Amino Acid Analysis Protocols (Methods in Molecular Biology, 159)

Amino acid analysis is a technique that has become commonplace in biotechnology, biomedical, and food analysis laboratories. This book describes a variety of amino acid analysis techniques and how each technique can be used to answer specific biological questions. The first two chapters in Amino Acid Analysis Protocols introduce the concepts, basic theory, and practice of amino acid analysis. The following chapters give detailed instructions on various methods and their applications. As highlighted, there are many different approaches to amino acid ana- sis, but in all cases the results depend heavily on the quality of the sample. Therefore a new way to desalt samples prior to hydrolysis is covered as an introductory chapter (Chapter 3), and most authors have devoted a section to sample preparation, especially to the collection and storage of bodily fluids. Some of the amino acid analysis methods described in this book are based on HPLC separation and analysis after precolumn derivatization. The precolumn derivatization techniques described use (a) 6-aminoquinolyl-N-hydro- succinimidyl carbamate (AQC) (Chapters 4 and 8); (b) 1-fluoro-2- dinitrophenyl-5-L-alanine amide (Marfey's reagent), which allows sepa- tion and analysis of enantiomeric amino acids (Chapter 5); (c) O-phthalal- hyde (OPA) (Chapters 6 and 10); (d) butylisothiocyanate (BITC) and benzylisothiocyante (BZITC) (Chapter 11); (e) phenylisothiocyanate (PITC) (Chapters 12 and 13); (f) ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-s- fonate (SBD-F) (Chapter 17); and (g) 9-fluorenylmethyl-chloroformate (FMOC-Cl) (Chapter 10).

Amino acid analysis is widely used in biotechnology, biomedical, and food analysis laboratories. Amino Acid Analysis Protocols constitutes a major collection of these indispensable analytical techniques, both classic and cutting-edge, of high utility for answering specific biological questions. Common methods include those based on HPLC or gas chromatography separation and analysis after precolumn derivatization. New techniques based on capillary electrophoresis separation, high-performance anion exchange chromatography, and mass spectrometry are also presented. Since results depend heavily on the quality of the sample, most contributors have devoted a section to sample preparation, particularly to the collection and storage of bodily fluids. A new method for desalting samples prior to hydrolysis is also provided. Each method is described in step-by-step detail to ensure successful experimental results, and contains helpful notes on pitfalls to avoid, and variations that enable the methods to be used with different systems.
Up-to-date and highly practical, Amino Acid Analysis Protocols offers analytical and clinical chemists, as well as a broad range of biological and biomedical investigators, a rich compendium of laboratory tools for the productive analysis of both common and uncommon amino acids.