Analytical Methodologies for Per/Poly Fluoroalkyl ... · PFAS Instrument Setup Background...

Analytical Methodologies for Per/Poly Fluoroalkyl Substances (PFAS) Measurement in Environmental Waters by LC/MS/MS

Tarun Anumol, Ph.D.Applications Development ScientistAgilent Technologies Inc.Wilmington, DE, USA

Contact: tarun.anumol@Agilent.com

Agilent ASTS-Houston

‘Fit for Purpose’ Analysis

8 August 20172

Considerations when selecting analytical technique to employ

8 August 2017 Agilent ASTS-Houston 3

I. Types of analytesII. Concentrations required (ppm,

ppb, ppt etc.)III. Type of matrix (drinking water,

surface water, wastewater, sludge, food etc.)

IV. Type of sample clean-up required V. Regulatory considerations (EPA

methods, EU & ASTM methods etc.)

Factors in deciding analytical method selection

Terminology

8 August 2017 Agilent ASTS-Houston 4

• Terminology of these compounds can be confusing but it is critical to understand the differences

TerminologyFluorinated Compounds

8 August 2017 Agilent ASTS-Houston 5

Any organic or inorganic compound that has a fluorine atom incorporated in it

TerminologyPer/PolyfluoroAlkyl Substances (PFAS)

8 August 2017 Agilent ASTS-Houston 6

R: Continuation of alkyl chain or functional group

When one or more Carbons (C) in an alkyl chain have the Hydrogen (H) replaced by Fluorine (F)

8 August 2017 Agilent ASTS-Houston 7

Terminology

Polyfluoroalkyl substance

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

SOME (but not all) H atoms linked to C in alkyl chain are substituted with F

Perfluoroalkyl substance

8 August 2017 Agilent ASTS-Houston 8

TerminologyPerfluoroalkyl acids (PFAAs)

• Perfluorocarboxylic acids (PFCAs)

• Perfluorosulfonic acids (PFSAs)

TerminologyPolyfluorinated compounds - Fluorotelomers

8 August 2017 Agilent ASTS-Houston 9

6:2 FTUCA

8:2 FTUCA

6-C alkyl chain: Hydrophobic 2-C: Hydrophilic

• Formed during the telomerization process in manufacturing

Per/Polyfluoroalkyl substancesUnique Properties

8 August 2017 Agilent ASTS-Houston 10

• Thermal & Chemical stability: grease-proof food packaging, stain repellents

• Zwitterionic properties: surfactants • Surface-tension lowering: fire-fighting foams

Per/Polyfluoroalkyl substancesToxicology & Regulations in US

8 August 201711

Toxicological data:

• PFAS exposure can have adverse effects to the neurological and reproductive systems

• Long chain (>7-C chain) PFASs are bio accumulative, and persistent• Long half-lives for longer chain PFAS• Very difficult to degrade the final products (PFCAs, PFSAs) in the environment

Regulatory Overview:

• USEPA has a public health guidance for PFOA and PFOS at 70 ng/L in drinking water• Many states have their own health guidelines ranging from 20-400 ng/L for PFOS,

PFOA, PFNA and PFHxA

Agilent ASTS-Houston

PFAS Analysis – LC Instrument SetupEliminate Background Contamination

8 August 2017 Agilent ASTS-Houston 12

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

8 August 2017 Agilent ASTS-Houston 13

PFAS Analysis – LC SetupEliminate Background Contamination

PFAS Instrument SetupBackground Contamination

8 August 2017 Agilent ASTS-Houston 14

Actual PFOS peak

5 ng/L standard

Mili-Q Water (‘Blank’)

Background PFOS from LC system

• The use of a Delay column is convenient when running multiple methods on the same instrument.

• “Delay” column and not a “Trapping” column. I.e. the background peak will be retention time separated but will be present.

