.g., Department of
Defense (DoD), Department of Energy (DOE), etc., are described in
TestAmerica Denver policy DV-QA-024P, QA/QC 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 in the project requirements.

9.1.3

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

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9.1.4

9.2

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-QA-0031. This isin addition to the corrective actions described in the following sections.

Initial Performance Studies
Before analyzing samples, the laboratory must establish a method detection limit(MDL). In addition, an initial demonstration of capability (IDOC) must be performedby each analyst on the instrument he/she will be using. On-going proficiency must be
demonstrated by each analyst on an annual basis. See Section 12 for more details
on detection limit studies, initial demonstrations of capability, and analyst training andqualification.

9.3

Batch Definition
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, processed using the sameprocedures and reagents within the same time period. Batches should be kept
together through the whole analytical process as far as possible, but it is notmandatory to analyze prepared extracts on the same instrument or in the same
sequence. The method blank must be run on each instrument that is used to analyze
samples from the same preparation batch. See QC Policy DV-QA-003P for further
details.

9.4

Method Blank (MB)One method blank must be processed with each preparation batch. The method
blank is processed and analyzed just as if it were a field sample.

9.5

9.4.1

The method blank for batches of aqueous samples consists of 250 mL of
HPLC grade water free of any of the analyte(s) of interest.

9.4.2

The method blank for batches of soil samples consists of 10 g of baked
Ottawa sand free of any of the analyte(s) of interest.

Laboratory Control Sample / Laboratory Control Sample Duplicate (LCS/LCSD)
At least one LCS must be processed with each preparation batch. The LCS is carriedthrough the entire analytical procedure just as if it were a sample. When an MS/MSD
pair is not available, an LCS and LCSD are required.
9.5.1

For aqueous sample batches, the LCS consists of 250 mL of HPLC grade
water free of any of the analyte(s) of interest.

9.5.2

For soil sample batches, the LCS consists of 10 g of baked Ottawa sandfree of any of the analyte(s) of interest.
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9.6

9.7

Matrix Spike/Matrix Spike Duplicate (MS/MSD)
9.6.1

One MS/MSD pair must be processed with each preparation batch. A
matrix spike (MS) is a field sample to which known concentrations oftarget analytes have been added. It is prepared in a manner similar to
the LCS, but uses a real sample matrix in place of the blank matrix. A
matrix spike duplicate (MSD) is a second aliquot of the same sample(spiked exactly as the MS) that is prepared and analyzed along with thesample and matrix spike. Some programs allow spikes to be reported for
project-related samples only. Samples identified as field blanks cannot
be used for the MS/MSD analysis.

9.6.2

If insufficient sample volume is available for MS/MSD, an NCM must be
written and a LCSD must be prepared.

Isotopically Labeled Internal Standard (IS)Every field sample and QC sample (i.e. method blank, LCS, LCSD, MS, and MSD) is
spiked with internal standard compounds.

9.8

Surrogate Standard
Every field sample and QC sample (i.e. method blank, LCS, LCSD, MS, and MSD) is
spiked with the surrogate standard compounds.

10.0

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-QA0031. 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

All glassware and positions must be rotated through both QC and field samples.
Do not use specific glassware or positions for the MB and LCS/LCSD.

10.4

All equipment IDs for any support equipment (pipettes, thermometers, etc.) usedmust be recorded in the batch record.

10.5

Water Sample Extraction for PFOA/PFOS, HFPO-DA, and Full List PFCs, notincluding FOSA

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10.5.1The procedure described here in Section 10.5 is for water samples that are
to be analyzed for PFOA or PFOS only (Prep method 3535 with analyticalmethod LCMS_PFOA or analytical method 8321A_PFC_Du), the full-list of
PFCs (Prep method 3535 with analytical method PFC), or HFPO-DA only
(prep method 3535 with analytical method 8321A_HFPO_Du). If the sample
requires the analysis of FOSA it will be logged in with the prep method
3535_FOSA and the procedure in Section 10.6 should be followed instead.

10.5.2

The routine sample volume used is 250 mL. Samples should be received
from the client in un-preserved HDPE plastic bottles. If samples arereceived in glass containers, then an NCM should be written to document
the deviation from the procedure.

10.5.3

Inspect the water sample for the presence of sediment. The sample must
be free of particulate matter before it is introduced into the SPE extraction
cartridge. Particulate matter will obstruct the media and cause the analysis
to fail. Samples should NOT be filtered through glass fiber filter paper prior
to extraction. PFOA and PFOS are known to adhere to glass so filtering thesample should only be performed after the extraction has been attemptedand has failed and only with client approval.

10.5.4

If the water sample contains visible sediment or particulate matter, makesure that all particulate matter has settled to the bottom of the sample
container and take care not to disturb or re-suspend the particulate matter.
If necessary, centrifuge the sample to pull the particulate down to thebottom of the container.

