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高效液相色谱-荧光检测法测定血清中的犬尿氨酸
引用本文:罗昔波,唐爱国,皮兰敢,肖乐东,敖翔,卜艳红,王瑞.高效液相色谱-荧光检测法测定血清中的犬尿氨酸[J].色谱,2008,26(1):60-63.
作者姓名:罗昔波  唐爱国  皮兰敢  肖乐东  敖翔  卜艳红  王瑞
作者单位:Department of Clinical Laboratory, the Second Xiangya Hospital of Central ;South University, Changsha 410011, China
基金项目:湖南省医药卫生科研项目 , 湖南省科技基金
摘    要:建立了高效液相色谱-荧光检测(HPLC-FLD)测定血清中犬尿氨酸(kynurenine,Kyn)含量的方法。采用Hypersil C8色谱柱(300 mm×6.0 mm,10 μm), 流动相为0.25 mol/L醋酸锌及50 mmol/L醋酸溶液(含3%乙腈),流速为1.5 mL/min, 荧光检测激发波长和发射波长分别为365 nm和480 nm。血清标本经5%(v/v)高氯酸溶液去除蛋白质后取上层清液直接进样, Kyn经流动相等度洗脱分离后,用FLD进行测定。研究结果表明Kyn保留时间约为8.3 min,线性范围为0.098~19.6 μmol/L,最低检出浓度为0.04 μmol/L,回收率为90.8%~96.2%,日内、日间测定的相对标准偏差均小于5%,苯丙氨酸、酪氨酸、色氨酸、5-羟色胺和犬尿喹啉酸等物质对犬尿氨酸的测定均无干扰。建立的方法简便、快速、灵敏、特异,适用于临床和科研应用。

关 键 词:高效液相色谱法  荧光检测  犬尿氨酸  人血清
文章编号:1000-8713(2008)01-0060-04
收稿时间:2007-06-18
修稿时间:2007年6月18日

Determination of kynurenine in serum using high performance liquid chromatography-fluorescence detection
LUO Xibo,TANG Aiguo,PI Langan,XIAO Ledong,AO Xiang,PU Yanhong,WANG Rui.Determination of kynurenine in serum using high performance liquid chromatography-fluorescence detection[J].Chinese Journal of Chromatography,2008,26(1):60-63.
Authors:LUO Xibo  TANG Aiguo  PI Langan  XIAO Ledong  AO Xiang  PU Yanhong  WANG Rui
Institution:Department of Clinical Laboratory, the Second Xiangya Hospital of Central ;South University, Changsha 410011, China
Abstract:A sensitive method of high performance liquid chromatography-fluorescence detection (HPLC-FLD) for the determination of kynurenine (Kyn) in serum was developed. A 20 microL supernatant of a serum sample deproteinized by 5% perchloric acid solution was assayed and separated on a Hypersil C8 column (300 mm x 6.0 mm, 10 microm) with an isocratic elution of 0.25 mol/L zinc acetate-50 mmol/L acetate containing 3% (v/v) acetonitrile. The fluorescence excitation and emission wavelengths were 365 nm and 480 nm, respectively. The limit of detection was approximately 0.04 micromol/L (signal-to-noise ratio of 3) and the linearity of the assay was from 0.098 micromol/L to 19.6 micromol/L. The relative standard deviations of intraday and interday determinations were 3.8% and 4.6%, respectively. The results indicated tryptophan (Trp), 5-hydroxytryptamine (5-HT), kynurenic acid (KYNA), phenylalanine (Phe), tyrosine (Tyr) and creatinine (Cr) had no interfering effects to the determination of Kyn. The method is of high accuracy, easy operation, satisfactory recoveries and good reproducibility, and can be used for routine analysis.
Keywords:high performance liquid chromatography (HPLC)  fluorescence detection (FLD)  kynurenine  human serum
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