Liu Jifeng, Cao Weidong, Qiu Haibo, Sun Xiuhua, Yang Xiurong, Wang Erkang
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, People's Republic of China.
Clin Chem. 2002 Jul;48(7):1049-58.
Capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)3(2+)]-electrogenerated chemiluminescence (ECL) detection is a promising method for clinical analysis. In this study, a method combining CE with Ru(bpy)3(2+) ECL (CE-ECL) detection that can be applied to amine-containing clinical species was developed, and the performance of CE-ECL as a quantitative method for determination of sulpiride in human plasma or urine was evaluated.
Sulpiride was separated by capillary zone electrophoresis in uncoated fused-silica capillaries [50 cm x 25 microm (i.d.)] filled with phosphate buffer (pH 8.0) and a driving voltage of +15 kV, with end-column Ru(bpy)3(2+) ECL detection. A platinum disc electrode was used as working electrode. Sulpiride in human plasma or urine samples (100 microL) was extracted by a double-step liquid-liquid extraction procedure, dried under nitrogen at 35 degrees C in a water bath, and reconstituted with 100 microL of filtered water. The extraction solvent was ethyl acetate-dichloromethane (5:1 by volume).
Under optimum conditions (pH 8.0 phosphate buffer, injection for 6 s at 10 kV, and +1.2 V as detection potential), separation of sulpiride was accomplished within 4 min. The calibration curve was linear over a concentration range of 0.05-25.0 micromol/L, and the limit of detection was 2.9 x 10(-8) mol/L for sulpiride. Intra- and interday CVs for ECL intensities were <6%. Extraction recoveries of sulpiride were 95.6-101% with CVs of 2.9-6.0%. The method was clinically validated for patient plasma and urine samples.
CE combined with Ru(bpy)3(2+) ECL is reproducible, precise, selective, and enables the analysis of sulpiride in human plasma and urine. It thus is of value for rapid and efficient analysis of amine-containing analytes of clinical interest.
采用三（2,2'-联吡啶）钌（II）[Ru(bpy)₃²⁺] - 电致化学发光（ECL）检测的毛细管电泳（CE）是一种很有前景的临床分析方法。在本研究中，开发了一种将CE与Ru(bpy)₃²⁺ ECL（CE - ECL）检测相结合的方法，该方法可应用于含胺类临床样本，并评估了CE - ECL作为测定人血浆或尿液中舒必利的定量方法的性能。
在未涂层熔融石英毛细管[50 cm×25 µm（内径）]中，以磷酸盐缓冲液（pH 8.0）为填充液，+15 kV的驱动电压，通过毛细管区带电泳分离舒必利，并采用柱端Ru(bpy)₃²⁺ ECL检测。使用铂盘电极作为工作电极。人血浆或尿液样本（100 µL）中的舒必利通过两步液 - 液萃取程序进行萃取，在35℃水浴中用氮气吹干，并用100 µL过滤水复溶。萃取溶剂为乙酸乙酯 - 二氯甲烷（体积比5:1）。
在最佳条件下（pH 8.0磷酸盐缓冲液，10 kV下进样6 s，检测电位为 +1.2 V），4分钟内完成舒必利的分离。校准曲线在0.05 - 25.0 µmol/L浓度范围内呈线性，舒必利的检测限为2.9×10⁻⁸ mol/L。ECL强度的日内和日间变异系数均<6%。舒必利的萃取回收率为95.6 - 101%，变异系数为2.9 - 6.0%。该方法在患者血浆和尿液样本中得到临床验证。
CE与Ru(bpy)₃²⁺ ECL相结合具有可重复性、精确性、选择性，能够分析人血浆和尿液中的舒必利。因此，它对于快速高效分析具有临床意义的含胺类分析物具有重要价值。
