Determination of Twenty Proteinogenic Amino Acids and Additives in Cultural Liquid by High-Performance Liquid Chromatography
А. Д. Аскретков, A. A. Klishin, Dmitry Zybin, Н. В. Орлова, A. V. Kholodova, N. V. Lobanova, Yu. A. Seregin
Abstract
Abstract An HPLC procedure was developed to determine twenty proteinogenic amino acids, ammonium, cystine, and alanyl-glutamine dipeptide in liquid culture media. 6-Aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) was used as a fluorescent label for the derivatization of amino acids with the subsequent HPLC separation on a reversed-phase column. Specificity, linearity, correctness, precision, analytical range, and limit of quantification were evaluated for each particular component in the procedure validation tests. The analytical range for amino acids was 10–800 μM (20–800 μM for tryptophan and cystine), and the limit of quantification was no higher than 5 μM (15 μM for tryptophan and 8 μM for cystine).