Hydrophilic Interaction Liquid Chromatography (HILIC) And Advanced Applications......Page 2
Chromatographic Science Series......Page 3
Hydrophilic Interaction Liquid Chromatography (hilic) And Advanced Applications......Page 7
Contents......Page 10
Preface......Page 14
Editors......Page 16
Contributors......Page 18
Contents......Page 22
1.1 Fundamental Features of Aqueous Normal- Phase Retention......Page 23
1.2.1 ANP 1......Page 24
1.2.2 ANP 2......Page 25
1.2.3 ANP 3......Page 27
1.3.2 Temperature Behavior......Page 28
1.3.3 Repeatability......Page 30
1.3.4 Re-Equilibration......Page 31
1.4.2 Organic Component of Mobile Phase......Page 32
1.4.3 Mobile Phase Additive......Page 33
1.4.4 Sample Solvent Additive......Page 34
1.4.5 Gradients......Page 35
1.5.1 Amino Acids......Page 36
1.5.2 Carbohydrates......Page 37
1.5.3 Organic Acids......Page 38
1.5.4 Nucleotides......Page 41
1.5.5 Other Analytes......Page 43
Acknowledgment......Page 46
References......Page 47
2.1 Introduction......Page 48
2.2 Theoretical Introduction and Definitions......Page 50
2.3 Experimental Design......Page 53
2.4.1 Relationship between Experimentally Determined K' and calculated log Dph3.0
......Page 55
2.4.2 Discussion of Assumpt ions Made to Formulate Hypothesis......Page 56
2.5 Conclusions......Page 64
References......Page 65
Contents......Page 68
3.2.1 Introduction......Page 69
3.2.3 Column Chemistry......Page 70
3.3.1 Introduction......Page 71
3.3.2 Column Chemistry......Page 72
3.3.3 Separation Mechanism......Page 73
3.3.4 Applications......Page 74
3.4 HILI C Behavior of Reversed- Phase/ Ion- Exchange Mixed- Mode Stationary Phases......Page 76
3.4.1 Retent ion Behavior of Xanthins, Nucleosides, an d Wate r-Soluble Vitamins on RP/WAX Mixed-Mode Columns in HILIC Mode......Page 77
3.4.2 Retention Behavior of Fungal Metabolite s on RP/WAX Mixed-Mode Columns in HILIC Mode......Page 78
3.4.3 HILIC Behavior Compa rison of RP/IEX BimodalMixed-Mode Columns......Page 79
3.5.1 Pharmaceutical Active Ingredient s and Counterions......Page 83
3.5.2 Melamine and Cyanuric Acid......Page 85
3.5.3 Buffer Salts for Biopharmaceutical Applications......Page 89
3.5.4 Determination of Ethanol Consumption Markers......Page 91
3.6 Concluding Remarks......Page 92
Acknowledgments......Page 93
References......Page 94
4.1 Introduction......Page 98
4.2 Polyphenolic Compounds......Page 99
4.3 Chromatography Challenges for Separation of Various PAs......Page 100
4.4 HILIC Conditions for Separation of PAs......Page 101
4.6 Retention Behavior of Oligomeric PA S tandards in HILI C......Page 102
4.7 Retention Mechanism of Oligomeric PAs in HILI C......Page 105
4.8 HILIC Separation of Other Polyphenolic Compounds......Page 107
References......Page 109
5.1 Introduction......Page 110
5.2.1 HILIC with Conventional Detectors......Page 111
5.2.2 HILIC Coupled to Mass Spectrometry......Page 116
5.3 Application of Hydrophilic Interaction Chromatography to the Analysis of Safety- Related Compounds in Foods......Page 117
References......Page 125
6.1 Introduction......Page 126
6.2 Paralytic Shellfish Poisoning Toxins......Page 129
6.2.1 HILIC -MS Determination of PSP Toxins......Page 130
6.2.2 Application to Plankton and Shellfish Samples......Page 141
6.3.1 HILIC -MS D ete rminat ion of DA......Page 143
6.3.2 Application to Shellfish Samples......Page 145
6.4.1 HILIC -MS Determination of Assorted Cyanobacterial Toxins......Page 147