10.5.5The sample may be extracted directly from the sample container, or the
sample can be transferred to a 250 mL HDPE bottle.
10.5.5.1 If the sample is transferred to a HDPE bottle, place the HDPE
bottle on a balance and tare. Then carefully pour the sample fromthe original container into the tared HDPE bottle, making sure that
if any sediment is present, it is not transferred to the bottle. Stoppouring when the balance read out is between 250 g to 275 g.
Record the balance readout as the initial sample volume in mL on
the benchsheet.
NOTE: Some clients specifically disallow decanting thesample. If this is the case and sediment is present, an
NCM must be written to document that the client
requirements could not be followed due to thesediment in the sample that would interfere with theanalysis.
10.5.5.2 If the sample is extracted directly from the original sample
container, weigh the full original sample bottle to the nearest gramusing a top loading balance, and record the weight on thebenchsheet. After the extraction, weigh the empty sample bottle,
and record the weight. Subtract the empty bottle weight from theCompany Confidential & Proprietary

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full bottle weight and record the difference as the sample volume
in mL.
NOTE: Some client specifically disallow aliquoting the sample
gravimetrically and instead require the sample to bealiquoted volumetrically. Because the PFCs are known
to adhere to glass and because of the possible crosscontamination that can occur with the use of Class A
graduated cylinders, samples for this procedure are
NOT aliquoted volumetrically. Prepare an NCM todocument why the Method Comments were notfollowed.
10.5.6

Prepare any requested MS/MSD and duplicate samples by measuring
additional sample aliquots. If the client has not requested any MS/MSD or
duplicate samples, then a sample should be chosen at random for the
MS/MSD as long as there is sufficient sample volume to do so.

10.5.7

Prepare MB and LCS samples by measuring 250 mL of HPLC grade water
into a 250 mL HDPE bottle. One MB should be performed per batch. At
least one LCS should be performed per batch. If there was insufficient
sample volume to perform a matrix spike duplicate (MSD) or a sample
duplicate, then a Laboratory Control Sample Duplicate (LCSD) must be
performed.

10.5.8Using a calibrated pipette, add 0.1 mL of the correct IS/Surrogate standard
to every sample and every MS/MSD, duplicate, MB and LCS/LCSD sample.
To determine the correct internal standard solution, check the prep andanalytical method logged and reference WI-DV-0009.
10.5.8.1 If the sample is logged for prep method 3535 with an analyticalmethod of LCMS_PFOA or an analytical method of
8321A_PFC_Du, then the analytes of interest include PFOA andPFOS only and 0.1 mL of the “PFOA/S_Sur/IS”, solution describedin Section 7.13 should be used.
10.5.8.2 If the sample is logged for prep method 3535 with an analyticalmethod of PFC, then the analytes of interest include additional
PFC compounds and the “PFC_Surr&IS”, solution described in
Section 7.14 should be used.
10.5.8.3 If the sample is logged for prep method 3535 with an analyticalmethod of 8321A_HFPO_Du, then the analytes of interest include
HFPO only and the “HFPO I.S.” solution described in section 7.16
should be used.
10.5.8.4 If the sample is logged for prep method 3535_FOSA, then theprocedure in Section 10.6 should be followed instead of thisprocedure.

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NOTE:

10.5.9The addition of all spikes and surrogates should be done
immediately following a review by a 2nd analyst. See WI-DV-0009.

Using a calibrated pipette, add 0.1 mL of the correct spike standard to every
MS/MSD and the LCS samples. To determine the correct spike standard
solution, check the prep and analytical method logged and reference WIDV-0009.
10.5.9.1 If the sample is logged for prep method 3535 with an analyticalmethod of LCMS_PFOA, or analytical method 8321A_PFC_Du
then the analytes of interest include PFOA and PFOS only and the“PFOA/S_Spike” solution described in Section 7.9.3 should be
used.
10.5.9.2 If the sample is logged for prep method 3535 with an analyticalmethod of PFC, then the analytes of interest include additional
PFC compounds and the “PFC-LCS-BR” solution described in
Section 7.11 should be used.
10.5.9.3 If the sample is logged for prep method 3535 with an analyticalmethod of 8321A_HFPO_Du, then the analytes of interest
includes HFPO only and the “HFPO Spike” solution described in
section 7.17 should be used.
10.5.9.4 If the sample is logged for prep method 3535_FOSA, then theprocedure in Section 10.6 should be followed instead of thisprocedure.

10.5.10 Cap and shake each sample to ensure the surrogate and spike compounds
are incorporated into the sample matrix. If necessary, centrifuge the sample
to separate any suspended sediment that is suspended.
10.5.11 Rinse each vacuum manifold port and line once with HPLC grade methanol
and once with HPLC grade water. Care should be taken not to use lines
made from PTFE.
10.5.12 Pre-condition the Strata X-AW 33u Polymeric Weak Anion cartridges (orequivalent) with 12 mL of HPLC grade methanol by filling the cartridge twotimes with HPLC grade methanol and drawing it through the cartridge using
a vacuum manifold.
10.5.13 Condition the SPE cartridge with 24 mL of HPLC grade water by filling the
cartridge four times and drawing the water through the cartridge using avacuum manifold. Fill the cartridge a fifth time and close the valve toprevent the water from dripping through and making sure the packing is wet.
10.5.14 After conditioning, cap each cartridge. Make sure the cap is clean andrinsed free of solvent with HPLC grade water.

