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对氨基酸的反相高效液相色谱的双荧光检测作了介绍。采用邻苯二甲醛 /巯基乙醇(OPA/MCE)与 9-芴甲基氯甲酸酯 (FMOC)柱前衍生 ,同时采用两台串联的荧光检测器对伯氨基酸和仲氨基酸进行分析 ;利用高半胱氨酸 (HCYS) ,高丝氨酸 (HSER) ,正缬氨酸 (NVAL) ,硫代脯氨酸(TPRO)四种内标消除了系统误差。同时 ,由样品各组分保留时间重现性的相对误差及标准氨基酸衍生物的回收率实验来确定该方法的准确性。结果表明 :此检测氨基酸方法灵敏度高、检出限高且重现性好 ,克服了单一衍生剂及单检测分析法的不足 ;消除了NH3 峰对色谱柱影响 ;样品保留时间重现性相对误差 <5 % ,标样回收率 >90 % ,说明该分析方法结果可靠
Dual-fluorescence detection of amino acids by reversed-phase high performance liquid chromatography was introduced. The primary and secondary amino acids were analyzed by OPA / MCE and FMOC pre-column derivatization with two tandem fluorescence detectors. Four internal standards of cysteine (HCYS), homoserine (HSER), norvaline (NVAL) and thioproline (TPRO) eliminated systematic errors. At the same time, the accuracy of this method was determined by the relative error of reproducibility of the retention time of each sample and the recovery experiment of standard amino acid derivatives. The results showed that this method has the advantages of high sensitivity, high detection limit and good reproducibility. It overcomes the deficiencies of single derivatization reagent and single detection assay, eliminates the influence of NH3 peak on the column, relative reproducibility of sample retention time <5%, standard recovery rate> 90%, indicating that the results of the analytical method is reliable