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3M ENVIRONMENTAL LABORATORY AR 226-0190 OES -- METHOD DETERMINATION OF PERFLUOROOCTANE SULFONATE (PFOS), PERFLUOROOCTANE 'SULFONYLAMIDE (PFOSA), AND PERFLUOROOCTANOATE (POAA) IN WATER BY LIQUIDSOLID EXTRACTION AND HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY (HPLC/MS/MS) Method Number: ETS-8-154.0 `Adoption Date: Author: Kristen J. Hansen/Harold O. Johnson Revision Date: Approved By: William K. Reagen, Kent R. Lindstrom William K. Reagen, Laboratory Management Date Kristen J. Hansen, Ph.D., Group Leader Date Kent R. Lindstrom, Technical Reviewer Date 04550 DenisonofOS, FOS, POA i Wate byLig Sld EtcandLCMSAS 1.0 SCOPE AND APPLICATION 11 TdheitsermmientahtoidonporfovPiedrefslucoolrloeoccttioann,eesxutlrfaocntaitoen,(aPnFdOSa)n,alPyteircfalluporroooccetdaunreesSuflofrotnhyelamide (PFOSAY), and Perfluorooctanoate (POAA) in groundwater, surface water, and drinking water samples 12 This method was prepared according to the EPA document, "Guidelines and Format for MbaestehdodinsptaortbeonPrthoeporseepdoratt"4M0etChFoRdoPafrAtna1l3y6siosr Pfoarrtth1e41D"et(esrcmeiRneafteiorneoncfePe18r.f1l)u,oarnodocitsane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water" (see Reference 18.2). 2.0 SUMMARY OF METHOD 21 Water samples are collected from a site of interest and shipped cold to an analytical facility. PFOS, PFOSA, and solid phase extraction (SPE) POAA are cartridges. extracted from 40mL water The compounds are eluted samples from the using Cie Cys cartridge, using methanol. Separation, identification, and measurement are accomplished by high- performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis using multiple response monitoring (MRM). `The concentrationof each identified component is measured by comparing the MS responseofthe quantitation ion produced by that compound to the MS responseofthe quantitation ion produced by the same compound in an extracted calibration standard (external standard). 30 DEFINITIONS 3.1 Analytical Sample--A portionofan extracted Laboratory sample prepared for analysis 32 Calibration Standard--A solution prepared from the Working Standard (WS) and extracted according to this method. The calibration standard solutions are used to calibrate the instrument response with respect to analyte concentration. 33 Duplicate Sample (DS)--A separate aliquotof a sample, taken in the analytical tloabtohraattoofrythaendfirasntaallyizqeudotsegpiavreataelmyeawsiuthreoidfentthicealprpercoicseidounreass.socAinaatleydsiwsitohf DlaSbsorcaotomrpyared procedures, but not with sample collection, preservation, or storage procedures. 34 Field Blank Control Sample (FB)--Type I water placed in a sample container in the laboratory and treated as a sample in all respects, including exposure to sampling site conditions, storage, preservation and determineiftest substances or other all analytical interferences procedures. are present The in the purposeof the FB field environment. is to 04551 MS Word 97 ETS8.1540 Page 20f17 DeterminationofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS 3.5 Field Duplicate (FD)--A sample collected in duplicate at the same time as the sample and placed under identical circumstances and treated exactly the same throughout field and laboratory procedures. Analysisof FD) compared to that of the first sample gives a measureofthe precision associated with sample collection, preservation and storage, as well as with laboratory procedures. 3.6 Field Matrix Spike (FMS)--A sample collected in duplicate to which known quantities of the target analytes are added in the field at the timeofsample collection. The FMS should be spiked at approximately S0-150%ofthe expected analyte concentration in the sample. `The FMS is analyzed to ascertain if any matrix effects, interferences, or stability issues may complicate the interpretationof the sample analysis 3.7 Field Spike Control Sample (FSCS)--An aliquot of typIe water to which known quantities of the target analytes are added in the field at the time of sample collection (at an appropriate concentration to be determined by the project lead). The FSCS is extracted and analyzed exactly like a sample to determine whether a lossofanalyte could be attributed to sample storage and/or shipment 38 Laboratory Control Sample (LCS)--An aliquot of type I water to which known quantitiesofthe target analytes are added in the laboratory. Two levels are included, one at the LOQ (approx. 