Agilent Technologie PFAS User Manual

Prepare to Meet the Challenges
of a Regulated PFAS Landscape

A Public Health Crisis
Decades in the Making

Per- and polyfluoroalkyl substances (PFAS) are man-made substances widely used
in industry and manufacturing because of their unique properties. These compounds
have been used for several decades in many applications like nonstick cookware,
stain-repellent clothing, food packaging materials, detergents, cleaning products,
and firefighting foams. The widespread use of these compounds has led to their
ubiquity in the environment. Studies have indicated that PFAS are also present
in most humans. Research on these compounds has identified them as being
persistent and bioaccumulative (especially PFAS with a carbon chain length >C7).
Toxic effects, including tumors and thyroid disruption, have also been attributed to
some of them.1 This has resulted in regulatory guidance for water and soil, as well
as accelerated monitoring and identification of these compounds.

1

US EPA. 2020. Basic Information On PFAS | US EPA.

Firefighting foams often contain PFAS to assist in dousing flames.

3

Regulatory Overview

There are >4,000 known PFAS compounds that have been created for commercial use. Regulatory guidance
and restrictions exist for just a few of them, leading to unrestricted use for the rest.

– In 2009, perfluorooctanesulfonic acid (PFOS) and its salts were listed as persistent organic pollutants

(POPs) under the Stockholm Convention. All parties are required to eliminate the release of these
compounds into the environment.

– The European Union (EU) Water Framework Directive lists PFOS as a priority hazardous substance that poses

a “significant risk to the aquatic environment.” It has established an annual average environmental quality
standard (AA-EQS) of 0.65 ng/L in inland surface waters, and an AA-EQS of 0.13 ng/L for other surface
waters (Directive 2013/39/EU). The EU drinking water directive aims to have routine monitoring of up to 20
PFAS compounds in drinking water from 2021.

– The UK Chemical Investigation Program requires measurement of PFOS and perfluorooctanoic acid (PFOA)

down to 0.09 ng/L. Similarly, in October 2018, the EU Parliament approved a proposal to recast its Drinking
Water Directive with revised limits for monitoring PFOS and PFOA in drinking water. The new limits are
100 ng/L, with combined PFAS concentrations not to exceed 500 ng/L (COM (2017) 753 1.2.2018).

– Several European countries, including those in the Nordic region, have guidance levels for PFAS in drinking

water and surface water. In Sweden, recommended levels for a sum of 11 PFAS in drinking water should
not exceed 90 ng/L.

– In the United States, the US EPA has established a drinking water health advisory level for PFOS and PFOA

at a combined 70 ppt (ng/L). Several states have their own advisories for PFOA, PFOS, and other PFAS—
such as perfluorononanoic acid (PFNA) and GenX—at the low ppt range. Other initiatives are also in the
works. These include the PFAS Action Act (Jan 2019), the US EPA PFAS Plan (Feb 2019), and the US EPA

Commitment to PFAS Drinking Water Standards (Feb 2019).

– Australia, China, and several other countries are implementing restrictions (or establishing maximum

amounts) for drinking water and receiving water regulations for PFOA, PFOS, and newer PFAS detected
at low ng/L to pg/L levels.

As newer PFAS are identified in the environment, and as more toxicological information becomes available,
further guidelines and regulations are almost certain.

4

USA

Europe

Worldwide regions where strict PFAS monitoring regulations are being enacted.

Current Standards and Consensus methods for PFAS analysis in the environment.

Japan

China

Australia

Method Matrix Tested No. of

EPA 533 Drinking water 25 Solid phase extraction Isotope dilution

EPA 537 Drinking water 14 Solid phase extraction Internal standard correction

EPA 537.1 Drinking water 18 Solid phase extraction Internal standard correction

EPA 8327 (draft) Surface water, groundwater,

wastewater influent and effluent

ASTM 7979 Surface water, groundwater,

wastewater influent and effluent

ASTM 7968 Soil and solids 21 Organic extraction

ISO/DIS 21675 Drinking water, sea water,

fresh water, wastewater (<0.2% solids)

Analytes

24 Dilute and shoot External calibration (isotope dilution also

21 Dilute and shoot External calibration (isotope dilution also

30 Solid phase extraction Internal standard correction

Sample Preparation
Procedure

with MeOH

Quantification Technique

allowed)

allowed)

External calibration

5

Sample Preparation Techniques to Maximize
PFAS Recoveries and Minimize Contamination

Due to their widespread use in clothing, protective gear, and consumer products, PFAS have been
found in human and environmental samples in all regions of the world. Use these best practices
to avoid contaminating your sample with PFAS during collection and storage.

6

Do:

– Wear well-washed lab coats and nitrile lab gloves.

– Use high-density polyethylene (HDPE) or polypropylene (PP) containers and caps as recommended

in US EPA and ASTM methods.

– Refrigerate samples below 6 ºC during storage.

Don’t:

– Wear personal care products (such as sunscreen and hand creams) during sampling.

– Wear waterproof clothing or shoes that may be lined with PFAS or stain-repellent material.

