柱前衍生RP-HPLC测定仙灵骨葆胶囊中16种氨基酸  

Determination of 16 Amino Acids in Xianling Gubao Capsule by RP-HPLC with Pre-column Derivatization

作　　者：杨卫灵 李朝婵[1] 田红红[2] 谭高好 柳立伟[1] 伍庆[1] 

机构地区：[1]贵州省山地环境信息系统与生态环境保护重点实验室,贵阳550001 [2]贵州师范大学生命科学院,贵阳550001

出　　处：《中国实验方剂学杂志》2015年第18期44-47,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家科技支撑计划项目(2011BAI13B04-01)

摘　　要：目的：建立柱前衍生RP-HPLC同时测定仙灵骨葆胶囊中16种氨基酸的方法。方法：以正亮氨酸为内标物,异硫氰酸苯酯（PITC）为柱前衍生剂,用高效液相色谱测定仙灵骨葆胶囊中水解氨基酸的量。采用Hypersil C18色谱柱（4.6 mm×250 mm,5μm）,以乙腈-0.1 mol·L^-1乙酸钠溶液-乙酸（7∶93∶0.05）为流动相A,乙腈-水（80∶20）为流动相B进行梯度洗脱,体积流量1 m L·min^-1,检测波长254 nm,柱温40℃。结果：33 min内可完成仙灵骨葆胶囊中16种氨基酸的分离测定,线性关系良好（r〉0.996 8）,回收率97.5%-100.7%,RSD〈5%。结论：该方法简便快速、准确可靠,可用于仙灵骨葆胶囊中氨基酸的含量测定。Objective： To develop a RP-HPLC method for the simultaneous determination of 16 amino acids in Xianling Gubao capsule. Method： With norleucine as the internal standard, hydrolyzed amino acids of Xianling Gubao capsule were derivatized with phenylisothiocyanate （PITC） and determined by RP-HPLC. Hypersil C18 column （4.6 mm ×250 mm, 5 μm） was eluted with mobile phase A acetonitrile-O. 1 mol ·L^-1 sodium acetate solution-acetic acid （7： 93： 0. 05） and mobile phase B acetonitrile-water （80：20） in a gradient mode at a flow rate of 1 mL ·min^-1. The detection wavelength was set at 254 nm and the column temperature was 40℃. Result： The 16 amino acids were separated within 33 min with a good linearity （r 〉 0. 996 8） within the range of the test concentration. The average recoveries ranged between 97.5% and 100.7% and the RSD values were less than 5%. Conclusion： The method is simple, quick, accurate and reliable and can be used for the determination of amino acids in Xianling Gubao capsule.

关 键 词：仙灵骨葆胶囊 柱前衍生 氨基酸 异硫氰酸苯酯 高效液相色谱 

分 类 号：R284.1[医药卫生—中药学]