利用反相高效液相色谱(RP-HPLC)分离小麦胚乳醇溶蛋白的研究  

Study on Separating Wheat Gliadin in Endosperm Cells by Reversed-phase High Performance Liquid Chromatorgraphy

作　　者：赵龙刚[1] 李士平[2] 宋希云[1] 王兆华[1] 张恩盈[1] 徐坤[1] 

机构地区：[1]莱阳农学院生命科学学院,山东青岛266109 [2]德州学院,山东德州253015

出　　处：《麦类作物学报》2007年第4期634-638,共5页Journal of Triticeae Crops

基　　金：学院博士创新基金(630408)

摘　　要：为了研究洗脱时间、柱温、流速、洗脱梯度、蛋白提取方法等色谱条件对反相高效液相色谱法(RP-HPLC)分离小麦胚乳醇溶蛋白的影响,对上述条件分别进行了对比试验。结果表明,利用RP-HPLC分离小麦胚乳醇溶蛋白的最佳实验方法为:通过0.05 mol/L NaCl→H2O→70%乙醇三步提取醇溶蛋白,在1.0mL/min的流速、60℃的柱温条件下,对上样醇溶蛋白进行洗脱,洗脱梯度为流动相B的体积比在55 min内由21%升至48%(V/V),最后通过210 nm紫外光检测洗脱组分的吸收值。重复三次的检验结果证实,本试验方法稳定可靠。In order to investigate grade and protein extraction method effects of elution time, column temperature, flow speed, elution on the separation of wheat gliadin in endosperm cells using reversed-phase high performance liquid chromatorgraphy, experiments involving the above factors were carried out. The results showed that the optimal method for separation of wheat gliadin in endosperm cells was that the protein was extracted by 0.05 mol/L NaCl→H2O→70% alcohol, eluted at flowing speed of 1.0 mL/min, column temperature of 60℃ and the concentration of elution B increased from 21% to 48% （V/V） within 55 minutes, and the absorbance of the eluted protein was checked at 210 nm. The experiments were conducted for three times, and the results showed that the optimal procedure were stable and reliable for separating wheat gliadin in endosperm cells.

关 键 词：小麦 醇溶蛋白 反相高效液相色谱法 

分 类 号：S512.1[农业科学—作物学] S331