THE HPLC SOLVENT GUIDE SECOND EDITION

PAUL C. SADEK

(Cf}\WILEY-^TyiNTERSCIENCE A John Wiley & Sons, Inc., Publication

CONTENTS

Preface Introduction Abbreviations, Acronyms, and Symbols

XIII

xv xvii

Chapter 1 Physical and Chemical Solvent Properties

1.1 UV Cutoff, 2 1.2 Absorbance Versus Wavelength Curves, 3 1.3 Reversed-Phase Solvents, 5

1.3.1 Mobile Phases and Mobile Phase Modifiers, 9 1.4 Normal-Phase Solvents, 11 1.5 System Peaks, 14 1.6 Lot-to-lot Solvent Variability, 16 1.7 Viscosity, 19 1.8 Miscibility and Solubility, 22 1.9 Buffers and Other Mobile Phase Additives, 25 1.10 Volatility, 29 1.11 Solvents: Instability, Reactivity, and Denaturants, 31

11.1 Ethers, 32 11.2 Chlorinated Alkanes, 34 11.3 Ethanol, 36 11.4 Water, 36 11.5 Acetone, 38

1.12 Sample Stability in Solvents, 39 1.13 Mobile Phase Interaction with Stationary Phase, 40 1.14 Inherent Contaminants, 41 1.15 Solvent Effects on Fluorescence, 42 1.16 Particulates and Solvent Filtering, 43 1.17 Manufacturers and Testing Protocols, 44

vi CONTENTS

Chapter 2 Method Optimization 45

2.1 Eluotropic Series and Solvent Strength Parameters, 46 2.1.1 Eluotropic Series, 46 2.1.2 Solvent Strength Parameters, 49 2.1.3 Mathematical Representation of Solvent Strength, 52

2.2 Triangulation and Other Algorithmic Methods, 53 2.3 Scout Gradients, 58 2.4 Window Diagrams, 60 2.5 Brute-force Method, 60 2.6 Other Optimization Techniques and Retention Parameters, 61

Chapter 3 Method Validation and Ongoing Performance Evaluation 63

3.1 Qualification Process, 64 3.1.1 Instrument Qualification, 64 3.1.2 Design Qualification, 65 3.1.3 Installation Qualification, 65 3.1.4 Operational Qualification, 66 3.1.5 Performance Qualification, 66

3.2 Method Validation, 67 3.2.1 Accuracy, 68 3.2.2 Precision, 68 3.2.3 Linearity, 69 3.2.4 Linear and Working Ranges, 71 3.2.5 Specificity, 72 3.2.6 Detection and Quantitation Limit, 72 3.2.7 Robustness, 73 3.2.8 Ruggedness, 73

3.3 System Suitability, 74

Chapter 4 Alcohols 75

4.1 Impurities, 81 4.2 General Analytes, 82

4.2.1 Simple Substituted Benzene Analytes, 82 4.2.2 Other Organic Compounds, 89 4.2.3 Organometallic Compounds and Metal-Ligand Complexes, 93 4.2.4 Summary, 96

4.3 Environmentally Important Analytes, 96 4.3.1 PAHs, Substituted PAHs, and Related Analytes, 96 4.3.2 Nitrated and Chlorinated Nonpesticide/Herbicide Pollutant

Analytes, 100 4.3.3 Pesticides, Herbicide, and Fungicides, 103

4.3.3.1 Pesticides, 103 4.3.3.2 Herbicides, 108 4.3.3.3 Fungicides, 111

4.3.4 Summary, 113

CONTENTS vii

4.4 Industrial and Polymer Analytes, 113 4.4.1 Surfactant and Additive Analytes, 113 4.4.2 Polymers and Polymer Additives, 115 4.4.3 Sunscreen Agents, 118 4.4.4 Dyes, 120 4.4.5 Other Industrial Analytes, 123 4.4.6 Summary, 124

4.5 Biological Analytes, 124 4.5.1 Carboxylic Acid Analytes, 124 4.5.2 Basic Amine Analytes, 130 4.5.3 Aflatoxins, Mycotoxins, and Other Toxic Analytes, 133 4.5.4 Vitamins and Related Analytes, 139

4.5.4.1 Water-soluble Vitamins and Related Compounds, 139

4.5.4.2 Fat-soluble Vitamins and Related Compounds, 140

4.5.5 Terpenoids, Flavonoids, and Related Compounds, 148 4.5.5.1 Terpenoids, 148 4.5.5.2 Flavanoids, 151 4.5.5.3 Caffeine and Related Compounds, 153 4.5.5.4 Other Compounds, 155

4.5.6 Analytes Derived from Oils and Fats, 169 4.5.7 Nucleotides, Nucleosides, and Related Analytes, 171 4.5.8 Other Analytes, 174 4.5.9 Summary, 178

