Ion-exchange chromatography for the determination of tryptophan

✍ Scribed by Edward J. Robel

• Publisher: Elsevier Science
• Year: 1967
• Tongue: English
• Weight: 445 KB
• Volume: 18
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(67)90098-x

✦ Synopsis

Luven (1) used column ion-exchange chromatography developed by Spackman, Stein, and Moore (2) for the estimation of tryptophan in alkaline hydrolyzates of crude plant proteins. The procedure was time consuming, and due to the analytical difficulties involved variable tryptophan values were obtained. Noltmann, Mahowald, and Kuby (3) used the chromatographic method for the determination of tryptophan in adenosine triphosphate-creatine transphosphorylase hydrolyzed with Ba (OH) 2. In this procedure, the loss of tryptophan from the hydrolyzate by adsorption on BaCO, necessitated an indirect determination for tryptophan by the use of a reference amino acid. Babin et al. ( 4) patterned the determination of tryptophan on the method of Noltmann et al. (3) and modified the accelerated chromatographic analysis developed by Benson and Patterson (5) in order that reference amino acids and tryptophan could be determined on the same chromatogram.

This paper describes a procedure for quantitative recovery of tryptophan from Ba (OH) 2 hydrolyzates and modifications of. the basic amino acid column ion-exchange chromatographic procedure of Spa&man, Stein, and Moore (2). The procedure and modifications provide a direct and rapid method for the analysis of tryptophan in crude and purified proteins.

MATERIALS AND METHODS