Document 3w0yL3LyjqRDgJQQvZRx2YV0

3M ENVIRONMENTAL LABORATORY ARLLG-c06Y BactCopy ofO Tw = 28 ooo = METHOD EH-2 A SPECTROMETRY (HPLC/MS/MS) DETERMINATION OF PERFLUOROOCTANE SULFONATE (PFOS), PERFLUOROOCTANE SULFONYLAMIDE (PFOSA), AND PERFLUOROOCTANOATE (POAA) IN WATER BY LIQUIDSOLID EXTRACTION AND HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY/TANDEM MASS Method Number: ETS-8-154.0 Adoption Date: 04/2/2000 Author: Kristen J. Hansen/Harold O. Johanson Revision Date: `Approved By: William K. Reagen, Kent R. Lindstrom dit William K. Reagen, Laboratory Management 2/08/0 Date fit fz Kristen J. Hansen, Ph.D., Group Leader tL Kent R. Lindstrom, Technical Reviewer 04/23/00 Date oulDate MS Wor9d7 ETS81540 Page tof17 `Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS CGC29'7 Be 1.R 0 SCOPEAANDAAPPLICATIONee 1.1 dTehtiesrmmientahtoidonporfovPiedrefslcuoolrloeoccttiaonn,e esxutlrfaocntaitoen,(PaFnOdSa)n,alPyetrifcalluoprroooccetdaunreeSsuflofrotnhye.lamide (PFOSAY), and `water samples. Perfluorooctanoate (POAA) in groundwater, surface water, and drinking 12 MTheitshomdesthtoodbewaPsropproespeadreadt a4c0coCrFdRingPatrott1h3e6EoPrAPadrotc1u4m1e"nt(,se"eGRuiedfeelriennecsea1n8.d1)F,oarnmdatisfor sbualsfeodnaitnep(arPtFOoSn)t,hePerrefplourtor"oMoecttahnoedsoulffAonnayllyasmiisdefo(rPtFhOeSDAe)t,eramnidnaPteirofnlouforPoeorcftlaunooraotoectane (POAA) in Water" (see Reference 18.2). 20 SUMMARY OF METHOD 21 fWaactileirtys.aPmFpOlSes, aPrFeOcSolAl,ecatnedd fPrOoAm Aasairteeeofxtirnatecrteesdtfarnodm s4h0imppLedwactoelrdstaomapnleansaulystiincgalC,q solid phase extraction (SPE) cartridges. The compounds are eluted from theC,,cartridge, using methanol. Separation, identification, and measurement are accomplished by high- performance liquid chromatography/tendem using multiple response monitoring (MRM). mass spectrometry (HPLC/MS/MS) analysis `rTehsepocnosnecoenfttrhaetiqounoanftietaacthioindieonntipfireodduccoemdpobnyetnhtatiscmoemapsouurneddtboytcheomMpSarirensgptohneseMoSf the (queaxntteimtaaltisotnanfdoanrdp)r.oduced by the same compound in an extracted calibration standard 30 DerNiTIONs 31 Analytical Sample--A portion of an 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 0labtohraatotofrtyhaendfirasntaallyizqeudotsegpiavreaatemleyawsiutrheoidfetnhtiecaplrepcrioscieodunraesss.ocAinaatleydswiisothflDabSosractoomrpyared procedures, but not with sample collection, preservation, or storage procedures. 34 lFaibeolrdaBtolraynaknCdotnrteratoeldaSsamapslaemp(lFeB)in--aTllypreespIecwtast,erinpcllaucdeidnginexapsoasmuprleetocosnatmapilnienrginsitthee conditions, storage, preservation and all analytical procedures. The purposeofthe FB is to determineiftest substances or other interferences are present in the field environment. MS Word97 ETS$-1540 Page20f17 DeterminationofPFOS, PFOSA, POA in Water by Liquid-Solid Extraction and LOMSMS ~~CGQZ 9S 35 FainedplldaDcuepdluincadteer(idFeDnt)i--caAl sciarmcpulmestcoalnlceecsteadndintdrueaptleidcaetxeaactttlyhethseasmaemteitmheraosugthheoustafmipellde and laboratory procedures. Analysis ofFD) compared to thatofthe first sample gives a `measureofthe precision associated `well as with laboratory procedures. with sample collection, preservation and storage, as 3.6 Field Matrix Spike (FMS)--A sample collected in duplicate to which known quantities osfhtouhledtbaregestpiakneadlyattesapaprreoaxdidmeadteilnyt5h0e -f1ie5l0d %atoftthehteimeexopefcstaemdpalnealcyotleleccotnicoenn.tTrhateioFnMSin.the sample. The FMS is analyzed to ascertainifany matrix effects, interferences, or stability issues may complicate the interpretationofthe sample analysis. 37 Field Spike Control Sample (FSCS)--An aliquotoftype I water to which known `quantitiesofthe target analytes are added in the field at the timeof sample collection (at aanndapapnraolpyrzieadteexcaocntcleyntlriaketiaonsatmopbleedteotdeertmeirnmeidnbeywhtheethperorjaecltoslseoadf)a.nTahleytFeScCoSulids beextracted attributed to sample storage and/or shipment. 