Sacramento
SOP No. WS-LC-0031, Rev. 2.1
Effective Date: 04/03/2019
Page No.: 1 of 27
NOTE: The method described in this document is the exclusive, confidential, proprietary, and secretproperty of Chemours, developed by TestAmerica Sacramento under contract to Chemours. Use of thisdocument and the information contained in it is restricted and governed by the terms of the
CONFIDENTIALITY RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD OF THIS DOCUMENT
OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR PART, FOR ANY PURPOSE WITHOUT THE
PRIOR WRITTEN CONSENT OF CHEMOURS.

Title: Determination of Table 3 Compounds by LC/MS/MS
Chemours Fluoroproducts

Approvals (Signature/Date):
Robert Hrabak
Technical Manager

Lisa Stafford
Quality Assurance Manager

03/27/2019
Date

04/01/2019
Date

03/27/2019
Joe Schairer
Date
Health & Safety Manager / Coordinator

Chris Williams
Laboratory Manager

03/28/2019
Date

Copyright Information:
This documentation has been prepared by TestAmerica Laboratories, Inc. and its affiliates (“TestAmerica”),
solely for their own use and the use of their customers in evaluating their qualifications and capabilities inconnection with a particular project. The user of this document agrees by its acceptance to return it to
TestAmerica upon request and not to reproduce, copy, lend, or otherwise disclose its contents, directly or
indirectly, and not to use it for any purpose other than that for which it was specifically provided. The user also
agrees not to give access to this document to any third parties including but not limited to consultants, unlesssuch third parties specifically agree to these conditions.
THIS DOCUMENT CONTAINS VALUABLE CONFIDENTIAL AND PROPRIETARY INFORMATION.
DISCLOSURE, USE OR REPRODUCTION OF THESE MATERIALS WITHOUT THE WRITTENAUTHORIZATION OF TESTAMERICA IS STRICTLY PROHIBITED. THIS UNPUBLISHED WORK BYTESTAMERICA IS PROTECTED BY STATE AND FEDERAL LAW OF THE UNITED STATES. IF
PUBLICATION OF THIS WORK SHOULD OCCUR THE FOLLOWING NOTICE SHALL APPLY:©COPYRIGHT 2019 TESTAMERICA LABORATORIES, INC. ALL RIGHTS RESERVED.

Uncontrolled
Facility Distribution No. ___________

Sacramento Bids Folder
Distributed To:_______________________NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

1.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 2 of 27

SCOPE AND APPLICATION
1.1.

This procedure describes the analysis of water samples for the following compounds
using liquid chromatography / tandem mass spectrometry (LC/MS/MS).

Table 3 Compound Name

Abbreviation

PFO5DAPEPAPMPAPFECA_G
Byproduct 1
Byproduct 2
PFMOAAPFO2HxAPFO3OAPFO4DAMeFOSA
EtFOSA
MetFOSE
EtFOSEDFSA
MMF
MTP
PPF AcidNVHOSPFECA BPES
HFPO-DA
Hydro-EVE Acid
EVE Acid
R-EVE
Byproduct 4
Byproduct 5
Byproduct 6

1.2.

CAS #

PFO5A
PEPA
PMPA
PFECA_G
PFESA BP1
PFESA BP2PFMOAAPFO2HxAPFO3DAPFO4DAMeFOSA
EtFOSAMeFOSE
EtFOSEDFSA
MMF
MTP
PPF AcidNVHOSPFECA BPES
HFPO-DA
Hydro-EVE Acid
EVE Acid
R-EVE
Byproduct 4
Byproduct 5
Byproduct 639492-91-6
NA13140-29-9
801212-59-9
29311-67-9749836-20-2
674-13-539492-88-1
39492-89-2
39492-90-5
31506-32-8
4151-50-2
24448-09-7
1691-99-2
422-67-3
1514-85-893449-21-9
422-64-0
1132933-86-8
151772-58-6
113507-82-7
13252-13-6
773804-62-9
69087-46-3
EVS1428
EVS1429
EVS1430
EVS1431

The working range of the method is listed below. The linear range can be extended bydiluting the extracts.
MatrixWater

Nominal Sample
Size

Reporting Limit

Working Range

2.5 mL

2-50 ng/L

2 ng/L – 500 ng/L

Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

2.

3.

4.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 3 of 27

SUMMARY OF METHOD
2.1.

Water samples are diluted 50:50 with methanol, filtered and analyzed by LC/MS/MS.

2.2.

The final 50:50 methanol:water extracts are analyzed by LC/MS/MS. PFAS are
separated from other components on a C18 column with a solvent gradient programusing 20 mM ammonium acetate/water and methanol. The mass spectrometer detector
is operated in the electrospray (ESI) negative ion mode for the analysis of PFAS.

DEFINITIONS
3.1.

PFAS: Per- and Polyfluorinated Alkyl Substances

3.2.

PTFE: Polytetrafluoroethylene (e.g., Teflon®)

3.3.

PP:

Polypropylene

3.4.

PE:

Polyethylene

3.5.

HDPE: High density polyethylene

3.6.

Further definitions of terms used in this SOP may be found in the glossary of theLaboratory Quality Assurance Manual (QAM).

INTERFERENCES
4.1.

PFAS have been used in a wide variety of manufacturing processes, and laboratory
supplies should be considered potentially contaminated until they have been tested andshown 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 in anarea separate from where samples are extracted.

4.3.

The use of PTFE in the procedure should be avoided or at least thoroughly tested
before use. Polypropylene (PP) or polyethylene (PE, HDPE) products may be used inplace of PTFE products to minimize PFOA contamination.

4.4.

Standards and samples are injected from polypropylene auto sampler vials with
polypropylene screw caps once. Multiple injections may be performed on Primers
when conditioning the instrument for analysis.
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

4.5.

5.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 4 of 27

Teflon-lined screw caps have detected PFAS at low concentrations. Repeated injection
from the same Teflon-lined screw cap have detected PFNA at increasing concentration
as each repeated injection was performed, therefore, it is best to use polypropylene
screw caps.

