Slippery when wet: Overview and comparison of Methods 537, ASTM D7579 and D7968 William Lipps, Brahm Prakash Shimadzu Scientific Instruments, Inc. Columbia MD August, 2017

ESI standard What is LCMSMS?

Liquid Chromatograph/Mass Spectrometer enables injection of water miscible phases and detection by mass 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 Polar Molecules No Derivatization Qualitative Quantitative Incomplete Separation 646 Pesticides in 10.5 minutes

LCMS Ionization introduces “molecular ion” into mass spectrometer ESI standard

A triple quadrupole is two mass spectrometers with a collision cell MRM Q1 Q3 CID Collision Cell M5+ M1+ M2+ M3+ M4+ m3+ m2+ m1+ SIM Fragment

The collision cell fragments the molecular ion Q1:SIM (at precursor mass) m/z 609 Q3:Scan Q2:Fragment Product ion spectrum of Reserpine 195 397 Structure of Reserpine 609 N m/z 195 O H N H H Q3 m/z O H O O O O O O m/z 397 O

Recent Improvements in LC result in sharp, narrow peaks and rapid run times Narrow peak widths result (as low as 1 second) Operating pressures as high as 19,000 psi 5 µM particle size 1.7 µM particle size

Modern Mass spectrometers combined with new LC technology enables high precision and short run times 6.25 6.30 6.35 6.40 6.45 6.50 6.55 6.60 6.65 6.70 6.75 6.80 0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00 Separation of 25 pesticides by mass

ESI standard Why run PFC’s

The EPA has issued a health advisory for Perfluorinated Compounds in drinking water  

What is a health advisory? Health Advisories (HAs) provide information on contaminants that can cause human health effects and are known or anticipated to occur in drinking water. EPA's HAs are non-enforceable and provide technical guidance to states agencies and other public health officials on health effects, analytical methodologies, and treatment technologies associated with drinking water contamination. https://www.epa.gov/dwstandardsregulations/drinking-water-contaminant-human-health-effects-information

Meet federal monitoring requirements Laboratories supporting public water systems must use EPA-approved analytical methods when analyzing samples to: Meet federal monitoring requirements Demonstrate compliance with drinking water regulations https://www.epa.gov/dwanalyticalmethods/learn-about-drinking-water-analytical-methods

Perfluorinated Compounds are measured by HPLC-MSMS using EPA or ASTM methods

Non- Drinking Water PFCs

Military bases with PFC detections

Wastewater and wastewater treatment

Land applied biosolids

Overview of Methods

SPE- Extraction Technique

What Method 537 is: Solid Phase Extraction DRINKING WATER method Internal Standard Calibration ESI LCMSMS

What Method 537 is not: “Direct Injection” “Isotope Dilution” Automated SPE Wastewater or Solid Waste Method

Method 537: Problems / Implementation Suggestions General: Variable, analyte dependent, recovery Method recommends use of a polypropylene transfer tube system to transfer the sample directly from the sample container to the SPE cartridge.  Must rinse sample bottle  must extract entire 250 ml

Method 537: Problems / Implementation Suggestions Laboratory and field blank contamination: Many lab supplies and equipment can contain PFAAs. Section 8.3.1 requires a field reagent blank (FRB), Sampler must open the shipped FRB in the field and pour the preserved reagent water into the empty shipped sample bottle and seal the FRB. Ensures PFAAs were not introduced into the sample during sample collection/handling or from preservatives and bottles.

Method 537: Problems / Implementation Suggestions Polypropylene (PP) vials/caps are necessary to prevent contamination of the sample from PTFE coated septa. PP caps do not re-seal, so evaporation occurs after injection. Multiple injections from the same vial are not possible. Suggest splitting extract before injection PFAAs can build up in the PTFE solvent transfer lines on the LC during inactivity (Polytetrafluoroethylene) Recommend replacing PTFE lines with PEEK (polyetheretherketone)tubing Recommend replacing PTFE solvent frits with stainless steel frits