6.4.2 Applications to Cyanobacterial Samples......Page 149
6.5 Conclusions......Page 151
References......Page 152
Contents......Page 154
7.2.1 Generalities......Page 155
7.2.2 Paralytic Shellfish Poisoning Toxins......Page 156
7.2.3 Tetrodotoxin and Analogs......Page 161
7.2.5 Concluding Remarks......Page 163
7.3.1 Estrogens......Page 164
7.3.2 Cytostatic Drugs......Page 167
7.3.4 Miscellaneous Pharmaceuticals......Page 168
7.3.5 Drugs of Abuse......Page 170
7.4 Pesticides......Page 171
7.4.2 Phenylurea Herbicides......Page 172
7.4.3 Quaternary Ammonium Salt Herbicides......Page 173
7.5.2 Oligosaccharides......Page 174
References......Page 175
Contents......Page 178
8.1.2 Antimicrobials in the Environment......Page 179
8.1.4 Analysis of Spectinomycin and Lincomycin......Page 181
8.1.5 Spectinomycin......Page 182
8.1.7 Objective......Page 183
8.2.3.1.1 Liquid Swine Manure......Page 184
8.2.4.1.1 Manure Liquid Component......Page 185
8.2.4.2.2 Manure Solids Component......Page 186
8.2.6.2 Mass Spectrometer......Page 187
8.3.2.1 Liquid Manure......Page 188
8.3.3 Mass Spectrometry......Page 189
8.3.4 Chromatography......Page 191
8.3.6 Application of Method to Analysis of Environmental Matrices......Page 194
References......Page 195
Contents......Page 198
9.1 Characteristics of Clinical Samples and Clinical Analysis......Page 199
9.2.2 Biomarker Profiling......Page 200
9.2.3.1.2 Free Metanephrines......Page 201
9.2.3.1.4 Methylmalonic Acid......Page 202
9.2.3.1.6 Ornithine......Page 203
9.2.3.1.8 Tobacco-Specific Nitrosamine Metabolites......Page 205
9.2.3.2.2 1,5-Anhydroglucitol......Page 207
9.2.3.2.3 Arginine, Synthetic Dimethylarginine, and AsymmetricDimethylarginine......Page 208
9.2.3.2.4 Cocaine and Metabolites......Page 209
9.2.3.2.6 Organophosphorus Nerve Agent Metabolites......Page 210
9.2.3.3.1 3-Nitrotyrosine, Tyrosine, Hydroxyproline, and Proline......Page 211
References......Page 212
10.1 Introduction......Page 224
10.2 Arginine and Its Dimethyl Derivatives......Page 225
10.3 MS Setup......Page 226
10.4.2 Effect of ACN Percentage on HILIC and Comparison between Organic-Rich and Water-Rich Mobile Phaseon Silica Column Chromatography......Page 228
10.4.3 Effect of TFA Percentage on the HILIC Method......Page 232
10.4.4 Effect of Column Temperature on the HILIC Method......Page 233
10.5 Plasma Method......Page 235
10.7 Method Development for the Analysis of CSF......Page 240
References......Page 243
11.1 Introduction......Page 246
11.2 Ionic Functional Groups in HILIC Separation......Page 248
11.2.1 Effect of Stationary Phase Composition......Page 249
11.2.2 Effect of Buffer Concentration (Ionic Strength)......Page 254
11.2.3 Effect of Column Temperature......Page 260
11.3.1 Effect of Mobile Phase Composition......Page 265
11.3.2 Effect of Column Temperature......Page 273
References......Page 277
Contents......Page 280
12.1 Introduction......Page 281
12.2 Trace Analysis of Genotoxic Impurities......Page 282
12.2.1 Alkyl Sulfonates......Page 284
12.2.2 Alkyl Halides......Page 288
12.3.2 (3S)-3-Morpholine methanol......Page 294
12.3.4 Organic Acids......Page 296
12.4.1 5-Fluorouracil in 5-Fluorocytosine......Page 297
12.4.2 Guanine in Acyclovir......Page 298
12.4.3 Epirubicin......Page 299
12.4.4 Chiral Compounds......Page 300
12.4.5 Carbamates......Page 301
12.4.6 Sodium Cromoglicate......Page 303
12.5 Counterion A nalysis......Page 304
