EPAMethod8331TetrazenebyReversePhaseHighPerformanceLiquid.pdf

《EPAMethod8331TetrazenebyReversePhaseHighPerformanceLiquid.pdf》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。

METHOD8331TETRAZENEBYREVERSEPHASEHIGHPERFORMANCELIQUIDCHROMATOGRAPHY(HPLC)1.0SCOPEANDAPPLICATION1.1Thismethodisintendedfortheanalysisoftetrazene,anexplosiveresidue,insoilandwater.Thismethodislimitedtousebyanalystsexperiencedinhandlingandanalyzingexplosivematerials.Thefollowingcompoundscanbedeterminedbythismethod:___________________________________________________________________________aCompoundCASNo___________________________________________________________________________Tetrazene31330-63-9___________________________________________________________________________aChemicalAbstractsServiceRegistrynumber1.2Tetrazenedegradesrapidlyinwaterandmethanolatroomtemperature.Specialcaremustbetakentorefrigerateorcoolallsolutionsthroughouttheanalyticalprocess.1.3Tetrazene,initsdryform,isextremelyexplosive.Cautionmustbetakenduringpreparationofstandards.1.4Theestimatedquantitationlimit(EQL)ofMethod8331fordeterminingtheconcentrationoftetrazeneisapproximately7µg/Linwaterandapproximately1mg/kginsoil.1.5ThismethodisrestrictedtousebyorunderthesupervisionofanalystsexperiencedintheuseofHPLC,skilledintheinterpretationofchromatograms,andexperiencedinhandlingexplosivematerials.Eachanalystmustdemonstratetheabilitytogenerateacceptableresultswiththismethod.2.0SUMMARYOFMETHOD2.1A10mLwatersampleisfiltered,elutedonaC-18columnusingionpairingreversephaseHPLC,andquantitatedat280nm.2.22gofsoilareextractedwith55:45v/vmethanol-waterand1-decanesulfonicacidonaplatformshaker,filtered,andelutedonaC-18columnusingionpairingreversephaseHPLC,andquantitatedat280nm.CD-ROM8331-1Revision0September1994 3.0INTERFERENCES3.1Nointerferencesareknown.Tetrazeneelutesearly,however,andifacomputingintegratorisusedforpeakquantification,thebaselinesettingmayhavetobesettoexcludebaselineaberrations.Baselinesettingisparticularlyimportantatlowconcentrationsofanalyte.4.0APPARATUSANDMATERIALS4.1HPLCsystem4.1.1HPLC-Pumpcapableofachieving4000psi.4.1.2100µLloopinjector.4.1.3Variableorfixedwavelengthdetectorcapableofreading280nm.4.1.4C-18reversephaseHPLCcolumn,25cmx4.6mm(5µm)(SupelcoLC-18,orequivalent).4.1.5Digitalintegrator-HP3390A(orequivalent)4.1.6Stripchartrecorder.4.2Otherapparatus4.2.1Platformorbitalshaker.4.2.2Analyticalbalance-+0.0001g.4.2.3Desiccator.4.3Materials4.3.1Injectionsyringe-500µL.4.3.2Filters-0.5µmMillex-SRand0.5µmMillex-HV,disposable,orequivalent.4.3.3Pipets-volumetric,glass,ClassA.4.3.4Scintillationvials-20mL,glass.4.3.5Syringes-10mL.4.3.6Volumetricflasks,ClassA-100mL,200mL.4.3.7Erlenmeyerflaskswithgroundglassstoppers-125mL.CD-ROM8331-2Revision0September1994 