Amino acid analysis of mammalian cell culture medium by liquid chromatography with UV and fluorescence detection and derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate

摘要: Most amino acids do not have a strong UV chromophore, which impedes detection by UV. They are similar in structure, which complicates chromatographic separation. Selective modification of the amine functional group of amino acids with a chromophore and/or fluorophore differentiates them from the matrix. Furthermore, the modification facilitates chromatographic separation because of the addition of a hydrophobic moiety that improves retention by reversed phase of the otherwise poorly retained amino acids. A powerful reagent for pre-column derivatization is 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) because of the stability of AQC-derivatized amino acids and the reproducibility of the derivatization reaction with both primary and secondary amino acids. 1The AQC reagent reacts with primary and secondary amino acids to form stable UV-active and fluorescent derivatives. In this method, manual …