25Pg/mL), the other at a concentration of approx. 100-250Pg/mL or another concentration to be determined by the project lead. The LCS is extracted and analyzed exactly ike a laboratory sample to determine whether the methodology is in control, and whether the laboratory is capable of making accurate measurements at the required method detection limit and higher 39 Laboratory Sample--A portion oaf sample received from the field for testing 3.10 Limit of Detection (LOD)--The lowest concentrationofan analyte that can be measured and reported with 99% confidence that the analyte concentration is greater than zero. The LOD can be determined in several ways, including signal-to-noise ratio and statistical calculations. 3.01 Limit of Quantitation (LOQ)--The lowest concentration (LLOQ) or highest concentration (ULOQ) that can be reliably achieved within the specified limitsofprecision and accuracy during routine operating conditions. cNoontcee:ntTrhaetiLonLiOsQscilscgteenderaaslltyhe5l-o1w0esttimneosn-tzheerLoOsDt.andFaorrdmian nthyeacnaalliybtreast,iotnhecuLrvLe.OQHoawnealvyetre, it may be nominally chosen within these tated guidelines to simplify data reporting. Sample LLOQS arc: `matrix-dependent. 3.12 Matrix Spike (MS)--An aliquot ofa sample, to which known quantitiesof target analytes are added in the laboratory. The MS is extracted and analyzed exactly like a laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The background concentrationsof the analytes in the sample matrix must be determined in a separate aliquot and the measured values in the MS corrected for background concentrations. 004552 MS Ward 97 ETS$1540 Page3of 17 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Salid Extraction and LOMSIMS 3.13 Method Blank--An aliquotoftype I water thati treated exactly like a laboratory sample including exposure to all glassware, equipment, solvents, reagents, internal standards, and surrogates that are used with other laboratory samples. The method blank is used to determineif test substances or other interferences are present in the laboratory environment, the reagents, or the apparatus. 3.14 Method Detection Limit (MDL) Determination--Oneofseveral processes that may be used to establish a LOD value. The statistically calculated minimum amountofan analyte that can be measured with 99% confidence that the reported value is greater than zero. "This term is usually associated with the EPA definition in 40 CFR Part 136 Appendix B. 3.15 Sample--A sample is a small portion collected from a larger quantity of material intended to represent the original source material. 3.16 Spiking Stock Standard (SSS)--A solution prepared from stock standards used to prepare the working standard. 3.07 Stock Standard (SS)--A concentrated solution ofa single analyte prepared in the laboratory with an assayed reference compound. 3.18 Working Standard (WS)--A solutionofseveral analytes prepared in the laboratory from Ss and diluted as needed to prepare calibration standards and other required analyte solutions 4.0 WARNINGS AND CAUTIONS 41 Health and Safety Warnings 4.1.1 The acute and chronic toxicityofthe standards for this method have not been precisely determined; however, each should be treated as a potential health hazard. 4.1.2 Unknown samples may contain high concentrationsof volatile toxic compounds. Sample containers should be opened in a hood and handled with gloves to prevent exposure. 4.13 The laboratory is responsible for maintaining a safe work environment and a current awarenessoflocal regulations regarding the handlingofthe chemicals used in this method. A reference file of material safety data sheets (MSDS) should be available to all personnel involved in these analyses. 