柱前衍生化-反相高效液相色谱法测定蛋白粉中氨基酸的含量  

RP-HPLC coupling with precolumn derivatization for determination of amino acid in protein power

作　　者：姜雯[1] 王棘[1] 

机构地区：[1]辽宁省食品药品检验所,沈阳110023

出　　处：《中国卫生检验杂志》2008年第2期280-282,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的：建立柱前衍生化-反相高效液相色谱方法测定蛋白粉中氨基酸的含量。方法：用6 mol/L HCl水解提取蛋白粉中总氨基酸,以异硫氰酸苯酯（PITC）为柱前衍生化试剂,采用外标法,梯度洗脱的方式分析。色谱柱为ODS（Kromasil 100-5 C18250×4.6 mm）,采用两种流动相进行梯度洗脱：流动相A为0.1 mol/L醋酸钠缓冲液（pH6.5）-乙腈（97：3）;流动相B为水-乙腈（1：4）,使用紫外检测器,检测波长为254 nm。结果：17种氨基酸在40 min内均可得到很好的分离;在10～200μg/ml的浓度范围内呈良好的线性关系（r=0.9991-0.9999）;平均回收率为89.1%～98.2%;相对标准偏差为0.36%～1.24%。结论：本方法分离效果好、灵敏、准确,可广泛用于含有多种氨基酸样品的分析。Objective：The method of RP-HPLC determination of amino acid in protein power Coupling with precolumn derivatization was developed.Methods：To extract total amino acids by the method of hydrolysis in 6 mol/L HCl and use phenylisothio cyanate（PITC） as the derivating agent with external standard method to analyze them by gradient elution.The analytical column was Φ 250×4.6 mm KromasilODS.Gradient elution was used with two mobile phase.The mobile phase A was 0.1 mol/L sodium acetate solution（pH6.5） and acetonitrile（93：7）.The mobile phase B was water and acetonitrile（20：80）.The detective wavelength was 254 nm.Results：Using this method,seventeen kinds of amino acids could be separated well in 40 min.Amino acids were linear over the range of 10～200 μg/ml（r=0.9991-0.9999）,the average recoveries were 89.1%～98.2%,and the RSD were 0.36%～1.24%.Conclusion：The method is sensitive,accurate,and reliable for the determination of amino acids in many other samples.

关 键 词：蛋白粉 氨基酸 反相高效液相 柱前衍生 

分 类 号：O657.72[理学—分析化学]