Document kmR237La3vdBr8rRkg3pryx2B

ARNG5 3M ENVIRONMENTAL LABORATORY METHOD ANALYSIS OF POTASSIUM PERFLUOROOCTANESULFONATE OR OTHER FLUOROCHEMICALS IN SERUM EXTRACTS USING HPLC-ELECTROSPRAY/MASS SPECTROMETRY Method Number: ETS-8-5.1 Author: Lisa Clemen, Robert Wynne Approved By. HE Laboratory Manager i / ater Ae Group Leader Hie A Clonee Technical Reviewer Adoption Date: 03/01/99 Revision Date: 4/3/71 "ec, Date 7/26/95 Date 04/2/31 Date 1.0 ScopeANDAPPLICATION 0000000 1.1 Scope: This method describes the analysisof serum extracts for fluorochemical surfactants using HPLC-electrospray/mass spectrometry. 1.2 Applicable Compounds: Fluorochemical surfactants or other fluorinated compounds, or other ionizable compounds. 1.3 Matrices: Rabbit, rat bovine, monkey, and human serum, or other fluids as designated in the validation report. Word 695 Analysisof SerEuTmS.Ex8t.r5a1ct Using ESMS Page 10f9 001012 2.0 SUMMARY OF METHOD 2.1 This method describes the analysis of fluorochemical surfactants extracted from serum or other fluids, using HPLC-electrospray/mass spectrometry, or similar system as appropriate. `fTlhueoraoncahleymsiiscails,pseurcfhoramsetdhebypemrofnliutoorroiocntgaanessiunlgfloenaitoen c(hPaFrOacSt)erainstiiocno,fma/zp=ar4t9i9c.ular Additionally, samples may be analyzedusing a tandem mass spectrometteor further verify the identity of a compound by detecting daughter ions of the parent ion. 3.0 DeemTIONS 3.1 Atmospheric Pressure Ionization (API): The Micromass Quattro I triple quadrupole sinytsetrefamcsesa.llTohwefsoer vinacrliuodues mbuettharoedsnootfliiomniitzeadtitoo:n EblyecuttriloiszpirnagyvIaorniiozuastisoonur(cEeSsD,),prAotbmeos,spahnedric Pressure chemical Ionization (API), Thermospray, etc. The ionization process in these techniques occurs at atmospheric pressure (i.c., not under a vacuum). 3.2 Electrospray Ionization (ES, ESI): a methodof ionization performed at atmospheric pressure, whereby ions in solution are transferred to the gas phase via tiny charged droplets. These charged droplets are produced by the applicationof a strong electrical field. 3.3 Mass Spectrometry, Mass Spectrometer (MS), Tandem Mass Spectrometer (MS/MS): `The API Quattro II triple quadrupole systems are equipped with quadrupole mass selective detectors. Tons are selectively discriminated by mass to charge ratio (m/z) and subsequently sdpeetceicftiecd.frAagsmienngtlaetiMoSn imnafoyrmbaetieomnp.loyed for ion detection or aseries (MS/MS) for more 3.4 Conventional vs. Z-spray probe interface: The latest models of Micromass Quattro II triple quadrupole systems (post 1998) utilize a "Z-spray" conformation. The spray emitted from a probe is orthogonal to the cone aperture. In the conventional conformation it is aimed directly at the cone aperture, after passing through 2 tortuous pathway in the counter melaeicnttreodnea.ncTehaoruegthhethseamceo.nfiHgouwreatvieorn,iZs-dsifpfrearyencto,mtphoenmeentthsodasnodfcoonpveernattiioonn,alclceoamnpinogn,enatnsd are not compatible with one another, but only with similar systems (i.., Z-spray components are compatible with some other Z-spray systems, etc.) 3.5 Mass Lynx Software: System software designed for the specific operationofthese Quattro 1 triple quadrupole systems. Currently MassLynx has Windows 95 and WindowsNT 4.0 versions. All versions are similar. For more details see the manual specific to the instrument (Micromass Quattro Il triple quadrupole MassLynx or MassLynxNT User's Guide). 