5.0 INTERFERENCES 51 During extraction and analysis, major potential contaminant sources are reagents and liquid-solid extraction devices. 52 All materials used in the analyses shall be demonstrated to be free from interferences under conditions of analysis by running method blanks 53 Teflon containing materials (e.g. caps, wash bottles) contain fluorocompounds which may cause interferences and should not be used during collection, storage, extraction, or analysisofthe samples. 04553 MS Word 97 ETS8-1540 Pagedof 17 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMS/MS 6.0 EQUIPMENT, SUPPLIES, AND MATERIALS Note: Brand names, suppliers, and part numbers are for illustrative purposes oly. Equivalent performance may be achieved using apparatus and materialsotherthan those specified here, but demonstrationofequivalent performance that meets the requirementsofthis methodisthe responsibility ofthe laboratory 6.1 Sampling Equipment 6.1.1 Sample collection bottles--LDPE (e.g, NalgeneTM) narrow-mouth bottles with screw cap. Note: Do not use Teflon bottles or Teflon lined caps. 6.1.2 Coolers for sample shipment 6.13 Ice for sample shipment. 6.1.4 Bottles must be lot-certified to be freeofartifacts by running Method blanks according to this method. 62 Laboratory Equipment (Extraction and Analytical) 62.1 Balance, analytical (display at least 0.0001g), Mettler. 62.2 Vacuum pump, BCchi 62.3 Visiprep vacuum manifold, Supelco. 6.2.4 Sep Pak Vac 6c (1g) tCis cartridges (part # WAT036795),Waters. 62.5 SOmL disposable polypropylene centrifuge tubes, VWR. 62.6 15mL disposable polypropylene centrifuge tubes, VWR. 62.7 Disposable micropipettes (50-100uL, 100-200uL), Drummond. 62.8 Class A pipettes and volumetric flasks, various 6.2.9 Hypercarb drop-in guard column (4mm) (part # 844017400), Keystone. 6.2.10 Stand-alone drop-in guard cartridge holder, Keystone. 62.11 125mL LDPE narrow-mouth bottles, Nalgene. 6.2.12 HPLC pump (LCI0AD), Shimadzu. 6.2.13 2mL clear HPLC vial kit (cat# 5181-3400), Hewlett Packard 6.2.14 Standard lab equipment (graduated cylinders, disposable tubes, etc.), various, 6.2.15 LC/MS/MS and HPLC systems, as described in section 10.1 63 Equipment Notes 6.3.1 In order to avoid contamination, the useofdisposable labware is highly recommended (tubes, pipettes, etc.) 6.32 Teflon or Teflon-lined containers or equipment, including Teflon-lined HPLC vials or caps for the HPLC auto sampler must not be used. 633 Type I water used during the sample and standard extraction should be filtered through a Hypercarb guard column using & HPLC pump. This water is referred to as "filtered type [ water", hereafter in this report 63.4 Itis necessary to check the solvents (methanol) for the presence of contaminants (especially POAA) by LC/MS/MS prior to use. Certain lot numbers have been found to be unsuitable for use. 6.3.5 Use disposable micropipettes or pipettes to aliquot standard solutions to make calibration standards and matrix spikes. 04554 MSWord 97 ETSS140 Pagesof17 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Sold Extraction and LOMS/MS 7.0 REAGENTS AND STANDARDS `Note: Suppliers and catalog numbers are for illustrative purposes only. Equivalent performance may be achieved using chemicals obtained from other suppliers. Do not usc a lesser grade of chemical than those listed 7.1 Chemicals 7.1.1 Methanol (MeOH), HPLC grade, JT Baker, Catalog No. JT9093-2, 7.12 Ammonium Acetate, Reagent grade, Sigma-Aldrich, Catalog No. A~7330. 7.13 Water, type I, prepared in-house. 7.14 Sodium Thiosulfate, Reagent grade, JT Baker. 72 Standards 7.2.1 Potassium perfluorooctane sulfonate (sce Attachment A, Figure 1). 7.22 Perfluorooctane sulfonylamide (see Attachment A, Figure 2). 7.23 Ammonium perfluorooctancate (see Attachment A, Figure 3) 7.3 Reagent Preparation 7.3.1 250mg/mL sodium thiosulfate solution (Extraction)--Dissolve 25g of sodium thiosulfate in 100mL reagent water. 7.32 40% methanol (Extraction)--Measure 400mL methanol and adjust the volume to 1.0L with reagent water. 