4.6 Amino Acid, Peptide, and Protein Analytes, 178 4.6.1 Amino Acid Analytes, 178 4.6.2 Peptide Analytes, 180 4.6.3 Protein Analytes, 181 4.6.4 Summary, 184

4.7 Pharmaceutical Analytes, 184 4.7.1 Drug Surveys and Screening Procedures, 184 4.7.2 Retention Mechanisms for Drug Compounds, 186 4.7.3 NSAIDs and Analgesie Drugs, 187 4.7.4 Antibiotic Drugs, 189 4.7.5 Anticancer Drugs, 192 4.7.6 Antiepileptic Drugs, 195 4.7.7 Steroidal Drugs, 196 4.7.8 Anthelmintics, 198 4.7.9 Illicit Drugs, 199 4.7.10 Antihistamines, 201 4.7.11 Anti-HIV Drugs, 202 4.7.12 Antianxiety and Antipsychotic Drugs, 202 4.7.13 Other Drug Analytes, 204 4.7.14 Summary, 213

viii CONTENTS

Chapter 5 Alkanes and Alkyl Aromatics 214

5.1 Impurities, 216 5.2 General Analytes, 221

5.2.1 Simple Substituted Aromatic Analytes, 221 5.2.2 Organometallics and Metal-Ligand Complexes, 226 5.2.3 Summary, 227

5.3 Environmentally Important Analytes, 227 5.3.1 PAHs, Substituted PAHs, and Related Analytes, 227 5.3.2 Nitrated and Chlorinated Nonpesticide/herbicide Pollutant

Analytes, 230 5.3.3 Pesticide, Herbicide, and Fungicide Analytes, 231 5.3.4 Summary, 233

5.4 Industrial and Polymer Analytes, 233 5.4.1 Surfactant and Additive Analytes, 233 5.4.2 Other Polymerie Analytes, 235 5.4.3 Fullerenes, 236 5.4.4 Summary, 237

5.5 Biological Analytes, 238 5.5.1 Carboxylic Acid Analytes, 238 5.5.2 Vitamins and Related Analytes, 239 5.5.3 Terpenoids, Flavonoids, Steroids, and Related Compounds, 242 5.5.4 Analytes Derived from Oils and Fats, 244 5.5.5 Other Analytes, 248 5.5.6 Summary, 250

5.6 Amino Acid and Peptide Analytes, 250 5.6.1 Summary, 250

5.7 Pharmaceutical Analytes, 252 5.7.1 Cardiac Glycosides, 252 5.7.2 NSAIDs and Analgesie Drugs, 253 5.7.3 Benzodiazepines, 253 5.7.4 Other Analytes, 254 5.7.5 Summary, 257

Chapter 6 Chlorinated Alkanes and Chlorinated Benzenes 258

6.1 Impurities, 260 6.2 General Analytes, 266

6.2.1 Simple Substituted Benzene Analytes, 266 6.2.2 Organometallics and Metal-Ligand Complexes, 269 6.2.3 Summary, 269

6.3 Environmentally Important Analytes, 269 6.3.1 PAHs, Substituted PAHs, and Related Analytes, 269 6.3.2 Nitrated and Chlorinated Nonpesticide/herbicide Analytes, 271 6.3.3 Pesticide and Herbicide Analytes, 271 6.3.4 Summary, 272

CONTENTS ix

6.4 Industrial and Polymer Analytes, 272 6.4.1 Surfactant and Additive Analytes, 272 6.4.2 Polymerie Analytes, 273 6.4.3 Fullerenes, 274 6.4.4 Summary, 275

6.5 Biological Analytes, 275 6.5.1 Carboxylic Acid Analytes, 275 6.5.2 Vitamins and Related Analytes, 275 6.5.3 Analytes from Fats and Oils, 277 6.5.4 Other Analytes, 278 6.5.5 Summary, 280

6.6 Amino Acid and Peptide Analytes, 280 6.7 Pharmaceutical Analytes, 281

Chapter 7 Ethers 285

7.1 Impurities, 287 7.2 General Analytes, 288

7.2.1 Simple Substituted Benzenes and Related Analytes, 288 7.3 Environmentally Important Analytes, 291

7.3.1 PAHs, Substituted PAHs, and Related Analytes, 291 7.3.2 Pesticide and Herbicide Residue Analytes, 292

7.4 Industrial and Polymer Analytes, 294 7.4.1 Surfactant and Additive Analytes, 294 7.4.2 Polymerie Analytes, 294 7.4.3 Fullerenes and Other Industrial Analytes, 296

7.5 Biological Analytes, 297 7.5.1 Carboxylic Acid Analytes, 297 7.5.2 Vitamins and Related Analytes, 298 7.5.3 Terpenoids, Flavonoids, Steroids, and Related Analytes, 300 7.5.4 Analytes Derived from Oils and Fats, 302 7.5.5 Other Analytes, 303

