Ultra–High Performance Liquid Chromatography and Its Applications

Explores both the benefits and limitations of new UHPLCtechnology

High performance liquid chromatography (HPLC) has been widelyused in analytical chemistry and biochemistry to separate,identify, and quantify compounds for decades. The science of liquidchromatography, however, was revolutionized a few years ago withthe advent of ultra–high performance liquid chromatography (UHPLC),which made it possible for researchers to analyze sample compoundswith greater speed, resolution, and sensitivity.

Ultra–High Performance Liquid Chromatography and ItsApplications enables readers to maximize the performance ofUHPLC as well as develop UHPLC methods tailored to their particularresearch needs. Readers familiar with HPLC methods will learn howto transfer these methods to a UHPLC platform and vice versa. Inaddition, the book explores a variety of UHPLC applicationsdesigned to support research in such fields as pharmaceuticals,food safety, clinical medicine, and environmental science.

The book begins with discussions of UHPLC method development andmethod transfer between HPLC and UHPLC platforms. It then examinespractical aspects of UHPLC. Next, the book covers:

Coupling UHPLC with mass spectrometry Potential of shell particles in fast liquid chromatography Determination of abused drugs in human biological matrices Analyses of isoflavones and flavonoids Therapeutic protein characterization Analysis of illicit drugs

The final chapter of the book explores the use of UHPLC in drugmetabolism and pharmacokinetics studies for traditional Chinesemedicine.

With its frank discussions of UHPLC′s benefits and limitations,Ultra–High Performance Liquid Chromatography and ItsApplications equips analytical scientists with the skills andknowledge needed to take full advantage of this new separationtechnology.

Preface vii

Contributors ix

1. UHPLC Method Development 1
Shujun Chen and Alireza Kord

2. Method Transfer Between HPLC and UHPLC Platforms 31
Gregory K. Webster, Thomas F. Cullen, and Laila Kott

3. Practical Aspects of Ultrahigh Performance LiquidChromatography 55
Naijun Wu, Christopher J. Welch, Theresa K. Natishan, Hong Gao,Tilak Chandrasekaran, and Li Zhang

4. Coupling UHPLC with MS: The Needs, Challenges, andApplications 95
Julie Schappler, Serge Rudaz, Jean–Luc Veuthey, and DavyGuillarme

5. The Potential of Shell Particles in Fast LiquidChromatography 133
Szabolcs Fekete and Jeno Fekete

6. UHPLC Determination of Drugs of Abuse in Human BiologicalMatrices 169
Fabio Gosetti, Eleonora Mazzucco, and Maria CarlaGennaro

7. UHPLC in the Analyses of Isoflavones and Flavonoids 197
Sylwia Magiera and Irena Baranowska

8. UHPLC for Characterization of Protein Therapeutics 235
Jennifer C. Rea, Yajun Jennifer Wang, and Taylor Zhang

9. UHPLC/MS Analysis of Illicit Drugs 253
Guifeng Jiang, Jason R. Stenzel, Ray Chen, and DiabElmashni

10. Ultra–High Performance Liquid Chromatography MassSpectrometry and Its Application 271
Zhili Xiong, Ying Deng, and Famei Li

Index 291

QUANYUN ALAN XU, PhD, is a Senior Research Scientist in the Pharmacology and Analytical Laboratory Facility at the University of Texas MD Anderson Cancer Center. Dr. Xu is the coauthor of Analytical Methods for Therapeutic Drug Monitoring and Toxicology as well as more than seventy scientific papers and book chapters.