7.3.3 100mM ammonium acetate solution (Analysis)--Weigh 7.71g of ammonium acetate and dissolve in 1.0Lofreagent water. Dilute the 100mM solution by a factor of 50 to make the 2mM ammonium acetate solution used for mobile phase A Note: Altemative volumes may be prepared as longa the atiosofthe solventot solute ratios are maintained. 7.4 Spiking Stock Standard (SSS) Preparation 7.4.1 100ug/mL each PFOS, PFOSA, and POAA SSSs--Weigh out 10mg of analytical standard (corrected for percent salt and purity--i.c., 10 mg CaF1:S0K purity 90% = 835mg CyF1780;-) and dilute to 100mL with methanol in a 100mL. volumetric flask. Transfer to a 125mL LDPE bottle. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 4:2C for a maximum period of 6 months from the dateofpreparation. 7.42 ug/ml mixed SSS--Add 1.0mL eachof the 100ug/mL SSSs (from 7.4.1) to a 100mL volumetric flask and bring up to volume with methanol 7.43 0.1pg/mL mixed SSS--Add 10.0mL of the 1.0ug/mL-mixed solution (from 7.4.2) 10 a 100mL volumetric flask and bring up to volume with methanol 7.44 0.0lug/mL mixed SSS--Add 10.0mLofthe 0.1pg/mL-mixed solution (from 7.4.3) t0 a 100mL volumetric flask and bring up to volume with methanol 7.45 Storage Conditions--Store all SSSs in a refrigerator in 125mL LDPE bottles at 422C for a maximum period of3 months from the dateofpreparation 004555 MS Word 97 ETS81540 Page Gof 17 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSIMS 7.5 Calibration Standards 7.5.1 100pg/mL each PFOS, PFOSA, and POAA stock standard solutions--Weigh out 10mgof analytical standard (corrected for percent salt and purity) and dilute 10 100mL with methanol in a 100mL volumetric flask. Transfer to a 125mL LDPE bottle. Prepare a separate solution for each analyte. Store solutions in a refrigerator at 442C for a maximum period of6 months from the date of preparation. 7.52 Iug/mL Working Standard--Add 1.0mL each of the 100ug/mL SS solutions (from 7.5.1) to a 100mL volumetric flask and bring up to volume with methanol 7.53 O.Ipg/mL Working Standard --Add 10.0mLofthe 1.0pg/mL mixed solution (from 7.5.2) to a 100mL volumetric flask and bring up to volume with methanol. 7.54 0.01pg/mL Working Standard --Add 10.0mLofthe 0.1 pg/mL mixed solution (from 7.5.3) to a 100mL volumetric flask and bring up to volume with methanol. 7.55 Storage Conditions--Store all WSs in a refrigerator (in 125mL LDPE bottles) at 422C for a maximum period of3 months from the dateofpreparation 7.56 Calibration Standard--Prepare a minimumoffive calibration solutions in filtered type I water according to the following table: `Concentration Volume of Final Calibration Standard ofWS, ug/ml, WS, ul Volume, mL 00 0 "0 0010 100 40 0.010 200 "0 0.010 400 "0 010 100 "0 010 200 "0 010 300 "0 010 400 "0 T2 aMyaybbeepprepraredotboo evxpetnenddetBheemlramnagebbeoyoonnd TSOSOOPPE.R Final Concentration of Calibration Standard, Pg/mL 0 2 50 100 250 500 750! 1000 Notes The absolute volumesofthe standards may be varied by the analyst as long as the correct proportionsofsolute to solvent are maintained. 7.57 The standards are processed through the extraction procedure (Section 9.0), identical to the laboratory samples. The extracted concentrationof the calibration standard is equal to 8X the initial concentration, due to the concentrationofthe standard during the extraction process. 7.58 Storage Conditions--Store all extracted calibration standards in 15mL. polypropylene tubes at 442C, for a maximum periodof two weeks from the date of preparation. 04556 MSWord 97 ETS8-1540 Page7ar17 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS 8.0 SAMPLE COLLECTION, PRESERVATION AND STORAGE Note: Sampling equipment, including automatic samplers, must be freeofTeflon tubing, gaskets, and other parts that may leach interfering analytes into the water sample. Automatic samplers that `composite samples over time should use refrigerated polypropylene sample containersifpossible. `Sample bottles should notberinsed before sample collection. 8.1 Tap Water--Open the tap and allow the system to flush until the water temperature (1510C) has stabilized (usually about two minutes). Adjust the flow to about 500mL/min and collect samples from the flowing stream. 