38 Laboratory Control Sample (LCS)--An aliquotoftyIpwateer 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 concentrationof approx. 100-250Pg/mL or another concentration to be determined by the project lead. The LCS is extracted and. analyzed exactly like a laboratory sample to determine whetherthe methodology is in control, and whether the laboratory is capableofmaking accurate measurements at the required method detection limit and higher. 39 Laboratory Sample--A portion ofa sample received from the field for testing. 310 Limit of Detection (LOD)--The lowest concentration ofan 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 31 Limit of Quantitation (LOQ)--The lowest concentration (LLOQ) or highest concentration (ULOQ) that can be reliably achieved within the specified limits of precision and accuracy during routine operating conditions. Ncoontcee:ntTrhaetiLonLOisQseilsegcteenderaaslltyhe5-lo1w0etstimneosnt-hzeerLoOsDt.anFdoarrdmianntyheancaallyitbersa,titohnecLurLvOe.QHaonwaelvyeter, it may be LLOQs nominally chosen within are matrix-dependent. these stated guidelines to simplify data reporting. Sample: 3.2 Matrix Spike (MS)--An aliquot ofasample, to which known quantitiesoftarget analytes are added in the laboratory. The MS is extracted and analyzed exactly likae laboratory sample to determine whether the sample matrix contributes bias to the analytical results. The background concentrationsofthe analytes in the sample matrix `must be determined in a separate aliquotandthe measured values in the MS corrected for background concentrations. MS Word 87 ETS81540 Page 30f17 `Determinationof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LOMSMS ~~ CGORZ99 3.13 sMaemtphloedinBclluadnikng--eAxnpoasluiqrueotto oaftllygplaIesswwaartee,retqhautipismetnreta,tesdolevxeancttsl,yrleiakgeenatsl,abionrtaetronraly standards, and surrogates that are used with other laboratory samples. The method blank iesnvuisreodntmoendte,tetrhmeirneeagieftnetsst, sourbtshteanacpepasroartuost.her interferences are present in the laboratory 3.14 Method Detection Limit (MDL) Determination--Oneof several 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 isgreaterthan 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 ofmaterial 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. 3.17 Stock Standard (SS)--A concentrated solutionof a single analyte prepared in the laboratory with an assayed reference compound. 3.18 Working Standard (WS)--A solutionofseveral analytes prepared in the laboratory from SSs and diluted as needed to prepare calibration standards and other required analyte solutions. 4.0 WARNINGS AND CAUTIONS 41 Health and Safety Warnings 4.11 The acute and chronic toxicityofthe standards for this method have not been precisely determined; however, each should be treated as2 potential health hazard. 41.2 Unknown samples may contain high concentrationsofvolatile toxic compounds. `Sample containers should be opened in a hood and handled with gloves to prevent exposure. 413 The laboratory is responsible for maintaining a safe work environment and a current awareness of local regulations regarding the handlingofthe chemicals used in this method. A reference file ofmaterial safety data sheets (MSDS) should be available to all personnel involved in these analyses. 50 INTERFERENCES 5.1 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 conditionsofanalysis by running method blanks. 53 Teflon containing materials (e.g. caps, wash bottles) contain fluorocompounds which may cause interferences analysisofthe samples. and should not be used during collection, storage, extraction, or MS Word 97 ETS 81540 Pagedo1f7 DeterminationofPEOS, PFOSA, POAA in Water by Liguid-Solid Extraction and LOMSMS CG 200 6.0 EQUIPMENT, SUPPLIES, AND MATERIALS `pNeortfeo:rBmraanncde nmaamyesb,esaucphpilieevresd,uasnidngpaarptpmaruamtbuesrasnadremaftoerriilallusstortahteirvetphuarnptohsoesseosnpleyc.ifEiqeudihvearlee,ntbut demonstrationofequivalent performance that meets the requirements of this method is the responsibiltyofthe laboratory. 