毛细管电泳指导手册 Capillary+Electrophoresis+Guidebook

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1、CHAPTER1FundamentalsofCapillaryElectrophoresisTheoryKevinD.Altria1.IntroductionThissectionwilldescribethefundamentaltheory,equations,anddefi-nitionsnecessarytocomprehendtheconceptsinvolvedincapillaryelec-trophoresis(CE).Thisisnotanexhaustivetreatment,butisconsideredsufficienttoc

2、omprehendandappreciatetheprinciplesofCE.Moredetailedtheoreticalbackgroundcanbeobtainedfromanumberofrefer-encebooks(I-6).DevelopmentsinthefieldofCEarereviewedindetailannuallyinthejournalAnalyticalChemistry.Forexample,the801paperspublishedin1992-l993wererecentlyreviewed(7).CEcanbe

3、broadlydescribedashigh-efficiencyseparationsofsampleionsinanarrowbore(25-100pm)capillarytubethatisfilledwithanelectrolytesolution.AtypicalschematicofaninstrumentsetupisshowninFig.1.Theprincipalcomponentsareahigh-voltagepowersupply,acapil-larythatpassesthroughtheopticalcenterofad

4、etectionsystemcon-nectedtoadataacquisitiondevice,asampleintroductionsystem,andanautosampler.Typically,theCEinstrumentiscontrolledbyapersonalcomputer.Thecapillaryisfirstfilledwiththerequiredbuffersolution.Samplesolution(typicallyl-20nL)isthenintroducedattheendofthecapillaryawayfr

5、omthedetector(usuallytheanode).Thecapillaryendsarethendippedintoreservoirscontaininghigh-voltageelectrodesandtherequiredbuffersolution.OneelectrodeisconnectedtoacableleadingtoFromMethodsmMolecularBology,Vol52CapdaryElectrophoresrsEd&dbyKAltrlaCopyrIghtHumanaPressInc,Totowa,NJ34A

6、ltrianHighvoltagesupplyEmptyvial0Fig.1.Typicalinstrumentalsetup.thehigh-voltageoutput,whereastheother(situatedatthedetectorendofthecapillary)isconnectedtoanearthingcable.Electrodesarecomposedofaninertmaterial,suchasplatinum.Applicationofavoltage(forexample,10-30kV)acrossthecapil

7、larycauseselectrophoreticandelectroendosmoticmovements(discussedlaterinthischapter)resultingintheionicspeciesinthesamplemovingalongthecapillaryandpassingthroughtheon-linedetector.Aplotofdetectorresponse(usuallyUVabsorbance)withtimeisgenerated,whichistermedanelectropherogram.1.1.

8、ElectrophoresisThisprocessisthemovementofsample