HPLC-ECD法测定大鼠血浆中同型半胱氨酸的含量

Determination of Homocysteine Level in Rats' Plasma by HPLC-ECD

目的:建立检测大鼠血浆中同型半胱氨酸(Hcy)含量的HPLC-ECD测定方法。方法:大鼠血浆中的Hcy经二硫苏糖醇还原,色谱系统采用ODS2反相柱,10%甲醇流动相分离,用电化学检测器测定其浓度。结果:Hcy浓度在0.1~100μmol.L-1范围内线性关系良好,相关系数为0.999 3(n=7);样品加样回收率为100.73%,RSD=2.628%;精密度RSD=2.3%,日内稳定性RSD=2.61%,日间稳定性RSD=5.62%。结论:HPLC-ECD法测定血浆中Hcy的含量,操作简便,灵敏度高,稳定性及重复性好,可用于大鼠血浆中Hcy含量的研究。

Objective：To establish a method for assay of homocysteine in rats plasma with High Performance Liquid Chromatography and Electrochemical Detector.Methods：Plasma homocysteine was reduced by dithiothreitol,separated on reverse phase ODS2 column with 10% methanol mobile phase and detected on ESA 5600 electrochemical detector.Results：The method showed excellent linearity over the concentration range 0.1～100 μmol·L^（-1） of Hcy（r=（0.999 3）,n=7）.The recoveries of analytes were from 96.79 %～104.40 %.The precisions,and the stability of the analytes meet the requirements.Conclusion：The established method show high sensitivity and good reproducibility,which make it suitable to determine Hcy level in rats plasma.