高效液相色譜概述高效液相色譜概述PharmaceuticalAnalysis講解

PharmaceuticalAnalysis高效液相色譜概述高效液相色譜概述ZhongWenwuOverviewofHighPerformanceLiquidChromatographyOverviewofHighPerformanceLiquidChromatographyDefinitionandcharacteristicsofhighperformanceliquidchromatographyThemainstructure,separationprincipleandrelatedtermsofhighperformanceliquidchromatographQualitativeandQuantitativeMethodsofHighPerformanceLiquidChromatographySummary目錄MasterSeparationprincipleandqualitativeandquantitativemethodsofhighperformanceliquidchromatographyFamiliarStructureandconditionsettingofhighperformanceliquidchromatographUnderstandHowtousehighperformanceliquidchromatographykeyanddifficultpointsDefinitionandcharacteristicsofhighperformanceliquidchromatographyDefinitionandcharacteristicsofhighperformanceliquidchromatographyHighperformanceliquidchromatography，HPLCDefinition:Themobilephaseistransportedathighpressure,usinghigh-efficiencystationaryphaseandhigh-sensitivitydetectorchromatographicanalysistechnologyforonlinedetectionDifference:ThedifferencebetweenHPLCandclassicliquidchromatographyisthatthefillerparticlesaresmallanduniform.Smallparticleshavehighcolumnefficiency,butwillcausehighresistanceandrequirehighpressuretotransportthemobilephase.DefinitionandcharacteristicsofhighperformanceliquidchromatographyAdvantagesofhighperformanceliquidchromatographyHighseparationefficiencyFastanalysisLowdetectionlimitWideselectionofmobilephaseColumncanbeusedrepeatedlyEasycollectionofeffluentcomponentsOperationautomationWiderangeofapplicationsThemainstructure,separationprincipleandrelatedtermsofhighperformanceliquidchromatographyMaincomponentsandseparationprincipleThemainstructure,separationprincipleandrelatedtermsofhighperformanceliquidchromatograph1.Mobilephasesystem2.Samplingsystem3.Columnseparationsystem4.Inspectionrecordsystem1.Liquid-solidadsorptionchromatography2.Liquid-liquiddistributionchromatography3.Ionexchangechromatography4.MolecularexclusionchromatographyAccordingtotheprincipleofcolumnseparationMainstructureofinstrument圖1.Mobilephasesystem2.進(jìn)樣系統(tǒng)injectionsystem3.Chromatographiccolumnseparationsystem4.DetectionandrecordingsystemThemainstructureofhighperformanceliquidchromatography1.高壓輸液系統(tǒng)：儲(chǔ)液器、泵

1.Highpressureinfusionsystem:liquidreservoirandpump2.進(jìn)樣系統(tǒng)：六通進(jìn)樣閥13/50002.Sampleinjectionsystem:six-waysampleinjectionvalve3.色譜分離系統(tǒng)：色譜柱3.Chromatographyseparationsystem:chromatographiccolumn4.檢測(cè)系統(tǒng)：紫外檢測(cè)器、熒光、電化學(xué)、蒸發(fā)

光散射、視差折光4.Detectionsystem:UVdetector,fluorescence,electrochemistry,evaporationLightscattering,parallaxrefraction5.數(shù)據(jù)記錄和處理系統(tǒng)5.Datarecordingandprocessingsystem高效液相色譜儀HighperformanceliquidchromatographyCommonHPLCinstrumentsShimadzuDionexWatersAgilentHPLCterminologyRetentiontimetRtR’ThepeakheighthHalfpeakwidthY1/2hPeakareaAn:numberoftheoreticalplatesR:separatingdegreeT:TrailingfactorQualitativeandquantitativemethodsforhighperformanceliquidchromatographyQualitativeanalysismethodQualitativemethod

1.Chromatographicidentification：Qualitativemethodofreservedvalue：tR,VR,ri,s2.Chemicalidentification3.Combineduseoftwospectra：GC-MS,GC-FIDQualitativeanalysismethod利血平標(biāo)準(zhǔn)品Standardreserpine1234ThepeakareaA1ofsubstance1withknownquantityM1wasdeterminedbychromatographyTheunknownsubstance1ofM1xwasdeterminedbychromatography,andthepeakareaofsubstance1wasA1xQuantitativeanalysismethod:externalstandardmethodQuantitativeanalysismethod3:ExternalstandardmethodExternalstandardmethodNote:bothmeasurementsmustbeofonesubstanceExternalstandardmethodSameinjectionvolume:Example:QCLiusedexternalstandardmethod,HPLCdetectionofabatchofreserpininedrugs.Theconcentrationofreserpinestandardsubstancesolutionwas0.05mmol/mL,andachromatographicpeakwithapeakareaof5000wasobtainedin15minutes.ThenthereserpinesolutionwasinjectedandthechromatographicpeakofAwasobtainedwiththepeakareaof6800atthesameretentiontime.Howmuchistheconcentrationofreserpineinthismedicine?summary1.QualitativeandquantitativemethodsforHIGHperformanceliqui