7.6 Amino Acid and Peptide Analytes, 305 7.7 Pharmaceutical Analytes, 305

7.7.1 Summary, 311

Chapter 8 Ketones and Esters 312

8.1 Impurities, 314 8.2 General Analytes, 316 8.3 Environmentally Important Analytes, 318 8.4 Industrial and Polymer Analytes, 319 8.5 Biological Analytes, 320 8.6 Pharmaceutical Analytes, 325

x CONTENTS

Chapter 9 Nitriles and Nitrogenous Solvents 327

9.1 Impurities, 328 9.2 General Analytes, 329

9.2.1 General Sample Solvent Considerations, 329 9.2.2 Simple Substituted Hydrocarbons and Benzene Analyte Retention

Studies, 332 9.2.3 Other Compounds, 337 9.2.4 Organometallic and Metal-Ligand Complexes, 341 9.2.5 Summary, 343

9.3 Environmentally Important Analytes, 343 9.3.1 Substitutes Benzenes and Related Analytes, 343 9.3.2 PAHs, Substituted PAHs, and Related Analytes, 344 9.3.3 Nitro-, Nitroso-, and Chlorinated Nonpesticide/herbicide

Pollutant Analytes, 348 9.3.4 Pesticide, Herbicide, and Fungicide Analytes, 350

9.3.4.1 Pesticides, 350 9.3.4.2 Herbicides, 363 9.3.4.3 Fungicides, 368

9.3.4 Summary, 369 9.4 Industrial and Polymer Analytes, 370

9.4.1 Surfactant and Additive Analytes, 370 9.4.2 Polymerie Analytes, 373 9.4.3 Dyes and Related Analytes, 375 9.4.4 Other Industrial Analytes, 378 9.4.5 Personal Care and Cosmetic Analytes, 379 9.4.6 Summary, 380

9.5 Biological Analytes, 381 9.5.1 Carboxylic Acid Analytes, 381 9.5.2 Basic Amine Analytes, 387 9.5.3 Aflatoxins, Mycotoxins, and Other Toxic Analytes, 395 9.5.4 Vitamins and Related Analytes, 401

9.5.4.1 Water-soluble Vitamins and Related Compounds, 401 9.5.4.2 Fat-Soluble Vitamins and Related Compounds, 402

9.5.5 Terpenoids, Flavonoids, and Other Naturally Occurring Analytes, 405 9.5.5.1 Terpenoids, 405 9.5.5.2 Flavanoids and Related Compounds, 408 9.5.5.3 Alkaloids and Related Compounds, 418 9.5.5.4 Fats, Oils, and Related Analytes, 425

9.5.6 Nucleotides, Nucleosides, and Related Analytes, 428 9.5.7 Sugars and Related Analytes, 430 9.5.8 Other Analytes, 433 9.5.9 Summary, 450

9.6 Amino Acid, Peptide, and Protein Analytes, 450

CONTENTS xi

9.6.1 Amino Acid Analytes, 450 9.6.2 Peptide Analytes, 452 9.6.3 Protein Analytes, 457 9.6.4 Summary, 459

9.7 Pharmaceutical Analytes, 459 9.7.1 Drug Surveys and Screening Procedures, 459 9.7.2 NSAIDs and Analgesie Drugs, 460 9.7.3 Antibiotic Drugs, 464 9.7.4 Anticancer Drugs, 476 9.7.5 Antiepileptic Drugs, 482 9.7.6 Anthelmintics, 485 9.7.7 Illicit and Related Drugs, 487 9.7.8 Antihistamines, 490 9.7.9 Antiretroviral Drugs, 492 9.7.10 Antidepressants, 496 9.7.11 Antibacterial Drugs, 499 9.7.12 Anesthetics, 503 9.7.13 Immunosuppressants, 504 9.7.14 Analgesics, 505 9.7.15 Antihypercholesterolemic and Antihyperlipidemic Drugs, 506 9.7.16 Antihypertension Drugs, 507 9.7.17 Antimalarials, 511 9.7.18 Steroids, 512 9.7.19 Antipsychotics, 515 9.7.20 Other Drug Analytes, 518

9.8 Summary, 527

Chapter 10 Water, Dimethyl Sulfoxide, and Common Acidic Modiners 531

10.1 General Considerations and Impurities, 531 10.1.2 Solvent Preparation, 532

10.2 Industrial Analytes, 535 10.3 Biological Analytes, 538

10.3.1 Carboxylic Acid Analytes, 539 10.3.2 Basic Amine Analytes, 540 10.3.3 Toxins, 541 10.3.4 Vitamins, 542 10.3.5 Nucleotides, Nucleosides, Amino Acids, and Peptides, 543 10.3.6 Terpenoids, Flavonoids, and Related Analytes, 545

10.4 Pharmaceutical Analytes, 546 10.5 Summary, 547 10.6 Dimethyl Sulfoxide, 549

References 552

Index 607