82 Ground Water--Purge the wellofstanding water using a pump or a bailer. Collect the sample directly from the pump or from the bailer 83 Surface Water--When sampling from an open body of water, fil the sample container `with water from a representative area 84 Sample Dechlorination--All samples should be iced or refiigerated at 4:2C and kept in the dark from the timeofcollection until extraction. Residual chlorine should be reduced by adding 200uL ofa 250mg/mL sodium thiosulfate solution to each water sample, FB, and FSCS (which may be placed in each bottle before leaving for the sampling site). 8.5 Holding Time (HT)-- Resultsof the time/storage studyofall target analytes showed that the three compounds are stable for 14 days in water samples when the samples are dechlorinated and stored as described in section 8.4 (see also reference 18.3). Therefore, laboratory samples must be extracted within 14 days and the extracts analyzed within 30 daysof sample collection.If the HT exceeds 14 days, great care is used when evaluating field spikes to avoid misrepresentationofthe sample concentration. 86 Field Blanks 8.6.1 Process a Field Blank Control Sample (FB) along with each sample set (samples collected from the same general sample site at approximately the same time). At the laboratory, prior to sample collection, il a sample container with filtered type water, seal, and ship the FB to the sampling site along with the empty sample containers. Return the FB to the laboratory with the filled sample bottles. 8.62 When sodium thiosulfate is added to samples, use the same procedure to preserve the FB. 8.7 Field Duplicates 8.7.1 Collect a Field Duplicate (FD) for every ten (10) samples collected or per each sampling set, ifess than 10 samples are collected. 8.7.2 Separate FDs must be collected for each typeofwater sample (ground, tap, etc.) collected. 8.7.3 Collect the FD immediately after the sample. 8.74 Preserve, store and ship FD using the same procedures as used for the samples 04557 MS Word 97 ETS$-1540 Pagesof 17 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LCMSIMS 88 Field Spike Control Sample (FSCS) 88.1 A Field Spike Control Sample (FSCS) must be prepared for each sample shipment. Ifmultiple coolers are used to ship a set of samples, each cooler must contain a FSCS. 8.82 At the laboratory, fill a sample container with 100mLoftype I water. Seal and ship to the sampling ite along with the empty sample containers and FB. 8.83 When sodium thiosulfate is added to samples, use the same procedure to add the same amounts to the FSCS 8.8.4 Seal and gently invert the FSCS to mix. Store and ship the FSCS using the same procedures as used for the samples. 04558 MS Ward 97 ETS8.1540 Page dof17 Determinationof PROS, PFOSA, POAA in Water by Liquid-Salid Extraction and LOMS/MS 9.0 EXTRACTION PROCEDURE 9.1 Extraction Scheme 9.11 Allow samples to equilibrate to room temperature. Thoroughly mix samples by `gently inverting the sample bottle. 9.12 Measure 40mL of sample into SOmL polypropylene centrifuge tubes (Spike the QC and Matrix spikes as required, replace lid and mix well. Note: * Samples may need to be prescreened to determine an appropriate matrix spike level (spically 50-150%of sample concenration). 9.13 Condition the Cys SPE cartridges (1g, 6mL) by passing 10mL methanol followed by SmL filtered typIe water (~2drop/sec). Do not let column run dry. Note: For the following steps, maintain a ~Idrop/sec flow rate. Do not allow thecolumnto run dry atany time. 9.1.4 Load the analytical sample onto the Cy SPE cartridge. Discard eluate. 9.5 Wash with ~SmL 40% methanol in water. Discard eluate. 9.1.6 Elute with ~SmL 100% methanol. Collect SmL of eluate into graduated 15mL polypropylene centrifuge tubes. This is the target elution fraction (final volume = Sm), 9.1.7 Analyze a portionof the target elution fraction eluent using negative electrospray HPLC/MS/MS (Section 10.2). Note: Final Samples are Vol. = SmL. concentrated by a factorof eight during the extraction; Initial Vol = 40mL 9.1.8 Samples are stable at room temperature for at least 24 hours. Analytical samples may be stored in a refrigerator at 442C until analysis. 