HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF AMINO ACIDS IN COPPER(II) COMPLEX FORM - APPLICATION TO VALINE FERMENTATION SAMPLES

摘要

Reversed-phase liquid chromatography with copper(II) ions was applied to develop a simple and rapid technique for the determination of branched-chain amino acids in fermentation media of industrial microorganisms. In situ detection of amino acids at 240 nm was facilitated by the formation of amino acid-copper complexes. The influence of methanol concentration and the presence of an ion-pair reagent in the eluent on the retention of amino acid-copper complexes and the selectivity of the column was investigated. The proposed technique was used for the detection of traces of the aliphatic amino acids norvaline, isoleucine, norleucine and homoleucine in valine fermentation media and for other tasks. Time of analysis less than 10 min and the possibility of detecting impurities at levels of about 0.1% of the main compound were achieved.