61 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. 6.1.3 Ice for sample shipment. 6.1.4 Bottles must be lot-certified to be free ofartifacts by running Method blanks `according to this method. 62 Laboratory Equipment (Extraction and Analytical) 62.1 Balance, analytical (display at least 0.0001), Mettler. 62.2 Vacuum pump, Biichi. 62.3 Visiprep vacuum manifold, Supelco. 62.4 Sep Pak Vac Gee (1g) (C, cartridges (par#t WAT036795),Waters. 6.2.5 S0mL disposable polypropylene centrifuge tubes, VWR. 62.6 15mL disposable polypropylene centrifuge tubes, VWR. 6.2.7 Disposable micropipette (50-1004L, 100-200uL), Drummond. 6.2.8 Class Apipettes and volumetric flasks, various. 62.9 Hypercarb drop-in guard column (mim) (part# 844017400), Keystone. 6.2.10 Stand-alone drop-in guard cartridge holder, Keystone. 6.2.11 125ml LDPE narrow-mouth bottles, Nalgene. 62.12 HPLC pump (LCI0AD), Shimadzu. 62.13 2mL clear HPLC vial kit (cat# 5181-3400), Hewlett Packard. 62.14 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 63.1 In order to avoid contamination, the useofdisposable labware is highly recommended (tubes, pipettes, (c.). 63.2 Teflon or Teflon-lined containers or equipment, including Teflon-lined HPLC vials or capsforthe HPLC auto sampler must notbe used. 63.3 Type water used during the sample and standard extraction should be filtered through a Hypercarb guard column usinag HPLC pump. This water isreferredto as "filtered type I water", hereafter in this report. 634 tis necessary to check the solvents (methanol) for the presenceofcontaminants (especially POAA) by LC/MS/MS prior to use. Certain lot numbers have been found to be unsuitable for use 635 Use disposable micropipettes or pipettes to aliquot standard solutions to make calibration standards and matrix spikes. MSWord 97 ETS.8.1540 Page sof17 DeterminationofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction snd LOMSMS 0.0 G30 7.0 REAGENTS AND STANDARDS ``Nmoatye:beSuapcphliiecvresdaunsdincagtcahleomgincuamlbseorbstaairneedfofrriollmusottrhaetrivseupppulripeorsse.sDoonlnyo.tEuqsueivaalleesnsterpegrrfaodermoafnce chemical than those listed. 71 Chemicals 7.11 Methanol (MeOH), HPLC grade, JT Baker, Catalog No. JT9093-2. 7.12 7.1.3 Ammonium Water, type Acetate, Reagent grade, I, prepared in-house. Sigma-Aldrich, Catalog No. A~7330. 7.14 Sodium Thiosulfate, Reagent grade,JT Baker. 72 Standards 7.2.1 7.22 PPeortfalsusoiruomocptearnfelusourlofoocntyalnaemsiudlefo(nsaeteeA(tsteaecAhtmteancthAm,enFtigAu,reFi2g).ure 1). 7.2.3 Ammonium perfluorooctanoate (see Attachment A, Figure 3). 73 Reagent Preparation 7.31 250mg/mL sodium thiosulfate solution (Extraction)--Dissolve 25gofsodium thiosulfate in 100mL reagent water. 732 40% methanol(Extraction) --Measure 400mL methanol and adjust the volume 10 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: Alternative are maintained. volumes may be prepared as long as the ratios ofthe solvent to solute ratios 74 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 C.F,SOK. purity 90% = 8.35mg C;F,,S0,-) and dilute to 100mL with methanol in a 100mL Volumetric flask. Transfer to a 125mL LDPE bottle. Prepare a separate solution for cach analyte. Store solutions in a refrigerator at 4:2C for a maximum period of6months from the date of preparation. 7.42 Ipg/mL mixed SSS--Add 1.0mL cachofthe 100pg/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 ofthe 1.0ug/mL-mixed solution (from 7.42) 10 a 100mL volumetric flask and bring up to volume with methanol. 