PFAS Analysis in theEnvironment
Agilent solutions to improve
productivity & robustness

Tarun Anumol, Ph.D.
Global Environment Industry Manager
Agilent Technologies Inc.
February 2019

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PFAS Solutions

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Terminology

Perfluoroalkyl substancePFOS

ALL H atoms linked to C in alkyl
chain are substituted with F

Polyfluoroalkyl substance
SOME (but not all) H atoms linked toC in alkyl chain are substituted with F

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PFOA

PFAS Classifications and Terminology>4000 PFAS compounds in commerce

Wang, Z et al. (2017). Environ. Sci. Technol. 51, 2508-2518.

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PFAS Solutions

ASMSPer/Polyfluoroalkyl substances (PFAS)
Unique Properties• Thermal & Chemical stability: grease-proof food packaging,stain repellents• Zwitterionic properties: surfactants• Surface-tension lowering: fire-fighting foams

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Regulatory Updates
What is going on?
Who are the agencies involved inregulating/monitoring PFAS in the US?
US Environmental Protection Agency (U.S.
EPA) – for federal complianceThis relates to federal status as a contaminant in drinking
water only.

State EPA’s & Department of Environmental
Protection (DEP) – for State complianceDepartment of Defense – for Military sites
Industrial companies – for individual discharge& remediation sites
Water Utilities – to meet discharge permits

EU Drinking Water Directive guidelines
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This bill will require PFAS to be monitored at all superfund
sitesRegulatory Guidance for PFAS in Drinking Water

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CompoundAgency/State

Concentration (ng/L)

Year

PFOA + PFOSEPA DWHA702016

PFOAMN35

2017

PFOS

MN

27

2017

PFOA

NJ

14 (40)2017 (2007)

GenXNC140

2017

PFOA/PFOS

UK

300 (Reg. 10)

2009

PFOS/PFHxS

AUS70

2016

PFOS

VIC EPA

0.23

2016(Proposed)

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Available Methods
Department of Defense – Military Testing
Draft method exp. In 2019 – SPE
Currently: must meet QSM ver. 5.1 orhigher

US EPAASTM 7979 (water); ASTM 7968
(solids)

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EPA 537/537.1(Oct 2018)
Drinking Water
ONLY

Draft EPA 8327
Non-potable Water(wastewater,groundwater) – 2019

EPA 533(expected 2020)

EPA 8328 (proposed)
Soils and solids

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Non-compliance
ResearchAcademia
Industrial Clients
Ex. Direct injection; online SPE

PFAS Analysis – LC Instrument Setup
Eliminate Background Contamination
Potential Contamination Sources• Solvents• Filtration apparatus• Teflon lined tubing

Application Note: Recommended Plumbing Configurations
for Reduction in Per/Polyfluoroalkyl Substance Backgroundwith Agilent 1260/1290 Infinity (II) LC’s, Agilent Application
Note (5991-7863EN)

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PFAS Analysis – LC Instrument SetupEliminate Background Contamination
PFOS

BeforeModification

AfterModification

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PFHxS

PFAS Instrument Setup
Background Contamination
5 ng/L standard• The use of a Delay column isconvenient when running
multiple methods on the same
instrument.

ActualPFOSpeak• “Delay” column and not a“Trapping” column. I.e. thebackground peak will beretention time separated but will
be present.

Mili-Q Water (‘Blank’)

Background
PFOS fromLC system

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PFAS Solutions

ASMS• The delay column does notaccount for any contamination
after the pump (the autosampler)

EPA Method 537/537.1
Analysis of PFAS in drinking water• Method is written ONLY for Drinking Water.
• 18 Analytes in EPA 537.1 (14 analytes in EPA 537)
• Method requires Solid Phase Extraction + LC/MS/MS• Agilent polystyrenedivinylbenzene (SDVB) SPE cartridge• 10 µL injection (96/4: MeOH/Water)• 37 min run time
• MDLs: <1-5 ng/L

https://cfpub.epa.gov/si/si_public_record_Re
port.cfm?dirEntryId=343042&Lab=NERL
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Analysis of PFASEPA 537+ analytes

PFOS(0.1-50 ng/mL)
Infinity II
HPLC Stack

17 PFASs at 0.5 ng/mL
spiked in extracted watersample

UltivoTriple QuadAnalysis of per/polyfluoroalkyl substances (PFASs)
in drinking water using the Agilent Ultivo triple
quadrupole LC/MS; 5991-8969EN
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PFOA
(0.1-50 ng/mL)

USEPA Method 537.1
Addition of 4 analytes in methods + PFBA & PFPeA with Ultivo LC-MS/MSPFTANMeFOSAA

PFUnA

11Cl-PF3OUdS

NEtFOSAAPFHpA

Adona
PFOA
PFTrDA

PFPeA

PFBA

PFBS

PFHxA
GenX
PFHxS

PFNA
9Cl- PFDA
PF3NOSPFOS

PFDoA

Agilent Zorbax Eclipse Plus C18, 3.0 x 50 mm, 1.8 um column
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PFAS Solutions

ASMSEPA Method 537.1 “Modified”
adding analytes, changes to SPE, lower MRLs etc.
Different SPE cartridge – better recoveries

More analytes; Lower DLs

Application note: 5994-0250EN
Application note: 5991-7951EN

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Solid Phase Extraction for Water
USEPA 537 vs Modified
150
EPA 537 (n=7)
Modified SPE (n=6)

10075

Spiking concentrations:50

Modified SPE: 4 ng/L in 250 mL
EPA 537: 5-21 ng/L in 250 mL

25Final extract: 1 mL ~96% MeOH

16PFOA
PFNAPFDAPFUdAPFDoPF A
Tr
DPF A
TeDA
PFBSPFHx
S
P
N-M FOSeFOSAN-E
AtF
OSAA

0PF
Hx
A
PF
HpA

Recovery (%)

