HPLC法测定莲子中还原型谷胱甘肽（GSH）和总巯基（-SH）含量

摘要

Reduced glutathione (GSH) and total thiols in Lotus Seed were determined by 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. Sample pretreatment and elution solutions were optimized in order to inhibit reduced sensitivity and serious tag of chromatographic signal for derivatized product 2-nitroso-5-thiol-benzoic acid (NTB), and selectively determined GSH and total thiols (-SH) in Lotus Seed. The results showed that the average amount of GSH and total thiols in Lotus Seed were 0.971 1μmol/g and 1.368 6μmol/g, the average recoveries of the methods were 101.02 % and 100.09 %, detection limits were 4.21μmol/L and 4.63μmol/L respectively.%利用5，5'-二硫双-2-硝基苯甲酸（DTNB）为衍生化试剂，采用细胞破壁、衍生化为一体的同步衍生化提取、HPLC法测定了莲子中GSH和总巯基（-SH）含量。本文对样品前处理过程和色谱流动相的选择分别进行了优化，有效的阻止了前处理过程中巯基氧化变性对测定灵敏度降低而导致的漏检及消除了衍生化产物3，5-硝基-3-巯基-苯甲酸（NTB）色谱峰拖尾所造成的干扰，并借此选择性测定了莲子中GSH和总巯基（-SH）含量。结果表明；三次测定莲子中GSH和总巯基（-SH）含量均值分别为0.9711μmol/g和1.3686μmol/g ，方法的回收率分别为101.02%和100.09%，检测限分别为4.21μmol/L和4.63μmol/L。