SAFETY
Employees must abide by the policies and procedures in the Corporate Safety Manual,
Sacramento Supplement to the CSM, and this document. All work must be stopped in theevent of a known or potential compromise to the health or safety of an associate. The situationmust be reported immediately to a supervisor, the EH&S Staff, or a senior manager.
5.1.

Specific Safety Concerns
Preliminary toxicity studies indicate that PFAS could have significant toxic effects. In
the interest of keeping exposure levels as low as reasonably achievable, PFAS and
PFAS samples must be handled in the laboratory as hazardous and toxic chemicals.

5.2.

5.1.1.

Exercise caution when using syringes with attached filter disc assemblies.
Application of excessive force has, upon occasion, caused a filter disc to burst
during the process.

5.1.2.

Laboratory procedures such as repetitive use of pipets, repetitive transferringof extracts and manipulation of filled separatory funnels and other glassware
represent a significant potential for repetitive motion or other ergonomic
injuries. Laboratory associates performing these procedures are in the best
position to realize when they are at risk for these types of injuries. Whenever asituation is found in which an employee is performing the same repetitive
motion, the employee shall immediately bring this to the attention of theirsupervisor, manager, or the EH&S staff. The task will be analyzed todetermine a better means of accomplishing it.

5.1.3.

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

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 themethod. 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
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 5 of 27

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.
(1)

(2)

Material

Hazards

Exposure Limit

Acetic Acid
(3-2-1)

Corrosive
PoisonFlammable

10 ppm-TWA
15 ppm-STEL

Methanol (2-3-0)

Flammable
PoisonIrritant

200 ppm (TWA)

Potassium
Hydroxide(3-0-1)

Corrosive Poison
Water Reactive

2 mg/m PEC/ceiling

Sulfuric Acid (1)(3-0-2)

Corrosive
Oxidizer
Dehydrator

1 mg/m3

3

Signs and Symptoms of Exposure
Contact with concentrated solution may cause serious
damage to the skin and eyes. Inhalation of concentrated
vapors may cause serious damage to the lining of thenose, throat, and lungs. Breathing difficulties may occur.
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 adefatting agent and may cause skin to become dry andcracked. Skin absorption can occur; symptoms may
parallel inhalation exposure. Irritant to the eyes.
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.
Contact with concentrated solution may cause serious
damage to the skin and eyes. Inhalation of concentrated
vapors may cause serious damage to the lining of thenose, throat, and lungs. Breathing difficulties may occur.

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

6.

EQUIPMENT AND SUPPLIES
6.1.

250 mL HDPE bottles with HDPE screw caps.

6.2.

Analytical balance capable of accurately weighing to the nearest 0.0001g, and checked
for accuracy each day it is used in accordance with WS-QA-0041.

6.3.

Auto-pipets capable of accurately dispensing volumes of 2.5 mL, 2.0 mL, and 0.5 mL,
0.2 µm GHP filters, and other equipment used to prepare standards and reagents.

6.4.

Syringe filter, Millipore Millex-HV 0.45 um, or equivalent. Do not use PTFE typefilters.

6.5.

2mL auto sampler vials, clear glass, Thermo Scientific Nation surestop vial, part no.
C5000-1, or equivalent.

6.6.

Vial caps, Thermo Scientific National AVCS blue cap, pre slit TEF/STL septa, part no.
C5000-55B or equivalent.

Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 6 of 27

6.7.

Eppendorf 1000uL epTIPS, part no. 022491954 or equivalent.

6.8.

1000uL Pipette: Eppendorf Research Plus

6.9.

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

6.10.

pH indicator paper, wide range.

6.11.

Glass fiber filter, Whatman GF/F, catalog number 1825 090 or equivalent.

6.12.

Liquid Chromatography/Tandem Mass Spectrometer (LC/MS/MS) – described below,
or equivalent, may be used for this method. The HPLC is equipped with a refrigerated
auto sampler, an injection valve, and a pump capable of variable flow rate. The use of acolumn heater is required to maintain a stable temperature throughout the analyticalrun. Data is processed using Chrom Peak Review, version 2.1 or equivalent.
6.12.1. SCIEX LC/MS/MS
6.12.1.1. This system consists of a Shimadzu HPLC interfaced with a SCIEX
5500 Triple Quad MS. The instrument control and data acquisition
software is SCIEX Analyst, version 1.6.3 or equivalent.
6.12.2. Shimadzu CTO-20AC HPLC equipped with 3 LC-20AD pumps and one
DGU-20 degassing unit or equivalent.
6.12.2.1. Phenomenex Gemini C18 3 um, 3.0 mm x 100 mm, Part No. 00D4439-Y0, or equivalent.
6.12.2.2. PFAS Isolator column, Phenomenex Luna C18 5 um, 50 mm x 4.6mm, part no. 00B-4252-E0 or equivalent. This is plumbed between
the UPLC pumps and auto sampler valve to minimize PFAS
background from the UPLC solvent lines and filters.
6.12.2.3. Diol Guard Cartridges, Part No. 820950-911 and High Perf.
ZORBAX Guard, Part No. 820999-901. This is plumbed in front of
the Phenomenex Gemini C18 3 um, 3.0 mm x 100 mm column.
6.13.

Preventive and routine maintenance is described in the table below

Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 7 of 27

HPLC/MS/MS Preventative Maintenance
As Needed:
Change pump seals.
Change in-line filters in auto sampler (HPLC).
Check/replace in-line frit if excessive pressure orpoor performance.
Replace column if no change following in-line fritchange.
Clean corona needle.
Replace sample inlet tube in APCI (10.1 cm).
Replace fused silica tube in ESI interface.
Clean lenses.
Clean skimmer.
Ballast rough pump 30 minutes.
Create all eluents in Reagent module, labelndeluent containers with TALS label and place 2label into maintenance log when put into use.
Semi-AnnuallyReplace rough-pump oil (4-6 months).
Replace oil mist and odor elements.
Replace activated alumina filter if applicable

7.

