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

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.