125▪ Recoveries calculated inReagent Water▪ Modified SPE cartridge:
SampliQ WAX cartridge (6 cc,150 mg)▪ All recoveries between 70-130 %June 16, 2019

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Solid Phase Extraction
USEPA 537 vs Modified
150150
EPA 537 (n=7)
Modified SPE (n=6)

Modified SPE (n=6)
125Recovery (%)

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PFPF

EPA 537 Analytes

B
PF A
Pe
A
FOSA
FH
EA
FO
EA
FD
E
4- A
2FT
6- S
2FT
8- S
2FT
6S
2FT
8- UA
2FTU
A
PFB
PF S
Pe
PF S
Hx
S
PFNS
PFDS

0PF
O
A
PF
NA
PFDA
PF
UdA
PFDo
PF A
Tr
DPF A
Te
DA
PFBSPFHxSP
N-M FO
SeF
OSAN-EAtFOSAA0HpA25HxA25PF

Recovery (%)

125

Additional PFAS including ASTMASMSEPA 8327 (draft) & ASTM 7979
Analysis of PFAS in ground, surface and wastewater• Method is for non-potable waters – surface water,groundwater, wastewater• EPA 8327 is proposed for release to public in Feb. It issimilar as ASTM 7979
• 28 Analytes• Meant for high throughput; very simple and quick sample
prep.
• Method using simple dilution & acidification before large
volume injection• 30 µL injection (50/50: MeOH/Water)• 21 min run time
• MRLs: 10 ng/L (range: 10-400 ng/L)

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ASMSSample Preparation Protocol
5 mL water sample• ASTM 7979

Add surrogate (internal standard) mix solution• EPA draft method 8327

Add 5 mL MeOH, mix

Filter through Acrodisc GxF/0.2µm GHP
Adjust pH to acidic with 10 µL acetic acid

LC-MS/MS analysis

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ASMSEPA 8327 (draft)
Agilent data from the inter-lab validation for this method
ALL THE ANALYTES DETECTED < REPORTING LIMITSURROGATES RECOVERIES BETWEEN 70-130% IN ALL SAMPLESSurrogate Recovery at 160 ng/L in Unknown Water Samples
140120%100806040200Application: 5994-0678ENUnknown Surface Water Sample 1
Unknown Surface Water Sample 3
Unknown Reagent Water Sample 2

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Unknown Surface Water Sample 2
Unknown Reagent Water Sample 1
Unknown Ground Water Sample

ASTM 7968/EPA method expected based on thisPFAS analysis in soil and solid matrices• Method is for soil and solid matrices• EPA method expected to be based on ASTM 7968.

• 21 Analytes• Method using simple dilution & acidification before large
volume injection• 30 µL injection (50/50: MeOH/Water)• 21 min run time
• MRLs: 25 ng/kg (range: 25-1000 ng/L for most)21

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ASTM 7968
PFAS analysis in soil and solid matrices
2g Soil Sample

Extract with 10 mL methanol : water at pH 9-10

Tumble on rotator for 1 hour, centrifuge

Syringe Filter

Acidify

Analyze by LC/MS/MS

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Analysis of PFOA & PFOS in Water by Online SPE

Agilent Flexcube
Online SPE

Application note: 5991-6115EN

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Quantification of expanded list of Emerging PFAS
Analysis with 6495 in Water & Soil
Bradley Clarke – RMIT University (>50 PFAS in 1method)

Webinar: www.chromatographyonline.com/lcgc_p/treatment
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Application: 5994-0919EN

Analysis of >50 PFAS in Water
Method Performance (1 uL injection of extract)30%>92% of PFAS tested have
RSDs <20% & MDLs <5 ng/L

1825%161420%

121015%810%

64Precision (%RSD)*

Method Detection Limits (ng/L)^

205%

200%

* Seven replicates at 5 ng/L spiked into 250 mL water samples; followed by SPE and injection of 1 uL onto 6495 LC-MS/MS; ^ based on USEPA 40 CFR Part 136 Appendix B Revision 225June 16, 2019

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Perfluorinated Alkyl Acids in Drinking Water by LC/TQ
Direct Aqueous Injection with Agilent 6495 LC MS/MS

System• 1290 Infinity LC coupled to a6495 LC-MS/MS• 14 PFAAs in 6 minMDLs:• PFOA, PFOS: <1 ng/L
• Others: 1-5 ng/L
• 75 µL aqueous injection
Matrices:• Drinking Water• Surface Water• Ground Water

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Suspect Screening and Untargeted Analysis• Agilent 6545 LC/Q-TOF• Custom PFAS database• MS/MS spectra and retention time data available for asubset of compounds

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Suspect Screening Personal Compounds Database
WWTP1 Final Effluent
6:2 FTS
Mass: 427.97458
Mass diff: -1.41 ppm
Score: 98.11
6:2 FTS ID’d in all samplesof WWTPStandard28

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Unknowns Screening Results

Effluent

Influent• Targeted: Only captured ~6% ofcompounds• Suspects: 10-16% of fluorinatedcompounds• Unknowns: 66-81% of fluorinatedcompounds• Large portion of fluorinated anioniccompounds in sample unknowns

60-70% of fluorine containing molecules in these samples is ‘unknown’29

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Ion-Mobility MS
The ultimate tool for true PFAS identification“Rapid Assessment of Isomeric Diversity inPFAS by Ion Mobility Spectrometry-Mass
Spectrometry (IMS-MS)”James Dodds & Erin Baker , NC StateMP 12730

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Thank You
Let’s continue our Discussion

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