Method Development in Analytical HPLC presents the essential information for understanding the process of developing an HPLC method of analysis. It includes foundational information related to HPLC, as well as discussion of sample types, the properties of analytes and matrices in the samples, and sample preparation. The core of the book describes the best ways for approaching method development in various types of HPLC and the criteria for method optimization and validation. This book provides clear guidance for adopting analytical methods from the literature and describes the development of original methods with selection of the suitable type of HPLC, of specific columns, mobile phase, and detection techniques with an emphasis on the use of mass spectrometry for detection, as well as optimization and validation of the chosen analytical method. The book includes useful details on method development for specific types of chromatography such as RP-HPLC, HILIC, ion exchange, size exclusion, and chiral. Method Development in Analytical HPLC also includes information about green chemistry in analytical methods, computer assisted method development, and other key contemporary aspects of the subject.
1. 1.7 Introduction to data processing in HPLC analysis - p. 39 2. 2. Retention and elution processes in high-performance liquid chromatography - p. 59 3. 2.1 Equilibrium types in HPLC - p. 59 4. 2.2 Intermolecular interactions involved in HPLC separations - p. 67 5. 2.3 A classification of analytical HPLC types - p. 81 6. 2.4 Influence of pH, ionic strength, and temperature on separation equilibria - p. 85 7. 3. Instrumentation in high-performance liquid chromatography - p. 95 8. 3.1 Overall description of an HPLC instrument - p. 95 9. 3.2 The separatory section of an HPLC instrument - p. 99 10. 3.3 Detectors in HPLC other than mass spectrometers - p. 108 11. 3.4 Mass spectrometers used as HPLC detectors - p. 122 12. 4. Performance and utilization of high-performance liquid chromatography separation and detection - p. 147 13. 4.1 Parameters related to the characterization of the chromatographic peak - p. 147 14. 4.2 Parameters related to peak separation in HPLC - p. 166 15. 4.3 Parameters for characterization of gradient separation in HPLC - p. 170 16. 4.4 Characterization of detector performance in HPLC - p. 177 17. 4.5 Parameters related to quantitation in HPLC - p. 193 18. 4.6 Utilizations of HPLC in chemical analysis - p. 198 19. 4.7 Utilization of HPLC in nontargeted type analysis - p. 208 20. 5. Samples in high-performance liquid chromatography analysis - p. 219 21. 6. Sample preparation for high-performance liquid chromatography analysis - p. 255 22. 6.1 The role of sample preparation for HPLC - p. 255 23. 6.2 Mechanical processing in sample preparation - p. 261 24. 6.3 Phase transfer techniques used in sample preparation - p. 265 25. 6.4 Liquid–liquid extraction in sample preparation - p. 272 26. 6.5 Liquid–solid extraction in sample preparation - p. 279 27. 6.6 Solid-phase extraction in sample preparation for HPLC - p. 284 28. 6.7 Sample derivatization - p. 288 29. 6.8 Automated sample preparation and on-line coupling with the HPLC - p. 294 30. 7. Chromatographic columns in high-performance liquid chromatography - p. 301 31. 7.1 Construction of an HPLC column - p. 301 32. 7.2 Columns with hydrophobic character - p. 310 33. 7.3 Columns with polar character - p. 328 34. 7.4 Chiral chromatographic columns - p. 331 35. 7.5 Columns used in ion exchange chromatography and related techniques - p. 337 36. 7.6 Chromatographic columns used in size exclusion - p. 344 37. 7.7 Other types of chromatographic columns - p. 346 38. 7.8 Care for the chromatographic column - p. 349 39. 8. Mobile phase and the role of solvents in high-performance liquid chromatography - p. 355 40. 9. Development of a high-performance liquid chromatography analytical method - p. 389 41. 9.1 General plan for adopting or developing an analytical method - p. 389 42. 9.2 Stages in developing a new HPLC analytical method - p. 394 43. 9.3 Development of an RP-HPLC analytical method - p. 399 44. 9.4 Development of an ion-pair HPLC analytical method - p. 431 45. 9.5 Development of an analytical method on polar columns - p. 438 46. 9.6 Development of an HPLC analytical method for the separation of chiral compounds - p. 447 47. 9.7 Development of an analytical method for ion chromatography - p. 460 48. 9.8 Development of an analytical method for size exclusion HPLC - p. 473 49. 9.9 Development of a method for immunoaffinity HPLC - p. 485 50. 9.10 Development of a method for mixed-mode HPLC - p. 489 51. 9.11 Development of a bidimensional HPLC analytical method - p. 495 52. 9.12 Additional issues to consider in developing an HPLC analytical method - p. 498 53. 10. Selection of the best options in developing a high-performance liquid chromatography analytical method - p. 511 54. 10.1 Improvement and optimization processes in an HPLC method - p. 511 55. 10.2 Trial-and-error in method development - p. 521 56. 10.3 Theoretical approaches used for prediction of various parameters in HPLC - p. 524 57. 10.4 Methodologies in HPLC optimization - p. 534 58. 10.5 Computer-assisted method development in HPLC - p. 543 59. 10.6 Selection between different HPLC methods - p. 544 60. 11. Method and data validation in high-performance liquid chromatography analysis - p. 553 61. Appendix 1 - p. 583 62. Index - p. 593
Method Development in Analytical HPLC presents the essential information for understanding the process of developing an HPLC method of analysis. It includes foundational information related to HPLC, as well as discussion of sample types, the properties of analytes and matrices in the samples, and sample preparation. The core of the book describes the best ways for approaching method development in various types of HPLC and the criteria for method optimization and validation. This book provides clear guidance for adopting analytical methods from the literature and describes the development of original methods with selection of the suitable type of HPLC, of specific columns, mobile phase, and detection techniques with an emphasis on the use of mass spectrometry for detection, as well as optimization and validation of the chosen analytical method. The book includes useful details on method development for specific types of chromatography such as RP-HPLC, HILIC, ion exchange, size exclusion, and chiral. Method Development in Analytical HPLC also includes information about green chemistry in analytical methods, computer assisted method development, and other key contemporary aspects of the subject.
