[高效液相色谱-荧光检测法快速分析血清中的色氨酸]

[Rapid analysis of tryptophan in serum by high performance liquid chromatography with fluorescence detection].

作者信息

Wang Qing-ping, Tang Ai-guo

机构信息

Department of Clinical Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.

出版信息

Se Pu. 2002 Jan;20(1):52-5.

PMID:12541620

Abstract

A sensitive, rapid and specific method for the determination of tryptophan(Trp) in serum by high performance liquid chromatography with fluorescence detection (HPLC-FLD) has been established. Serum sample was precipitated by 5% perchloric acid solution and centrifuged to remove protein and then assayed by HPLC-FLD. The operating conditions were Nova-Pak C18 column (3.9 mm i.d x 150 mm, 4 microns), 5 mmol/L KH2PO4 as the mobile phase at a flow rate of 1.0 mL/min. The fluorescence detector was operated at lambda ex 254 nm and lambda em 338 nm. The method was proved to be linear in the range of 0.49 mumol/L-490.00 mumol/L with a regression coefficient of 0.9999. The minimum detection limit was 0.10 mumol/L. The recoveries were 97.2%-98.6%. The intra-batch and inter-batch RSDs were 2.0% and 3.2% respectively. The detection time of serum Trp was within 5 min after injection. The results suggest that this method is simple, fast, accurate and convenient.

摘要

建立了一种采用高效液相色谱-荧光检测法（HPLC-FLD）测定血清中色氨酸（Trp）的灵敏、快速且特异的方法。血清样本用5%高氯酸溶液沉淀，离心去除蛋白质后，采用HPLC-FLD进行测定。操作条件为Nova-Pak C18柱（内径3.9 mm×150 mm，4微米），以5 mmol/L磷酸二氢钾作为流动相，流速为1.0 mL/min。荧光检测器在激发波长254 nm和发射波长338 nm下运行。该方法在0.49 μmol/L - 490.00 μmol/L范围内呈线性，回归系数为0.9999。最低检测限为0.10 μmol/L。回收率为97.2% - 98.6%。批内和批间相对标准偏差分别为2.0%和3.2%。血清Trp的检测时间在进样后5分钟内。结果表明该方法简单、快速、准确且方便。