Chromatography Principles and Instrumentation

281 Pages·2017·5.9 MB·english

by  Mark F. Vitha

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CHROMATOGRAPHY
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CHROMATOGRAPHY
Principles and Instrumentation
Mark F. Vitha
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LibraryofCongressCataloging-in-PublicationData:
Names:Vitha,MarkF.
Title:Chromatography:principlesandinstrumentation/MarkF.Vitha.
Description:Hoboken,NewJersey:JohnWiley&Sons,Inc.,[2017]|Includes
bibliographicalreferencesandindex.
Identifiers:LCCN2016011576|ISBN9781119270881(cloth:alk.paper)|ISBN
9781119270898(pdf:alk.paper)|ISBN9781119270904(epub:alk.paper)
Subjects:LCSH:Chromatographicanalysis.|Gaschromatography.|Liquid
chromatography.
Classification:LCCQD79.C4V582017|DDC543/.8–dc23LCrecordavailableat
http://lccn.loc.gov/2016011576
Aboutthecover:Atwo-dimensionalliquidchromatography(LC×LC)separationofmaizeseedextract.
Groundseedwasextractedusingmethanolmixedwithaphosphatebuffer(pH5.7)followedbysolidphase
extractionpriortoanalysis.ThefirstdimensionseparationusedaZorbaxSB-C3columnwhileacarbon-clad
core-shellsilicacolumnwasusedfortheseconddimension.Additionalexperimentaldetailsaboutthe
extractionandchromatographicsystemcanbefoundinHuang,Y.;Gu,H.;Filgueira,M.;Carr,P.W.
J.Chromatogr.A,1218,2011,2984–2995andFilgueira,M.;Huang,Y.;WittK.;CastellsC.;Carr,P.W.AnalChem.
83,2011,9531–9539.Theauthorthanksthecreatorsofthisimagefortheirpermissiontousethisimageand
MarceloFilgueirafortheworkheputintoitforthisbook.
Typesetin10/13ptPalatinoLTStd-RomanbySPiGlobal,Chennai,India
PrintedintheUnitedStatesofAmerica
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CONTENTS
PREFACE ix
1. FundamentalsofChromatography 1
1.1 Theory 1
1.1.1 ComponentSeparation 3
1.1.2 RetentionFactor 6
1.1.3 Separation 11
1.1.4 ResolutionandTheoreticalPlates 13
1.2 BandBroadening 20
1.2.1 Diffusion 21
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1.2.3 BroadeninginOpenTubeswithNoStationaryPhase
andNoRetention 24
1.2.4 BroadeninginOpenTubeswithaStationaryPhase 28
1.2.5 BroadeninginaPackedColumn 34
1.2.6 PuttingItAllTogether 43
1.2.7 PracticalConsequencesofBroadeningTheory 45
1.3 GeneralResolutionEquation 47
1.4 PeakSymmetry 51
1.5 KeyOperatingVariables 51
1.6 Instrumentation 53
1.7 PracticeofTheTechnique 53
1.7.1 Quantitation 53
1.7.2 InternalStandardsandtheMethodofStandardAdditions 55
1.8 EmergingTrendsandApplications 55
1.9 Summary 55
Problems 56
References 59
FurtherReading 59
2. GasChromatography 61
2.1 TheoryofGasChromatographicSeparations 61
2.1.1 GCColumnsandPartitioning 64
2.2 KeyOperatingVariablesthatControlRetention 64
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vi CONTENTS
2.2.1 AdjustingRetentionTime:Temperature 65
2.2.2 AdjustingRetentionTime:TemperatureProgramming 67
2.2.3 AdjustingRetentionTime:MobilePhaseFlowRate 69
2.2.4 AdjustingRetentionTime:TheColumnandtheStationary
Phase 72
2.2.5 AdjustingRetentionTime:Summary 78
2.2.6 MeasuresofRetention 78
2.3 GasChromatographyInstrumentation 82
2.3.1 CarrierGasSupply 83
2.3.2 TheInjectionPortandtheSoluteInjectionProcess 83
2.3.3 Oven/ColumnCompartment 97
2.3.4 Detectors 98
2.4 AMoreDetailedLookatStationaryPhaseChemistry:KovatsIndices
andMcreynoldsConstants 111
2.4.1 KovatsRetentionIndices 111
2.4.2 StationaryPhaseSelection 120
2.5 GasChromatographyinPractice 124
2.5.1 SyringeWashing 124
2.5.2 ControlsandBlanks/GhostPeaks 124
2.5.3 Autosamplers 125
2.5.4 GCSepta 125
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2.5.5 QualitativeAnalysis 126
2.5.6 QuantitativeAnalysis 126
2.5.7 Derivatization 128
2.5.8 High-SpeedGC 128
2.5.9 TandemGC 129
2.5.10 MicrofabricatedGC 129
2.6 A“Real-World”ApplicationofGasChromatography 131
2.6.1 GCandInternationalOilTrading 131
2.7 Summary 136
Problems 137
References 143
FurtherReading 144
3. LiquidChromatography 145
3.1 ExamplesofLiquidChromatographyAnalyses 145
3.2 ScopeofLiquidChromatography 147
3.3 HistoryofLC 148
3.3.1 ModernPackingMaterials 149
3.4 ModesofLiquidChromatography 152
3.4.1 NormalPhaseLiquidChromatography(NPLC) 152
3.4.2 Reversed-PhaseLiquidChromatography(RPLC) 154
3.4.3 Ion-ExchangeChromatography(IEX) 165
3.4.4 HydrophilicInteractionChromatography(HILIC) 173
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CONTENTS vii
3.4.5 SizeExclusionChromatography(SEC) 175
3.4.6 AffinityChromatography 178
3.5 HPLCInstrumentation 180
3.5.1 TheProportioningValve 181
3.5.2 MixingChamber 181
3.5.3 Pumps 181
3.5.4 Injection 183
3.5.5 TheColumnandParticles 185
3.5.6 GuardColumns 187
3.5.7 Detectors 188
3.6 SpecificUsesofandAdvancesinLiquidChromatography 201
3.6.1 ChiralSeparations 202
3.6.2 Preparative-ScaleChromatography 207
3.6.3 Ultra-HighPerformanceLiquidChromatography(UHPLC)
forHigh-SpeedSeparations 212
3.6.4 Tandem-ColumnLiquidChromatography 216
3.6.5 Two-DimensionalLiquidChromatography(2D-LC) 218
3.7 ApplicationofLC–AnalysisofPharmaceuticalCompoundsin
Groundwater 224
3.7.1 Sampling 225
3.7.2 AnalysisMethodfor21Antibiotics–SamplePretreatment 225
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3.7.3 UseofInternalStandardsandOtherQualityAssurance
Issues 227
3.7.4 LCAnalyses 228
3.7.5 MassSpectrometricSelectedIonMonitoringDetection 228
3.7.6 Results 229
3.8 Summary 230
Problems 230
References 232
SOLUTIONS 237
Index 263
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