麦胚谷胱甘肽提取与含量测定方法研究

旨在建立用反相高效液相色谱(RP-HPLC)法同时检测小麦胚中谷胱甘肽(glutathione)还原型(G-SH)和氧化型(G-S-S-G)含量的方法.为了更高效地提取谷胱甘肽,在水作溶剂条件下,考察了不同浸泡时间、超声时间、料液比、超声温度、溶剂pH和沉淀剂比例对谷胱甘肽提取效率的影响.并在2种色谱柱上,考察了5种流动相下谷胱甘肽含量测定结果.最终确定最适提取条件为:提取溶剂pH4.0,料液比1∶30,浸泡1h后在0℃下超声10min,沉淀剂比例为1∶2.0；色谱条件为:Fusion-RP柱(150×4.6mm,4μm,8nm)；流动相:水(0.08％辛烷磺酸钠+0.24％磷酸二氢钠,以磷酸调pH2.5)-乙腈(体积比为82∶8)；检测波长为210nm;流速为1.0mL·min-1；柱温为25℃.测定了19份不同来源麦胚中谷胱甘肽的含量,发现谷胱甘肽含量差异显著,最高达180.71mg·(100g)-1.

Study on the Methods of Extracted the Glutathione and Determined Contents from Wheat Germ

This study is to establish the RP-HPLC method to determine the content of GSH and GSSG simultaneously.Using distilled water as solvent,the effect of the time of soak,time of ultrasonic extraction,material liquid ratio,temperature of ultrasonic extraction,solvent pH,and precipitating agent proportion on the the extracted efficiency of Glutathione were determined.The contents of glutathione were determined by using 2 chromatogram columniations and 5 mobile phases.Finally,the optimal conditions were determined.The solvent pH was 4.0,the material liquid ratio was 1∶30,the time of soak was 1 h,ultrasonic extraction 10 min in 0℃,and precipitating agent proportion was 1∶2.0.Chromatogram conditions were Fusion-RP column(4.60×150 mm,4 μm,8 nm)at 25 ℃,the mobile phase were both a mixture of 0.08% octanesulfonic acid sodium salt and 0.24%Sodium dihydrogen phosphate in water and acetonitrile(82∶8,V/V),adjusted to pH 2.5 with phosphoric acid.The UV detection wavelength was set at 210 nm.The flow rate was 1.0 mL·min-1.The glutathione contents from 19 wheat germs were determined.The glutathione contents had significant difference.The maximal content was 180.71 mg·(100 g)-1.