4.0 WARNINGS AND CAUTIONS 41 Health and Safety Warnings: 4.11 `Uesmeplcoayustiaonvowlittahgetohfeavpoplrtoaxgiemcaatbelelsyf5o0r0t0heVoplrtosb.e. When engaged, the probe 4.1.2 When handling samplesor solvents wear appropriate protective gloves, eyewear, and clothing. Analysis ofSeErTuSm.8E.xt5r.a1ct Using ESMS Page 2069 001013 42 Cautions: 42.1 DIfothneotbaocpkerparteesssuorleveenxtcpeeudmsps40a0bobavre, ctahpeaHciPt1y1o0f040w0ilbairni(ti5a8t0e0auptsio)mabtaiccksphruetssduorwen.. 422 Do not run solvent pumps to dryness. 5.0 INTERFERENCES 5.1 Tstoormaigneiomrizaenyinptaerrtfoefriennsctersuwmhenetnataniaolnytzhiantgcsoammeplsesin,ctoenftlaocntswhiotuhldthneotsabmepulseeodrfeoxrtrsaacmtp.le 6.0 EQuipment eee 6.1 Equipment listed below may be modified in order to optimize the system. Document any `modifications in the raw dataas method deviations. 6.1.1 Micromass Quattro II triple quadrupole Mass Spectrometer equipped with an electrospray ionization source 6.1.2 HP1100 low pulse solvent pumping system, solvent degasser, column compartment, and autosampler 7.0 SUPPLIES AND MATERIALS 7.1 Supplies 7.11 High purity grade nitrogen gas regulated to approximately 100psi (House air system) 7.1.2 HPLC analytical column, specifics to be determined by the analyst and documented in the raw data. 7.1.3 Capped autovials or capped 15 mL centrifuge tubes. 8R .0 EAGENTSANDSTAND0A00R 000D00S 00 81 Reagents 8.1.1 Methanol, HPLC grade or equivalent 8.1.2 Milli-QTM water, all water used in this method should be Milli-QTM water or equivalent, and may be provided by a Milli-Q TOC Plus system or other vendor 8.1.3 Ammonium acetate, reagent grade or equivalent 82 Standards 8.2.1 Typically two method blanks, two matrix blanks, and eighteen matrix standards are prepared during the extraction procedure. See ETS-8-4.1. 9.0 SAMPLE HANDLING 9.1 Fresh matrix standards are prepared with each analysis. Extracted standards and samples are stored in capped autovials or capped 15 mL centrifuge tubes until analysis. Analysis ofSerEuTmS.E8x.t5r.a1ct Using ESMS Pag3e 0f9 001014 9.2 Ifanalysis will approximately be 4 Cd,eloaryeatd,roexotmratcetmepdersattaunrdea,rdusntainldansaalmypsliesscacnanbebeperrefforirgmeerda.ted at 0.0 QUALITY ConTrOL 10.1 Solvent Blanks, Method Blanks and Matrix Blanks 10.1.1 Seoalcvhebnattbclhatnoksd,etmeertmhionde bcloanntkasmiannadtimoantroirxcbalrarynokvsera.re prepared and analyzed with 10.1.2 Analyze amethod blank and a matrix blank prior to each calibration curve. 