血浆同型半胱氨酸高效液相色谱法测定

High-performance Liquid Chromatographic Method for Quantitative Assay of Homocysteine in Plasma

目的 建立高效液相色谱荧光检测法测定血浆同型半胱氨酸 (HCY)的方法。方法 血浆HCY经三丁基磷还原后 ,与巯基特异结合的荧光物质SBD F进行衍化反应。衍化反应产物经反向液相色谱进行分离 ,用荧光检测器测定。结果 所建立的方法HCY浓度在 0～ 45 μmol L范围内与绝对峰面积比呈线性关系 ,回收率为 92 .13%～ 10 8.0 4% ,样内差异变异系数范围 0 .5 %～ 3.1% ,日内差变异系数范围 1.6%～ 3.8% ,日间差变异系数范围 2 .8%～ 4.6%。结论 高效液相色谱 (HPLC)荧光检测法测定血浆HCY ,方法灵敏度高 ,稳定性及重复性好 。

Objective To establish a method for assay of homocysteine in plasma with high-performance liquid Chromatography.Methods plasma homocysteine was reduced by tributylphosphine and derivatized by fluorogenic thoil-specific reagent SBD-F.Derivatives of homocysteine were separated by fluorescence spectrophotometer.Results Calibration curve was linear in the range of 0～45 μmol/L for homocysteine.The recovery was 92.13%～108.04%.Intra-sample coefficient of variation was from 0.5 to 3.1%.Intra-and inter-day coefficients of variation were from 1.6% to 3.8% and from 2.8% to 4.6%.Conclusion The method that we have established has high sensitivity and good reproducibility and may be applied routine clinical use and scientific research.