Chromatography: Basic Principles, Sample Preparations and Related Methods

223 Pages·2013·6.633 MB·English

by  Elsa Lundanes, Léon Reubsaet, Tyge Greibrokk

Elsa Lundanes, Léon Reubsaet and Tyge Greibrokk
Chromatography
Basic Principles, Sample Preparations
and Related Methods

ElsaLundanes
L(cid:1)eonReubsaet
TygeGreibrokk
Chromatography
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Elsa Lundanes, Leon Reubsaet and Tyge Greibrokk
Chromatography
Basic Principles, Sample Preparations and Related Methods
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V
Contents
Preface XIII
1 GeneralConcepts 1
1.1 Introduction 1
1.2 MigrationandRetention 2
1.2.1 General 2
1.2.2 MobileandStationaryPhases 3
1.2.3 Chromatograms 3
1.2.4 RetentionFactor 3
1.3 BandBroadening 5
1.3.1 EddyDiffusion 6
1.3.2 LongitudinalDiffusion 6
1.3.3 ResistancetoMassTransfer 7
1.3.4 CombinedBandBroadeninginaColumn 8
1.3.5 BandBroadeningoutsidetheColumn 9
1.4 MeasuringColumnEfficiency 9
1.4.1 PlateNumbers 9
1.4.2 CouplingColumns 10
1.4.3 PlateHeight 10
1.4.3.1 ReducedPlateHeight 11
1.4.4 EffectivePlateNumber 11
1.4.5 Asymmetry 11
1.5 Resolution 11
1.5.1 IncreasingtheResolution 13
1.6 PeakCapacity 13
1.7 Two-DimensionalSystems 13
1.8 IncreasedPerformance 14
References 15
2 GasChromatography 17
2.1 Introduction 17
2.2 MobilePhase/CarrierGas 17
2.3 InjectionSystems 19
VIjContents
2.3.1 PackedColumnInjector(EvaporationInjector) 20
2.3.2 InjectionSystemsforCapillaryColumns 21
2.3.2.1 SplitInjection 21
2.3.2.2 SplitlessInjection 22
2.3.2.3 On-ColumnInjection 22
2.3.2.4 Large-VolumeInjectors 23
2.3.2.5 HeadspaceTechniques 23
2.4 Columns 24
2.4.1 PackedColumns 25
2.4.2 OpenTubularColumns 25
2.5 Detectors 26
2.5.1 Introduction 26
2.5.2 ThermalConductivityDetector 28
2.5.3 FlameIonizationDetector 28
2.5.4 Nitrogen–PhosphorusDetector 30
2.5.5 ElectronCaptureDetector 31
2.5.6 MassSpectrometry 32
2.5.6.1 PositiveIonization 33
2.5.6.2 NegativeIonization 33
2.5.6.3 GasChromatography–MassSpectrometry(GC–MS)Interfacing 33
2.5.7 OtherDetectors 35
2.5.7.1 TheFlamePhotometricDetector 35
2.5.7.2 TheChemiluminescentDetector 35
2.5.7.3 TheElectrolyticConductivityDetector 35
2.5.7.4 ThePhotoionizationDetector 35
2.5.7.5 TheAtomicEmissionDetector 36
2.5.7.6 FourierTransformInfraredDetector 36
2.6 StationaryPhases 36
2.6.1 GSC–AdsorptionChromatography 36
2.6.2 GLC–PartitionChromatography 37
2.6.2.1 Matrix 37
2.6.2.2 ChoosingtheStationaryPhase 37
2.6.2.3 TypesofStationaryPhasesinGLC 38
2.6.2.4 StationaryPhase(Film)Thickness 40
2.6.2.5 Temperature 41
2.7 Two-DimensionalSeparations 42
2.8 QualitativeandQuantitativeAnalyses 43
2.9 Derivatization 44
References 46
3 High-PerformanceLiquidChromatography(HPLC) 47
3.1 Introduction 47
3.2 SolventsandSolventDelivery 47
3.2.1 Maintenance 49
3.2.2 Automation 50
ContentsjVII
3.3 Injection 50
3.3.1 Techniques 50
3.3.1.1 ConstantVolumeInjection 50
3.3.1.2 VariableVolumeInjection 51
3.3.1.3 VolumesandPrecision 51
