高效液相色谱-电化学检测法测定鼠脑微透析液中谷胱甘肽和半胱氨酸

采用高效液相色谱法，用电化学检测手段成功地测定了沙土鼠脑纹状体微透析液内的谷胱甘肽（ＧＳＨ）和半胱氨酸（Ｃｙｓ）。检测前用电化学自清洗方式能有效地提高玻璃碳电极的检测水平，使其具有较好的重现性和回收率。Ｃｙｓ和ＧＳＨ的回收率分别为９１．４％和８７．３％。电化学检测的工作电压为０．９Ｖ，用外标法定量。由微透析探针获取的纹状体透析液Ｃｙｓ和ＧＳＨ的浓度分别为６８．５９和６５．５４ｎｍｏｌ／Ｌ，验证了方法的实用性和可靠性。

Quantitative Determination of Extracellular Glutathione and Cysteine in Gerbil Brain by Microdialysis and High Performance Liquid Chromatography with Electrochemical Detection

A method for the determination of glutathione and cysteine in microdialysate by HPLC with electrochemical detector has been described. The column used was Hypersil ODS, 5 microns, 200 mm x 4.6 mm i.d. and mobile phase was 50% 0.2 mol/L KCl, 10.6% 0.2 mol/L HCl, 39.4% methanol and 0.1 mmol/L EDTA (pH 2.0) with a flow rate of 1 mL/min. The microdialysis probe (membrane: pc, length: 10 mm, diameter: 0.5 mm/CMA) was inserted into the right striatum of gerbil at the following coordinates: 1.5 mm anterior and 1.5 mm lateral to bregma, and 3.5 mm below dura. Electrochemical pretreatment of a glassy carbon electrode used as a working electrode in an electrochemical detector has been found to enhance the analytical capability of detector for the determination of glutathione and cysteine. The quantitative determination was made with an electrochemical detector at 0.9 V and by external standard method. The method gave good recoveries and reproducibilities. The average recoveries of two analytes were 91.4% and 87.3% respectively. Extracellular fluid glutathione and cysteine concentrations in the striatum of gerbil brain were determined by the microdialysis and chromatography described in this study. The contents of Cys and GSH in microdialysate of gerbil striatum were 65.54 +/- 12.96 nmol/L and 68.59 +/- 13.23 nmol/L. The results were comparable with those reported in previous works.