– Use sample collection apparatus that may contain polytetrafluoroethylene (PTFE) or other

plastics containing PFAS.

– Use aluminum foil to cover openings of sample containers, as PFAS can be transferred from foil.

Proper sample cleanup and concentration
are essential to robust, accurate, and reliable
analysis. As the world’s chromatography leader,
Agilent supports your efforts with innovative LC
columns, solid phase extraction (SPE) cartridges,
vials, and filters manufactured to demanding
specifications. All are tested under strict
conditions for reliable analysis of PFAS.

7

Extracting PFAS from water

Several regulatory methods, including EPA 537 and
533, call for extracting PFAS from drinking water
using SPE cartridges followed by LC/TQ analysis.
Typically, a weak anion exchange (WAX) cartridge is
suggested due to its ability to extract both shorter­and longer-chain PFAS with good recoveries as
done in EPA 533 and ISO methods. EPA 537 uses an
Agilent Bond Elut LMS cartridge, which provides high
recoveries for medium- and long-chain PFAS.

From instruments, columns, and supplies, to
fast, worldwide delivery, to decades of method
development expertise—Agilent supports your
entire workflow for PFAS testing.

Average recovery is between 40% and 70%
RSD is greater than 19%

All 30 PFAS were recovered, and US EPA 537 compounds had recoveries
between 70 and 130% with RSD <15% for both water qualities. These
results comply with US EPA method QA/QC requirements. In addition,
only 4 of the 30 compounds had recoveries below 70%, but all 4 were
above 40%. The 14 compounds in the US EPA method produced
acceptable recovery using the Agilent WAX SPE car tridge. The 16 other
PFAS, including compounds on the ASTM list, also had good recoveries
and can be analyzed with this method.

Name LC Grade Water

Average
Recovery

EPA 537 Compounds

PFBS 85 14 99 1

PFDA 101 5 95 6

PFDoA 86 3 88 2

PFHpA 105 10 101 3

PFHxS 97 15 102 1

PFHxA 104 8 107 2

PFNA 100 5 104 3

PFOS 92 13 94 3

PFOA 102 10 106 2

PFTrDA 91 3 103 15

PFUdA 100 6 102 3

PFTrDA 91 3 103 15

N-MeFOSAA 84 11 79 10

N-EtFOSAA 84 9 89 3

Additional Compounds

PFDoA 86 3 88 2

PFTeDA 96 10 86 8

FOSA 57 14 67 20

FHEA 107 6 90 8

FOEA 61 13 52 14

FDEA 56 21 58 15

PFHpPA 47 27 46 14

4-2 FTS 91 10 91 13

6-2 FTS 87 16 100 6

8-2 FTS 104 10

6-2 FTUA 118 8 106 3

8-2 FTUA 96 11 78 13

PFPeS 97 15 104 2

PFHpS 83 11 83 3

PFNS 93 12 91 8

PFDS 85 4 81 6

LC Water
RSD (%)

Tap Water
Average
Recovery

101 10

Tap Water
RSD (%)

Want a closer look?

Read application note 5994-0250EN: Extraction of Per/Polyfluoroalkyl Substances in Water Using Agilent Offline
Solid Phase Extraction

8

N MeFOSAA

5 µg/L spike

Recovery (%)

Acquisition time (min)

Extracting PFAS in biota

PFAS compounds are readily absorbed into animal
and human tissue. Since these substances have
been used in industry for many years, their existence
in human blood and serum—as well as presence
in fish, salmon, and other wildlife—is pervasive.
Research has shown that longer chain PFAS (>C7)
have the potential to bioaccumulate, increasing both
the need and urgency to test and analyze for PFAS
levels in biota and biological fluids.

125

100

20 µg/L spike

75

50

25

SPE and supported liquid extraction (SLE) can be
time consuming and complicated to perform on
biological samples. Agilent Captiva EMR—Lipid
makes it easy to remove interferences, particularly
phospholipids, in minutes without PFAS loss. Its
pass-through format is fast, repeatable, and delivers
a clean extract with minimal ion suppression,
extending column life and reducing the frequency
of MS cleaning.

0

PFBA

6:2 FTA

PFPeA

PFHxA

PFBS

8:2 FTA

PFHpA

PFOA

PFHxS

PFNA

10:2 FTA

PFDA

N EtFOSAA

PFOS

PFUdA

PFDoA

PFTrDA

PFTeDA

PFAS recovery fell between 70 and 130% with most compounds having
an extraction recovery of >90%. The full procedure for extraction with
Agilent Captiva EMR—Lipid is detailed in application note 5991-8656EN,
and shows how this simple technique can be performed in minutes.

Protein precipitation
EMR-Lipid cleanup

Relative abundance

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

Chromatographic baseline of EMR—Lipid phospholipid cleanup
compared to protein precipitation. We reduced MS cleaning and
increased column life without sacrificing PFAS recovery.

Want a closer look?

Read application note 5991-8656EN: Analysis of Per- and Polyfluoroalkyl Substances (PFAS) in Biological Fluid Using a Novel
Lipid Removing Sorbent and LC/TQ

For Research Use Only. Not for use in diagnostic procedures.

9