Daily (When in use)
Check solvent reservoirs for sufficient level ofsolvent.
Verify that pump is primed, operating pulse free.
Check needle wash reservoir for sufficient solvent.
Verify capillary heater temperature functioning.
Verify vaporizer heater temperature.
Verify rough pump oil levels.
Verify turbo-pump functioning.
Verify nitrogen pressure for auxiliary and sheathgasses.
Verify that corona and multiplier are functioning.

Annually
Vacuum system components including fans and fan
covers.
Clean/replace fan filters, if applicable.

REAGENTS AND STANDARDS
7.1.

Reagent grade chemicals shall be used in all tests whenever available. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications of theCommittee on the Analytical Reagents of the American Chemical Society, where suchspecifications are available. Other grades may be used, provided it is first ascertainedthat the reagent is of sufficiently high purity to permit its use without lessening theaccuracy of the determination.

7.2.

Acetic acid, glacial

7.3.

Ammonium acetate (20 mM in water): Prepared by weighing 1.509g of ammonium
acetate and dissolving in 1L of water. The resultant solution is filtered through a
0.22um filter before use. This solution is volatile and should be replaced every 7 daysor sooner.

7.4.

Methanol

7.5.

Potassium hydroxide (KOH), 0.4% in water: Prepared by weighing 16g of potassiumhydroxide and dissolving in 4L of water.

7.6.

Sulfuric Acid (H2SO4), concentrated, reagent grade
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 8 of 27

7.7.

Water, Nanopure or Millipore, must be free of interference and target analytes

7.8.

Standards
7.8.1.

Most analytes were provided by Chemours, as high purity solids (96% or
greater) or as certified solutions. The solid stock material is stored at room
temperature or as specified by Chemours. The FOSA and FOSE analytes werepurchased from a commercial vendor, Wellington Labs or equivalent.

7.9.

Calibration Standards
The stock solution is prepared by diluting the appropriate amounts of the stock analytes
into water to create a solution at 100 ppb. The stock solution should be stored in arefrigerator when not in use. An aliquot is diluted 2X with methanol to produce a 50
ppb standard in a 50:50 methanol: water matrix; this standard is then used for spiking
and to prepare dilutions for the calibration curve. All standards are filtered in the same
way as the samples.

7.10.

Initial Calibration (ICAL) Levels (ng/mL)

Note: Not all calibration points are used for all analytes.
Table 3Compound

CS-1

CS-2

CS-3

CS-4

CS-5

CS-6

CS-7

CS-8

CS-9

TAF
PEPA
PMPA
PFECA_G
Byproduct 1
Byproduct 2
PFMOAAPFO2HxAPFO3OAPFO4DAMeFOSA
EtFOSA
MetFOSE
EtFOSEDFSA
MMF
MTP
PPF Acid

0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001

0.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.00250.0025

0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005

0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01

0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025

0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05

0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10

0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25

0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50

Company Confidential & Proprietary

CS10
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

8.

9.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 9 of 27

Table 3Compound

CS-1

CS-2

CS-3

CS-4

CS-5

CS-6

CS-7

CS-8

CS-9

NVHOS
PFECA BPES
HFPO-DA
Hydro-EVE Acid
EVE Acid
R-EVE
Byproduct 4
Byproduct 5
Byproduct 6

0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001
0.001

0.00250.00250.00250.0025

0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005
0.005

0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01
0.01

0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025
0.025

0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05
0.05

0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10

0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25
0.25

0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50
0.50

0.00250.00250.00250.00250.00250.0025

CS10
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00
1.00

7.11.

Initial Calibration Verification Standard (ICV)For these analytes, a second analyst will prepare a second source standard from thesame source as the ICAL to produce an ICV. The recommended concentration of the
ICV standard should be in the mid-range of the calibration curve. The concentration
may be adjusted if the initial calibration levels are changed or altered.

7.12.

LCS/Matrix Spike Solution, 1 ng/mL
The spike solution is prepared by using the solution described in Section 7.10. Thissolution contains each analyte at a concentration of 1 ng/mL in methanol.

SAMPLE COLLECTION, PRESERVATION, AND STORAGE
8.1.

Water samples are collected in pre-cleaned 250 mL HDPE containers. Other containers
may also be suitable. Samples are chilled to 0 - 6°C for shipment to the laboratory.

8.2.

Samples are logged in following normal laboratory procedures and are stored underrefrigeration at 0 - 6°C. Water samples should be analyzed within 28 days ofcollection.

QUALITY CONTROL
9.1.

Initial Demonstration of Capability (IDOC)
The initial demonstration and method detection limit (MDL) studies described in
Section 13 must be acceptable before analysis of samples may begin.

9.2.

Batches are defined at the sample preparation step. Batches should be kept together
through the whole analytical process as far as possible, but it is not mandatory toCompany Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 10 of 27

analyze prepared extracts on the same instrument or in the same sequence. Refer to the
QC program document (WS-PQA-003) for further details of the batch definition.
9.2.1.

9.3.

9.4.

The quality control batch is a set of up to 20 samples of the same matrix
processed using the same procedure and reagents within the same time period.
The quality control batch must contain a method blank (MB), a laboratory
control sample (LCS), a matrix spike (MS) and sample duplicate (Dup). Eachsample requires a MS and Dup. Laboratory generated QC samples (MB,LCS) do not count toward the maximum 20 samples in a batch. Field QC
samples are not included in the batch count.

One method blank (MB, laboratory reagent blank) must be extracted with every
process batch of similar matrix, not to exceed twenty (20) samples. For aqueous
samples, the method blank is an aliquot of laboratory reagent water. The method blankis processed in the same manner and at the same time as the associated samples.
Corrective actions must be documented on a Non-Conformance memo, and thenimplemented when target analytes are detected in the method blank above the reporting
limit. Re-extraction of the blank, other batch QC and the affected samples are required
when the method blank is deemed unacceptable. See policy WS-PQA-003 for specificacceptance criteria.
9.3.1.

If the MB produces a peak within the retention time window of any of theanalytes, determine the source of the contamination and eliminate theinterference before processing samples.

9.3.2.

The method blank must not contain any analyte at or above ½ the reporting
limit, or at or above 10% of the measured concentration of that analyte in theassociated samples, whichever is higher.

