(U)HPLC Method Development Unveiled
“Any sufficiently advanced technology is indistinguishable from magic,”
Arthur C. Clarke famously said.
This sentiment is particularly relevant in the realm of (U)HPLC method development, where skilled specialists are often regarded as modern-day magicians.
But is this perception justified? Can a regular QC HPLC chemist swiftly and effectively master the intricacies of HPLC method development?
These questions have intrigued me for a long time, leading me to undertake the creation of this book. The result of that endeavor is now in your hands.
This book is a guide, but it will only be as useful as the effort and practice you put into it. Now you can try; now you can dare. I wish you good luck on this journey and all the best.
Sincerely yours,
The Author
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Table of Contents
1. Three step workflow of HPLC method development
1.1. Three factors influencing chromatographic resolution and the corresponding steps in HPLC method development
1.2. Optimal retention factor range and optimal HPLC separation selectivity
2. LC (Liquid Chromatography) modes
2.1. What is LC mode?
2.2. Exclusion LC modes
2.3. Adsorption LC modes
2.4. Forecasting analyte behavior in any LC mode using specialized diagrams
2.5. Mixed LC modes
3. Fine-tuning retention and HPLC separation selectivity
3.1. Retention models
3.2. Retention model of RP mode
3.3. Retention model of NP mode
3.4. Retention model of IC mode
3.5. Retention model of HILIC mode
3.6. Dynamic modification for mixed-mode separations. Ion pairing mode
3.7. Retention model of CT mode
3.8. Comprehensive overview of contemporary HPLC stationary phase chemistries
3.9. Switching modes on and off. Fine-tuning mode balance in mixed modes
4. Selecting an appropriate LC mode
4.1. Advantages and disadvantages of RP chromatography
4.2. Advantages and disadvantages of HILIC chromatography
4.3. Advantages and disadvantages of Ion Chromatography
4.4. Advantages and disadvantages of CT chromatography
5. Specificity and robustness verification during HPLC method development
6. Qualification of RP, HILIC and IC HPLC columns
6.1. Design of tests for HPLC column
6.2. Conventional HPLC packing efficiency tests
6.3. A specialized two-parameter test designed to ascertain the chemistry of all types of reversed-phase HPLC packing materials
6.4. Evaluating the chemical inertness of RP packings
6.5. A specialized two-parameter test designed to ascertain the chemistry of all types of HILIC and silica-based ion-exchange HPLC packing materials
7. Principles governing chiral HPLC
7.1. Primary types of chiral stationary phases
7.2. Patterns in chiral HPLC
7.3. Structural characteristics of challenging chiral compounds
7.4. Enhancing the selectivity of chiral separation in NP mode
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