1. 1.7 Introduction to data processing in HPLC analysis - p. 39 2. 2. Retention and elution processes in high-performance liquid chromatography - p. 59 3. 2.1 Equilibrium types in HPLC - p. 59 4. 2.2 Intermolecular interactions involved in HPLC separations - p. 67 5. 2.3 A classification of analytical HPLC types - p. 81 6. 2.4 Influence of pH, ionic strength, and temperature on separation equilibria - p. 85 7. 3. Instrumentation in high-performance liquid chromatography - p. 95 8. 3.1 Overall description of an HPLC instrument - p. 95 9. 3.2 The separatory section of an HPLC instrument - p. 99 10. 3.3 Detectors in HPLC other than mass spectrometers - p. 108 11. 3.4 Mass spectrometers used as HPLC detectors - p. 122 12. 4. Performance and utilization of high-performance liquid chromatography separation and detection - p. 147 13. 4.1 Parameters related to the characterization of the chromatographic peak - p. 147 14. 4.2 Parameters related to peak separation in HPLC - p. 166 15. 4.3 Parameters for characterization of gradient separation in HPLC - p. 170 16. 4.4 Characterization of detector performance in HPLC - p. 177 17. 4.5 Parameters related to quantitation in HPLC - p. 193 18. 4.6 Utilizations of HPLC in chemical analysis - p. 198 19. 4.7 Utilization of HPLC in nontargeted type analysis - p. 208 20. 5. Samples in high-performance liquid chromatography analysis - p. 219 21. 6. Sample preparation for high-performance liquid chromatography analysis - p. 255 22. 6.1 The role of sample preparation for HPLC - p. 255 23. 6.2 Mechanical processing in sample preparation - p. 261 24. 6.3 Phase transfer techniques used in sample preparation - p. 265 25. 6.4 Liquid–liquid extraction in sample preparation - p. 272 26. 6.5 Liquid–solid extraction in sample preparation - p. 279 27. 6.6 Solid-phase extraction in sample preparation for HPLC - p. 284 28. 6.7 Sample derivatization - p. 288 29. 6.8 Automated sample preparation and on-line coupling with the HPLC - p. 294 30. 7. Chromatographic columns in high-performance liquid chromatography - p. 301 31. 7.1 Construction of an HPLC column - p. 301 32. 7.2 Columns with hydrophobic character - p. 310 33. 7.3 Columns with polar character - p. 328 34. 7.4 Chiral chromatographic columns - p. 331 35. 7.5 Columns used in ion exchange chromatography and related techniques - p. 337 36. 7.6 Chromatographic columns used in size exclusion - p. 344 37. 7.7 Other types of chromatographic columns - p. 346 38. 7.8 Care for the chromatographic column - p. 349 39. 8. Mobile phase and the role of solvents in high-performance liquid chromatography - p. 355 40. 9. Development of a high-performance liquid chromatography analytical method - p. 389 41. 9.1 General plan for adopting or developing an analytical method - p. 389 42. 9.2 Stages in developing a new HPLC analytical method - p. 394 43. 9.3 Development of an RP-HPLC analytical method - p. 399 44. 9.4 Development of an ion-pair HPLC analytical method - p. 431 45. 9.5 Development of an analytical method on polar columns - p. 438 46. 9.6 Development of an HPLC analytical method for the separation of chiral compounds - p. 447 47. 9.7 Development of an analytical method for ion chromatography - p. 460 48. 9.8 Development of an analytical method for size exclusion HPLC - p. 473 49. 9.9 Development of a method for immunoaffinity HPLC - p. 485 50. 9.10 Development of a method for mixed-mode HPLC - p. 489 51. 9.11 Development of a bidimensional HPLC analytical method - p. 495 52. 9.12 Additional issues to consider in developing an HPLC analytical method - p. 498 53. 10. Selection of the best options in developing a high-performance liquid chromatography analytical method - p. 511 54. 10.1 Improvement and optimization processes in an HPLC method - p. 511 55. 10.2 Trial-and-error in method development - p. 521 56. 10.3 Theoretical approaches used for prediction of various parameters in HPLC - p. 524 57. 10.4 Methodologies in HPLC optimization - p. 534 58. 10.5 Computer-assisted method development in HPLC - p. 543 59. 10.6 Selection between different HPLC methods - p. 544 60. 11. Method and data validation in high-performance liquid chromatography analysis - p. 553 61. Appendix 1 - p. 583 62. Index - p. 593