9.3.3.

If there is no target analyte greater than the RL in the samples associated withan unacceptable method blank, the data may be reported with qualifiers. Suchaction should be taken in consultation with the client.

9.3.4.

Re-extraction and re-analysis of samples associated with an unacceptable
method blank is required when reportable concentrations are determined in thesamples.

9.3.5.

Refer to WS-PQA-003 for further details of the corrective actions.

A laboratory control sample (LCS) must be extracted with every process batch ofsimilar matrix, not to exceed twenty (20) samples. The LCS is an aliquot of laboratory
matrix (e.g. water for aqueous samples) spiked with analytes of known identity andCompany Confidential & Proprietary

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RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 11 of 27

concentration. The LCS must be processed in the same manner and at the same time as
the associated samples. The control limits for the LCS are 70-130% for most analytes.
The FOSA and FOSE analytes control limits for the LCS are 50-150%.

10.

9.5.

A matrix spike (MS) must be prepared for each sample with every process batch ofsimilar matrix, not to exceed twenty (20) samples. An MS is an aliquot of a selected
field sample spiked with analytes of known identity and concentration. The MS must
be processed in the same manner and at the same time as the associated samples. The
control limits for the MS are 70-130% for most analytes. The control limits for the
FOSA and FOSE analytes for the MS are 50-150%. Spike recovery outside this range
does not necessitate reprep, but results should be flagged for further investigation due
to matrix effects. These actions must be documented on a Non-Conformance memo.

9.6.

A sample duplicate (Dup) must be prepared for each sample with every process batchof similar matrix, not to exceed twenty (20) samples. Duplicate preps of the same
sample must have a %RPD of < 25% at mid- and high- range concentrations and <50% at the minimum reporting limit

9.7.

Initial calibration verification (ICV) –A second source standard is analyzed with theinitial calibration curve. The concentration should be at the mid range of the curve. The
control limits for the ICV are 70-130% for most analytes. The FOSA and FOSE
analytes control limits for the ICV are 50-150%.

9.8.

Corrective actions for the ICV include: Rerun the ICV.
 Remake or acquire a new ICV.
 Evaluate the instrument conditions.
 Evaluate the initial calibration standards.
 Rerun the initial calibration.

CALIBRATION
10.1.

For details of the calculations used to generate the regression equations, and how to use
the factors generated by these equations, refer to SOP CA-Q-P-003 “Calibration
Curves and Selection of Calibration Points”.

10.2.

Routine instrument operating conditions are listed in the table in Section 11.3.

10.3.

Instrument Tuning

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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 12 of 27

Instrument tuning is done initially when the method is first developed and thereafter as
needed to maintain the sensitivity and selectivity of the method. Tuning is done byinfusing each individual compound into the mobile phase using a tee fitting at a point
just before the entrance to the electrospray probe. The responses for the parent anddaughter ions for each compound are observed and optimized for sensitivity andresolution. Mass assignments are reviewed and calibrated if necessary. The mass
assignments must be within  0.5 amu of the values shown in the table in Section 11.3.
10.3.1. Once the optimal mass assignments (within ±0.5 amu of true) are madeimmediately following the initial tune, the lowest level standard from theinitial calibration curve is assessed to ensure that a signal to noise ratio greaterthan 10 to 1 (S/N > 10:1) is achieved for each PFAS analyte. The first levelstandard from the initial calibration curve is used to evaluate the tune stability
on an ongoing basis. The instrument mass windows are set initially at ± 0.5
amu of the true value; therefore, continued detection of the analyte transition
with S/N > 10:1 serves as verification that the assigned mass remains within ±0.5 amu of the true value.
10.4.

A new calibration curve must be generated after major changes to the system or when
the continuing calibration criteria cannot be met. Major changes include, but are notlimited to, new columns or pump seals. A new calibration is not required after minor
maintenance.

10.5.

With the exception of the circumstances delineated in policy CA-Q-P-003, it is not
acceptable to remove points from a calibration curve. At least seven points must be
included in the calibration curve. Average Response Factor and linear fit calibrations
require a minimum of five points, whereas Quadratic (second order) calibrations
require a minimum of six points.

10.6.

A fixed injection volume is used for quantitation purposes and is to be the same forboth the sample and standards.

10.7.

All units used in the calculations must be consistently uniform, such as concentrationin ng/mL.

10.8.

Initial Calibration
10.8.1. Ten analytical standards of different analyte concentrations are used to generate
the curve. Each standard is injected once to obtain the peak response for eachanalyte at each concentration. These standards define the working range of theanalysis.

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RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 13 of 27

10.8.1.1. All ten standards are used to evaluate the calibration.
10.8.1.2. Calibration is by average response factor, linear fit, or by quadratic
fit. Quadratic fit is used for the analyte if the response is non-linear.
10.8.1.3. For average response factor, analytes must be within 70-130% of
their true value for each calibration standard. For average response
factor (RFa), the relative standard deviation (RSD) for allcompounds must be < 50% for the curve to be valid.
10.8.1.4. For linear fit, the intercept of the line must be less than ½ thereporting limit, and the coefficient of determination (r2) must be
greater than or equal to 0.990 for the curve to be considered valid(or the correlation coefficient (r) > 0.995).
10.9.

Calibration Curve Fits
10.9.1. Linear regression or quadratic curves may be used to fit the data to acalibration function. Detailed descriptions and formulas for each fitting typecan be found in SOP CA-Q-P-003, “Calibration Curves and Selection of
Calibration Points”.

10.9.2. The linear curve uses the following function:y  bx  cEquation 1
Where:
y

=

x
b
c

=
=
=

× Response of Analyte (area)
concentration
slope
intercept

10.9.3. The quadratic curve uses the following function:
Equation 2y  ax 2  bx  c

Where y, x, b, and c are the same as above, and a = curvature.
10.9.4. Evaluation of Calibration Curves
The following requirements must be met for any calibration to be used:
Response must increase with increasing concentration.

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contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 14 of 27



The absolute value of the intercept of a regression line (linear or nonlinear) at zero response must be less than the reporting limit.

