Amino Acid Analysis of Submicrogram Hydrolyzate Samples

Abstract

Understanding the mechanisms by which diseases occur is a complex process, and often involves the isolation and characterization of minute quantities of rare proteins from living tissue. Amino acid analysis is a vital tool for the elucidation of the structure of such molecules, but today’s researchers must operate at or beyond the utmost limits of conventional methods. In February, 1984 (1) we reported on a new approach to precolumn derivatization and analysis of amino acids which appeared to satisfy all the needs of protein chemists. The approach was based upon the formation of a phenylthiocarbamyl (PTC) derivative of the amino acids (2). The method was shown to be rapid, efficient, sensitive, and specific for the analysis of primary and secondary amino acids in protein hydrolyzates. The method allows for the rapid, bonded phase separation with ultraviolet (UV) detection at 254 nm of the common amino acids with 12 minute analysis time and a one picomole sensitivity for a standard amino acid sample.