高效液相色谱法测定人血浆中同型半胱氨酸浓度

目的建立柱前衍生化反相高效液相色谱-荧光检测法测定血浆中同型半胱氨酸(Hcy)浓度的方法。方法以tris-(2-carboxylethyl)-phosphine hydrochloride(TCEP)为还原剂,以7-fluorbenzo-2-oxa-1,3-diazole-4-sulfonate(SBD-F)为衍生剂,色谱柱为Xterra RP18柱,柱温35℃,流动相为甲醇︰磷酸二氢钠缓冲液(pH值3.0)=3︰97,激发波长380 nm,发射波长510 nm,外标法定量。结果 Hcy浓度在1.95～125μmol/L范围内线性关系良好(r=0.999 8)。日内和日间相对标准偏差均<7%,方法回收率为103.2%～111.9%。结论此方法准确、灵敏、快速,是一种适合于实验室研究和临床检测血浆中Hcy浓度的方法。

Determination of Homocysteine Concentration in Plasma by High Performance Liquid Chromatography

Objective To establish a high performance liquid chromatography and fluorescence detection method for determining homocysteine(Hcy) concentration in human plasma.Methods Hcy was reduced by tris-(2-carboxylethyl)-phosphine hydrochloride(TCEP),then derived by 7-fluorbenzo-2-oxa-1,3-diazole-4-sulfonate(SBD-F).Xterra RP18 column was used;the mobile phase was composed of methanol-phosphate buffer solution(pH 3.0)(3︰97);detection wavelength was set at λex 380 nm and λex 510 nm.Results A linearity was obtained from 1.95 to 125 μmol/L(r=0.999 8) for Hcy.The recovery rates were between 103.2% and 111.9%.The intra-day and inter-day precisions were less than 7%.Conclusion The method is accurate,sensitive and rapid for the determination of Hcy concentration in human plasma.