There should be no carryover at or above 1/2 of the reporting limit
after a high CAL standard.
If these criteria are not met, instrument conditions and standards will bechecked, and the ICAL successfully repeated before continuing.
10.9.5. Weighting of Calibration Points
In linear and quadratic calibration fits, the points at the lower end of thecalibration curve have less absolute variance than points at the highconcentration end of the curve. This can cause severe errors in quantitation at
the low end of the calibration. Because accuracy at the low end of the curve isvery important for this analysis, it is preferable to increase the weighting of
the lower concentration points. 1/concentration or 1/x weighting is
encouraged. Visual inspection of the line fitted to the data is important inselecting the best fit.
10.10. Initial Calibration Blank (ICB)
10.10.1. Immediately following the ICAL, a calibration blank is analyzed that consistsof an injection of 50:50 methanol:water blank.
10.10.2. The result for the calibration blank must be less than the reporting limit.
10.10.3. If the ICB is greater than the reporting limit then the source of contaminationmust be identified and any necessary cleaning completed, and then the
instrument should be recalibrated.
10.10.4. The instrument blank must be < ½ the RL.
10.11. Initial Calibration Verification (ICV)
10.11.1. Following the ICAL and the ICB, an ICV standard prepared by anotheranalyst than the ICAL standards is analyzed. This ICV standard is a mid-range
standard.
10.11.2. The recovery for the ICV must meet the appropriate following criteria:
10.11.2.1. The recovery for the ICV standard must be equal to or within 70130% for all analytes. The FOSA and FOSE analyte control limits
for the ICV are 50-150%.

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Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 15 of 27

10.11.3. See Section 9.7 for corrective actions in the event that the ICV does not meetthe criteria above.
10.12. Continuing Calibration Verification (CCV) – At the beginning of a run, the end of arun, and after every 10 samples are analyzed a CCV must be injected to determine if
the calibration is still valid. The exception is after an acceptable curve and ICV are run
10 samples can be analyzed before a CCV is required. The CCVs are usually at the
mid level range of the curve. The curve and ICV do not need to be run every day. To
start an analytical run a CCV can be analyzed and if it meets acceptance criteria a run
can be started.
10.12.1. The recovery for the CCV standards must be equal to or within 70-130% for
all analytes. The FOSA and FOSE analyte control limit for the CCV are 50150%.
11.

PROCEDURE
11.1.

One-time procedural variations are allowed only if deemed necessary in theprofessional judgment of a supervisor to accommodate variation in sample matrix,
chemistry, sample size, or other parameters. Any variation in procedure shall becompletely documented using a Non-Conformance Memo (NCM). The NCM process
is described in more detail in SOP WS-QA-0023. The NCM shall be filed in the project
file and addressed in the case narrative.
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.

11.2.

Water Sample Preparation
11.2.1. All samples are tested for pH when being prepared to ensure that they areneutral (pH 6-8). If necessary, dilute solutions of KOH or H2SO4 are used toadjust the pH accordingly.
11.2.2. For each sample prepared a sample duplicate (Dup) and matrix spike (MS) arealso prepared. The sample and sample duplicate are prepared at a 2X dilution
in methanol (2.5 mL of sample and 2.5 mL of methanol) to produce a 50:50
methanol:water matrix. The MS is prepared with 2.5 mL of sample, 2.0 mL
methanol, and 0.5 mL of the Matrix Spike solution (Section 7.12), for a total
volume of 5.0 mL.
11.2.2.1. Vortex the sample, duplicate, and MS after the dilutions are made.

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contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 16 of 27

11.2.2.2. Any subsequent dilutions are made with a 50:50 methanol:watermix.
11.2.3. Prepare a 5mL aliquot (2.5 mL DI water and 2.5 mL methanol) for the method
blank.
11.2.4. Prepare the LCS with 2.5 mL of DI water, 2.0 mL methanol, and 0.5 mL of
the LCS Spike Solution (Section 7.12).
11.2.5. All samples are filtered with a 0.2 µm GHP filter prior to analysis.
11.2.6. Measure 1 mL of each sample using an Eppendorf pipette and pour into alabeled 2.0 mL injection vial. Archive the rest of the extracts for re-injection
and dilution.
11.2.7. Seal the vial with a polypropylene screw cap
Note: Teflon lined caps can not be used due to detection of low level
concentration of PFAS
11.3.

Instrument Analysis
Suggested operating conditions are listed in Tables 1-3 for the SCIEX LCMS system.
Table 1 - Recommended Instrument Operating Conditions
HPLC Conditions (Shimadzu HPLC)

Column (Column temp = 45C)

Phenomenex Gemini 3 μm C18 110Å, 100 X 3.0 mm

Mobile Phase Composition

A = 20 mM Ammonium Acetate in Water

Gradient Program

Injection Size

B = Methanol

Time

%A

%B

Flow Rate - mL/min

0
0.50
5.50
5.60

90
75
75
35

10
25
25
65

0.30
0.30
0.30
0.60

9.40
9.50
15.00
15.10
15.15
15.20

5
1
1
90
90
90

95
99
99
10
10
10

0.60
0.60
0.60
0.60
0.60
0.30

18.00
90
10
Maximum pressure limit = 7,500 psi

0.30

500 L (fixed amount throughout the sequence).

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SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 17 of 27

Table 1 - Recommended Instrument Operating Conditions
HPLC Conditions (Shimadzu HPLC)
Column (Column temp = 45C)

Phenomenex Gemini 3 μm C18 110Å, 100 X 3.0 mm

Mobile Phase Composition

A = 20 mM Ammonium Acetate in Water

Run Time~18 minutes

B = Methanol

Mass Spectrometer Interface Settings (SCIEX 5500)
MS Interface Mode
Ion Spray Voltage (kV)
Entrance Potential (V)Declustering Potential (V)Desolvation Temp
Curtain Gas
Collision Gas

ESI Negative Ion. Minimum of 10 scans/peak.
4.5
5
25
550ºC
35 psi
8 psi

Table 2 - Recommended Instrument Operating Conditions
Mass Spectrometer Scan Settings (SCIEX 5500)

