大白菜种子蛋白分离的高效液相色谱法的研究

该文首次利用高效液相色谱法时大白菜种子的水溶性蛋白质进行了分析,确定了适宜的色谱分析条件:选用径向加压型Nova-Pak C_(18)反相色谱柱(规格为8mm (?).d.×10cm length,填充颗粒为球形,直径4μm,Waters Co.)以甲醇水溶液进行洗脱,种子中各蛋白组分得到了比较好的分离。洗脱时采用混合梯度洗脱方式:0-20分钟,从9%甲醇水溶液(A)到35%甲醇 0.02%三氧乙酸(B)的线性梯度洗脱;20-45分钟,以B洗脱液进行等强度洗脱.流速均为0.8ml/min。洗脱后的各组分用紫外光/可见光检测器在280nm处进行监测。实验结果还表明:不同提取液对色谱分离的结果有一定的影响,用0.1%巯基乙醇水溶液提取时,色谱峰的分离效果较好,主要组分的色谱峰比较显著,因而比较适于大白菜种子的分析。

Optimization of Hple Conditions for the Separation of Seed proteins of Chinese Cabbage

In this article, the optimization of separation conditions for water-soluble proteins in Chinese Cabbage seeds using high performance liquid chomatography (HPLC) was described. All the main protein components were well separated on a radial type Nova-Pak C18 column (10cm length x 8mm i. d., 4um spherical particles, Waters Co.) with the following programme: 0-20 min., a linear gradient elution from 9% methanol (eluent A) to 35% methanol 0.02% trifluoric acid TFA] (eluent B), and then maintain at B from 20 to 45 min., The flow speed is 0. 8 ml/min. The eluate was monitored at 280nm with a multi-channel UV/VIS detector. The effects of various extraction solvents were also compared and 0. 1% mercaptoethanol proved to be most suitable for this purpose.