中性毛细管电泳电化学发光法检测血清中的同型半胱氨酸和还原型谷胱甘肽

Neutral capillary electrophoresis with electrochemiluminescence sensing for determination of homocysteine and reduced glutathione assay in human serum

生物硫醇(BT)是动脉粥样硬化等心脑血管疾病的独立危险因子。采用中性毛细管电泳电化学发光法(CE-ECL)检测血清中2种心脑血管疾病BT类标志物,同型半胱氨酸(DL-Hcy)和还原型谷胱甘肽(L-GSH)。基于DL-Hcy和L-GSH对石墨烯/联吡啶钌的电化学发光信号增敏作用,考察了缓冲溶液pH、分离电压、进样时间对DL-Hcy和L-GSH分离的影响。结果显示:DL-Hcy和L-GSH在0.1 mol/L磷酸盐缓冲溶液(pH 7.0)中经10 s进样、12 kV电压分离,于600 s内获得理想的分离检测。DL-Hcy的检测线性范围为5~200μmol/L,线性方程为y=643.9+12.8x(R^(2)=0.9817),检出限为3.2μmol/L;L-GSH的检测线性范围为10~200μmol/L,线性方程为y=144.3+8.1x(R^(2)=0.9831),检出限为8.3μmol/L。加标回收率为96.1%~106.0%,测定值的相对标准偏差均小于9.6%。中性CE-ECL方法可实现血清中DL-Hcy和L-GSH等2种生物硫醇的高效分离检测。

Biothiol is an independent risk factor for cardiac-cerebral vascular disease such as atherosclerosis.The neutral capillary electrophoresis matrixed with electrochemiluminescence(CE-ECL)detection method was proposed for the simultaneous determination of two vital biothiols biomarkers of cardiovascular disease,based on the sensibilization behavior of homocysteine(DL-Hcy)and reduced glutathione(L-CSH)for ECL signal of graphene/bipyridinium ruthenium.The effects of buffer pH,separation voltage and injection time on the separation of DL-Hcy and L-GSH were optimized.The DL-Hcy and L-CSH were successfully separated by 10 s injection with the separation voltage of 12 kV in 0.1 mol/L phosphate buffer(pH 7.0)within 600 s.The linear relationships revealed sensitive ECL intensity response in the concentration range of 5-200μmol/L for DL-Hcy(y=643.9+12.8x,R^(2)=0.9817)and 10-200μmol/L for L-GSH assay(y=144.3+8.1x,R^(2)=0.9831).The detection limits were 3.2μmol/L for DL-Hcy and 8.3μmol/L for L-CSH,with the relative standard deviations less than 9.6%,and the recoveries were in the range of 96.1%-106.0%.The proposed CE-ECL method was successfully employed to detect DL-Hcy and L-CSH in human serum.