在这里，描述了同时检测细胞培养基中通常存在的20种氨基酸的逐步方案。该方案通过使用邻苯二甲醛和9-芴基甲基氯甲酸酯作为衍生剂的两步柱前衍生策略促进了一级和二级氨基酸的检测。通过反相色谱分离具有不同疏水性的衍生氨基酸。通过使用338和262nm的可变波长检测器在单个工作流程中同时检测氨基酸。该方案适用于哺乳动物和细菌细胞培养基质，具有柱前衍生自动化的选项。
Herein, a step-by-step protocol for simultaneous detection of 20 amino acids commonly present in cell culture media is described. The protocol facilitates detection of both primary and secondary amino acids through a two-step precolumn derivatization strategy using ortho-phthalaldehyde and 9-fluorenylmethyl chloroformate as derivatizing agents. The separation of derivatized amino acids with varying hydrophobicity is achieved through reverse-phase chromatography. The amino acids are simultaneously detected in a single workflow through the use of Variable Wavelength Detector at 338 and 262 nm. The protocol is applicable for both mammalian and bacterial cell culture matrices with an option for automation of precolumn derivatization.