CompoundReaction
(MRM)

Dwell
(sec)

Ent
.
Pot
.
(V)

PFMOAA

179 > 84.9

0.011

-10

-20

-35

-5

6.13

PMPA
PFO2HxA

229 > 185
245 > 85278.9 >
234.9
310.9 > 85
463 >
262.9
378.9 >
184.9
376.9 > 85
443 >.
146.9
442.9 > 85
512 > 169
526 > 169616 > 59630 > 59
174.9 > 81
139 > 51
175 > 97
163 >

0.011
0.011

-10
-10

-12
-24

-15
-25

-5
-5

10.12
11.49

0.011

-10

-10

-20

-5

11.79

0.011

-10

-26

-55

-5

12.20

0.011

-10

-32

-160

-5

12.53

0.011

-10

-16

-87

-5

12.62

0.011

-10

-34

-10

-5

12.73

0.011

-10

-30

-143

-5

12.18

0.011
0.011
0.011
0.011
0.011
0.011
0.011
0.011
0.011

-10
-7
-7
-5
-5
-10
-10
-10
-10

-38
-37
-37
-70
-70
-25
-12
-22
-16

-65
-75
-75
-20
-20
-18
-45
-45
-35

-5
-15
-15
-10
-10
-9
-5
-9
-13

13.22
12.84
13.00
4.94
5.04
5.78
5.77
5.66
5.84

PEPA
PFO3OA
Byproduct 2
PFECA GPFO4DA
Byproduct 2
PFO5DAMeFOSA
EtFOSA
MetFOSE
EtFOSEDFSA
MMF
MTP
PPF AcidCol.
Energy(V)

Declu.
Pot.
(V)

Cell
Exit
Pot.
(V)

Typ RT (Min)Company Confidential & Proprietary

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Table 2 - Recommended Instrument Operating Conditions
Mass Spectrometer Scan Settings (SCIEX 5500)

CompoundNVHOSPFECA BPES
HFPO-DA
Hydro-EVE Acid
EVE Acid
R-EVE
Byproduct 4
Byproduct 5
Byproduct 6

Reaction
(MRM)
118.9
297 > 135
295 > 201314.9 >
135
329 >
284.9
427 >
282.9
407 >
262.9
405 > 217440.9 >
241
439 > 343
397 > 217

Dwell
(sec)

Ent
.
Pot
.
(V)

Col.
Energy(V)

Declu.
Pot.
(V)

Cell
Exit
Pot.
(V)

Typ RT (Min)

0.011
0.011

-10
-10

-34
-24

-210
-25

-7
-7

11.16
12.05

0.011

-10

-34

-200

-7

11.91

0.011

-10

-10

-65

-7

12.25

0.011

-10

-12

-45

-5

12.46

0.011

-10

-12

-45

-5

12.79

0.011

-10

-24

-65

-35

11.07

0.011

-10

-34

-200

-7

11.13

0.011
0.011

-10
-10

-36
-34

-210
-210

-7
-7

11.15
12.47

Table 3
Table 3 Compounds

Typical Native RT (minutes)

PFMOAAPMPAPFO2HxAPEPA
PFO3OA
Byproduct 2
PFECA GPFO4DA
Byproduct 1
PFO5DAMeFOSA
EtFOSA
MetFOSE
EtFOSEDFSA
MMF
MTP
PPF Acid
NVHOS

6.13
10.12
11.49
11.79
12.20
12.53
12.62
12.73
12.18
13.22
12.84
13.00
4.94
5.04
5.78
5.77
5.66
5.84
11.16

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PFECA BPES
HFPO-DA
Hydro-EVE Acid
EVE Acid
R-EVE
Byproduct 4
Byproduct 5
Byproduct 6

12.05
11.91
12.25
12.46
12.79
11.07
11.13
11.15
12.47

11.3.1. Tune and calibrate the instrument as described in Section 10.
11.3.2. A typical run sequence is as follows:Start ICAL (10 points), If ICAL is not required start with a CCV. (TheICB and ICV are not required.)

ICB

ICV

Methanol blankwater blankMethod blank
LCS
Sample 1
Sample 1 DupSample 1 MS
Sample 2
Sample 2 DupSample 2 MS
up to 10 samples (includes sample duplicates and MS in the count)CCV
up to 10 more samples

CCV
12.

CALCULATIONS
12.1.

If the concentration of the analyte ions exceeds the working range as defined by thecalibration standards, then the sample must be diluted and reanalyzed. It may be
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necessary to dilute samples due to matrix. Matrix spike samples that exceed theworking range when the unspiked sample is in the range are reported without further
dilution and flagged appropriately.
12.2.

Qualitative Identification
12.2.1. The analyte RT must be within  0.3 minutes of the ICV and CCV standards.

12.3.

The ICAL established in Section 10 is used to calculate concentrations for the extracts.

12.4.

Extract concentrations are calculated as below. Each equation applies to a different
calibration model, as noted.

Equation 3yCF

Equation 4

Concentration, ng/mL 

Equation 5 b  b 2  4 a (c  y )Concentration, ng/mL 
2aConcentration, ng/mL 

Where:
y =
CF =
a =
b =
c =
12.5.

y c
b

Area (analyte)
Calibration Factor (average response factor model only)
curvatureslope
intercept

Water Sample Result Calculation:

Concentration, ng/L 

Equation 6

CexVt
Vo

Where:
Cex
Vt
Vo

12.6.

=
=
=

Concentration measured in sample extract (ng/mL)
Volume of total extract (mL)
Volume of water extracted (L)Raw data, calibration summaries, QC data, and sample results are reviewed by the
analyst. These must also be reviewed thoroughly by a second qualified person. See theCompany Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 21 of 27

Data Review Policy (WS-PQA-0012). These reviews are documented on the Data
Review Checklist.
13.

14.

METHOD PERFORMANCE
13.1.

The group/team leader has the responsibility to ensure that this procedure is performedby an associate who has been properly trained in its use and has the required expertise.

13.2.

