摘要
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 a method which is suitable for detection of seleno amino acid in selenium protein. Methods Selenium protein samples,with the method of acid deproteinization,was conducted for protein denaturation,under acid condition lead to protein precipitation,sediment was seperated by taking use of protease,trypsin,protease K chain selenium protein enzymolysis sample; 5 mmol / L ammonium citrate and 2% methanol solution as mobile phase,using citric acid or ammonia precise to adjust the mobile phase pH value（ pH = 4. 9）,ion exchange column was used to separate selenium cysteine（ SeCys）,selenomethionine（ Se Met） such as selenium generation amino acid; ICP-MS was used to detect its content,and use the methodology to validate results. Results As for the established method,the detection limit was 3. 5 μg / kg,11. 2 μg / kg; Linear range was in Se of 0 μg / L-200 μg / L; Standard addition recovery was 75%-120%. Conclusion The experiment has proved that the detection method is suitable for food nutrition fortifier selenium in selenium protein amino acid analysis,the establishment of this method has great significance to the development and utilization of food composition analysis and food.
出处
《中国卫生检验杂志》
CAS
2015年第5期636-638,641,共4页
Chinese Journal of Health Laboratory Technology
