摘要
目的建立RP-HPLC法同时测定女贞叶乙醇提取物中齐墩果酸与熊果酸的方法。方法通过系统地考察温度、pH值、甲醇-乙腈混合有机相中甲醇的量对齐墩果酸、熊果酸保留时间和分离度的影响,最终确定HPLC色谱条件为大连依利特KromasilC18(250mm×4.6mm,5μm)色谱柱;甲醇-乙腈-10mmol/L乙酸铵(55:27:18)为流动相,体积流量为1mL/min,检测波长为205nm,柱温20℃。结果齐墩果酸与熊果酸分别在0.300～3.00μg、0.692～6.92μg线性关系良好;加样平均回收率分别为101.4%、100.2%(n=9)。结论低温、中性pH、甲醇-乙腈混合有机相有利于齐墩果酸与熊果酸的分离。优化后的色谱方法分离度高,重现性好,可用于女贞叶乙醇提取物中齐墩果酸与熊果酸的测定。
Objective To establish a RP-HPLC method for simultaneous quantitative determination of oleanolic acid and ursolic acid in ethanol extract of Ligustrurn lucidum Ait. leaves. Methods Temperature, pH value and proportion of methanol and acetonitrile in organic phase were systematically studied on the influence of retention and resolution between oleanolic acid and ursolic acid. The optimized chromatographic condition was Kromasil C18 column （250 mm×4. 6 mm, 5 μm）; methanol-acetonitrile-10 mmol/L ammonium acetate （55 ： 27 ： 18） as mobile phase with flow rate 1.0 mL/min. The detection wavelength was 205 nm and column temperature was 20 ℃. Results The linear ranges of oleanolic acid and ursolic acid were 0. 300-3.00 μg, 0. 692-6.92 μg respectively; the average recoveries were 101.4%, 100. 2% （n=9） respectively. Conclusion Low temperature, neutral pH and mixed organic phase with methanolacetonitrile can improve resolution of oleanolic acid and ursolic acid. The optimized method can achieve better resolution and reproducibility, and is suitable to determine the content of oleanolic acid and ursolic acid in ethanol extract of Ligustrum lucidum Ait. leaves.
出处
《现代药物与临床》
CAS
2010年第3期210-214,共5页
Drugs & Clinic
关键词
高效液相色谱
女贞叶乙醇提取物
齐墩果酸
熊果酸
HPLC
ethanol extract of Ligustrum lucidum Ait. Leaves
oleanolic acid
ursolic acid