102 Matrix Spikes 10.2.1 Mreactorviexrsypeifkfeisciaernecyp.repared and analyzed to determine the matrix effect on the 10.2.2 Mreactorviexrsypfiokreedaucphliacnaatleystea.re prepared and analyzed to measure the precision and the 10.2.3 `Amnianlyizmeumaomfat2risxpsikpeiskepearnbdatmcaht.rix spike duplicate per forty samples, with a 10.2.4 Mthaetriinixtisaplickaeliabnrdatmiaontrciuxrvsep.ikAeddduiptliiocnaatle scpoinkceenctornacteinotnrsatwiilolnsfamllaiyn ftahlle mid-range of in the low- rangeofthe initial calibration curve. 10.3 Continuing Calibration Verifications 10.3.1oCfotnhteincuailnigbrcaatliiobnrcautrivoen.verifications are analyzed to verify the continued accuracy 1032 oAfnoanleyzpeear bmaitdc-hr.ange calibration standard after every tenth sample, with a minimum 11.0 CALIBRATION AND STANDARDIZATION 11.1 Aavnearlaygzeeotfhteweoxtsrtaacntdedarmdatcrurivxesstawinldlarbdespplroitotredtobaynldinfeoalrlorewgirnegsesaiocnh(syet=omfyext+rabc)t,s.weTihgheted 1/x, not forced through zero, using MassLynx or other suitable software. 11.2 Isfttahnedacrudrcvuerdvoe(eisfnnoetcmeseseatryr)eqaunidrermeeanntasl,yzpee.rform routine maintenance or reextract the 11.3 Fusoer pthuerplooswesenodfoafcctuhreaccayliwbhraetnioqnuacnutrivteatriantghelrotwhalnevtehlesoffuallnarlayntgee,ofittmhaeystbaendnaercdescusravrey.to Ecaxlaimbprlaeti:onwchuervneactotnesmipsttiinngg tooftqhuaentsittaantdearadpsprforxoimma5tpeplby t1o0 1p0p0bopfpbanraaltyhteer,tgheannerthaetefuall rangeofthe curve (5 ppb to regression weightingofhigh 1c0o0n0cepnptbr)a.tiTohnisstawnidllarrdesd.uce inaccuracy attributed to linear Analysis ofSeErTumSE8xt5ra1ct Using ESMS Page dof? 001015 20 Proceoures 12.1 Acquisition Set up 12.1.1 Click on start button in the Acquisition Control Panel. Set up a sample list. Assign a filename using MO-DAY-last digitofyear-sample for acquiring, and type in sample descriptions. number, assign a method (MS) 12.1.2 To create a method click on scan button in the Acquisition control panel and select SIR (Single lon Recording) or MRM. Set Ionization Mode as appropriate and mass 10 499 or other appropriate masses. A full scan is usually collected along with the SIRs. Save `product ion acquisition method. If MS/MS fragmentation information may binesctorlulmecetnetds. arSeeeemMpilcoyreodm,asasdditional MassLynx GUIDE TO DATA ACQUISITION for additional information and MRM (Multiple Reaction Monitoring). 12.1.3 Typically the analytical batch run sequence begins with a setofextracted matrix standards and ends with a setofextracted matrix standards. 12.1.4 Samples are analyzed with a continuing calibration check injected after every tenth sample. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered samples but may be included as such. 12.2 Using the Autosampler 12.2.1 Set up sample tray according to the sample list prepared in Section 12.1.1. 12.2.2 Set-up the HP1100/autosampler at the following conditions or at conditions the analyst considers appropriate for optimal response. Record actual conditionsinthe instrument logbook: 12.2.2.1 Sample size = 10 pL injection 12.2.2.2 Inject/sample = 1 1122.22..22.43 CSoylcvleenttirmaem=p =13.5 minutes Time 2.0mM Ammonium acetate (T0min 90% 10% | 12.2.2.5 Pressthe "Start" button. 