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10.5.15 Using the tubing that has been rinsed with HPLC grade methanol and HPLC
grade water, connect the cartridge to the sample container. The tubing
should NOT be made of PTFE. If the extraction is being performed directlyfrom the sample container care should be taken if there are solids that havesettled to the bottom of the bottle. If necessary, use a centrifuge to pull
sediment to the bottom of the sample container and clip the tubing so thatthe end is not resting on the bottom, but suspended above the solids.
Secure the bottles to each other with rubber bands in groups of 3 or 4 sothat as the sample is drawn out of the bottle the weight of the tubing does
not cause them to tip over.
10.5.16 Turn on the vacuum and open the valves slowly so that the sample is drawn
through the cartridge at a rate of approximately 4 mL /minute. The solution
should come out of the cartridge as individual drops. If the sample comesout of the cartridge in a stream instead of drops, the extraction rate is too
fast. The extraction should take approximately 1 hour to complete, howevera slower drip rate is not detrimental to the extraction.
10.5.17 A cartridge is considered clogged if a flow rate of 4 mL/min cannot be
achieved. If the cartridge clogs during sample loading, a second cartridge
can be used for the sample and then extracts are combined. Alternatively,
measure the amount of sample successfully extracted, and use that volume
for the extraction constant. If this approach is used, then the surrogate andspike volumes must be corrected for the new initial volume. See Section
11 on how to calculate the actual surrogate and spike volumes.
10.5.18 As the sample level in the container drops, rinse the walls of the container
with HPLC water. If a clip was used to keep the end of the line off of thebottom of the sample bottle and away from solids, lower it at this point in an
attempt to extract the entire sample. If there is so much sediment thatrinsing will cause the sediment to clog the cartridge, write an NCM stating
the rinse could not be performed. After all of the sample and the water used
to rinse the container has gone through the line, close the valve and remove
the line and the cap from the cartridge. If the valve was not closed before
the cartridge went dry, re-hydrate the packing as soon as possible withreagent water. It is important not to leave the cartridge packing dry, but do
not leave any water remaining above the glass frit that sits on top of thecolumn packing.
10.5.19 After all samples have passed through the cartridge and valves have beenclosed and all lines and caps have been removed from the cartridges, rinse
each cartridge once with 6 mL of the 2% formic acid solution described in
Section 7.8. Open the valves and draw the formic acid solution through withthe vacuum. Do not allow the cartridge to go dry, but do not leave any
solution remaining above the glass frit that sits on top of the column
packing.
10.5.20 Rinse each cartridge with HPLC Grade methanol by filling the cartridges
once with 6 mL of methanol and opening the valves and drawing it throughwith the vacuum. Do not allow the cartridge to go dry, but do not leave any
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solution remaining above the glass frit that sits on top of the column
packing.
10.5.21 Turn off the vacuum and remove the manifold lid. Rinse the needles on theunderside of the manifold lid with a lab tissue that has been wetted with
HPLC grade methanol. Care should be taken not to cross-contaminate thesamples during this step. Place it to the side on a clean lab tissue. Placethe vial holder inside the manifold and place 8 mL HDPE vials that havebeen calibrated to 5 mL inside the manifold on top of the vial holder.
Replace the manifold lid and be sure that each valve needle is positioned
inside a vial.
10.5.22 Using a bottle-top pump add 4 mL of the 10% ammonium hydroxide inmethanol solution described in Section 7.6 to each cartridge. Turn on thevacuum pump while the valves on the manifold are still closed, then quickly
open and close each valve to create a vacuum in the cartridges. Turn off
the vacuum pump and open the manifold valve to break the vacuum in the
manifold. Open the valves to allow the ammonium hydroxide to drip
gravimetrically.
10.5.23 After the solution has stopped dripping, reapply the vacuum to ensure thatthe last portion of solution is collected. This is a very important step.
10.5.24 If two cartridges were used, transfer all extracts into one collection vial andevaporate down to approximately 4 mL using an N-Evap. Continue to thenext step.
10.5.25 Add HPLC grade water to the extract in the sample collection vial until themeniscus reaches the 5.0-mL mark.
10.5.26 The water sample extracts are ready for analysis. See SOP DV-LC-0012
for details on the analysis.
10.5.27 See Section 14.0 for proper disposal of all waste.
10.6

Water Sample Extraction for FOSA by 3535_FOSA
10.6.1

Inspect the water sample for the presence of sediment. The sample must
be free of particulate matter before it is introduced into the SPE extraction
cartridge. Particulate matter will obstruct the media and cause the analysis
to fail. Samples should NOT be filtered through glass fiber filter paper prior
to extraction. PFCs are known to adhere to glass so filtering the sample
should only be performed after the extraction has been attempted and has
failed and only with client approval.

