Chapter 6 HPLC

TransmittalNo.96-E1(9/96)FormFDA2905a(6/92)600–1SECTION600PesticideAnalyticalManualVol.ITableofContentspagedate601:GeneralInformation601A:Principles601-11/94601B:ModesofOperation601-21/94Liquid-SolidChromatography601-21/94Liquid-LiquidChromatography601-31/94BondedPhaseChromatography601-31/94IonExchangeChromatography601-41/94SizeExclusionChromatography601-41/94601C:InstrumentationandApparatus601-51/94BasicComponents601-51/94HPLCSystemPlumbing601-71/94601D:SolventsandReagents601-101/94PotentialProblems601-101/94SpecificSolvents601-121/94Chapter1RegulatoryOperationsChapter2GeneralAnalyticalOperationsandInformationChapter3MulticlassMRMsChapter5GLCChapter4SelectiveMRMsChapter6HPLCTransmittalNo.96-E1(9/96)FormFDA2905a(6/92)600–2PesticideAnalyticalManualVol.ISECTION600pagedateReagentBlanks601-141/94SafetyPrecautions601-141/94601E:SamplePreparation601-141/94SampleCleanup601-141/94SampleFiltration601-141/94SampleSolventDegassing601-151/94ChoiceofSampleSolvent601-151/94601F:ReferenceStandards601-151/94StockSolutions601-151/94WorkingStandardSolutions601-161/94Storage601-161/94References601-161/94602:Columns602A:ColumnSelection602-11/94602B:AnalyticalColumns602-11/94Liquid-SolidChromatography602-21/94BondedPhases602-21/94IonExchange602-31/94IonPair602-41/94SizeExclusion602-41/94602C:ColumnEvaluation602-51/94602D:ColumnSpecifications602-61/94602E:AnalyticalColumnProtection602-81/94Filters602-81/94Precolumns602-91/94GuardColumns602-91/94602F:ColumnMaintenanceandTroubleshooting602-91/94ColumnCare602-91/94ColumnEvaluationbyInjectionof602-101/94TestMixturesColumnStorage602-111/94ColumnRegeneration602-111/94References602-121/94603:MobilePhaseSelection,Preparation,andDelivery603A:MobilePhaseSelection603-11/94NormalPhaseChromatography603-11/94ReversePhaseChromatography603-31/94IonExchangeChromatography603-41/94IonPairChromatography603-51/94SizeExclusionChromatography603-51/94GradientElutioninHPLC603-51/94603B:MobilePhasePreparation603-61/94FilteringSolvents603-61/94DegassingSolvents603-61/94TransmittalNo.96-E1(9/96)FormFDA2905a(6/92)600–3SECTION600PesticideAnalyticalManualVol.IpagedatePreparationofMultisolventMobilePhases603-71/94SolventReservoirs603-71/94603C:MobilePhaseDeliverySystems603-81/94Pumps603-81/94GradientProgrammingSystems603-101/94603D:MaintenanceandTroubleshooting603-101/94ProblemswithPumps603-101/94References603-131/94604:InjectionSystems604A:InjectionValves604-11/94604B:AutomaticInjectors604-31/94604C:Operation,Maintenance,Troubleshooting,604-31/94andRepairofInjectionValvesReferences604-41/94605:Detectors605A:UV/VISAbsorbanceDetectors605-21/94FixedWavelengthUVDetectors605-31/94VariableWavelengthUVDetectors605-41/94Solvents605-41/94PerformanceCharacteristics605-41/94MultichannelorPhotodiodeArrayDetectors605-51/94Applications605-51/94Problems,Maintenance,and605-51/94Troubleshooting605B:FluorescenceDetectors605-61/94DetectorDesign605-61/94Solvents605-71/94PerformanceCharacteristics605-71/94ParameterAdjustments605-81/94Applications605-81/94DetectorMaintenance605-81/94605C:ElectrochemicalDetectors605-81/94ConductivityDetectors605-91/94AmperometricandCoulometricDetectors605-91/94PerformanceCharacteristics605-101/94Applications605-111/94605D:PhotoconductivityDetectors605-111/94Apparatus605-111/94PerformanceCharacteristics605-111/94Applications605-131/94605E:MassSpectrometricDetectors605-131/94605F:DerivatizationforDetectionEnhancement605-141/94ComparisonofPre-andPost-Column605-141/94DerivatizationTransmittalNo.96-E1(9/96)FormFDA2905a(6/92)600–4PesticideAnalyticalManualVol.ISECTION600pagedatePost-ColumnReactorDesign605-141/94References605-161/94606:ResidueIdentificationandQuantitation606A:Residueidentification606-11/94Co-chromatography606-11/94UseofAlternativeColumns606-21/94SpectrometricConfirmation606-21/94606B:Quantitation606-31/94Reference606-31/94607:QualityAssuranceandTroubleshooting607A:LiquidChromatographMonitoringand607-11/94PerformanceTesting607B:TroubleshootingfromChromatograms607-21/94608:BibliographyGeneralTexts608-11/94Columns608-11/94Detectors608-21/94Troubleshooting608-21/94ApplicationtoPesticides608-21/94Figures601-aChromatographicSeparationTechniques601-11/94601-bHPLCModesofOperation601-21/94601-cGuidetoSelectionofHPLCMode601-51/94601-dBlockDiagramofHPLCSystem601-61/94601-eColumnOutletFittings601-71/94601-fLowDeadVolumeFitting601-71/94601-gStandardInternalFitting601-81/94601-hInternalThreadLowDeadVolumeFitting601-81/94602-aCalculationofColumnPerformanceParameters602-51/94603-aReciprocatingPump603-81/94603-bGradientSystemI603-91/94603-cGradientSystemII603-91/94603-dGradientSystemIII603-101/94604-aExternalLoopInjector:Six-PortInjectionValve604-11/94604-bInternalLoopInjector604-21/94605-aUV/VISDetector605-31/94605-bVariableWavelengthUV/VISDetector605-41/94605-cFluorescenceHPLCDetector605-69/96605-dThreeElectrodeElectrochemicalDetector605-101/94605-ePost-ColumnReactors605-151/94TransmittalNo.96-E1(9/96)FormFDA2905a(6/92)600–5SECTION600PesticideAnalyticalManualVol.IpagedateTables602-a:HPLCColumnSpecificati