• The delay column does not account for any contamination after the pump (the autosampler)

250 mL sample collected in Polypropylene Bottles with 5 g/L Trizma (to quench residual chlorine)

Solid Phase Extraction and evaporation to final 1 mL extract in ~96/4 MeOH/Water

Analysis by LC-MS/MS in Negative Electrospray mode for 14 analytes (9 PFCAs, 3 PFSAs, 2 FOSAAs) – 37 min analysis (10 µL injection)

EPA 537: Perfluorinated Alkyl Acids in Drinking Water by SPE-LC/MS/MS

8 August 2017 Agilent ASTS-Houston 15

Confidentiality Label

Solid Phase ExtractionUSEPA 537 vs Modified

8 August 2017 Agilent ASTS-Houston 16

Recoveries calculated in Reagent Water

Modified SPE cartridge: Agilent SampliQ WAX cartridge (6 cc, 150 mg)

All recoveries between 70-130 %

Spiking concentrations:

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

Final extract: 1 mL ~95% MeOH

PFHxA

PFHpA

PFOA

PFNA

PFDA

PFUdA

PFDoA

PFTrDA

PFTeDA

PFBS

PFHxS

PFOS

N-MeF

OSAA

N-EtFOSAA 0

25

50

75

100

125

150EPA 537 (n=7)Modified SPE (n=6)

Rec

over

y (%

)

Agilent ASTS-Houston

Typical LC/TQ Lab Layout

6 feet1.83 meters

8 August 201717

Quick Change Ion Injector

Fast Ion Injector exchange without system venting

Ion Injector Tool VacShield

Easy to use, increased throughput, less downtime for maintenance

8 August 2017 Agilent ASTS-Houston 18

8 August 2017 Agilent ASTS-Houston 19

Easy Change Detector Assembly

Analysis of PFAS in Drinking Water with UltivoEPA Method 537

8 August 2017 Agilent ASTS-Houston 20

Analysis of 17 PFASs in drinking water with <0.1 ng/mL DLs in extract (includes all comps in EPA 537)

Analysis of PFAS in Drinking Water with UltivoEPA Method 537

8 August 2017 Agilent ASTS-Houston 21

PFOS - 7 Levels, 7 Levels Used, 14 Points, 14 Points Used, 0 QCs

Concentration (ng/

-1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

Res

pons

es

3x10

-0.2

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2.2

2.4

2.6

2.8

3

y = 147.003489 * x + 20.563484 R^2 = 0.99890136 Type:Linear, Origin:Ignore, Weight:NonePFOA - 7 Levels, 7 Levels Used, 14 Points, 14 Points Used, 0 QCs

Concentration (ng/

-1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21

Res

pons

es

4x10

-0.5

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

5

5.5

6

y = 3046.084833 * x + 1326.038272 R^2 = 0.99894706 Type:Linear, Origin:Ignore, Weight:None

PFOS PFOA

Calibration Curves from 0.1-20 ng/mL for PFOA and PFOS

Post-spike recoveries of 17 PFASs in a water sample extracted according to EPA 537 with Agilent SampliQ WAX cartridges

Ultivo Triple Quad LC/MS

• All 17 PFASs had linear calibration curves, with R2 > 0.99

• Overall recoveries were between 70–125% for both spiking levels

• %RSD was 0.3–10.8 %for all compounds at both spiking levels (1.0 μg/L and 5.0 μg/L)

Analysis of 30 PFASs in Drinking WaterAgilent 1260 Infinity II + 6470 Triple Quadrupole LC-MS

8 August 2017 Agilent ASTS-Houston 22

Application Note: Analysis of Per/Polyfluoroalkyl Substances in Water Using an Agilent 6470 Triple Quadrupole LC/MS (5991-7951EN)

Chromatogram depicting MRM transitions of 30 PFASs post-spiked in a drinking water extract at 1 µg/L

Analysis of 30 PFASs in Drinking WaterAgilent 1260 Infinity II + 6470 Triple Quadrupole LC-MS