10.6.2

If the water sample contains visible sediment or particulate matter, makesure that all particulate matter has settled to the bottom of the sample
container and take care not to disturb or re-suspend the particulate matter.
If necessary, centrifuge the sample to pull the particulate down to theCompany Confidential & Proprietary

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bottom of the container.
10.6.3The sample may be extracted directly from the sample container, or the
sample can be transferred to a 250 mL HDPE bottle.
10.6.3.1 If the sample is transferred to a HDPE bottle, place the HDPE
bottle on a balance and tare. Then carefully pour the sample fromthe original container into the tared HDPE bottle, making sure that
if any sediment is present, it is not transferred to the bottle. Stoppouring when the balance read out is between 250 g to 275 g.
Record the balance readout as the initial sample volume in mL on
the benchsheet.
NOTE: Some client requirements specifically disallow
decanting the sample. If this is the case and sedimentis present, an NCM must be written to document thatthe client requirements could not be followed due to thesediment in the sample that would interfere with theanalysis.
10.6.3.2 If the sample is extracted directly from the original sample
container, weigh the full original sample bottle to the nearest gramusing a top loading balance, and record the weight on thebenchsheet. After the extraction, weigh the empty sample bottle,
and record the weight. Subtract the empty bottle weight from thefull bottle weight and record the difference as the sample volume
in mL.
NOTE: Some client specifically disallow aliquoting the sample
gravimetrically and instead require the sample to bealiquoted volumetrically. Because the PFCs are known
to adhere to glass and because of the possible crosscontamination that can occur with the use of Class A
graduated cylinders, samples for this procedure are
NOT aliquoted volumetrically. Prepare an NCM todocument why the Method Comments were notfollowed.

10.6.4

Prepare any requested MS/MSD and duplicate samples by measuring
additional sample aliquots. If the client has not requested any MS/MSD or
duplicate samples, then a sample should be chosen at random for the
MS/MSD as long as there is sufficient sample volume to do so. When an
MS/MSD pair is not available, an LCS and LCSD are required.

10.6.5Prepare MB and LCS samples by measuring 250 mL of HPLC grade water
into a 250 mL HDPE bottle. One MB should be performed per batch. At
least one LCS should be performed per batch. If there was insufficient
sample volume to perform a matrix spike duplicate (MSD) or a sample
duplicate, then a Laboratory Control Sample Duplicate (LCSD) should be
performed.
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NOTE:

The addition of all spikes and surrogates should be done immediately
following a review by a 2nd analyst. See WI-DV-0009.

10.6.6Using a calibrated pipette, add 1.0 mL of the “FOSA8-IS-SPK” solution
described in Section 7.14.3 to all samples and all QC samples.

10.6.7

Using a calibrated pipette, add 1.0 mL of the “FOSA-H2O-LCS” solution
described in Section 7.12 to every MS/MSD and to the LCS samples.

10.6.8Cap and shake each sample to ensure the surrogate and spike compounds
are incorporated into the sample matrix. If necessary, centrifuge the sample
to separate any suspended sediment that is suspended.

10.6.9Rinse each vacuum manifold port and line once with HPLC grade methanol
and once with HPLC grade water. Care should be taken not to use lines
made from PTFE.

10.6.10 Pre-condition the Waters tC18 SPE cartridge (or equivalent) with 12 mL of
HPLC grade methanol by filling the cartridge two times with methanol anddrawing it through the cartridge using a vacuum manifold.
10.6.11 Condition the SPE cartridge with approximately 24 mL of HPLC grade waterby filling the cartridge four times and drawing the water through the cartridge
using a vacuum manifold. Fill the cartridge a fifth time and close the valve
to prevent the water from dripping through and making sure the packing iswet.
10.6.12 After conditioning, cap each cartridge. Make sure the cap is clean andrinsed free of solvent with HPLC grade water.
10.6.13 Using the tubing that has been rinsed with HPLC grade methanol and HPLC
grade water, connect the cartridge to the sample container. The tubing
should NOT be made of PTFE. If the extraction is being performed directlyfrom the sample container care should be taken if there are solids that havesettled to the bottom of the bottle. If necessary, use a centrifuge to pull
sediment to the bottom of the sample container and clip the tubing so thatthe end is not resting on the bottom, but suspended above the solids.
Secure the bottles to each other with rubber bands in groups of 3 or 4 sothat as the sample is drawn out of the bottle the weight of the tubing does
not cause them to tip over.
10.6.14 Turn on the vacuum and open the valves slowly so that the sample is drawn
through the cartridge at a rate of approximately 4 mL/minute. The solution
should come out of the cartridge as individual drops. If the sample comesout of the cartridge in a stream instead of drops, the extraction rate is too
fast. The extraction should take approximately 1 hour to complete, howevera slower drip rate is not detrimental to the extraction.
10.6.15 A cartridge is considered clogged if a flow rate of 4 mL/min cannot be
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achieved. If the tube clogs during sample loading, a second cartridge canbe used for the sample and then extracts are combined. Alternatively,
measure the amount of sample successfully extracted, and use that volume
for the extraction constant. If this approach is used, then the surrogate andspike volumes must be corrected for the new initial volume. See Section
11 on how to calculate the actual surrogate and spike volumes.
10.6.16 As the sample level in the container drops, rinse the walls of the container
with HPLC water. If a clip was used to keep the end of the line off of thebottom of the sample bottle and away from solids, lower it at this point in an
attempt to extract the entire sample. If there is so much sediment thatrinsing will cause the sediment to clog the cartridge, write an NCM stating
the rinse could not be performed.
10.6.17 After all of the sample and the water used to rinse the container has gone
through the line, close the valve and remove the line and the cap from the
cartridge. If the valve was not closed before the cartridge went dry, rehydrate the packing as soon as possible with reagent water. It is important
not to leave the cartridge packing dry, but do not leave any water remaining
above the glass frit that sits on top of the column packing.
10.6.18 It is important not to let the cartridge go dry at this time, but to leave water inthe cartridge.
10.6.19 After every sample and every QC sample has been loaded and all of thevalves are closed and the caps removed, open all valves and pull a vacuum
on all of the cartridges for 3 minutes to remove the remaining sample and to
dry the cartridge. Care should be taken that the samples are only dried for
3 minutes.
10.6.20 Turn off the vacuum, close all of the valves, and remove the manifold lid.
Rinse the needles on the underside of the manifold lid with HPLC grade
methanol and wipe them with a clean lab tissue. Place it to the side on aclean lab tissue. Place the vial holder inside the manifold and place 8 mL
HDPE vials that have been calibrated to 5 mL inside the manifold on top of
the vial holder. Replace the manifold lid and be sure that each valve ispositioned inside a vial.
10.6.21 Add 4 mL of HPLC grade methanol to each cartridge. Turn on the vacuum
pump while the valves on the manifold are still closed, then quickly open
and close each valve to create a vacuum in the cartridges. Turn off thevacuum pump open the manifold valve to break the vacuum in the manifold.
Open the valves to allow the methanol to slowly drip gravimetrically(approximately one drop in 5 seconds.).
10.6.22 After the methanol has stopped dripping, reapply the vacuum to ensure thatthe last portion of methanol is collected. This is a very important step.
10.6.23 If two cartridges were used, transfer all extracts into one collection vial andCompany Confidential & Proprietary

