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中国临床药理学与治疗学 ›› 2011, Vol. 16 ›› Issue (5): 549-552.

• 临床药理学 • 上一篇    下一篇

异硫氰酸苯酯柱前衍生化RP-HPLC法测定人血浆中10种氨基酸的浓度

杨智, 阳利龙, 祝文兵, 曹靖, 何周康   

  1. 湖南省儿童医院药剂科, 长沙 410007,湖南
  • 收稿日期:2011-03-19 修回日期:2011-04-06 发布日期:2011-07-08
  • 通讯作者: 何周康,男,本科,副主任药师,主要从事儿科临床药学工作。Tel:13907317886 E-mail:hnhospital2007@126.com
  • 作者简介:杨智,女,硕士,药师,主要从事儿科临床药学工作。Tel:15974168150 E-mail:yangzhi4837@126.com

Simultaneous determination of 10 amino acids in serum by reversed phase high performance liquid chromatography with pre-column phenylisothiocyanate derivatization

YANG Zhi, YANG Li-long, ZHU Wen-bing, CAO Jing, HE Zhou-kang   

  1. Children's Hospital of Hunan Province, Changsha 410007, Hunan,China
  • Received:2011-03-19 Revised:2011-04-06 Published:2011-07-08

摘要: 目的: 建立一种异硫氰酸苯酯柱前衍生化反相高效液相色谱同时测定人血浆中10种氨基酸的方法。方法: 以正亮氨酸为内标物,异硫氰酸苯酯为柱前衍生剂,Venusil-AA柱为分析柱,柱温 40 ℃,采用二元梯度洗脱,254 nm 波长处检测。结果: 氨基酸血浆浓度在 15.6~250 μmol/L 时,其峰面积与内标物峰面积的比值和氨基酸血浆浓度的线性相关系数,均大于 0.99;10种氨基酸的加样回收率在 96.3%~103.4%;日间和日内变异系数均在 4.8%以下。应用该方法对小儿血浆中氨基酸含量进行了测定,取得了满意的结果。结论: 该法简便、稳定, 并且分离效果好,可用于人血浆中氨基酸浓度的检测。

关键词: 高效液相色谱法, 柱前衍生化, 异硫氰酸苯酯, 正亮氨酸, 氨基酸

Abstract: AIM: To establish a reversed-phase high performance liquid chromatographic method for simultaneous determination of 10 amino acids in serum.METHODS: The sample was mixed with norleucine as the internal standard,then derivatized with phenylisothiocyanate(PITC)and analyzed on a Venusil-AA column at 40 ℃,using gradient elution with detection at 254 nm.RESULTS: The correlation coefficients between the ratios of peak area of amino acid to that of the internal standard and the amino acid concentrations were all above 0.99 in the linear range 15.6-250 μmol/L.The recoveries of amino acids added were 96.3%-103.4%,with-in and between day precision were less than 4.8%.The method was applied to analyze free amino acids in infants plasma and good results were obtained.CONCLUSION: The method is simple, stable, the separation of amino acids is satisfactory.It can be used for determination of amino acids in human plasma.

Key words: HPLC, Precolumn derivatization, Phenylisothiocyanate, Norleucine, Amino acid

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