Evaluation of liquid chromatographic analysis of nutritionally important amino acids in food and physiological samples

Abstract

Significant advances have been made in standardizing methods for amino acid analysis of foods. The methods included standardized hydrolysis of the food proteins followed by separation and quantitation of the released amino acids by ion-exchange chromatography (IEC). IEC is still the main method in use. Its use is, however, being replaced by the faster higher-performance liquid chromatographic (HPLC) methods of derivatized amino acids. The HPLC separation of precolumn phenylisothiocyanate (PITC) derivatives has been adapted for rapid analysis of all amino acids in protein hydrolysates (12 min) and nutritionally important amino acids in deproteinized physiological samples (20 min). The inter-laboratory variability of the PITC derivatization method has not been determined although the intra-laboratory variation of the HPLC method was found to be similar to that of IEC. When similar hydrolytic conditions were used in preparing protein hydrolysates, amino acid results obtained with the PITC derivatization method were generally in close agreement with those obtained by IEC. There is, however, room for improvement in the HPLC analysis of amino acids in physiological samples.