3.3.2 DilutionandRefocusing 51
3.3.2.1 InjectionVolumeRelatedtoSolventElutionStrength 51
3.3.2.2 TimedInjection 52
3.3.2.3 Carryover 52
3.3.2.4 CombinationwithSolid-PhaseExtractors 52
3.3.3 CalculationofMaximumInjectionVolumes 53
3.3.4 CalculatingtheDilutionoftheAnalyteintheColumn 54
3.4 Columns 54
3.4.1 PackedColumns 54
3.4.1.1 ColumnDimensionsandMaterials 54
3.4.1.2 EffectonDetection 55
3.4.1.3 SolventSaving 55
3.4.1.4 ColumnEfficiency 56
3.4.1.5 ColumnLifetime 57
3.4.1.6 PeakShapes 57
3.4.1.7 FlowandBackpressure 58
3.4.1.8 ConventionalTotallyPorousParticles 58
3.4.1.9 Core–ShellParticles 58
3.4.1.10 Ultrahigh-PressureLC(UHPLCorUPLC) 59
3.4.2 MonolithicColumns 59
3.4.3 MicrochipColumns 60
3.4.4 OpenTubularColumns 61
3.4.5 TemperatureControl 61
3.4.6 PreparativeLCandFlashChromatography 63
3.5 StationaryPhasesandTheirPropertiesinHPLC 64
3.5.1 Normal-PhaseMaterialsforAdsorptionChromatography 64
3.5.1.1 SeparationPrinciples 64
3.5.1.2 Silica 65
3.5.1.3 Alumina,Titania,andZirconia 65
3.5.1.4 SilicawithBondedPolarFunctionalGroups 66
3.5.1.5 HydrophilicInteractionLiquidChromatography(HILIC) 67
3.5.1.6 CarbonMaterials 68
3.5.2 Reversed-phaseMaterials 68
3.5.2.1 SeparationPrinciples 68
3.5.2.2 Retention 69
3.5.2.3 TheSolvationParameterModel 70
3.5.2.4 Silica-basedReversed-phaseMaterials 71
3.5.2.5 HybridMaterialsandHydrosilatedMaterials 72
3.5.2.6 OrganicPolymer-basedMaterials 72
3.5.2.7 IonPairChromatographyonReversed-PhaseColumns 72
VIIIjContents
3.5.2.8 HydrophobicInteractionChromatography 73
3.5.3 IonExchangeMaterials 73
3.5.3.1 Elution 74
3.5.3.2 Retention 74
3.5.4 Chromatofocusing 74
3.5.4.1 IonChromatographyforInorganicIons 75
3.5.5 SizeExclusionMaterials 76
3.5.5.1 SeparationPrinciples 76
3.5.5.2 Materials 76
3.5.5.3 MobilePhases 77
3.5.6 MaterialsforChiralSeparations 77
3.5.6.1 SeparationPrinciple 77
3.5.6.2 Materials 78
3.5.7 AffinityMaterials 78
3.5.7.1 SeparationPrinciple 78
3.5.7.2 AffinityMaterialsforChromatographyandMicroarrays 79
3.6 Detectors 80
3.6.1 UVDetection 81
3.6.1.1 SomeCommonChromophores 82
3.6.1.2 ChoosingtheRightWavelength 82
3.6.1.3 FlowCells 82
3.6.1.4 FilterPhotometricDetection 83
3.6.1.5 SpectrophotometricDetection 83
3.6.1.6 DiodeArrayDetectors 83
3.6.2 MassSpectrometricDetection 85
3.6.2.1 ElectrosprayIonization 86
3.6.2.2 AtmosphericPressureChemicalIonization 88
3.6.2.3 AtmosphericPressurePhotoionization 89
3.6.2.4 InductivelyCoupledPlasmaIonization 90
3.6.2.5 MassAnalysis 91
3.6.2.6 TheQuadrupoleMassAnalyzers 91
3.6.2.7 TheIonTrapAnalyzers 92
3.6.2.8 TheTime-of-FlightAnalyzers 92
3.6.2.9 TheFTMSAnalyzers 93
3.6.2.10 FragmentationinMassSpectrometry 94
3.6.3 FluorescenceDetection 95
3.6.3.1 FilterFluorimeters 97
3.6.3.2 Spectrofluorimeters 97
3.6.3.3 ChemiluminescenceDetection 97
3.6.4 ElectrochemicalDetection 98
3.6.4.1 AmperometricDetection 98
3.6.4.2 CoulometricDetector 99
3.6.5 LightScatteringDetection 100
3.6.6 RefractiveIndexDetection 100
3.6.7 OtherDetectors 102