Yao Xiao, Wang Yuting, Chen Gang
Department of Chemistry, Fudan University, Shanghai 200433, People's Republic of China.
Biomed Chromatogr. 2007 May;21(5):520-6. doi: 10.1002/bmc.787.
A method based on capillary electrophoresis with electrochemical detection has been employed for the separation and determination of homocysteine, cysteine, reduced glutathione, ascorbic acid and uric acid. Effects of several important factors such as the acidity and concentration of the running buffer, separation voltage, injection time and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 500 microm diameter platinum disk electrode at a working potential of +1.05 V (vs saturated calomel electrode). The five analytes were well separated within 10 min in a 50 cm long fused silica capillary at a separation voltage of 18 kV in a 100 mm phosphate buffer (pH 7.8). The relation between peak current and analyte concentration was linear over about 3 orders of magnitude with the detection limits (S/N = 3) ranging from 0.83 to 2.58 microm. The proposed method was successfully applied to determine cysteine, reduced glutathione, ascorbic acid and uric acid in human whole blood and rat brain tissues with satisfactory assay results and should find a wide range of bioanalytical applications.
一种基于毛细管电泳-电化学检测的方法已被用于分离和测定同型半胱氨酸、半胱氨酸、还原型谷胱甘肽、抗坏血酸和尿酸。研究了运行缓冲液的酸度和浓度、分离电压、进样时间和检测电位等几个重要因素的影响,以获得最佳条件。检测电极是直径为500微米的铂盘电极,工作电位为+1.05 V(相对于饱和甘汞电极)。在100毫米磷酸盐缓冲液(pH 7.8)中,于18 kV的分离电压下,在50厘米长的熔融石英毛细管中,5种分析物在10分钟内得到了很好的分离。峰电流与分析物浓度之间的关系在约3个数量级范围内呈线性,检测限(S/N = 3)为0.83至2.58微摩尔。该方法成功应用于测定人全血和大鼠脑组织中的半胱氨酸、还原型谷胱甘肽、抗坏血酸和尿酸,测定结果令人满意,应具有广泛的生物分析应用前景。