12.6 Conclusions and Future Prospects......Page 306
References......Page 308
13.1.1 HILIC and HILIC Stationary Phases......Page 312
13.1.2 Overview of UPLC and HILIC -UPLC Applications......Page 314
13.2.1 Introduction......Page 316
13.2.2 Experimental......Page 317
13.2.3.1 Univariate Method Development......Page 318
13.2.3.2 Multivariate Method Optimization......Page 321
References......Page 325
14.1 Introduction......Page 330
14.2 Eff ect of Organic S olvent......Page 333
14.3 Effect of Acetonitrile Content on Retention with Different Stationary Phases......Page 337
14.4 Effect of pH......Page 342
14.5 Effect of Buffer Concentration with Various Stationary Phases......Page 347
14.6 Temperature Effect with Various Stationary Phases......Page 353
14.7 Application Results and Method Validation......Page 359
References......Page 362
15.1 Introduction......Page 366
15.2.1.1 HILIC-Column Screening......Page 368
15.2.1.2 Mobile-Phase Components......Page 369
15.2.1.2.1 Mobile-Phase Strength......Page 371
15.2.1.2.2 Mobile-Phase pH......Page 374
15.2.1.2.3 Ionic Strength of Mobile Phase......Page 375
15.2.1.3 Column Temperature......Page 377
15.2.2 Method Validation......Page 379
15.3.1.1 Alcohol as a Weak Eluent......Page 381
15.3.1.2 Combined ZIC and HILIC Modes......Page 382
15.3.1.3 Type of Acid Additive......Page 383
15.3.1.4 Type of Buffer and Ionic Strength......Page 386
15.3.2 Method Validation......Page 389
References......Page 390
16.1 Introduction......Page 394
16.2 Analysis of Pharmaceutical Ingredients......Page 395
16.2.4 SCG in Ophthalmic Solution......Page 396
16.3 Retention Mechanism......Page 398
16.3.1.1 Choice of Organic Solvent......Page 399
16.3.1.2 Use of Alternate Solvent......Page 400
16.3.2 Mobile-Phase pH and Ion Exchange......Page 401
16.3.3 Ionic Strength......Page 407
16.3.4 Stationary Phase......Page 408
16.3.5 Diluent and Injection Volume......Page 412
16.3.6 Validation of Methods......Page 413
16.4 Advantages......Page 414
References......Page 418
17.1 Introduction......Page 422
17.2 Polar Stationary Phases for HILI C......Page 423
17.3 Selectivity of Various Polar Stationary Phases in HILIC......Page 427
17.4 Retentivity of Various Polar Stationary Phases in HILIC......Page 434
17.5 Factors Affecting Retention and Selectivity in HILIC......Page 436
17.6 Separation of Positional Isomers in HILI C......Page 443
17.7 Conclusions......Page 444
References......Page 445
Contents......Page 448
18.2 Application of HILI C- MS / MS in the Field of Pharmacokineticsβ Bioequivalence Studies......Page 449
18.4 Pharmacokinetics S tudies of Donepezil, Loratadine, and Cetirizine......Page 450
18.6 Pharmacokinetic Study of the Peptide Drug Taspoglutide......Page 455
18.7 HILIC- ESI - MS / MS Method for the Quantitation of Polar Metabolites of Acrylamide in Human Urine......Page 457
18.8 HILIC- MS / MS Technique f or Sensitive Monitoring of the Changes of Urinary E strogen Conjugates......Page 458
18.9 Combination of HILIC- MS and RPLC- MS for Profiling Polar Urine Metabolites......Page 459
References......Page 462
19.1 Introduction......Page 466
19.2.1 Chemicals and Materials......Page 468
19.2.3 Physiochemical Parameters......Page 472
19.3.1 Optimal pH......Page 473
19.3.3 Gradient Method Development......Page 477
19.3.5 Separation Results......Page 478
19.3.6 Method Validation......Page 482
Acknowledgments......Page 485