Method Detection Limit
The laboratory must generate a valid method detection limit for each analyte ofinterest. The MDL must be below the reporting limit for each analyte. The procedurefor determination of the method detection limit is given in 40 CFR Part 136, Appendix
B, and further defined in SOP WS-QA-0006 and policy WS-PQA-003. MDLs areavailable in the Quality Assurance Department.

13.3.

Initial Demonstration of Capability (IDOC)
Each analyst performing this procedure must successfully analyze four LCS QC
samples using current laboratory LCS control limits. IDOCs are approved by the
Quality Assurance Manager and the Technical Director. IDOC records are maintainedby the QA staff in the central training files.

13.4.

The laboratory must generate a valid method detection limit for each analyte ofinterest. The MDL must be below the reporting limit for each analyte. The procedurefor determination of the method detection limit is given in 40 CFR Part 136, Appendix
B, and further defined in WS-QA-0006 and policy WS-PQA-003.

POLLUTION PREVENTION
14.1.

All waste will be disposed of in accordance with Federal, State and Local regulations.

14.2.

Standards and reagents are purchased and prepared in volumes consistent withlaboratory use to minimize the volume of expired standards and reagents requiring
disposal.

14.3.

Where reasonably feasible, technological changes have been implemented to minimize
the potential for pollution of the environment. Employees will abide by this method
and the policies in Section 13 of the Corporate Safety Manual for “Waste Management
and Pollution Prevention.”

14.4.

Do not allow waste solvent to vent into the hoods. All solvent waste is stored in capped
containers unless waste is being transferred.
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

14.5.

15.

16.

17.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 22 of 27

Transfer waste solvent from collection cups (tri-pour and similar containers) to jugs
and/or carboys as quickly as possible to minimize evaporation.

WASTE MANAGEMENT
The following waste streams are produced when this method is carried out:
15.1.

Assorted test tubes, autovials, syringes, filter discs and cartridges. Dump the solid
waste into a yellow contaminated lab trash bucket. When the bucket is full or after nomore than one year, tie the plastic bag liner shut and put the lab trash into thehazardous waste – landfill steel collection drum in the H3 closet. When the drum is fullor after no more than 75 days, move it to the waste collection area for shipment.

15.2.

Waste Methanol. Collect the waste solvents in tripours during use. Empty the tripours
into a 1-liter to 4-liter carboy at the fume hood. When the carboy is full, or at the end ofyour shift, whichever comes first, empty the carboy into the steel flammable solvent
drum in the H3 closet. When full to no less than six inches of the top, or after no morethan 75 days, move the steel flammable solvent drum to the waste collection area forshipment.

15.3.

Aqueous acidic waste from the LCMS instrument contaminated with methanol. This iscollected in a 1-gallon carboy at the instrument. When the carboy is full, or after nomore than one year, it is emptied into the blue plastic HPLC collection drum in the H3
closet. When the drum is full to between two and six inches of the top, or after no morethan 75 days, move it to the waste collection area for shipment.

15.4.

Autovials contaminated with methanol. As the autovials are removed from the
instrument after analysis, they are collected in open containers at the instrument. After
all autovials are removed, the open container must be dumped into a closed satellite
collection container in a fume hood, as the punctured septa in the autovial can allow
methanol and other contaminants to evaporate into the atmosphere. The satellite
collection containers are transferred to the waste disposal area when full or after nomore than one year, where they are disposed through the vial eater.

REFERENCES
16.1.

A. Petlick edit of J Boyle, “Determination of Table 3 Compounds by LC/MS/MS
Chemours Fluoroproducts Analytical Method”, 4/3/2018.

16.2.

A. Petlick, “Determination of Table 3 Plus Compounds by LC/MS/MS Chemours
Fluoroproducts Analytical Method”, 1/10/2019

METHOD MODIFICATIONS
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

17.1.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 23 of 27

Modifications from the reference methods are detailed below:
17.1.1. Analytical columns are specific to TestAmerica’s inventory.
17.1.2. The LCMS system is specific to TestAmerica’s fleet.
17.1.3. Addition of MeFOSA, EtFOSA, MeFOSE, and EtFOSE to parameter list.
17.1.4. The ICAL consists of just 10 calibration points and is not analyzed daily (witheach sequence).
17.1.5. The Reporting Limits (RL) have been lowered from 200 ppt to 2-50 ppt byusing a large volume injection technique.

18.

ATTACHMENTS
18.1.

19.

None

REVISION HISTORY
19.1.

WS-LC-0031, Revision 2.1, Effective 04/03/2019
19.1.1. Section 1.1 updated table with current analyte list.
19.1.2. Section 1.2 revised reporting limit to “2-50 ng/L”.
19.1.3. Added Section 6.12.2.3, “Diol Guard Cartridges, Part No. 820950-911 andHigh Perf. ZORBAX Guard, Part No. 820999-901. This is plumbed in frontof the Phenomenex Gemini C18 3 um, 3.0 mm x 100 mm column.”
19.1.4. Section 7.10 updated table with current analyte list and table values.
19.1.5. Section 7.10 added note, “Not all calibration points are used for all analytes.”
19.1.6. Section 9.7 added, “The control limits for the ICV are 70-130% for mostanalytes. The FOSA and FOSE analytes control limits for the ICV are 50150%.”
19.1.7. Section 10.9.2 updated Equation 1.
19.1.8. Section 10.11.2.1 added, “The FOSA and FOSE analyte control limits for the
ICV are 50-150%.”Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 24 of 27

19.1.9. Section 10.12.1 added, “The FOSA and FOSE analyte control limit for the
CCV are 50-150%.”
19.1.10. Section 11.3 tables 1 and 2 updated with current recommended instrument
operating conditions.
19.1.11. Section 11.3 table 3 updated with current analyte list and typical native RT.
19.1.12. Section 11.3.2 removed, “(all samples have a 50:50 methanol:water blank run
after it)” and all instances of “50:50 water:methanol blank”.
19.1.13. Section 12.4 revised to, “Extract concentrations are calculated as below. Eachequation applies to a different calibration model, as noted.”
19.1.14. Section 12.4 added Equation 3.
19.1.15. Section 12.4 revised, “x=concentration” to “CF=Calibration Factor (average
response factor model only)”
19.1.16. Section 16.2 added, “A. Petlick, “Determination of Table 3 Plus Compounds
by LC/MS/MS Chemours Fluorproducts Analytical Method”, 1/10/2019”.
19.1.17. Section 17.1.5 revised “2-10 ppt” to “2-50 ppt”.
19.1.18. Throughout SOP updated number of calibration points from “seven” to “ten”.
19.1.19. Editorial changes.
19.2.

