高效液相-荧光检测法快速测定血清中的色氨酸

 利用色氨酸具有自然荧光的特点 ,建立了测定血清中色氨酸浓度的高效液相 荧光检测分析方法。血清标本经高氯酸沉淀蛋白后取上清液进行分析 ,柱为Nova PakC18柱 ,流动相为 5mmol/L磷酸二氢钾水溶液 ,流速为 1mL/min ,激发波长为 2 5 4nm ,荧光检测波长为 338nm。该法检测色氨酸浓度的线性范围为 0 4 9μmol/L～ 4 90 0 0 μmol/L ,最低检测限为 0 10 μmol/L ,回收率为 97 2 %～ 98 6 % (平均值为 97 9% ) ,批内、批间相对标准偏差均小于 5 % ,苯丙氨酸、酪氨酸、5 羟色胺和 5 羟吲哚乙酸等物质对该法均无干扰。

Rapid Analysis of Tryptophan in Serum by High PerformanceLiquid Chromatography with Fluorescence Detection

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.