目的建立一种适用于检测硒蛋白中硒代氨基酸的有效方法。方法硒蛋白样品采用酸脱蛋白质的方法,在酸性条件下使蛋白质变性沉淀,取沉淀物运用蛋白酶K、胰蛋白酶、链蛋白酶酶解硒蛋白样品;在5 mmol/L的柠檬酸铵中加2%的甲醇溶液作为流动相,并用柠檬酸或氨水精确调节流动相的p H值(p H=4.9),用离子交换柱分离硒代半胱氨酸(Se Cys)、硒代蛋氨酸(Se Met)等硒代氨基酸,通过ICP-MS检测其含量,并运用方法学验证实验结果。结果所建立的硒蛋白中硒代氨基酸检测方法,检出限分别为3.5μg/kg、11.2μg/kg,线性范围(以Se计)为0μg/L~200μg/L,加标回收率为75%~120%。结论经实验证明,该检测方法适用于食品营养强化剂硒蛋白中硒代氨基酸的分析,此方法的建立对食品的成分分析和食品的开发利用具有重要意义。 Objective To establish an effective method for detecting seleno-amino acids in selenoproteins. Methods The samples of selenoprotein were deproteinized by acid and the proteins were denatured and precipitated under acidic conditions. The samples were treated with proteinase K, trypsin, and streptinase to digest the selenoprotein samples. In 5 mmol / L ammonium citrate, 2 % Methanol solution as the mobile phase, and the pH value of the mobile phase (p H = 4.9) was accurately adjusted with citric acid or aqueous ammonia, Se Cys, Se Met ) And other seleno amino acids were detected by ICP-MS, and the experimental results were verified by methodological methods. Results The detection limits of selenoproteins in selenoproteins were 3.5μg / kg and 11.2μg / kg, respectively. The linear ranges (calculated by Se) ranged from 0μg / L to 200μg / L and the recoveries were 75 % ~ 120%. Conclusions The experiment proved that the method is suitable for the analysis of selenoprotein in food nutrition fortifier selenoprotein. The establishment of this method is of great significance to the composition analysis of food and the development and utilization of food.