7.4.4 0.01pg/mL mixed SSS--Add 10.0mLofthe 0.1pg/mLmixed solution (from 7.43) to 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 442C for a maximum periodof 3 months from the dateofpreparation. MS Word 97 ETS8-1540 Page 6of17 DetentionofPROS, PFOSA, POAA in Water byLiquidSold Extraction sd LOMSMS 0.60302 75 Calibration Standards 7.5.1 o1u0t01u0gm/gmoLfeaancalhyPtiFcOaSl,stPaFndOaSrAd,(caornrdecPteOdAfoAr spetrocceknsttsaalntdaanrdd psuorlituyt)iaonnds--dWilelitgeh to 100mL with methanol in a 100mLvolumetric flask. Transfer to a 125mL rLeDfiPigEerbaottotrlea.tP4re4pa2reCafosrepaamraatxeismoulmutipoenrifoodr oeafc6hmaonnaltyhtse.frStoomrethseolduattieoonsf in a preparation. 752 Ipg/mL Working Standard--Add 1.0mL each ofthe 100ug/mL SS solutions (fom 7.5.1) to a 100mL volumetric flaskandbring up to volume with methanol. 753 0.1pg/mL Working Standard --Add 10.0mL of the 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 0.1pg/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 date of preparation. 7.56 Calibration Standard--Prepare a minimumoffive calibration solutions in filtered type I water according to the following table: `Concentration OfWS, pg/mL Volume of WS, uL. Final Calibration Standard Volume, mL CaliFbirnaatliCoonnScteanntdraartdi,onPgof/mL 0 ow we 0010 100 "0 2 0010 200 "0 50 0010 400 "0 100 0.10 100 " 250 0.10 200 " 500 010 300 40 7501 0.10 400 40 10002 2TTVMobheapprepeyareedoooextend teemrangge bbeeyonnd O 750PL g/L. Note: The absolute volumes of the standards may be varied by the analyst as long as the correct p7r.o5p.o7rtioTnshoefssotlauntdeartdossaorlevepnrtoacreesmsaeidnttahirneodu.gh the extraction procedure (Section 9.0), identical to the laboratory samples. The extracted concentrationofthe calibration standard is equal to 8X the initial concentration, due to the concentrationof the standard during the extraction process. 7.58 Storage Conditions--Store all extracted calibration standards in 15mL `polypropylene tubes at 4:2C, for a maximum periodoftwo weeks from the dateof preparation. MSWord 97 ETS8-1540 Page7of17 DeterminationofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LMsts CG 0303 8.0 SAMPLE COLLECTION, PRESERVATION AND STORAGE. gNaostkee:tsS,aamnpdloitnhgerepqauritpsmetnhta,t imnacyluldeiancghaiunttoemrafteriicngsaamnpalleyrtse,s miunsottbhee wfareteeorfsaTmepflleo.n tAuubtionmga,tic samplers that composite samples over time should use refrigerated polypropylene sample containers ifpossible. Sample bottles should not be rinsed 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 500mLmin 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 baler 83 Surface Water--When sampling from an open bodyofwater, fill the sample container with water from a representative area. 84 Sample Dechlorination--All samples should be iced or refrigerated at 442C and kept in the dark from the timeofcollection until extraction. Residual chlorine should be reduced by adding 2004Lof a 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.). 85 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. Ifthe 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 cach sample set (samples collected from the same general sample site at approximately the same time). At the laboratory, prior to sample collection, fill a sample container with filtered type I 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. 87 Field Duplicates 8.7.1 Collecat Field Duplicate (FD) for every ten (10) samples collectedorper each sampling set, ifless than 10 samples are collected. 8.7.2 Separate FDs must be collected for each typeofwater sample (ground, tap, etc.) collected. 87.3 Collect the FD immediately afterthe sample. 8.74 Preserve, store and ship FD using the same procedures as used for the samples. MSWord 97 ETS8.1540 Page Sof 17 Determination of FOS, PFOSA, POAA in Water byLiquid Said Extraction and LCMsvs C0030 88 Field Spike Control Sample (FSCS) 88.1 AField Spike Control Sample (FSCS) mustbeprepared for cach sample shipment. Ifmultiple coolers are used to ship a setofsamples, each cooler must contain a FSCS. 882 Atthe laboratory, fll a sample container with 100mLof type I water. Seal and ship to the sampling site 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. 