WS-LC-0031, Revision 2.0, Effective 11/15/2018
19.2.1. Section 1.2 revised table with current reporting limit and working range.
19.2.2. Removed Section 6.5, “300 µL auto sampler vials, polypropylene, with
polypropylene screw caps, Waters PN 1860004112, or equivalent.”
19.2.3. Added Section 6.5, “2mL auto sampler vials, clear glass, Thermo Scientific
Nation surestop vial, part no. C5000-1, or equivalent.”
19.2.4. Added Section 6.6, “Vial caps, Thermo Scientific National AVCS blue cap,
pre slit TEF/STL septa, part no. C5000-55B or equivalent.”
19.2.5. Added Section 6.7, “Eppendorf 1000uL epTIPS, part no. 022491954 orequivalent.”
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 25 of 27

19.2.6. Added Section 6.8, “1000uL Pipette: Eppendorf Research Plus.”
19.2.7. Section 7.8.1 added, “The FOSA and FOSE analytes were purchased from acommercial vendor, Wellington Labs or equivalent.”
19.2.8. Section 7.10 revised table with current initial calibration levels.
19.2.9. Section 7.12 revised spike solution concentration to 1 ng/mL.
19.2.10. Section 8.2 added, “Water samples should be analyzed within 28 days ofcollection.”19.2.11. Section 9.4 added, “for most analytes. The FOSA and FOSE analytes control
limits for the LCS are 50-150%.”
19.2.12. Section 9.5 added, “for most analytes. The control limits for the FOSA andFOSE analytes for the MS are 50-150%.”
19.2.13. Section 10.4 removed, “with each analytical sequence or”.
19.2.14. Section 10.5 revised to, “With the exception of the circumstances delineatedin policy CA-Q-P-003, it is not acceptable to remove points from a calibration
curve. At least seven points must be included in the calibration curve.
Average Response Factor and linear fit calibrations require a minimum of five
points, whereas Quadratic (second order) calibrations require a minimum of
six points.”
19.2.15. Section 10.8.1 revised to, “Seven analytical standards of different analyte
concentrations are used to generate the curve. Each standard is injected once toobtain the peak response for each analyte at each concentration. Thesestandards define the working range of the analysis.”19.2.16. Section 10.8.1.3 revised to, “For average response factor (RFa), the relative
standard deviation (RSD) for all compounds must be < 50% for the curve to
be valid.”
19.2.17. Section 10.10.1 revised to, “Immediately following the ICAL, a calibration
blank is analyzed that consists of an injection of 50:50 methanol:water blank.”
19.2.18. Added Section 10.11.2.1, “The recovery for the ICV standard must be equal toor within 70-130% for all analytes.”
19.1.

Removed Section 10.12, “Continuing Calibration Verification (CCV)
Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 26 of 27

Analyze a CCV after every 10 samples to determine if the calibration is stillvalid. The CCVs are usually at the mid-level range of the curve.”
19.1.1. Section 11.2.6 revised to, “Measure 1 mL of each sample using an Eppendorf
pipette and pour into a labeled 2.0 mL injection vial. Archive the rest of theextracts for re-injection and dilution.”19.1.2. Section 11.2.7 added note, “Teflon lined caps can not be used due to detectionof low level concentration of PFAS.”
19.1.3. Updated Table 1 with current instrument operating conditions.
19.1.4. Updated Table 2 with current Instrument operating conditions.
19.1.5. Updated Table 3 with current typical native retention times.
19.1.6. Section 11.3.2 updated run sequence to one currently in use.
19.1.7. Added Section 17.1.4, “The ICAL consists of just 7 calibration points and is notanalyzed daily (with each sequence).”19.1.8. Added Section 17.1.5, “The Reporting Limits (RL) have been lowered from
200 ppt to 2-10 ppt by using a large volume injection technique.”19.1.9. Editorial changes.
19.2.

WS-LC-0031, Revision 1.0, Effective 08/16/2018
19.2.1. Section 1.1, updated table with four compounds: “MeFOSA, EtFOSA,MeFOSE, and EtFOSE”.
19.2.2. Section 7.10, updated table with four compounds: “MeFOSA, EtFOSA,MeFOSE, and EtFOSE”.
19.2.3. Updated Table 2 with four compounds: “MeFOSA, EtFOSA, MeFOSE, and
EtFOSE”.
19.2.4. Updated Table 3 with four compounds: “MeFOSA, EtFOSA, MeFOSE, and
EtFOSE”.
19.2.5. Added Section 17.1.3, “Addition of MeFOSA, EtFOSA, MeFOSE, and
EtFOSE to parameter list.”

Company Confidential & Proprietary

NOTE: The method described in this document is the exclusive, confidential,
proprietary, and secret property of Chemours, developed by TestAmerica
Sacramento under contract to Chemours. Use of this document and the information
contained in it is restricted and governed by the terms of the CONFIDENTIALITY
RESTRICTION with which it was transmitted. DO NOT COPY, EXTRACT,DUPLICATE, SUMMARIZE OR OTHERWISE CREATE A SEPARATE RECORD
OF THIS DOCUMENT OR THE INFORMATION CONTAINED IN IT, IN WHOLE OR
PART, FOR ANY PURPOSE WITHOUT THE PRIOR WRITTEN CONSENT OF
CHEMOURS.

SOP No. WS-LC-0031, Rev.2.1
Effective Date: 04/03/2019
Page No.: 27 of 27

19.2.6. Editorial changes.

Company Confidential & Proprietary