Bepaling van aminozuren met hoge druk vloeistofchromatografie. II Derivatisering met dabsylchloride

Abstract

Several pre-column derivatization methods have been reported for the determination of amino acids using high performance liquid chromatography (HPLC). the present report describes the derivatization procedure of amino acids with dabsylchloride and the HPLC analysis of dabsyl-amino acids for a number of applications in our Institute. The dabsyl derivatives show a number of advantages relative to other derivatives, like:- a very good stability of the dabsylderivatives ; - a good reproducibility ; - a good limit of detection of about 3 pmol ; - a complete HPLC separation for all 20 amino acids ; - specific detection at a wavelength of 436 nm. In our Institute, the here described pre-column derivatization with dabsylchloride was applied to a variety of projects in which amino acids are involved, like total amino acids analysis of protein hydrolysates for several applications, monitoring of the consumption of amino acids in animal cell cultures for the production of biologicals, determination of glycine in preparations of anti-thymocyt globulins, the ratio between tryptophan and four other hydrophobic amino acids in the study of pre-menstrual syndrome and the determination of thyronine residues in the thyroid for toxicological investigations. In conclusion, the pre-column derivatization of amino acids with dabsylchloride appears to be a good and practical method with a number of distinct advantages.<br>