高效排阻液相色谱法分析大豆7S和11S球蛋白的分子质量和纯度

Molecular Weight and Purity of Soy 7S,11S Globulin Analyzed by Size Exclusion-High-Performance Liquid Chromatography

本文采用SE-HPLC法定量分析大豆7S和11S球蛋白的分子质量和纯度。首先,对SE-HPLC的色谱条件进行优化,最佳的SE-HPLC色谱条件是:洗脱液(水/乙腈)70/30,洗脱液流速1.0 m L/min,进样量10μL。其次,绘制分子质量分析的标准曲线,分子质量标准曲线方程为Log(M)=-0.222 1 x+3.878 9,线性相关系数为R^2=0.996 9。以10倍信噪比确定定量限,得到7S(y=299.59x+3.771 2,R^2=0.979 2)和11S(y=197.08x+4.148 5,R^2=0.986 1)纯度分析标准曲线方程。最后根据分子质量和纯度分析的标准曲线方程计算大豆7S和11S球蛋白的分子质量和纯度。将所提纯的7S和11S样品的分子质量与7S和11S标品特征峰分子质量进行比对,表明所提纯样品为7S和11S。本试验所制得7S和11S的纯度分别达到86.3%和92.0%。

In this paper, the molecular weight and purity of soy 7S, llS globulin were quantificationally analyzed by SE - HPLC. Firstly, the optimization of the SE - HPLC chromatographic conditions were carried out, and the optimal SE - HPLC chromatographic conditions were： eluant （water/aeetonitrile） 70/30, flow rate of eluant 1.0 mL/min, sample size 10μL. Secondly, the standard curve of molecular weight analysis was drawn, and the standard curve equation of molecular weight was Log （M） = - 0. 2221x ＋ 3. 878 9, and the linear correlation coefficient was RE =0.996 9. With the 10 times signaltonoise ratio quantitative limit, the standard curve equation of 7S molecular purity （y =299.59x ＋3. 7712, R2 =0.979 2） and llS molecular purity （y = 197.08x ＋4. 1485, R2 = 0. 986 1 ） were obtained. Finally, this study provided a quick identification method of the molecular weight and purity of soy 7S, llS globulin. The purity of 7S or llS was respectively 86.3% and 92.0%.