8.84 Seal and gently invert the FSCS to mix. Store and ship the FSCS using the same procedures as used for the samples. MS Word 97 ETS8-1540 Page9of17 Determination of PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LoMsMs C0305 9.0 EXTRACTION PROCEDURE 9.1 Extraction Scheme 9.1.1 Allow samples to equilibrate to room temperature. Thoroughly mix samples by gently inverting the sample bottle. 9.12 Measure 40mLofsample into SOmL polypropylene centrifuge tubes (Spike the QC and Matrix spikes as required, replace lid and mix well). N(toytpei:ca*llSyaSm0p-l1es50m%aoyfsneaemdpltoebceonpcreenstcrraeteinone)d.to determine an appropriate matrix spike level 9.1.3 Condition the C,, SPE cartridges (1g, 6mL) by passing 10mL methanol followed by SmL filtered type I water (~2drop/sec). Do not let column run dy. dNroytea:t aFnoyr ttihmee.following steps, maintain a ~1dropisec flow rate. Do not allow the column to run 9.14 Load the analytical sample onto the C,, SPE cartridge. Discard eluate. 9.5 Wash with ~SmL 40% methanol in water. Discard eluate. 9.1.6 Elute with ~SmL 100% methanol. Collect SmLofeluate into graduated 15mL. `polypropylene centrifuge tubes. This is the target elution fraction (final volume = Sm). 9.1.7 Analyze a portionofthe target elution fraction eluent using negative electrospray HPLC/MS/MS (Section 10.2). Note: Samples are concentrated by a factorof eight during the extraction; Initial Vol = 40mL Final Vol. = SmL. 9.1.8 Samples are stable at room temperature for at least 24 hours. Analytical samples may be stored in a refrigerator at 44:2C until analysis. 9.19 StandardizationofCy, SPEcolumns--Ifpoor recoveries are observed, it may be necessary to standardize the C, SPE columns in the followingmanner before analyzing samples. 9.1.9.1 Use a standard with an analyte concentration between 1000 and 4000 Py/mL. Follow the extraction scheme as outlined from steps 9.1.1 t0 9.1.6, except, collectthe 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 Sm of 100% methanol. 9.19.3 Analyze all threefractionsby HPLC/MS/MS. Ifthe target fraction contains a minimum of 85%ofthe respective analytes, it may be considered acceptable. 9.19.4 Ifthe wash contains significant standard (>15%), ither the wash volume. or percentage of MeOH should be decreased. 9.19.5 Ifthe post-clution fraction contains significant standard (>15%), the target elution volume should be increased. MS Ward 97 ETS.8-1540 Page 10017 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction snd LOMsMs -- C00306 10.0 CALIBRATION AND STANDARDIZATION (ANALYTICAL SETUP) NToontge:asOtthheermientshtordumcernittesrimaaayrebmeetu.seTdhaendoptehreateoqrumiupsmtenotptainmdizcoenadnitdidonoscummaeyntbethveercyqudiipffmeernetnt as and 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 Note: A 4x 10mm Hypercarb drop-in guard cartridge (Keystone, part # 844017400) is attached. othna-tlimnaeyabfetritnhethpeumrogbeivlaelpvehaasnedabnedf/oorreHtPheLsCamspyslteemi.njector port fo trap any residue contaminants HPLC Column: Genesis C, (Jones Chromatography), 2.1mm x 50mm, 4um Column Temperature: 35C Injection Volume: 15uL Mobile Phase (A): 2mM Ammonium Acetate in filtered type I water (See 7.3.1) Mobile Phase (B): Methanol HPLC Gradient Program: Time, Percent Mobile min Phase A 0.0 60 04 60 10 10 70 10 75 0 90 0 95 60 135 60 140 60 Percent Mobile Phase B 40 "0 % 90 100 100 "0 40 4 Flow Rate, mL/min 03 03 03 03 03 04 04 04 03 Note: Other HPLC gradientsmaybe used as long as the method criteia are met. MS Word 97 ETS81540 Page 110f17 Detceminatonof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction snd Lonstvis C 00307 Tt may be necessary to adjust the HPLC gradient in order to optimize instrument performance. Columns with different dimensions (e.g. 2. 