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evaporate down to approximately 4 mL using a N-Evap. Continue to thenext step.
10.6.24 Add HPLC grade water to the extract in the sample collection vial until themeniscus reaches the 5.0 mL mark.
10.6.25 The water sample extracts are ready for analysis. See SOP DV-LC-0012
for details on the analysis.
10.6.26 See Section 14.0 for proper disposal of all waste.
10.7

Soil Sample Extraction
Note:

If samples are logged for prep method PFC_Leach and analytical method
8321A_HFPO_Du, do not follow the procedure described in 10.7. Instead,follow the procedure described in section 10.8.

10.7.1Visually inspect soil samples for homogeneity.

10.7.2

Weigh a representative 10-11 gram amount of each soil into a 125-mL
polypropylene wide-mouth bottle. Weigh additional sample amounts for thematrix spike and sample duplicate. Use baked Ottawa sand for the method
blank and LCS sample matrix

10.7.3Using a calibrated pipette, add 0.4 mL of the correct internal standard to
every sample and every MS/MSD, duplicate, MB and LCS/LCSD sample.
To determine the correct internal standard solution, check the analyticalmethod logged and reference WI-DV-0009.
10.7.3.1 If the sample is logged for analytical method LCMS_PFOA thenthe analytes of interest include PFOA and PFOS only and the“PFOA/S_Sur/IS” solution described in Section 7.13 should be
used.
10.7.3.2 If the sample is logged for analytical method PFC, then theanalytes of interest include additional PFC compounds and the“PFC_Surr&IS”, solution described in Section 7.14 should be
used.

NOTE:

The addition of all spikes and surrogates should be done immediately
following a review by a 2nd analyst. See WI-DV-0009.

10.7.4Using a calibrated pipette, add 0.4 mL of the correct spike solution to the
MS/MSD samples and any LCS or LCSD. To determine the correct spikesolution, check the analytical method logged and reference WI-DV-0009.
10.7.4.1 If the sample is logged for analytical method LCMS_PFOA thenthe analytes of interest include PFOA and PFOS only and theCompany Confidential & Proprietary

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used.
10.7.4.2 If the sample is logged analytical method PFC, then the analytes
of interest include additional PFC compounds and the “PFC-LCSBR” solution described in Section 7.11 should be used.
10.7.5Using a calibrated bottle top pump or a Class A graduated cylinder, add
exactly 3 mL of 0.67 N NaOH described in Section 7.4 and allow the sample
to soak for 30 minutes.

10.7.6Using a calibrated bottle top pump or a Class A graduated cylinder, add
exactly 10 mL of HPLC grade methanol to each field sample and QC
sample.

10.7.7

Tumble each sample at room temperature for 24 hours.

NOTE:

Make sure that tape is placed over the label to prevent it from being rubbed
off.

10.7.8Add 1 mL of the 2.4 M HCl solution described in Section 7.5 to all samples
and gently mix. Measure the pH of the extraction solution with wide range
pH paper. If the pH is greater than 7, add an additional 1 mL of 2.4 M HCl.
Repeat the HCl additions until the pH measures less than 7. Note howmuch acid is added to each sample.

10.7.9Using a calibrated pipette, or a Class A graduated cylinder, add theappropriate amount of HPLC grade methanol to adjust the extract solution
to 20 mL. For samples that required only 1 mL of 2.4 M HCl solution in
Section 10.7.8 above, 5.6 mL of HPLC grade methanol is needed to reach a
final volume of 20 mL. For LCS and MS/MSD samples the final volume willbe 20.4 mL.

