Multidimensional Liquid Chromatography: Theory and Applications in Industrial Chemistry and the Life Sciences

471 Pages·2008·39.198 MB·English

MULTIDIMENSIONAL
LIQUID
CHROMATOGRAPHY
MULTIDIMENSIONAL
LIQUID
CHROMATOGRAPHY
Theory and Applications in Industrial
Chemistry and the Life Sciences
Edited by
STEVENA. COHEN
LifeSciencesR&D,WatersCorporation
Milford,MA01757,USA
MARK R. SCHURE
TheoreticalSeparationScienceLaboratory
RohmandHaasCompany
Springhouse,PA19477-0904,USA
A JOHN WILEY & SONS, INC., PUBLICATION
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LibraryofCongressCataloging-in-PublicationData:
Multidimensionalliquidchromatography:theoryandapplicationsinindustrialchemistryandthelife
sciences/editedByStevenA.Cohen,MarkR.Schure.
p. cm.
Includesindex.
ISBN978-0-471-73847-3(cloth)
1. Liquidchromatography.2. Chemicalengineering.3. Chemistry,Technical.
4. Biochemistry. I.Cohen,StevenA.,1953-II.Schure,MarkR.;1952-
QD79.C454M852007
543’.84–dc22 2007041576
PrintedintheUnitedStatesofAmerica
10987654321
CONTENTS
Foreword xiii
Preface xv
Contributors xvii
1 Introduction 1
1.1 PreviousLiteratureWhichCoversMDLC 4
1.2 HowthisBookisOrganized 5
References 6
PARTI THEORY 9
2 ElementsoftheTheoryofMultidimensional
LiquidChromatography 11
2.1 Introduction 11
2.2 PeakCapacity 13
2.3 Resolution 17
2.4 Orthogonality 19
2.5 Two-DimensionalTheoryofPeakOverlap 21
2.6 Dimensionality,PeakOrdering,andClustering 23
2.7 TheoryofZoneSampling 24
2.8 DilutionandLimitofDetection 26
2.9 ChemometricAnalysis 27
v
vi CONTENTS
2.10 FutureDirections 28
References 30
3 PeakCapacityinTwo-DimensionalLiquidChromatography 35
3.1 Introduction 35
3.2 Theory 37
3.3 Procedures 41
3.4 ResultsandDiscussion 42
3.5 Conclusions 49
Appendix3AGenerationofRandomCorrelatedCoordinates 50
Appendix3BDerivationofLimitingCorrelationCoefficientr 54
References 56
4 DecodingComplex2DSeparations 59
4.1 Introduction 59
4.2 Fundamentals:TheStatisticalDescriptionofComplex
MulticomponentSeparations 62
4.3 Decoding1Dand2DMulticomponentSeparations
byUsingtheSMOPoissonStatistics 68
4.4 DecodingMulticomponentSeparationsbythe
AutocovarianceFunction 74
4.5 Applicationto2DSeparations 78
4.5.1 ResultsfromSMOMethod 81
4.5.2 Resultsfrom2DAutocovariance
FunctionMethod 84
4.6 ConcludingRemarks 88
Acknowledgments 88
References 88
PARTII COLUMNS,INSTRUMENTATIONANDMETHODS
DEVELOPMENT 91
5 InstrumentationforComprehensiveMultidimensional
LiquidChromatography 93
5.1 Introduction 93
5.2 Heart-CuttingVersusComprehensiveMode 95
5.3 ChromatographicHardware 97
5.3.1 Valves 97
5.4 CEInterfaces 104
5.4.1 GatedInterfaceforHPLC–CE 104
5.4.2 MicrofluidicValvesforOn-Chip
MultidimensionalAnalysis 105
CONTENTS vii
5.5 ColumnsandCombinations 106
5.5.1 ColumnSystems,Dilution,andSplitting 108
5.6 Detection 109
5.7 ComputerHardwareandSoftware 109
5.7.1 SoftwareDevelopment 110
5.7.2 ValveSequencing 111
5.7.3 DataFormatandStorage 113
5.8 ZoneVisualization 115
5.8.1 ContourVisualization 115
5.8.2 2DPeakPresentation 117
5.8.3 ZoneVisualizationinSpecificChemical(pI)Regions 117
5.8.4 ExternalPlottingPrograms 117
5.8.5 DifferencePlots 118
5.8.6 Multi-channelData 118
5.9 DataAnalysisandSignalProcessing 119
5.10 FutureProspects 120
References 121
