正交试验优化HPLC测定齐墩果酸与熊果酸的色谱条件

为建立简单、快速的分离测定齐墩果酸与熊果酸的HPLC方法,实验采用L16(44)的正交试验设计表,对HPLC分离测定齐墩果酸与熊果酸的主要影响因素(甲醇与水相的体积比、水相中乙酸和三乙胺的体积分数以及流动相流速)进行了优化.结果表明,以体积比为90∶10的甲醇与含有0.5%乙酸及0.01%三乙胺的水相混合溶液为流动相,流速为0.8 mL/min,齐墩果酸和熊果酸可以在16 min内得到有效分离,其工作曲线的线性范围对于齐墩果酸为2.5～340.0μg/mL,对于熊果酸为5.0～340.0μg/mL.将方法应用于女贞子和山楂中齐墩果酸和熊果酸含量的测定,相对标准偏差均小于2.0%,加样回收率均在95%～105%之间.

Study on the Conditions of HPLC Method for the Separation and Determination of Oleanolic and Ursolic Acid in the Orthogonal Experiment

To develop a rapid and convenient method for the separation and determination of oleanolic acid(OA) and ursolic acid(UA) by HPLC,the main factors influencing HPLC,including the volumetric ratio of methanol and aqueous phase in the mobile phase,the percent of acetic acid and triethylamine in the aqueous phase,and the flow rate of the mobile phase,were optimized with the L16(44) orthogonal experimental design table.Ursolic acid and oleanolic acid could be separated and determined effectively within 15 min when a mixture of MeOH and aqueous solution consisting of 0.5% acetic acid and 0.01% triethylamine with the volumetric ratio of 90∶ 10 was employed as the mobile phase with a flow rate of 0.8 mL/min.The linear ranges were 2.5—340.0 μg/mL for oleanolic acid and 5.0—340.0 μg/mL for ursolic acid with good correlation.The method was applied satisfactorily for the determination of oleanolic acid and ursolic acid in Ligustrum lucidum Ait.and hawthorn with the relative standard deviation less than 2.0%,and the recovery rate ranging from 95% to 105%.