8 August 2017 Agilent ASTS-Houston 23

Sensitivity (0.08 ng/L equil. In water) Precision

Analysis of Environmental Contaminants by LC-MS/MSIon Suppression Comparison by Sample Prep Method

Conventional SPEMass on Column :150 pg

Online SPEMass on Column :150 pg

Direct InjectionMass on Column :150 pg

8 August 2017 Agilent ASTS-Houston 24

Agilent 6495 Triple Quadrupole LC/MSUltimate Sensitivity, Extreme Reproducibility

8 August 2017 Agilent ASTS-Houston 25

8/8/2017Agilent 6495 QQQ LC/MS System

Agilent Jet Stream Hexabore Capillary Dual Ion Funnel

• Thermal gradient focusing• Efficient desolvation • Creates an ion rich zone

• Six capillary inlets• Samples x10 times more

ion rich gas

• Removes the gas but captures the ions

• Removes neutral noise

Agilent 6495 LC/TQ

Perfluorinated Alkyl Acids in Drinking Water by LC/TQCollaborative work with Suffolk County Water Authority

• 1290 Infinity LC coupled to a 6490 LC-MS/MS

• 14 PFAAs in 6 min

System

• PFOA, PFOS: <1 ng/L• Others: 1-10 ng/L• 80 µL aqueous injection

MDLs:

• Drinking Water• Surface Water• Ground Water

Matrices:

8 August 2017 Agilent ASTS-Houston 26

2x10

-0.1

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

Cpd 9: PFHpA: -ESI MRM Frag=380.0V CF=0.000 DF=0.000 CID@13.0 (362.90000 -> 169.00000) Cal Std ppb 2.d

1.66 2.02

1 1

Counts (%) vs. Acquisition Time (min)0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2

Analysis of PFOA/PFOSDirect Injection; Rapid Analysis (1290 II UHPLC + 6495 LC-MS/MS)

8 August 2017 Agilent ASTS-Houston 27

PFOS

PFOA

10 ng/L standard

• Use Multi-wash function!!!• PFOS sticks to needle and seat especially

with long worklists

Targeted Analysis LC-MS/MS vs LC-Q/TOF

8 August 2017 Agilent ASTS-Houston 28

PFHxSPFOA

PFOS

PFHxSPFOA

PFOS0

20

40

60LC-MS/MSLC-QTOF

Con

cent

ratio

n (n

g/L)

Influent Effluent

Collaborative project with RMIT University, Melbourne (Timothy Coggan ; Prof. Bradley Clarke)

Agilent 6550 LC-Q/TOF

Suspect ScreeningPersonal Compounds Database and Library with Spectra• Custom PFAS database with 106 compounds• MS/MS spectra and Retention time data available for a subset of compounds

8 August 2017 Agilent ASTS-Houston 29

Untargeted Analysis

8 August 2017 Agilent ASTS-Houston 30

Untargeted AnalysisIdentification of Novel PFAS

8 August 2017 Agilent ASTS-Houston 31

Inclusion list for targeted MS/MS and structural elucidation using fragment information using software tools (Molecular structure correlator)

Conclusions

Background contamination of PFASs is prevalent in ALL systems but can be fixed through systematic elimination of fluoropolymers in the LC.The Ultivo Triple Quad LC/MS offers significantly more sensitivity than

EPA 537 analysis while using a smaller footprint through several innovative technologies.The sensitive analysis of several newer classes of PFASs is possible

using the Ultivo and 6470.Rapid analysis of PFOA/PFOS and PFAAs can be performed by direct

large volume aqueous injection using the 6495 with <1 ng/L MDLs.Identification, screening and quantification of ‘emerging’ PFASs has

been shown using the Agilent LC-QTOF

8 August 2017 Agilent ASTS-Houston 32

References

8 August 2017 Agilent ASTS-Houston 33

5991-8156EN

5991-7951EN 5991-7863EN

5991-5669EN