ASTM D7979 is a “Direct Inject” Method Shimadzu app note No.C81

Separates PFCs with LC then measures by MSMS

Eliminates reagent contamination with a delay column

SPE direct inject Method EPA 537 D7979 D7968 Validation UCMR3 (ORD) Single Lab (Region V and ASTM) Single lab Extraction SPE 250 ml  1 ml Dilute 5 ml with 5 ml MeOH, Filter, direct inject Extract 2 g with 10 ml (1+1) MeOH Holding time Extract in 14 analyze in 28 days 5g/L Trisma ≤ 6°C 28 days ≤ 6°C

EPA 537 Extraction procedure 250 ml 250 ml Surrogate 10 ml/Minute 4 ml MeOH Elute

EPA 537 Extraction procedure LCMS-8040 1 ml 96% MeOH Internal Standard Evaporate

ASTM D7979 Extraction procedure 5 ml Sample Surrogate 5 ml MeOH

ASTM D7979 Extraction procedure LCMS-8060 10 µL Acetic Acid

ASTM D7968 Extraction procedure 20 µL NH4OH 2 g Sample Surrogate 10 ml (1+1) MeOH

ASTM D7968 Extraction procedure LCMS-8060 Tumble 1 hour 50 µL Acetic Acid

Comparison of method quantitation EPA 537 D7979 D7968 Analytes 14 + 3 surrogates and 3 internal standards 21 + 9 surrogates Quantitation Range 5 – 15 ng/L (at 1/250) Next slides Quantitation IS ES

Comparison of matrices tested Method EPA 537 D7979 D7968 Matrix Drinking Water Water and sludge (≤ 0.2% Solids) Soil Matrices Cincinnati tap, ground, surface Reagent water, Chicago river water, POTW effluent, POTW influent, 2 POTW with overflow Ottawa sand, Frederick sand, Annapolis clay, Vicksburg clay, Vicksburg silt

Quantitation range for D7979 – water (direct inject) Analyte Reporting Ranges (ng/L) PFTreA, PFTri, PFDoA, PFUnA, PFDA, PFNA, PFechS, PFOA, PFHxS, PFHpA, PFHxA, FOUEA, FHpPA, FHUEA 10 - 400 PFOS 15 - 400 PFBS 30 - 400 PFPeA, PFBA 50 - 2000 FHEA, FOEA 300 - 8000 FDEA 200- 8000

Quantitation range for D7968 – soil (2g/10ml) Analyte Reporting Ranges (ng/Kg) PFTreA, PFTriA, PFDoA, PFUnA, PFDA, PFNA, PFechS, PFOA, PFHxS, PFHpA, PFBS, FOUEA, FHpPA, FHUEA 25 – 1000 PFOS, PFHxA 50 - 1000 PFPeA, PFBA 125 – 5000 FHEA 600 – 20,000 FOEA 750 – 20,000 FDEA 500 – 20,000

Shimadzu LCMS-8060 ASTM D7979 (5 ng/L)

Comparison of Method calibration criteria EPA 537 D7979 D7968 Calibration ≥ 5 points 5 – 9 points Force through origin yes no Fit RSE ≤ 30% Asymmetry 0.8 – 1.5 Not required Confirmation ion if available

Comparison of Method batch QC criteria EPA 537 D7979 D7968 Batch size ≤ 20 samples ≤ 30 samples Blank ≤ 1/3 MRL ≤ ½ “RLCS” MRL 50 – 150 % 35 – 150% IS < 50% Area Drift Not required Surrogate recovery 70 – 130 % RLCS = Reporting Limit Control Standard

Comparison of sample specific batch QC criteria Method EPA 537 D7979 D7968 MS/MSD recovery 70 – 130 % RPD ≤ 30 %

Summary for the three US methods EPA 537 ASTM D7979 ASTM D7968 Drinking Water Wastewater Sludge and Solids

Triple Quad LCMS Products 2015 ASTM 2013 2-3 times more sensitive Scanning Speed 30,000 u/sec Polarity Switching Rate 5 msec 60000 Signal to Noise Ratio: 1 pg of Reserpine 2012 Scanning Speed: 15,000 u/sec Polarity Switching Rate: 15 msec 10000 2010 EPA 537 2000

ASTM D7979 ASTM D7968 EPA 537 Throughput Equipment, Standard and Labor Cost

Thank you wclipps@shimadzu.com www.ssi.shimadzu.com