6 MethodDevelopmentinComprehensiveMultidimensional
LiquidChromatography 127
6.1 Introduction 127
6.2 PreviousWork 128
6.3 ColumnVariables 130
6.4 MethodDevelopment 130
6.4.1 TheCardinalRulesof2DLCMethodDevelopment 132
6.5 PlanningtheExperiment 143
6.6 GeneralCommentsonOptimizingthe2DLCExperiment:
Speed–ResolutionTrade-off 143
Acknowledgment 144
References 144
7 MonolithicColumnsandTheir2D-HPLCApplications 147
7.1 Introduction 147
7.2 MonolithicPolymerColumns 148
7.2.1 StructuralPropertiesofPolymerMonoliths 148
7.2.2 ChromatographicPropertiesof
PolymerMonolithicColumns 150
7.2.3 Two-DimensionalHPLCUsingPolymerMonoliths 152
7.3 MonolithicSilicaColumns 153
7.3.1 Preparation 154
7.3.2 StructuralPropertiesofMonolithicSilicaColumns 154
7.3.3 ChromatographicPropertiesof
MonolithicSilicaColumns 156
viii CONTENTS
7.4 PeakCapacityIncreasebyUsingMonolithicSilicaColumns
inGradientElution 158
7.5 2DHPLCUsingMonolithicSilicaColumns 159
7.5.1 RP-RP2DHPLCUsingTwoDifferentColumns 161
7.5.2 RP–RP2DHPLCUsingTwoSimilarColumns 164
7.5.3 IonExchange–Reversed-Phase2DHPLC
UsingaMonolithicColumnforthe2nd-D 166
7.5.4 IEX-RP2DHPLCUsingaMonolithicRP
CapillaryColumnforthe2nd-D 168
7.6 SummaryandFutureImprovementof2DHPLC 171
References 171
8 UltrahighPressureMultidimensionalLiquidChromatography 177
8.1 Background:MDLCintheJorgensonLab 177
8.1.1 CationExchange–SizeExclusion 178
8.1.2 AnionExchange–ReversedPhase 180
8.1.3 CationExchange–ReversedPhase 181
8.1.4 SizeExclusion–ReversedPhase 183
8.2 OnlineVersusOff-LineMDLC 188
8.3 MDLCUsingUltrahighPressureLiquidChromatography:
BenefitsandChallenges 189
8.3.1 AnIntroductiontoUHPLC 190
8.3.2 UHPLCforLC(cid:1)LC:HighSpeed
VersusHighPeakCapacity 191
8.3.3 LC(cid:1)UHPLCforSeparationsofIntactProteins 191
8.4 ExperimentalDetails 193
8.4.1 Instrumentation 193
8.4.2 DataAnalysis 194
8.4.3 ChromatographicConditions 195
8.4.4 Samples 196
8.5 ResultsandDiscussion 196
8.6 FutureDirectionsforUHP-MDLC 202
References 203
PARTIII LIFESCIENCEAPPLICATIONS 205
9 Peptidomics 207
9.1 StateoftheArt—WhyPeptidomics? 207
9.2 StrategiesandSolutions 208
9.3 SummaryandConclusions 218
References 218
CONTENTS ix
10 ATwo-DimensionalLiquidMassMappingTechnique
forBiomarkerDiscovery 221
10.1 Introduction 221
10.2 MethodsforSeparatingandIdentifyingProteins 223
10.2.1 pI-BasedMethodsofSeparation 223
10.2.2 Chromatofocusing-AColumnBasedpHSeparation 225
10.2.3 NonporousSeparationofProteins 226
10.2.4 Electrospray-TimeofFlight-MassSpectrometry 228
10.2.5 MALDIPeptideMassFingerprinting 229
10.2.6 DataAnalysisandRecombination 230
10.3 Applications 230
10.3.1 ProteomicMappingandClustering
ofMultipleSamples—Applicationto
OvarianCancerCellLines 230
10.3.2 2DLiquidMassMappingofTumorCellLine
SecretedSamples,Applicationto
Metastasis-AssociatedProteinProfiles 233
10.3.3 IdentificationAnnotationandDataCorrelationin
MCF10HumanBreastCancerCellLines 235
10.4 SummaryandConclusions 237
Acknowledgments 238
References 238
11 CoupledMultidimensionalChromatographyandTandemMass
SpectrometrySystemsforComplexPeptideMixtureAnalysis 243
11.1 SCX-RP/MS/MS 245
11.2 SCX/RP/MS/MS 248
11.3 MudPIT 251
11.4 AlternativeFirstDimensionApproaches 254
11.5 Conclusion 255
References 255
12 DevelopmentofOrthogonal2DLCMethods