12.3 Instrument Set-up 12.3.1 Refer to ETS-9-24.0 for more details. 12.3.2 Check the solvent level in reservoirs and refillif necessary. Avsof SearEavesns Using ESS Page sats 001016 12.3.3 tChheetcikp.thTehestatiipnlsehssousltdeelbecafpliatllwairtyhatnothjeagegneddoefdtghese.prIofbteh. eUtsipeiasnfoeuynedpiteocebeto check unsatisfactory, disassemble the probe and replace the stainless steel capillary. 123.4 OSebtseHrPvLeCdroppulmetpstcoo"mOinn"g.oSuettoftthehefltoiwpotfo t1h0e-p5r0ob0eu.L/Amlilnowortoaseqaupiplriobprraitaetfeor approximately 10 minutes. 12.3.5 aTruomunodnthtehetinpitorfotgheen.prAobefi.neRmeiasdtjusshtoutlhed tbiep eoxfptehlelepdrowbiethifnnoonmiitsrtogiesnolbesaekrivnegd 123.6 cThhaenignestinruomrednetrutsoeosptthiemsiezepatrhaemreetseprosnsaet:the following settings. These settings may 12.3.6.1 Drying gas 250-400 liters/hour 12.3.6.2 ESI nebulizing gas 10-15 lters/hour 123.63 HPLC constant flow mode, flow rate 10 -- 500 uL/min 123.6.4 HPrPesLsCuries <op4e0r0abtairng(cTohrirsecptalry.a)meter isnot set, itis a guide to ensure the 123.7 fCuarrtehferu.llCyognuniedcettthheepvroolbteagientcoabtlheesotpoentihnegp.roIbnes.ert probe until it will not go any 123.8 tParipnetdtihnetotutnhee piangset,ruwmietnhtiltosgp.arameters, and store it in the studybinderwith a copy 12.3.9 tUhseinangatlhyesicsroofsbsi-oflloogwiccoaulnmtaetrreilceecs.trode in the ES/MS source is recommended for 12:3.10CMlaiscskLoynnxstavretrbsiuotntso,nsicnetahpepArcoqpuriisaitetiMoansCsoLntyrnoxlUPSanEeRl'(SthiGsUmIaDyE)v.arPyraesmsonthge start button." Ensure start and end sample number includes allsamplesto be analyzed. 13.0 DATA ANALYSIS AND CALCULATIONS 13. Calculations: 13.1.4 Calculate matrix spike percent recoveries using the following equation: %Recovery= Observed Result - Background Result x 100 Expected Result 13.15 Calculate percent difference using the following equation: % Difference = Expected CoEnxcp.ec-tCeadlcCuolnact.ed Cone. x 100 13.1.6 Calculate actual concentrationofPFOS, or other fluorochemical, in matrix (ng/mL): (Init(inaglofVoPlFuOmSecoaflmea.tfrrioxm(smtdL.)C+urmvLe xofDiluSutriroognatFeaScttoarn)dardx) Lug _ 1000 ng Final Volume (mL) Analysisof SeErTuSm8E-x5tr.a1ct Using ESIMS Page 6of9 001017 4.0 METHOD PERFORMANCE. 14.1 `MmeattrhioxdspDeectifeicct.ioPnlLeiasmeitsc(eMEDTLS)-8a-n4d.1L,imAitttoafcQhumaennttitBa,tifoonr(aLlOisQt)ingaorfe mceutrhreondt,vaanlaildyattee,dand MDL and LOQ values, 142 Solvent Blanks, Method Blanks, and Matrix Blanks 14.2.1 SToowlevsetntstblaanndkasr,d minetthheodcablliabnrkasti,oanncdurmvae:trix blanks values are must be below the 143 Calibration Curves 14.3.1 The r* value for the calibration curve must be 0.980 or better. 144 Matrix Spikes 14.4.1 Mcoantcreinxtrsaptiikoen.percent recoveries are must be within + 30%of the spiked 145 Continuing Calibration Verifications 145.1 cCoonncteinnturaitnigocna.libration verification percent recoveries must be + 30%ofthe spiked 14.6 `Ifcpreriftoerrmieadloisntetdhienstyhsitsemmetahnoddsapmeprlfeosrmraenacnealsyezcetdioonriostnh'termeatct,imoansinatsedneatnecremminaeydbbey the analyst. Document all actions in the appropriate logbook. 