9.19 Standardization of Cis SPE columns--If poor recoveries are observed, it may be necessary to standardize the Ci SPE columns in the following manner before analyzing samples. 9.1.9.1 Usea standard with an analyte concentration between 1000 and 4000 Py/mL. Follow the extraction scheme as outlined from steps 9.1.1 10 9.1.6, except, collect the eluate fraction separately (approx. SmL), as well as the target elution fraction. 9.192 After step 9.1.6, collect a post-elution fraction by, eluting with an additional SmL of 100% methanol. 9.19.3 Analyze all three fractions by HPLC/MS/MS. If the target fraction contains a minimumof 85%ofthe respective analytes, it may be considered acceptable. 9.1.9.4 Ifthe wash contains significant standard (>15%), ither the wash volume or percentage of MeOH should be decreased. 9.19.5 Ifthe post-elution fraction contains significant standard (>15%), the target elution volume should be increased. 04559 MSWord97 ETS81540 Page 100f17 DeterminationofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LCMSIMS 10.0 CALIBRATION AND STANDARDIZATION (ANALYTICAL SETUP) lNoontgesasOtthheermeitnhstordumcernittseimaaayrebmeetu.seTdhaenodptehreateoqrumiupmsetnotpatnimdiczoenadnitdiodnoscummaeyntbethveereyqudiipffmeernetntaansd settings used. 10.1 Establish the LC/MS/MS system and operating conditions equivalent to the following: Mass Spec: Micromass Quattro Ultima (Micromass) Interface: Electrospray (Micromass) Mode: Electrospray Negative, Multiple Response Monitoring (MRM) Harvard infusion pump (Harvard Instruments), for tuning Computer: COMPAQ Professional Workstation AP200 Software: Windows NT, MassLynx 3.3 HPLC: Hewlett Packard (HP) Series 1100 HP Quat Pump HP Vacuum Degasser HP Autosampler HP Column Oven oNnoltien:eAaf4txr t1he0mpumrgHeypvearlcvearabnddrboepfroirnegtuhaersdacmaprlteriidngjeec(tKoerypsotrotnet,optarratp #an8y44re0s1i7d-u4e0c0o)nitamaitntaancthsed that may be inthe mobile phase and/or HPLC system. HPLC Column: Genesis Cs (Jones Chromatography), 2.1mm x SOmm, um Column Temperature: 35C Injection Volume: 1SuL. Mobile Phase (A): 2mM Ammonium Acetate in filtered type Iwater (See 7.3.1) Mobile Phase (B): Methanol HPLC Gradient Program: Time, Percent Mobile min Phase A 00 60 04 60 10 10 70 10 75 0 90 0 95 60 135 60 14.0 60 Percent Mobile Phase B 40 40 9 % 100 100 4 40 40 FlmoLw/mRaitne, 03 03 03 03 03 04 04 04 03 Note: Other HPLC gradients may be used as long as the method criteia are met. 04560 MS Word 97 ETS81540 Page 1 af 17 Determinationof PROS, PFOSA, POAA in Water by Liquid-Sold Extraction and LC/MS/MS It may be Columns necessary to with different adjust the HPLC dimensions (e.g. gradient in order 2.1mm x 30mm) to optimize instrument performance. and columns from different `manufacturers (Keystone Betasil Cys etc.) may be used. Tons Monitored: Analyte | PriA mary lon POAA 413.0 PFOS 499.0 PFOSA| 4980 Product Tonnm 169.0 9.0 780 P ApprP oximE ate 50 52 58 Other product ions may be chosen at the discretionofthe analyst, although m/z 99 is suggested for PFOS. Useofthe suggested primary ion is recommended. Retention times may vary slightly, on a day-to-day basis, depending on the batch of mobile phase etc. Drift in retention times is acceptable within an analytical run, as long as the drift continues through the entire analysis and the standards are interspersed throughout the analytical run. 10.2 Tune File Parameters 10.2.1 The following values are provided as an example. Actual values may vary from instrument to instrument, Also, these values may be changed from time to time in order to optimize for greatest sensitivity. Analyte | Dwell, sec POAA 02-04 PFOS 02-04 PFOSA 02-04 Collision Energy, ev 10-25 30-60 20-50 Cone, V 20-30 50-80 30-60 Source Capillary Hexapole | Aperture | Hexapole 2 Source Block Temp. Desolvation Temp. - Set 2.56-3.5kV. 0sv 02v 08V 100-150C 250-400C 04561 MSWord 97 ETS 8.1540 Page 120117 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS Analyzer LM Res| HMRes 1 Energy | Entrance Exit LMRes2 HM Res2 IEnergy 2 Multiplier Gas Flows Cone Gas Desolvation Pressures Gas Cell Set 12.0-15.0V 120-15.0V 07v av wv ov nov 10v 650v Set 150L/he 700L/hr Set 3.0e-3mbar 1A0savnicaLQuautyCo0 wrm 00e0r 0 ILL Analytical resultsof the FB, FMS, FD, and FSCS should be evaluated at the conclusion of the study to help interpret the data quality of samples data. Analytical results for these control/duplicate samples must be reported with the sample data 04562 MS Wor9d7 ETS#1540 Page 130r17 Determinationof PROS, PFOSA, POAA in Water by Liquid-Soid Extraction and LC/MS/MS 12.0 ANALYTICAL PROCEDURE 12.1 Sample Analysis 12.1.1 Set up analysis sample queue. 