毛细管区带电泳法测定还原型谷胱甘肽中的有关物质

目的: 建立测定还原型谷胱甘肽中的有关物质的毛细管区带电泳方法。方法: 采用未涂层熔融石英毛细管(60 cm×75 μm,有效长度50 cm);以50 mmol.L^-1的磷酸二氢钠溶液(pH1.8),添加10％甲醇为运行缓冲液;运行电压为20 kV,柱温25℃;检测波长200 nm;压力进样3.45 kPa。结果: 还原型谷胱甘肽中4个已知杂质半胱氨酰甘氨酸、γ-谷氨酰半胱氨酸、氧化型谷胱甘肽和半胱氨酸,杂质峰/内标峰校正峰面积精密度(RSD)分别为0.57％,1.24％,0.58％,1.56％(n=5),最低检测浓度分别为0.0263,0.0234,0.0099,0.0257 mg.mL^-1;在0.005~0.2 mg.mL^-1范围内与各杂质峰/内标峰校正峰面积呈现良好线性(r分别为0.9996,0.9992,0.9998,0.9983);结论: 方法快速、简单且准确,可用于还原型谷胱甘肽中有关物质的测定。

Objective: To establish a CZE method for determination of related substances in reduced glutathione. Methods: A uncoated fused capillary(60 cm×75 μm,50 cm to the detector) was used. The electrophoresis buffer was 50 mmol.L^-1sodium dihydrogen phosphate(pH1.8),add 10% methanol. The separation voltage was 20 kV, column temperature 25℃, detection wavelength 200 nm,pressure injection 3.45 kPa. Results: The precision of Cys-Gly,γ-Clu-Cys, GSSG, Cys was good (RSD= 0.57%,1.24%,0.58%,1.56%, respectively;n=5). The detection limits were 0.0263,0.0234,0.0099,0.0257 mg·mL^-1, respectively. The calibration curve of these compounds was linear in the range of 0.005-0.2 mg·mL^-1(r=0.9996,0.9992,0.9998,0.9983, respectively). Conclusion: The proposed method can be applied to the determination of related substances in reduced glutathione.