1mm x 30mm)and columns from different manufacturers (Keystone Betasil C,, etc.) may be used. Tons Monitored: Analyte | Primary Ion Productlon pASpPprPoExiNmTatEeR PoAA| 4130 169.0 50 PFOS 499.0 9.0 52 PFOSA| 498.0 78.0 58 Other product ions may be chosen at the discretion ofthe analyst, although m/z 99 is suggested for PFOS. Useofthe suggested primary ion is recommended. Retention times may vary slightly, on aday-to-day basis, depending on the batch ofmobile 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. 102 Tune File Parameters 1021 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 1 Hexapole 2 Source Block Temp. Desolvation Temp. Set 2.56-3.5kV 05v 02v 08v 100-150C 250-400C MS Word97 ETS 8.1540 Page 12017 DeterminationofPROS, PFOSA, POAA in Water byLiquid Sold Exracion snd Lomsrvs. C00308 Analyzer LMRes1 HMRes | IEnergy 1 Entrance Exit LMRes2 HM Res 2 IEnergy 2 Multiplier Set 12.0-15.0V 120-15.0V ov 2v v 1.0v 1.0v 10v 650V Gas Flows Cone Gas Desolvation Set 150L/hr 700L/hr Pressures Gas Cell Set 3.0-3mbar 11.0 ANALYTICAL QUALITY CONTROL 111 Analytical resultsofthe FB, FMS, FD, and FSCS should be evaluated at the conclusion ofthestudyto help interpret the data qualityofsamples data. Analytical results for these. controliduplicate samples must be reported with the sample data. MS Word97 ETS81540 Page 130617 Determination ofPROS, PFOSA, POAA in Water byLiuid-Sid Exrscton snd Less. 00309 12.0 ANALYTICAL PROCEDURE 121 S12a.m1p1le ASneatluypsiansalysis sample queue. 12.12 Inject the same aliquot (between 5-25)ofeach standard, analytical sample, 12.1.3 rAelclosvaermyp,lecsonsthoolwietncg. ainrteostphoensLeCf/oMrSo/neMSorsmyostreema.nalytes above the response of the highest, active calibration curve level must be diluted and reanalyzed. 122 Calibration Curve 12.2.1 Starting with the standard of lowest concentration, inject the same size aliquot (between 10-25yL)ofeach extracted calibration standard according to Section t1h2e.1staannddatradb.ulUastee tlhieneraerssptoannsdear(dpecaukrvheesigfhotr oquraanxteiat)atvieornsugsentehreactoendcfeonrtzcaatcihon in analyte by linear regression with 1/x weightingofpeak arca versus calibration smtuasntdabred2c0o.n9c9en0tr(a7t2i0o.n9.80T)h,eIfccoarlreilbartaitoinoncoreefsfuilctisefnatll(ro)uftosridtehethceasleiblriamtiitosn, ctuhrevnes arpeparnoaplryizaetde. steps must be taken to adjust instrument operation and the standards 12.22 Curve--The measured value for cach curve point mustbe within 30% of Hthiegohreotirclalowvaplouienstswmheany cbuerdveeaicstievvaatleudatteodaocvheirevaertahnegsee acrpiptreoripar,iabtuet taontahcecdeapttaa.ble curve must contain at least ive active curve pois. 12.23 CshoonutlidnubienagnaCluyrzvedeeVveerrifyi5c-at1i0oinnj(eCctCiVon)s--.MTihde-anaanldytleolwe-vleelvemleacsaluirberdatiinonthcehecks (raCnCgeV, sdahtoaulcdolbleecwtietdhsinub+se3q0ue%notftothtehoereltaisctaplavsasliunegs.CICfVCCsVhoulfdulnootutbseiduesoefd.this Only dat collected between acceptable CCV or the inital curve can be use. 13.0 DATA ANALYSIS AND CALCULATIONS 13.1 Calculate the analytical sample (extract) conceniration from the standard curve using the following equation: `Extract Concepn/mt=rPeaakatr(esialoipnoet)ernce,pt) 132 Calculate the percent recovery ofthe FSCS using the following equation: FSCS YeYeE,== (SCoCnCeSS.cacododnneccd,n,PPPEgg///mmmLLL))) 133 Caleulate the percent recovery ofthe MSs usingth following equation: MSoreo, - MSconc,Py/mL-SampleConc.PgmL) (Concadded,Pg/mL) MS Wor9d7 ETSs1540 Page 14or17 DeterminationofPFOS, PFOSA, POAA in Water by Liquid-Solid Extraction snd Loss C0 0310 14.0 METHOD PERFORMANCE PARAMETERS Note: Any method performance parameters that are not achieved must be considered in the `valuationofthe data. Nonconformance to any specified parameters must be described and discussed in any reportingofthe data. 141 Linearity--Linear standard curves for quantitation generated for each analyte by linear regression with 1/x weightingofpeak area versus calibration standard concentration. The correlation coefficient () for the calibration curves must be 20.990 (P20.980). 