4.4Preparation4.4.1PrepareallmaterialsasdescribedinChapter4forvolatileorganics.5.0REAGENTS5.1HPLCgradechemicalsshallbeusedinalltests.ItisintendedthatallreagentsshallconformtothespecificationsoftheCommitteeonAnalyticalReagentsoftheAmericanChemicalSociety,wheresuchspecificationsareavailable.Othergradesmaybeused,provideditisfirstascertainedthatthereagentisofsufficientlyhighpuritytopermititsusewithoutloweringtheaccuracyofthedetermination.5.2General5.2.1Methanol,CH3OH-HPLCgrade.5.2.2Organic-freereagentwater-Allreferencestowaterinthismethodrefertoorganic-freereagentwater,asdefinedinChapterOne.5.2.31-Decanesulfonicacid,sodiumsalt,C10H21SO3Na-HPLCgrade.5.2.4Aceticacid(glacial),CH3COOH-reagentgrade.5.3StandardSolutions5.3.1Tetrazene-StandardAnalyticalReferenceMaterial.5.3.2Stockstandardsolution-Drytetrazenetoconstantweightinavacuumdesiccatorinthedark.(Tetrazeneisextremelyexplosiveinthedrystate.Donotdrymorereagentthanisnecessarytopreparestocksolutions.)Placeabout0.0010g(weighedto0.0001g)intoa100-mlvolumetricflaskanddilutetovolumewithmethanol.Invertflaskseveraltimesuntiltetrazeneisdissolved.Storeinfreezerat-10EC.Stocksolutionisabout100mg/L.Replacestockstandardsolutioneveryweek.5.3.3Intermediatestandardsolutions5.3.3.1Preparea4mg/Lstandardbydilutingthestocksolution1/25v/vwithmethanol.5.3.3.2Pipet0.5,1.0,2.0,5.0,10.0,and20.0mLofthe4mg/Lstandardsolutioninto6separate100mLvolumetricflasks,andmakeuptovolumewithmethanol.Pipet25.0mLofthe4mg/Lstandardsolutionintoa50mLvolumetricflask,andmakeuptovolumewithmethanol.Thisresultsinintermediatestandardsofabout0.02,0.04,0.08,0.2,0.4,0.8,2and4mg/L.5.3.3.3Coolimmediatelyonpreparationinrefrigeratororicebath.CD-ROM8331-3Revision0September1994 5.3.4Workingstandardsolutions5.3.4.1Inject4mLofeachoftheintermediatestandardsolutionsinto6.0mLofwater.Thisresultsinconcentrationsofabout0.008,0.016,0.032,0.08,0.16,0.3,0.8and1.6mg/L.5.3.4.2Coolimmediatelyonpreparationinrefrigeratororicebath.5.5QCspikeconcentratesolution5.5.1Drytetrazenetoconstantweightinavacuumdesiccatorinthedark.(Tetrazeneisextremelyexplosiveinthedrystate.Donotdryanymorethannecessarytopreparestandards.)Placeabout0.0011g(weighedto0.0001g)intoa200-mlvolumetricflaskanddilutetovolumewithmethanol.Invertflaskseveraltimesuntiltetrazeneisdissolved.Storeinfreezerat-10EC.QCspikeconcentratesolutionisabout55mg/L.Replacestockstandardsolutioneveryweek.5.5.2Preparespikingsolutions,atconcentrationsappropriatetotheconcentrationrangeofthesamplesbeinganalyzed,bydilutingtheQCspikeconcentratesolutionwithmethanol.Coolonpreparationinrefrigeratororicebath.5.6Eluent5.6.1Tomakeabout1literofeluent,add2.44gof1-decanesulfonicacid,sodiumsaltto400/600v/vmethanol/water,andadd2.0mLofglacialaceticacid.6.0SAMPLECOLLECTION,PRESERVATION,ANDHANDLING6.1SeetheintroductorymaterialtothisChapter,OrganicAnalytes,Section4.1.6.2Samplesmustbecollectedandstoredinglasscontainers.Followconventionalsamplingprocedures.o6.3Samplesmustbekeptbelow4Cfromthetimeofcollectionthroughanalysis.7.0PROCEDURE7.1SamplePreparation7.1.1FiltrationofWaterSamples7.1.1.1Placea10mLportionofeachwatersampleinasyringeandfilterthrougha0.5µmMillex-HVfilterunit.Discardfirst5mLoffiltrate,andretain5mLforanalysis.CD-ROM8331-4Revision0September1994 