14.7 Iffodoattnaotaerdeotno tbaeblreespoarntdeddiwshceunsspeedrifnotrhmeantecxetcorfitthereiarehpaovret.not been met, the data must be 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 pSiapmepttleeweaxsttreacitswdaisstpeosaenddinflbarmomkaebnlgelsaoslsvceonnttaisindeirsspolsoecdatiendhiingthheBlTaUborcaotnotrayi.ners, and glass 160 Recoros meet 16.1 Eheaacdherpaogrehgaennderwartiettdefn ooarn tshteudpyagmeu:ststhuadvyeotrhperfoojlecltowniunmgbienrf,oramcaqutiisointiionnclmuedtehdodei,ther in the iannatleygsrta.tion method, sample name, extraction date, dilution factor (ifapplicable), and 1622 Parpipnrtoptrhieattuensetpuadgyef,olsdaemr.plCeolpisyt, tahnedseacpqaugiessitainond mtaeptehoindtofrthoemiMnasstrsuLmyennxt rtuonilnocg.lude in the 163 Plot the store in tchaelisbtruadtyiofnoclduerrv.e by linear regression, weighted 1/x, then print these graphs and 16.4 Print data and store integration in the study summary, folder. integration method, and chromatograms, from MassLynx, Analysis ofSeErTuSm8E:x5t.ra1ct Using ESMS Page 70r9 001018 165 SAtutmamcahrmieznetdaAtafoursianngesxuaitmapbllee osoffatwsaurmem(aErxcyeslpr5e.a0d)sahenedt.store in the study folder, see 16.6 aBnadckloucpateiloencotfrboanicckduaptaetloecatprpornoipcridaattae. medium. Record in study notebook the file name 17.0 TABLES, DIAGRAMS, FLOWCHARTS, AND VALIDATION DATA 17.1 Attachment A: ETS-8-5.1 Data summary spreadsheet. 18.0 REFERENCES 18.1 cFAoCmTp-oMu-n4d.s1f,ro"EmxtSrearcutmiofnoorfAPnaoltyassissiuUmsiPnegrfHlPuLoCro-oEclteacntersoulsfpornaayt/eMoarsOstShpeercFtlruoomreotcrhyemical 182 TEoTnSi-z9a-t2i4o.n0/,Ma"sOspeSrpaetcitornomaentderMaQiunattetnroanIcTetorfitplheequMaidcrruopmoalessSyAsttmeomssp"heric Pressure: 183 The validation report associated with this method is ETS-8-4.0 & 5.0-V-1. 19.0 AFFECTED DOCUMENTS 19.1 ECToSm-8p-o4u.n1,ds"EfxrtoramcStieornuomfPfootraAsnsaliyusmisPeUrsfilnugorHoPoLcCta-nEelseucltfroonastperaoyr/OMtahsesrSFpleucotrroocmheetmriyc"al 200 Revisions Revision Number, 1 Section6.12 ClarificationRoefasHoPn1F1o0r0Rseyvsitseimoncomponents pSleoctttiionng 1l1i.n1earAvreegrraegsesoifontawnodcaudrdveesd,tnhoet1s/txawnedairgdhtvianlguoesf,tahreecuusreved.for Section Section 1172..12.C2.h4anClgaerdififcraotmioanotftascohlvmeennBttratmopA.. Revision Date 04/02/99 Analysis ofSerEumTSE8xt5ra1ct Using ESMS Page 8of9 001019 Laboratory Study # STetsutdyMaterial MMeatthrowdFRienvaisSioolnvent AinnasltyrtuimceanltESqouftiwpamreenVterSsyisotnem Number FRi-lSeqnuaamree:d Value: SloIpnet:ercept DDaattee ooffAExntarlaycstiisoAnn/aAnlaylsytst ae vm Taar] mreer [Tear Siope: Taken from Tne egreeion cqueion `Group/Dose: Taken from the study folder. Sample#: Taken from the study folder. iCotnacelntVroaltuimone ((umgL/)m:L)T: Taakfeinkomfroehem stthneudMyasolsdLeyrnx integration summary. DFiilnuatliCoonnFca,ct(ogr/:mLT)a:kenCaflrcoulmattheedsbtyuddyivfiodldienrg, the nitsvolume romtheconcentration tachmenAt:SummarySpreadshectAnalysisofSerEuTmSE8xt5r1actUsing ES/MS Page9of9 001020