142 Calibration Curve Standards--The measured value for each curve point must be within 30%oftheoretical 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 at 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%oftheoretical values. IfCCV fall outsideofthis range, data collected subsequent to the last passing CCV should not be used. Only data collected between acceptable CCV can be used. 144 Limit of Detection (LOD)--The lowest calibration standard with a peak area at least 2X the peak areaofthe 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 area ofthe LLOQ must be at least 2X that of the extraction blank. By definition, the measured valueofthe LLOQ must be within 30% ofthe theoretical value. 14.6 Matrix Spikes--Matrix spike percent recoveries must be within 30%ofthe spiked concentration. 14.7 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. compliantifno 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 Demonstrationof Specificity--Specificity is demonstrated by chromatographic retention time (within 3%ofstandard) and the mass spectral responseofunique product ions generated from a characteristic primary ion. 14.11 Documentation 14.1.1 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. 14.1.2If data are to be reported when performance criteria have not been met, the data must be footnoted on tables and discussed in the text of the report MSWord97 ETS 8.1540 Page150117 DeterminationofPFOS, PFOSA, POAA inWater by Liquid-Solid Extraction snd LOMsvs 00311 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Sample extract waste and flammable solvent is discarded in high BTU containers, and glass pipette waste i discarded in broken glass containers located in the laboratory. 160 RECORDS 16.1 Each page generated for a study must have the following information included, either in the header or hand-written on the page: study or project number, acquisition method, 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 cuarsdvesceribsed in.this method, then print these graphs and store in the study folder. 16.4 Print data integration summary, integration method, and chromatograms, from MassLyn, and store in the study folder. 165 Summarize data using suitable software (MS Excel 97)and store in the study folder. 166 Back up electronic data to appropriate medium. Record in study notebook the file name `and location ofbackup electronic data. 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 Protection Agency, Office of Science and Technology Office of Water, Washington, D.C. Draft 1996. 182 "MethodofAnalysis for the Determinationof Perfluorooctane sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc, State College, Pennsylvania, January 2000. 183 Validation report for the "Method ofAnalysis for the DeterminationofPerfluorooctane: sulfonate (PFOS), Perfluorooctane sulfonylamide (PFOSA), and Perfluorooctanoate (POAA) in Water", E. Wickremesinhe and J. Flaherty, Study Number 023-002, Centre Analytical Laboratories, Inc., State College, Pennsylvania, (Approval pending) 19.0 REVISIONS Revision Number. Reason For Revision Revision Date MSWord 97 ETS $1540 Page 16017 Decrminaion ofPEOS, FFOSA, POAA in Water byLiquidSold Bescon sna Lenses 000312 Figure 1: PFOS Chemical Name ~~ = Molecular ion = Perfluorooctane sulfonate 499 (CFSOY) { CgF1 Te 0 PFOS Note: Standardsare made from the salt, potassium perfluorooctane sulfonate [CgF17SO3K], m/w 538. Figure 2: PFOSA Chemical Name ~~ = Perfluorooctane sulfonylamide Molecular ion = 498 (CiFySONHy) CgF1 wn NH o PFOSA Figure 3: POAA Chemical Name ~~ = Molecular ion = Perfluorooctanoate 413(CFsC00) C7F1sCO" POAA Note: Standards are made from the salt, ammonium perfluorooctanoate [C7F1 SCOONHg), m/w 431 MS Word 97 Attachment A: ETS.8:1540 Page 170f17 Determinationof PFOS, PFOSA, POAA in Water by Liquid-Solid Extraction and LMsvs C30 31.3