12.1.2 Inject the same aliquot (between 5-25uL)of each standard, analytical sample, recovery, control etc. into the LC/MS/MS system. 12.1.3 All samples showing a response for one or more analytes above the response of the highest, active calibration curve level must be diluted and reanalyzed 122 Calibration Curve 12.2.1 Starting vith the standard of lowest concentration, inject the same size aliquot (12b.1etwaenedn t1ab0ul-a2te5t)heorfeesapcohnseext(rpaecatkedheciaglihbtroatriaornesst)avnedrarsdusatchceorcdoinncgenttoraSteicotnioinn the standard. Use linear standard curves for quantitation generated for each analyte by linear regression with 1/x weighting ofpeak arca versus calibration standard concentration. The correlation coefficient () for the calibration curves must be 20.990 (120.980).Ifcalibration results fall outside these limits, then appropriate reanalyzed. steps must be taken to adjust instrument operation and the standards 12.2.2 Curve--The measured value for each curve point must be within 30% of tHhiegohreotirclalowvaplouienstswhmeany cbuerdveeaicstievvaatleudatteodaocvheirevaertahnegsee acrpiptreoripar,iabtuet oantahcecedapttaa.ble curve must contain at least five active curve points 12233 Continuing Curve Verification (CCV)--Mid- and low-level calibration checks should be analyzed every 5-10 injections. The analyte level measured in the CVs sdahtoaulcdolbleecwtietdhisnub+se3q0u%eontfttohetohreeltiactalpavsasluiensg. CICf VCCshVoulfldlnoouttsbieduesoedf.thOisnlryandgaet,a collected between acceptable CCV or the initial curve can be used 13.0 DATA ANALYSIS AND CALCULATIONS 13.1 Calculate following tehqeuaatniaolny:tical sample (extract) concentration from the standard curve using the Extract Concentration, pgm, =(Paree a ina tercek pt) (slope) 13.2 Calculate the percent recovery of the FSCS using the following equation FSCS %rec, =L((5CFo0SncC5.Scaocodnndcc.e,d.,PPPgfg//mmmLLl)) .) 13.3 Caloulate the percent recoveryof the MSs using the following equation: MS%rec,rec =M(MSSconcc,o(PCgon/nmccLadSSd.aaemmd,pPpPllgeeg/CCmooL/nn)ce,m,PPg/L/mml.L)) 04563 MS Ward97 ETS4-1540 Page 14f17 DeteirasionofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMS/MS 4M0eTHODPERFORMANCEPARAMETERS Note: Any method performance parameters that are not achieved must be considered in the evaluationofthe data. Nonconformance to any specified parameters must be described and discussed in any reportingofthe data 14.1 Linearity--Linear standard curves for quantitation generated for each analyte by linear regression with 1/x weighting of peak area versus calibration standard concentration. The correlation coefficient (r) for the calibration curves must be 20.990 (20.980). 142 Calibration Curve Standards--The measured value for each curve point must be within 30%of theoretical values when curve is evaluated over a range appropriate to the data. High or low points may be deactivated to achieve these criteria, but an acceptable curve must contain a least five active curve points. 143 CCV Performance--Mid and low level calibration checks to be analyzed every 5-10 injections. The analyte level measured in the CCV should be within + 30%of theoretical values. IfCCV fall outsideof this range, data collected subsequent to the last passing CCV should not be used. Only data collected between acceptable CCV can be used. 14.4 LimitofDetection (LOD)--The lowest calibration standard with a peak area a least 2X the peak areaof the extraction blank that can be measured at a concentration greater than zero 145 Limits of Quantitation (LOQ)--The lower LOQ (LLOQ) is the lowest non-zero active standard in the calibration curve; the peak areaof the LLOQ must be at least 2X that of the extraction blank. By definition, the measured value of the LLOQ must be within 30% ofthe theoretical value. 