目的 采用6-氨基喹啉-N-(羟基琥珀酰亚胺基)氨基甲酸酯为柱前衍生化试剂,建立了一种柱前衍生高效液相色谱-荧光检测法用于测定血浆中17种氨基酸.方法 使用岛津Inertsil C18柱(250 mm×4.6 mm,5 μm)分析;流动相A为醋酸钠-乙腈混合溶液,流动相B为水∶甲醇∶乙腈(1∶1∶3),梯度洗脱;流速为1.0 mL·min-1;荧光检测,激发波长为235 nm,发射波长为395 nm;柱温为35 ℃.结果 17种氨基酸在0.012～0.36 mmol·L-1线性范围内相关系数均为0.999以上;日内、日间精密度良好(RSD均小于5%),加样回收率为80.0%～112.5%,RSD均小于6.9%(n=3).结论 该衍生化反应可瞬间完成,衍生化产物稳定,测定方法准确、方便、快速,适用于临床定量测定人体血浆中多种氨基酸.%Objective A revcrscd-phase high performance liquid chromatographic method with pre-column derivatization for the determination of amino-acids in human plasma was established. Method Amino-acids was derivatized with 6-aminoquinolyl-N-hydrox-ysuccinimicly carbamate (AQC). The separation was carried out on an Inertsil C18 column (250 mm×4. 6 mm,5 μm) with a gradient clution mode. The cluatc was monitored by a fluorescence detector at an excitation wavelength of 235 nm and an emission wavelength of 395 nm. The mobile phase consisted of A (sodium acetate solution),B ( water, acctonitrile and methanol). Result The linear range was from 0. 012 to 0. 36 mmol · L-1. The intra-day and inter-day relative standard deviations (RSD) for amino-acids were less than 5%(n = 3). The recovery was between 80. 0%-112. 5% with RSD of 6. 9%. Conclusion The reaction of derivatization was fast and the derivatives were stable. The developed method is accurate, simple, fast, and suitable for the clinical measurement.
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