Mendoza Jorge, Soto Paula, Ahumada Inés, Garrido Tatiana
Departamento de Química Inorgánica y Analítica, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Olivos 1007, Casilla 233, Santiago, Chile.
Electrophoresis. 2004 Mar;25(6):890-6. doi: 10.1002/elps.200305759.
A rapid method of capillary zone electrophoresis is described to determine the oxidized (GSSG) and reduced (GSH) form of glutathione in plant tissue. In order to separate both analytes in a fused-silica capillary, the pH and composition of the electrolyte solution were optimized. The electrolyte composition was 100 mmol/L, borate 25 mmol/L Tris, and 0.2% w/v metaphosphoric acid (MPA), pH 8.2. Some instrumental conditions used to run the samples were hydrostatic injection for 30 s, 30 kV applied voltage, and UV detection (185 nm) at 25 degrees C. Linearity and useful range obtained for the calibration curves were optimum, with correlation coefficients about 0.999 in the 0-120 micromol/L range. The migration time was highly reproducible, less than 5 min being afforded to run a sample. Electrolyte buffer and samples required a careful pH control for optimal separation of both analytes. This aspect constitutes a critical analytical step when acids are used in the procedure for sample preparation. Simultaneous analysis of GSH and GSSG may provide a useful tool for comparative studies of plants in order to select those species with a potential capacity for detoxification from toxic elements or those appearing promising from phytoremediation for these elements.
本文描述了一种快速的毛细管区带电泳方法,用于测定植物组织中谷胱甘肽的氧化形式(GSSG)和还原形式(GSH)。为了在熔融石英毛细管中分离这两种分析物,对电解质溶液的pH值和组成进行了优化。电解质组成为100 mmol/L硼酸盐、25 mmol/L Tris和0.2%(w/v)偏磷酸(MPA),pH值为8.2。运行样品时使用的一些仪器条件为:静水压进样30 s、施加电压30 kV以及在25℃下进行紫外检测(185 nm)。校准曲线的线性和有效范围最佳,在0 - 120 μmol/L范围内相关系数约为0.999。迁移时间具有高度重现性,运行一个样品所需时间不到5分钟。电解质缓冲液和样品需要仔细控制pH值,以实现两种分析物的最佳分离。当在样品制备过程中使用酸时,这一方面构成了关键的分析步骤。同时分析GSH和GSSG可为植物的比较研究提供有用工具,以便选择那些具有从有毒元素解毒潜在能力的物种,或那些在植物修复这些元素方面有前景的物种。