饮料中提取酪蛋白的方法选择 - 食品安全质量检测学报.docVIP

饮料中提取酪蛋白的方法选择 孙姗姗1，李婷婷2，覃 玲，张会亮，梁瑞强，曹 进1* （1．中国食品药品检定研究院, 北京 100050；2.酒泉市食品检验检测中心，甘肃 735000） 摘要： 目的 探索种沉淀蛋白质的方法在提取饮料酪蛋白过程中对其复溶率的影响，建立超高效液相色谱-三重四极杆串联质谱鉴别饮料中酪蛋白亚型及相应含量的测定最佳前处理方法。方法 选取乳粉、以乳粉或牛乳为原料的饮料、添加酪蛋白或酪蛋白酸钠的饮料作为测试样品，用pH=8.5的Tris-HCl溶液溶解，采用等电点沉淀法、有机相（乙腈）沉淀法、有机相（丙酮）沉淀法和有机相（乙醇）沉淀法提取样品中的蛋白质，低温离心后对沉淀的蛋白质进行复溶，用考马斯亮蓝法分别测定沉淀前和复溶后样品溶液中可溶性蛋白质的含量，计算类酪蛋白的复溶率。结果 乳粉和类饮料中的酪蛋白经等电点沉淀法提取后，乳粉的类酪蛋白复溶率在64.95%66.20%之间，饮料的类酪蛋白复溶率在69.73%84.95%之间，均优于其他种沉淀方法。αs-酪蛋白标准物质和β-酪蛋白标准物质的复溶率分别为70.16%和87.17%，且经高分辨质谱确证其复溶后的主成分为酪蛋白。另外对45种含酪蛋白饮料中的类酪蛋白含量进行了测定，其含量在0.1042.36 mg/g (mg/mL)之间。结论 等电点沉淀法提取类酪蛋白的复溶率较其他种沉淀方法相对稳定，关键词：酪蛋白；等电点沉淀；饮料 Selection of extraction method for casein in beverages SUN Shan-Shan1, LI Ting-Ting2, QIN Ling, ZHANG Hui-Liang, LIANG Rui-Qiang, CAO Jin1* （1. National Institute of Food and Drug Control, Beijing 100050, China; 2. Food Test Center of Jiuquan, Gansu 735000, China） ABSTRACT: Objective To select the optimized pretreatment method for identifying the subtype of casein and quantitating the content in beverages using triple-quadruplole mass spectrometer. The redissolution percentages during the protein extraction process as important value to evaluate the effect of 4 kinds of precipitation methods. Methods The testing samples were milk powder, beverage with milk or milk powder as raw materials and beverage with casein or sodium caseinate, which were firstly dissolved in Tris-HCl (pH=8.5) buffer solution. Then the proteins of samples were precipitated using isoelectric precipitation, acetonitrile precipitation, acetone precipitation and ethanol precipitation methods followed by the redissolution after low temperature centrifuge. Finally, the protein contents were determined with Bradford method and the corresponding redissolution percentages were calculated. Results Isoelectric precipitation was checked to extract casein in beverages. The redissolution percentages of casein from milk powder range from 64.95% to 66.20%, and the redissolution percentages of casein from th