Analysis of amino acids in human serum by isocratic reverse-phase high-performance liquid chromatography with electrochemical detection
Analysis of amino acids in human serum by isocratic reverse-phase high-performance liquid chromatography with electrochemical detection
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DOI：
10.1016/s0021-9673(00)01206-1
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发表时间：
2001-04-13
影响因子：
4.1
通讯作者：
Krasnova, IN
中科院分区：
文献类型：
--
作者：
Tcherkas, YV;Kartsova, LA;Krasnova, IN
建立了一种简便、灵敏、重现性好的等度高效液相色谱法测定人血清中氨基酸的方法。该方法包括用甲醇沉淀血清蛋白，然后用邻苯二甲醛-2-巯基乙醇或邻苯二甲醛-亚硫酸钠柱前衍生氨基酸。衍生物的HPLC分离使用具有等度移动的相系统和电化学检测（+0.75 V）的ODS柱进行。对于所有氨基酸，响应在5-300 μ M范围内呈线性。该方法允许定量测定谷氨酸、天冬酰胺、丝氨酸。谷氨酰胺、组氨酸、牛磺酸、丙氨酸、精氨酸、甲硫氨酸、异亮氨酸、鸟氨酸、亮氨酸、苯丙氨酸、赖氨酸和色氨酸，浓度低至0.5-5.0 pmol（信噪比=2）。用此方法测定了健康人和缺血性脑卒中患者血清中氨基酸的含量。(C)2001爱思唯尔科技有限公司。保留所有权利。
A simple, sensitive and reproducible isocratic high-performance liquid chromatography (HPLC) method has been developed for the determination of amino acids in human serum. The method involves precipitation of the serum proteins with methanol followed by pre-column derivatization of amino acids with o-phthalaldehyde-2-mercaptoethanol or o-phthalaldehyde-sodium sulfite. HPLC separation of the derivatives was performed using an ODS column with an isocratic mobile phase system and electrochemical detection (+0.75 V). The response was linear over the range 5-300 muM for all amino acids. The method allows quantitative determination of glutamic acid, asparagine, serine. glutamine, histidine, taurine, alanine, arginine, methionine, isoleucine, ornithine, leucine, phenylalanine, lysine and tryptophan at concentrations as low as 0.5-5.0 pmol (signal-to-noise ratio=2). Using this method, the levels of amino acids in serum from healthy donors and patients with ischemic stroke were determined. (C) 2001 Elsevier Science B.V. All rights reserved.