NOTE:

If more than 1 mL of HCl was added in Section 10.7.9 then add lessmethanol so that the final volume remains 20 mL, (20.4 mL for LCS and
MS/MSD samples). For example, if it required 2 mL of HCl to achieve a pH
less than 7, then only 4.6 mL of HPLC grade MeOH should be added.

10.7.10 Shake briefly to mix.
10.7.11 The samples may be centrifuged to help facilitate the filtration. Withdraw a
5-mL aliquot of the supernatant liquid and filter through a 0.45-m PVDF
syringe filter into a labeled 10 mL HDPE vial.
10.7.12 The soil sample extracts are ready for analysis. See SOP DV-LC-0012 fordetails on the analysis.
10.7.13 See Section 14.0 for proper disposal of all waste.

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10.8

Soil Sample Extraction for analytical method 8321A_HFPO_Du
Note: These samples are prepared with a final volume of 50 mL.
10.8.1Visually inspect soil samples for homogeneity.

10.8.2

Weigh a representative 10-11 gram amount of each soil into a 125-mL
polypropylene wide-mouth bottle. Weigh additional sample amounts for thematrix spike and sample duplicate. Use baked Ottawa sand for the method
blank and LCS sample matrix.

10.8.3Using a calibrated pipette, add 1.0 mL of HFPO I.S. (described in section
7.16) to every sample and every MS/MSD, duplicate, MB, and
LCS/LCSD/LLCS sample.

NOTE:

The addition of all spikes and surrogates should be done immediately
following a review by a 2nd analyst. See WI-DV-0009.

10.8.4Using a calibrated pipette, add 1.0 mL of HFPO Spike (described in section
7.17) to the MS/MSD samples and any LCS or LCSD. Add 0.1 mL of HFPO
Spike to any LLCS.

10.8.5

Cap the bottles and allow the standards to settle into the sample matrix.
Gently shake the bottles to mix the spike into the matrix.

10.8.6Using a calibrated bottle top pump or a Class A graduated cylinder, add
enough volume of the 5% NH4OH solution described in section 7.7 to bringthe extract volume to 45 mL. For example, add 43 mL to any LCS, MS, andadd 44 mL to the MB, field samples, and LLCS.

10.8.7

Seal the bottles and vortex to completely mix the soil and the extraction
solvent. Typically, 10-15 seconds on the vortex is sufficient, but dependingon the sample matrix, some samples may require more time.

10.8.8

Tumble each sample at room temperature for 18-24 hours.

NOTE:

Make sure that tape is placed over the label to prevent it from being rubbed
off.

10.8.9Add 5 mL of the 2% Formic Acid solution described in 7.8 to all samples andgently mix. This brings the volume to 50 mL.

10.8.10 Shake briefly to mix.
10.8.11 The samples may be centrifuged to help facilitate the filtration. Withdraw a
5-mL aliquot of the supernatant liquid and filter through a 0.45-m PVDF
syringe filter into a labeled 10 mL HDPE vial.

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10.8.12 The soil sample extracts are ready for analysis. See SOP DV-LC-0012 fordetails on the analysis.
10.8.13 See Section 14.0 for proper disposal of all waste.
10.9

MaintenanceNot applicable for this procedure.

10.10 Troubleshooting
When dealing with unusual sample matrices, consult QA or a supervisor.
11.0

Data Analysis and Calculations
Volume of Surrogate or Spike Extracted = (VSA ) x (VE ) ÷ (VI)
Where:
VSA = Volume of Spike or Surrogate originally added.
VE = Volume of Sample that was extracted through the cartridge
VI = Volume of Sample that was originally spiked
Example: 0.1 mL of surrogate standard was added to a 253 mL sample. During
the extraction, the cartridge clogged and only 233 mL of sample wasactually extracted.
Volume of Surrogate Extracted = 0.1 mL x 233 mL ÷ 253 mL= 0.092 mL
Therefore the initial volume on the benchsheet should be entered as
233 mL and the volume of surrogate should be entered as 0.092 mL.

12.0

Method Performance
12.1

Method Validation
Because this method has not been validated by an authoritative standards setting
body, such as the U.S. EPA or ASTM, TestAmerica Denver conducted a series ofmethod validation studies before writing this SOP. The validation studies werebased on the guidelines of the U.S. EPA Pesticide Program. These studies aredescribed in detail in the TestAmerica Denver documents “Method Validation Study
for Analysis of Perfluorooctanoic Acid in Waters by High Performance Liquid
Chromatography / Tandem Mass Spectrometry (HPLC/MS/MS)” and “Method

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Validation Study for Analysis of Ammonium Perfluorooctanate in Soil Matrices byHigh Performance Liquid Chromatography / Mass Spectrometry (HPLC/MS/MS).”
12.2

The method detection limit (MDL) is the lowest concentration that can be detected fora given analytical method and sample matrix with 99% confidence that the analyte ispresent. The MDL is determined according to the laboratory’s MDL policy in DV-QA005P. MDLs reflect a calculated (statistical) value determined under ideal laboratory
conditions in a clean matrix, and may not be achievable in all environmental matrices.
The laboratory maintains MDL studies for analyses performed; these are verified atleast annually unless method or program requirements require a greater frequency.