7.1.2ExtractionandFiltrationofSoilSamples7.1.2.1Determinationofsample%dryweight-Incertaincases,sampleresultsaredesiredbasedondry-weightbasis.Whensuchdataisdesired,aportionofsampleforthisdeterminationshouldbeweighedoutatthesametimeastheportionusedforanalyticaldetermination.WARNING:Thedryingovenshouldbecontainedinahoodorvented.Significantlaboratorycontaminationmayresultfromaheavilycontaminatedhazardouswastesample.7.1.2.1.1Immediatelyafterweighingthesampleforextraction,weigh5-10gofthesampleintoataredcrucible.Determinethe%dryweightofthesamplebyodryingovernightat105C.Allowtocoolinadesiccatorbeforeweighing:%dryweight=gofdrysamplex100gofsample7.1.2.2Weigh2gsoilsubsamplesinto125mLErlenmeyerflaskswithgroundglassstoppers.7.1.2.3Add50mLof55/45v/vmethanol-waterwith1-decanesulfonicacid,sodiumsaltaddedtomakea0.1Msolution.7.1.2.4Vortexfor15seconds.7.1.2.5Shakefor5hrat2000rpmonplatformshaker.7.1.2.6Placea10mLportionofeachsoilsampleextractinasyringeandfilterthrougha0.5µmMillex-SRfilterunit.Discardfirst5mLoffiltrate,andretain5mLforanalysis.7.2SampleAnalysis7.2.1AnalyzethesamplesusingthechromatographicconditionsgiveninSection7.2.1.1.Undertheseconditions,theretentiontimeoftetrazeneis2.8min.Asamplechromatogram,includingothercompoundslikelytobepresentinsamplescontainingtetrazene,isshowninFigure1.7.2.1.1ChromatographicConditionsSolvent:0.01M1-decanesulfonicacid,inacidicmethanol/water(Section5.5)Flowrate:1.5mL/minInjectionvolume:100µLUVDetector:280nmCD-ROM8331-5Revision0September1994 7.3CalibrationofHPLC7.3.1InitialCalibration-Analyzetheworkingstandards(Section5.3.4),startingwiththe0.008mg/Lstandardsandendingwiththe0.30mg/Lstandard.Ifthepercentrelativestandarddeviation(%RSD)ofthemeanresponsefactor(RF)foreachanalytedoesnotexceed20%,thesystemiscalibratedandtheanalysisofsamplesmayproceed.Ifthe%RSDforanyanalyteexceeds20%,recheckthesystemand/orrecalibratewithfreshlypreparedcalibrationsolutions.7.3.2ContinuingCalibration-Onadailybasis,inject250µLofstockstandardinto20mLwater.Keepsolutioninrefrigeratoruntilanalysis.Analyzeintriplicate(byoverfillingloop)atthebeginningoftheday,singlyaftereachfivesamples,andsinglyafterthelastsampleoftheday.Compareresponsefactorsfromthemeanpeakareaorpeakheightobtainedoverthedaywiththeresponsefactoratinitialcalibration.Ifthesevaluesdonotagreewithin10%,recalibrate.8.0QUALITYCONTROL8.1RefertoChapterOneforspecificqualitycontrolprocedures.8.2Priortopreparationofstocksolutions,methanolshouldbeanalyzedtodeterminepossibleinterferenceswiththetetrazenepeak.Ifthemethanolshowscontamination,adifferentbatchofmethanolshouldbeused.8.3MethodBlanks8.3.1Methodblanksfortheanalysisofwatersamplesshouldbeorganic-freereagentwatercarriedthroughallsamplestorageandhandlingprocedures.8.3.2Methodblanksfortheanalysisofsoilsamplesshouldbeuncontaminatedsoilcarriedthroughallsamplestorage,extraction,andhandlingprocedures.9.0METHODPERFORMANCE9.1Method8331wastestedinalaboratoryoveraperiodoffourdays.Spikedorganic-freereagentwaterandstandardsoilwereanalyzedinduplicateeachdayforfourdays.TheHPLCwascalibrateddailyaccordingtotheproceduresgiveninSection7.1.MethodperformancedataarepresentedinTables1and2.10.0REFERENCES1.Walsh,M.E.,andT.F.Jenkins,"AnalyticalMethodforDeterminingTetrazeneinWater,"U.S.ArmyCorpsofEngineers,ColdRegionsResearch&EngineeringLaboratory,SpecialReport87-25,1987.CD-ROM8331-6Revision0September1994 