将荧光试剂2,3-二甲醛基喹喔啉首次应用到氨基酸的高效液相色谱分离分析中.考察了该荧光试剂与伯胺氨基酸衍生反应的pH值、比例、反应时间等影响因素,确定衍生反应的最佳条件为:100 mmol/L硼砂缓冲液(pH 9.5),二巯基乙醇、衍生试剂与氨基酸的比例为9∶3∶1,室温下反应5 min.采用C18色谱柱,经二元梯度洗脱,荧光检测,17种伯胺蛋白氨基酸得到较好分离.氨基酸的浓度在0.01 ～1.00 mmol/L范围内与其衍生物峰面积呈较好的线性关系,氨基酸衍生物的检出限为0.02～0.07 μg/L.应用该方法对麦冬中游离氨基酸的含量进行测定,结果满意.此方法快速,高效,灵敏度高,精密度好.%A fluorescence reagent quinoxaline-2, 3-dicarboxaldehyde was firstly applied in the sepa-ration of amino acids by high performance liquid chromatography ( HPLC). The reaction of primary a-mine amino acids with fluorescence reagent was studied in detail. The best conditions for the reac-tion; buffer; pH 9. 5 100 mmol/L borate, a mole ratio of 2-mercapto ethanol, fluorescence reagent and amino acid: 9 : 3 : 1, reaction time: 5 min at room temperature. Under the optimal condi-tions , seventeen derivatives were detected with fluorescence detector after separation on a C18 column by gradient elution. The calibration curves were linear over the range of 0. 01 - 1. 00 mmol/L for all derivatives, with detection limits of 0. 02 - 0. 07 μg/L. This method was simple, rapid and sensi-tive , and was used for the measurement of the concentration of amino acid in ophiopogon of the Chi-nese traditional medicines with satisfactory results.
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