12.3

Demonstration of Capabilities
All personnel are required to perform an initial demonstration of proficiency (IDOC)
on the 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.

12.4

12.3.1

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.

12.3.2

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

12.3.3

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.

12.3.4

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

12.3.5Further 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 by anassociate who has been properly trained in its use and has the required experience.
A new analyst must be working under supervision prior to approval of the IDOC.
Documentation that a new analyst is performing under supervision must be enteredinto the batch record (View Batch Information) until that analyst’s IDOC has been
approved by the QA Manager (or designee). See requirements for demonstration ofanalyst proficiency in SOP DV-QA-0024.

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13.0

14.0

Pollution Control
13.1

Solid phase extraction used for water samples greatly reduces the amount ofsolvent used compared to liquid/liquid extraction.

13.2

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

Waste Management
14.1

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 the Environmental Health and Safety
Manual, and DV-HS-001P, “Waste Management Program.”

14.2

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

NOTE:

15.0

14.2.1

Methanol waste – Flammable Solvent (C )14.2.2

Water Samples, post extraction and all other aqueous waste generated bythis procedure is collected and turned into the Waste Coordinator forincineration under waste stream PFOA.

14.2.3

Soil Samples, post extraction and all other solid waste generated by thisprocedure such as disposable pipette tips and extraction bottles arecollected and turned into the Waste Coordinator for incineration under waste
stream PFOA.

14.2.4

Expired Chemicals/Reagents/Standards – Contact Waste CoordinatorRadioactive and potentially radioactive or mixed waste must be segregated from
non-radioactive waste as appropriate. Contact the Waste Coordinator for proper
management of radioactive or potentially radioactive waste generated by thisprocedure.

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

16.0

Method Modifications:
The SPE procedure is modified from SW-846 Method 3535A in order to achieve theoptimum recovery for the analytes of interest.

17.0

TablesTable 1: Perfluorinated Hydrocarbons (PFCs)
Table 2: Isotopically Labeled PFCs
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Table 3: Surrogate Compounds
18.0

Revision History
18.1

Revision 9, dated 8 December 2017
 Changed all references to PFC-LCS to the new standard name, PFCLCS-BR.
 Added section 7.7 to include 5% Ammonium Hydroxide reagent;
renumbered subsequent entries in section 7.
 Added 7.16 and 7.17 to include HFPO-DA standards.
 Added detail to 10.5.1 to include HFPO-DA.
 Added 10.5.8.3 and 10.5.9.3 (and renumbered subsequent sections) toinclude standards used for HFPO-DA.
 Added Note to section 10.7 to redirect to 10.8 for HFPO-DA soils.
 Added section 10.8 to outline procedure developed for extracting HFPODA from soils.

18.2

Revision 8, dated 31 October 2016
 Added initial paragraph to section 3.0 to reference the QAM for general
analytical and QA/QC terms
 Added initial paragraph to section 6.0 regarding the documentation of
support equipment IDs
 Added section 6.3 on Software and Hardware
 Updated sections 6.2.1, 7.6, 7.9, 7.11.1, 7.12.1, 7.13.1, 7.14.1, 10.3.12,
10.3.20, 10.4.10, 10.4.21, 10.5.6, 10.5.9 to reflect the change from using
LCMS grade methanol to HPLC grade Methanol. Per the company, HPLC
grade is now clean enough to use for preparation and analysis.
 Removed section 7.2 regarding LCMS grade MeOH used in the making ofstandards, conditioning of SPE cartridges, the extraction of water samples
for FOSA and the extraction of soil samples.
 Updated section 7.1 with information regarding HPLC grade MeOH used in
the making of standards, conditioning of SPE cartridges, the extraction ofwater samples for FOSA, and the extraction of soil samples.
 Updated section 9.1 and it’s subsections to reflect current practices andconsistent verbiage
 Removed “At least” from section 9.4 to reflect that one MB is performed foreach batch Added numbering to sections 9.6.1 and 9.6.2
 Added LCS/LCSD requirements when no MS/MSD to sections 9.5, 9.6.2,
10.5.7, 10.6.4.
 Added section 10.3 comment regarding the rotation of glassware andpositions Added section 10.4 regarding the documentation of support equipment IDs

18.3

Revision 7, dated 31 October 2015
 Removed Confidentiality Restriction Note boxes

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








Updated Sections 12.2 and 12.3 to reflect consistent information asprovided in other SOPs.
Section 6.1 was revised to include a centrifuge in the equipment list.
Added instructions to the solid phase extraction procedure to cap andshake water samples after the addition of the surrogate and spikestandards.
Added instructions to the solid phase extraction procedure to use centrifuge
to separate solids from the sample to prevent the cartridge from clogging.
Also instructed analysts to write an NCM if sediment prevents them fromrinsing the sample container.
Revised the solid phase procedure to clarify that it is important to re-hydrate
the cartridge packing as soon as possible if the cartridge goes dry.
Revised the pre-conditioning of the FOSA cartridge to be 24 mL instead of
48 mL of water to bring the procedure in line with other SPE methods.