14.6 Matrix Spikes--Matrix spike percent recoveries must be within 30%of the spiked concentration. 147 Solvent Blanks, Method Blanks, and Matrix Blanks--Values must be below the Towest non-zero active standard in the calibration curve. Matrix blanks are considered compliant if no test substance is detected above the LOD for that analyte. 148 Reproducibility--Reproducibilityof the method is defined by the resultsof the matrix spikes and matrix spike duplicates. The MS/MSD should be reproducible to within 20%. 149 UseofConfirmatory Methods--None 14.10 Demonstration of Specificity--Specificiy is demonstrated by chromatographic retention time (within 3%ofstandard) and the mass spectral response of unique product ions generated from a characteristic primary ion. 14.11 Documentation 14.111 Ifcriteria listed in this method performance section are not met, maintenance may be performed on the system and samples reanalyzed, or other actions taken as determined by the analyst. Document all actions in the appropriate logbook. 4.11.2 If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the textofthe report. 04564 MSWord 97 ETS81540 Pa15gofe17 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 gSlaamspslpeipeextttreacwtaswtaesties dainsdcafrldaemdmainblberoskoelnvegnltasissdciosnctaaridneedrsinlohciagthedBiTnUthceonltaabionraetrosr,y.and 160 RECORDS 16.1 tEhaechhepaadgeer ogrenhearnadt-ewdriftotreansotnudtyhemupasgte:havsetutdhyeofrolplroowjiecntg innufmobremra,tiaocnquiinsciltuidoend,meetihtohedr,in integration method, sample name, extraction date, dilution factor (if applicable), and analyst 162 Print the tune page, sample list, and acquisition method from MassLynx to include in the appropriate study folder. Copy these pages and tape into the instrument run log. 163 Plot the calibration curves as described in this method, then print these graphs and store in the study folder. 16.4 Print data and store integration in the study summary, folder. integration method, and chromatograms, from MassLyns, 165 Summarize data using suitable software (MS Excel 97) and store in the study folder. 16.6 Back up electronic data to appropriate and locationofbackup electronic data. medium. Record in study notebook the file name 17.0 ATTACHMENTS 17.1 Attachment A: Figures--Fluorochemical Compounds 18.0 REFERENCES 18.1 "Guidelines and Format for Methods to be Proposed at 40 CFR Part 136 or Part 141", U.S. Environmental Water, Washington, Protection D.C. Draft Agency, 1996. Officeof Science and Technology Office of 182 P"eMreftlhuoordooocftAannaelyssuilsfofnoyrltahmeidDeet(ePrFmOiSnAat)i,onaonfdPPeerrfflluuoorrooooccttaannoeastuelf(oPnOatAeA()PFinOSW)a,ter", E. Wickremesinhe and Inc, State College, J. Flaherty, Study Number 023-002, Pennsylvania, January 2000. Centre Analytical Laboratories, 18.3 sVuallfiodnaattieon(PrFepOoSr)t,fPorertfhleu"orMoeotchtoandeofsuAlnfaolnyysliasmifdoer (thPeFODSetAe)r,miannadtiPoenrfolfuoPreoroflcutoarnoooactteane. A(nPaOlyAtAi)calinLWaabotreart"o,riEe.s,WiIcnck.,reSmteastienChoellaengeJd,.PFelnanhseyrltvya,niSat,ud(yApNpurmobvaelr 023-002, pending) Centre 19.0 REVISIONS Revision `Number. Reason For Revision Revision Date MS Word 97 ETS$-1540 604565 Page 160f17 Determination of PFOS, PFOSA, POAA inWaterby Liquid-Solid Extraction and LCMS/MS Figure 1: PFOS Chemical Name = Molecular ion = Perfluorooctane sulfonate 499 (CFS) f CgF1 Te o PFOS Note: Standards are made from the salt, potassium perfluorooctane sulfonate [C,Fy,SOSK], mv 538. Figure 2: PFOSA Chemical Name ~~ = Perfluorooctane sulfonylamide Molecular ion = 498 (CiFiiSONHy) CgF1 + ---- NHp o PFOSA Figure 3: POAA Chemical Name ~~ = Molecular ion = Perfluorooctanoate 413(GFC00) C7F15CO" POAA Note: Standards are made from the salt, ammonium perfluorooctanoate [C:F\COONHL], mw 31 04566 MS Word 97 Attachment A: ETS-8-1540 Page 170617 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extractionand LOMS/MS