Amino Acid Analysis Protocols

2000. Auflage 2000
ISBN: 978-0-89603-656-7
Verlag: Humana Press

Buch, Englisch, Band 159, 265 Seiten, Format (B × H): 160 mm x 237 mm, Gewicht: 581 g

Reihe: Methods in Molecular Biology

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).

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Cooper, Catherine

Fachgebiete

• Naturwissenschaften Chemie Analytische Chemie
• Naturwissenschaften Chemie Organische Chemie Biochemie
• Naturwissenschaften Biowissenschaften Biochemie (nichtmedizinisch)

Inhaltsverzeichnis

Amino Acid Analysis.- Amino Acid Analysis, Using Postcolumn Ninhydrin Detection, in a Biotechnology Laboratory.- Purification of Proteins Using UltraMacro Spin Columns or ProSorb Sample Preparation Cartridges for Amino Acid Analysis.- Amino Acid Analysis Using Precolumn Derivatization with 6-Aminoquinolyl-N-Hydroxysuccinimidyl Carbamate.- Amino Acid Analysis by High-Performance Liquid Chromatography after Derivatization with 1-Fluoro-2,4-Dinitrophenyl-5-L-Alanine Amide (Marfey's Reagent).- The Analysis of Amino Acids Using Precolumn Derivatization, HPLC, and Electrochemical Detection.- Anion Exchange Chromatography and Integrated Amperometric Detection of Amino Acids.- Ion-Pair Chromatography for Identification of Picomolar-Order Protein on a PVDF Membrane.- Capillary Gas Chromatographic Analysis of Protein and Nonprotein Amino Acids in Biological Samples.- Measurement of Blood Plasma Amino Acids in Ultrafiltrates by High-Performance Liquid Chromatography with Automatic Precolumn O-Phthaldialdehyde Derivatization.- Determination of Amino Acids in Foods by Reversed-Phase High-Performance Liquid Chromatography with New Precolumn Derivatives, Butylthiocarbamyl, and Benzylthiocarbamyl Derivatives Compared to the Phenylthiocarbamyl Derivative and Ion Exchange Chromatography.- Amino Acid Measurement in Body Fluids Using PITC Derivatives.- Determination of Proteins, Phosphatidylethanolamine, and Phosphatidylserine in Lipid-Rich Materials by Analysis of Phenylthiocarbamyl Derivatives.- Analysis of O-Phosphoamino Acids in Biological Samples by Gas Chromatography with Flame Photometric Detection.- Determination of Sulfur Amino Acids, Glutathione, and Related Aminothiols in Biological Samples by Gas Chromatography with Flame Photometric Detection.- Capillary Electrophoretic Determination of 4-Hydroxyproline.- Total Plasma Homocysteine Analysis by HPLC with SBD-F Precolumn Derivatization.- Determination of Early Glycation Products by Mass Spectrometry and Quantification of Glycation Mediated Protein Crosslinks by the Incorporation of [14C]lysine into Proteins.