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).

中文翻译：

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高效液相色谱法测定培养液中的二十种蛋白氨基酸和添加剂

摘要

开发了HPLC方法，以确定液体培养基中的二十种蛋白氨基酸，铵，胱氨酸和丙氨酰-谷氨酰胺二肽。使用6-氨基喹啉基-N-羟基琥珀酰亚胺基氨基甲酸酯（AQC）作为荧光标记，用于氨基酸衍生化，随后在反相柱上进行HPLC分离。在程序验证测试中针对每个特定组件评估了特异性，线性，正确性，精密度，分析范围和定量限。氨基酸的分析范围为10–800μM（色氨酸和胱氨酸为20–800μM），定量限不超过5μM（色氨酸为15μM，胱氨酸为8μM）。