References......Page 486
20.1 Introduction......Page 490
20.2 Protein Modifications: Glycosylation......Page 492
20.3 Troubles in Glycoproteome Analysis......Page 493
20.4 Enrichment Methods for Glycoproteome......Page 494
20.5 HILIC: Mechanism and Material......Page 495
20.6 HILIC: From Glycomics to Glycoproteomics......Page 498
20.7 Glycoproteomics by HILIC and Mass Spectrometry......Page 499
20.8 Application to Biomarker Discovery......Page 503
References......Page 506
Contents......Page 512
21.1.2 Techniques for Separation and Detection of Carbohydrates......Page 513
21.1.3 HILIC in the Separation of Carbohydrates......Page 515
21.2.1 Use of Different Stationary Phases......Page 519
21.2.2 Mobile-Phase Composition......Page 520
21.2.3 Temperature and Other Variables......Page 526
21.2.4 Sample Preparation Strategy......Page 529
21.2.5 Detection of Carbohydrates......Page 530
21.2.6 Short Outline for the Development of an HILIC Method......Page 532
21.3.1 HILIC Separation of Mono-, Di-, and Oligosaccharidesby Cyclodextrin Columns in Combinat ion with UV an d RI Detection......Page 533
21.3.2 HILIC Separation by an Amide-80 Columnand MS Analysis of Carbohydrates from Plants......Page 534
21.3.3 HILIC Separation by a ZIC -HILIC Column and MS Analysisof Tryptic Glycopeptides from Human Immunoglobulin......Page 535
21.3.5 Separation of Glucosinolate s by a ZIC -HILIC Column and Detection by Absorbance......Page 536
21.3.6 Separation of GAGs by an Amide-80 Column Combined with MS Analysis......Page 538
References......Page 541
Contents......Page 544
22.1.1 Importance of Glycosylat ion in Biological Systems and How Variation Can Lead to Disease......Page 545
22.1.2 Types of Glycosylation and Glycan Patterns......Page 546
22.1.3.2 MALDI -MS......Page 548
22.2.1 Glycoprote in Enrichment from Complex Solutions......Page 549
22.2.1.1 Lectin Affinity Chromatography......Page 550
22.2.1.3 Other Separation Techniques......Page 551
22.2.2.1 Hydrophilic Interaction Liquid Chromatography......Page 552
22.2.2.3 Hydrazine Chemistry......Page 555
22.2.2.5 Others......Page 556
22.3 Examples of Glycoprotein Characterization Using HILIC......Page 557
22.3.2 Characterization of Glycoforms......Page 558
22.3.3 Characterization of Glycosylation Site Occupancy......Page 559
22.3.4 Full Characterization of Glycoproteins......Page 561
References......Page 563
Contents......Page 572
23.2 Background......Page 573
23.3.1 HILIC SPE for Sample Prepa rat ion of Glycan s an d Glycopept ides......Page 575
23.3.1.1 Selective Enrichment of Glycopeptidesfrom Purified/Semi-Purified Glycoprotein......Page 576
23.3.1.2 Enrichment and Desalting of Released Glycans......Page 578
23.3.1.4 Enrichment of Glycopeptides from Complex Mixture Using HILIC SPE......Page 580
23.3.2.1 HILIC Separation of Glycans with Off-Line MS Detection......Page 584
23.3.2.2 HILIC Separation of Glycopeptides with Off-Line MS Detection......Page 585
23.3.2.3 Online HILI C-MS of Glycans......Page 586
23.3.2.4 Online HILI C-MS of Glycopeptides......Page 587
23.4.1.2 Strong Cation Exchange......Page 588
23.4.3.1 Early Applications of HILI C in Phosphoprotein/Peptide Analysis......Page 589
23.4.3.2 Application in Phosphoproteomics......Page 590
23.4.4 Electrostatic Repulsion Hydrophilic Interaction Chromatography......Page 591
23.4.5 Multiphosphopeptide Enrichment Prior to HILIC Fractionation......Page 592
References......Page 593