高效液相色谱法测定香铃草子中还原型谷胱甘肽和总巯基含量

Dertermination of Reduced Glutathione and Total Thiols in Hibiscustrionum L.Seeds by High Performance Liquid Chromatography

以5,5′-二硫双-2-硝基苯甲酸为衍生化试剂,使用细胞破壁、衍生化为一体的同步衍生化法,采用高效液相色谱法测定了香铃草子中还原型谷胱甘肽(GSH)和总巯基(-SH)含量。结果表明:香铃草子中GSH和总巯基含量均值分别为4.07μmol/g和6.06μmol/g。总巯基含量中扣除GSH含量大致为巯基蛋白含量,达1.99μmol/g,说明香玲草子富含GSH和巯基蛋白。优化色谱条件下GSH和总疏基的回收率分别为100.43%和101.79%,检测限分别为5.31μmol/L和6.18μmol/L。

Reduced glutathione and total thiols in Hibiscus trionum L.Seeds were determined by high performance liquid chromatography（HPLC）assay utilizing 5,5′-dithio-bis（2-nitrobenzoic acid）（DTNB,Ellman＇s reagent）as the derivatizing agent with concomitant cell rupture during the sample pretreatment.The results indicated that the contents of reduced glutathione and total thiols determined in the Hibiscus trionum L.Seeds were 4.07μmol/g and 6.06μmol/g,respectively.The difference between total thiols and reduced glutathione（1.99μmol/g）implies the content of protein thiols,indicating that Hibiscus trionum L.Seeds is abundant with reduced glutathione and thiol proteins.The HPLC conditions were optimized,and the recoveries of the methods under the optimized condition are100.43% and 101.79%,detection limits are 5.31μmol/L and 6.18μmol/L for reduced glutathione and total thiol detections respectively.