2.Walsh,M.E.,andT.F.Jenkins,"AnalyticalMethodforDeterminingTetrazeneinSoil,"U.S.ArmyCorpsofEngineers,ColdRegionsResearch&EngineeringLaboratory,SpecialReport88-15,1988.11.0SAFETY11.1StandardprecautionarymeasuresusedforhandlingotherorganiccompoundsshouldbesufficientforsafehandlingoftheanalytestargetedbyMethod8331.CD-ROM8331-7Revision0September1994 FIGURE1CD-ROM8331-8Revision0September1994 TABLE1.METHODPERFORMANCE,WATERMATRIX______________________________________________________________________________SpikeAvg%RecoveryConc.Average(µg/L)ReplicateDay1Day2Day3Day4%Recovery______________________________________________________________________________0.00Replicate10.00.00.00.0%RecoveryNANANANANAReplicate20.00.00.00.0%RecoveryNANANANANA7.25Replicate18.97.87.49.4%Recovery122108102130116Replicate26.69.98.56.7%Recovery911371179210914.5Replicate114.614.613.814.6%Recovery1011019510199Replicate214.814.114.115.2%Recovery102979810510029Replicate131.830.030.828.7%Recovery11010310699105Replicate229.529.730.430.7%Recovery10210210510610472.5Replicate171.173.675.773.9%Recovery98102104102101Replicate271.271.370.771.6%Recovery9898989998145Replicate1140.6143.8144.7142.1%Recovery97991009898Replicate2138.5140.8140.9136.9%Recovery9697979496290Replicate1289.4288.5291.0289.8%Recovery10099100100100Replicate2282.0284.2281.9282.5%Recovery9798979797725Replicate1737.6707.2714.3722.0%Recovery102989910099Replicate2700.2695.8714.2716.3%Recovery9796999997OVERALL102______________________________________________________________________________CD-ROM8331-9Revision0September1994 TABLE2METHODPERFORMANCE,SOILMATRIX________________________________________________________________________________SpikeAvg%RecoveryConc.Average(µg/L)ReplicateDay1Day2Day3Day4%Recovery________________________________________________________________________________0.00Replicate10.00.00.00.0%RecoveryNANANANANAReplicate20.00.00.00.0%RecoveryNANANANANA1.28Replicate10.60.90.61.0%Recovery4973487461Replicate21.20.70.80.7%Recovery92566356672.56Replicate11.41.51.61.6%Recovery5658616159Replicate21.52.01.41.3%Recovery59795650615.12Replicate12.93.02.92.9%Recovery5758565657Replicate23.03.03.53.1%Recovery585969606112.8Replicate17.87.67.88.1%Recovery6159616361Replicate28.08.47.78.2%Recovery626660646325.6Replicate117.216.717.417.3%Recovery6765686867Replicate216.716.817.617.2%Recovery6566696767OVERALL62________________________________________________________________________________CD-ROM8331-10Revision0September1994 METHOD8331TETRAZENEBYREVERSEPHASEHIGHPERFORMANCELIQUIDCHROMATOGRAPHY(HPLC)CD-ROM8331-11Revision0September1994