forSeparationofPeptides 261
12.1 Introduction 261
12.2 PreviousWork 263
12.3 DevelopingOrthogonal2DLCMethods 264
12.3.1 LCSelectivityforPeptides:ExperimentalDesign 264
12.3.2 Investigationof2DLCOrthogonality
forSeparationofPeptides 266
12.3.3 GeometricApproachtoOrthogonalityin2DLC 271
12.3.4 Practical2DLCConsiderationsinProteomeResearch 275
12.3.5 EvaluationofSelected2DLCMS/MSSystems 276
x CONTENTS
12.3.6 PeakCapacityin2DLC-MS/MS 280
12.3.7 ConsiderationsofConcentrationDynamicRange 282
12.4 Conclusions 284
Acknowledgment 284
References 284
13 MultidimensionalSeparationofProteinswithOnlineElectrospray
Time-of-FlightMassSpectrometricDetection 291
13.1 Introduction 291
13.2 ChromatographicParameters 293
13.3 AnalyteDetectionandSubsequentAnalysis 293
13.4 BuildingaMultidimensionalProteinSeparation 294
13.4.1 SelectionofanIon-Exchange–Reversed-Phase
SeparationSystemforProtein-LevelSeparations 295
13.4.2 ChromatographicSorbentConsiderations 295
13.4.3 ChromatographicBehaviorofProteins 296
13.5 ComprehensiveMultidimensionalChromatographicSystems 296
13.6 Coupling2DLCwithOnlineESI–MSDetection 299
13.6.1 InteractionsbetweentheTwoDimensionsof
Chromatography(StepVs.Linear) 304
13.6.2 RecognizingIncreasedSelectivityin2DLCSeparations 306
13.7 ExpandingMultidimensionalSeparationsintoa“Middle-Out”
ApproachtoProteomicAnalysis 308
13.8 FutureDirectionsinProteinMDLC 311
13.8.1 ProteinChromatography 312
13.8.2 MSAnalysisofProteins 313
13.8.3 DataInterpretation 314
13.9 Conclusion 314
References 315
14 AnalysisofEnantiomericCompoundsUsingMultidimensional
LiquidChromatography 319
14.1 OnlineAchiral-ChiralLC-LC 320
14.2 Applications 323
14.2.1 AnalysisofEnantiomersinPlasmaandUrine 323
14.3 AminoAcids 328
14.3.1 PhysiologicalFluidsorTissues 328
14.3.2 InFood,Beverages,andOtherProducts 333
14.4 OtherApplications 334
14.4.1 AnalysisofEnantiomersfromPlant
andEnvironmentalSources 334
14.5 MiscellaneousApplications 336
14.6 Conclusion 338
References 339
CONTENTS xi
PARTIV MULTIDIMENSIONALSEPARATION
USINGCAPILLARYELECTROPHORESIS 345
15 Two-DimensionalCapillaryElectrophoresisforthe
ComprehensiveAnalysisofComplexProteinMixtures 347
15.1 Introduction 347
15.2 PreviousWork 348
15.2.1 MiniaturizedIEF/SDS-PAGE 348
15.2.2 One-DimensionalCapillaryElectrophoresis
forProteinAnalysis 349
15.3 Two-DimensionalCapillarySeparationsforAnalysisof
PeptidesandProteins 352
15.3.1 CapillaryLiquidChromatographyCoupled
withCapillaryElectrophoresisforAnalysisof
UnlabeledPeptidesandProteins 352
15.3.2 Two-DimensionalCapillaryElectrophoresisfor
AnalysisofProteins 352
15.3.3 High-SpeedTwo-DimensionalCapillaryElectrophoresis 356
15.3.4 TheAnalysisofaSingleFixedCell 358
15.4 Conclusions 360
15.5 Abbreviations 360
References 360
16 Two-DimensionalHPLC–CEMethods
forProtein/PeptideSeparation 365
16.1 Introduction 365
16.2 Off-lineVersusOnline 366
16.3 HPLCFractionation 366
16.4 2DHPLC–CE 367
16.5 CE–MSDetection 368
16.6 Applications 370
16.7 ConcludingRemarks 380
Acknowledgment 381
References 381
PARTV INDUSTRIALAPPLICATIONS 385
17 MultidimensionalLiquidChromatographyin
IndustrialApplications 387
17.1 Introduction 387
17.2 PrinciplesofMultidimensionalLiquidChromatography
asAppliedtoPolymerAnalysis 390