18.4

Revision 6, dated 13 October 2014
 Revised Section 6.2.4 and 10.3.11 to reflect current cartridge in use andadded “or equivalent” to enable laboratory obtain cartridges from anothervendor or change the cartridge to obtain equivalent performance.
 Updated Section 8 to revise HT for water to reflect validation study
performed by TestAmerica Denver and to be consistent with Section 8 in
the analytical SOP, DV-LC-0012.
 Added new section 12.2 to address MDLs/verification of MDLs.
 Replaced 4C  2C with 0-6C for consistency with current EPA practiceand laboratory control limits on refrigerator temperatures.
 Formatting and editorial changes throughout.

18.5

Revision 5, Dated 04 March 2014
 The procedure was revised to remove the 3 hour sonication step in the soil
extraction. A new MDL study and IDOC were performed prior to thechange.
 Troubleshooting and Maintenance Sections were added per DoD QSM 5.0.
 Section 9.1 was revised to state the procedure meets all criteria for DoD
QSM 5.0 unless otherwise stated.
 Sections 9.4, 9.5, 9.6, 9.7, and 9.8 were revised to remove acceptancecriteria and corrective actions. This information can be found in theanalytical SOP.
 Section 10.1 was revised to clarify that any deviations from this procedure
discovered after the work has been performed must be documented in an
NCM.
 Section 10.4.2 was revised to give a range for the initial sample mass.
 Section 10.4.11 was revised to place the centrifuge step before theinstructions to withdraw the supernatant.

18.6

Revision 4, Dated December 10, 2013
 Confidential information included
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18.7

Revision 3, Dated March 4, 2013
 Annual Technical Review
 Section 2.3 was revised to better explain that the soil extracts are brought
up to final volume with methanol.
 Section 10.2.18 was revised to better explain that the cartridge packing
cannot be allowed to go dry, but excess water should not be left in the
cartridge.
 Section 10.2.21 was revised to give guidance on how to avoid crosscontamination during the rinsing of the manifold lid.
 Section 10.4.5 was removed as the 0.4 mL of IS and 0.4 mL of spikesolution readily absorbs into the sample and therefore capping and shaking
was not necessary.
 Section 10.4.6 and 10.4.7 (now 10.4.5 and 10.4.6) were revised to matchcurrent practice of using a calibrated bottle-top pump instead of amechanical pipette to add the NaOH and the MeOH.

18.8

Revision 2.1, Dated January 31, 2012
 Removed references to method “8321_PFC_Du” from the soil extraction
procedure as the extraction required is no longer per this procedure, but pera client specific QAS.
 Section 2.3 was corrected to state that soil samples are adjusted to a pH
less than or equal to 7.
 Section 6.1 was corrected to state that the balance is capable of measuring± 0.1g.
 Section 6.2 was corrected to state that the Waters Oasis WAX cartridges
are 150 mg.
 Sections 7.6, 7.7, and 7.8 were revised to instruct analysts to add acid andbase to water instead of adding water to the acids and base.
 Updated Section 9.1 and 10.1 to reflect the current NCM process.
 Added clarification to Sections 10.2.17 and 10.3.17 on how to calculate thevolume of surrogate and spike extracted when a cartridge clogs. Added
Section 11 to include the formula required to do the calculation.

18.9

Revision 2, dated January 21, 2011 The procedure was revised to disallow the use of glass volumetrics in thealiquotting of samples. This was done to prevent loss of target analytes as
the target analytes are known to adhere to glass.
 The procedure was revised to change the final extract volume for soil
extracts from 50 mL to 20 mL. The volume of spike, internal standards, andsurrogate standards used in the extractions were changed to maintain the
spike concentration in the extract.
 The internal standard used to quantitate the concentration of FOSA has
been changed from MeFOSA to 13C8 FOSA.
 The extraction procedure in Section 10.3 for the extraction of FOSA wasrevised extensively. This was done to compensate for matrix issues arisingfrom the high pH required in the old procedure.
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Earlier revision histories have been archived and are available upon request.

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Table 1 – Perfluorinated Hydrocarbons (PFCs)
NAME
Perfluorodecane sulfonate
Perfluorohexane sulfonate
Perfluorobutyric acid
Perfluoropentanoic acid
Perfluorohexanoic acid
Perfluoroheptanoic acid
Perfluorononanoic acid
Perfluorodecanoic acid
Perfluoroundecanoic acid
Perfluorododecanoic acid
Perfluorotridecanoic acid
Perfluorotetradeconoic acid
Perfluorobutane sulfonate
Perfluorooctanoic sulfonate
Perfluorooctanoic acid
Perfluorooctane sulfonamide

ABBREVIATION
PFDS
PFHxS
PFBA
PFPA
PFHxA
PFHpA
PFNA
PFDA
PFUnA
PFDoA
PFTriA
PFTeA
PFBS
PFOS
PFOA
FOSA

Table 2 – Isotopically Labeled PFCs
NAME
13C4 PFOA
13C4 PFOS
13C4 PFBA
13C4 PFHxA
18O2 PFHxA
13C5 PFNA
13C2 PFDA
13C2 PFUnA
13C2 PFDoA
13C8 FOSA
Table 3 – Surrogate Compounds
NAME
13C8 PFOA
13C8 PFOS

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