湘西桐叶药材熊果酸及齐墩果酸的HPLC法测定

HPLC Determination of Ursolic Acid and Oleanolic Acid in Paulownia Leaves from Xiangxi

建立一种高效、准确、稳定的桐叶熊果酸和齐墩果酸测定方法,测定湖南省湘西地区桐叶药材中熊果酸和齐墩果酸含量.采用高效液相色谱(HPLC)法,色谱条件依次为,色谱柱:TADE-PAKAF-C18柱(250 mm×4.6 mm,5μm);流动相:乙腈-甲醇-0.5%醋酸铵溶液(体积比61∶18∶21);流速:1 mL/min;检测波长:210 nm;柱温:35℃;进样量:10μL.结果表明:齐墩果酸与熊果酸分离度为1.73,均符合要求;熊果酸和齐墩果酸对照品分别在0.4976~9.9520μg/mL和0.1261~2.5230μg/mL范围内线性关系良好(r均为0.9999);熊果酸、齐墩果酸平均加样回收率分别为99.93%,98.97%;湘西产桐叶含熊果酸4.54~7.07 mg/g,含齐墩果酸2.50~3.20 mg/g.HPLC法分离效果好、灵敏度高、准确度高,是检测桐叶中熊果酸、齐墩果酸含量的科学有效手段.

An efficient, accurate and stable method for the determination of ursolic acid and oleanolic acid has been established.Then the contents of ursolic acid and oleanolic acid in Paulownia leaves in Xiangxi were determined.High performance liquid chromatography(HPLC) method was adopted, and the chromatographic conditions were as follows:(1) chromatographic column: TADE-PAK AF-C18 column(250 mm×4.6 mm, 5 μm);(2) mobile phase: acetonitrile-methanol-0.5% ammonium acetate solution(Volume ratio 61∶18∶21);(3) flow rate: 1 mL/min;(4) detection wavelength: 210 nm;(5) column temperature: 35 ℃;(6) sample size: 10 μL.The results showed that the separation degree of oleanolic acid and ursolic acid was 1.73,which met the requirements.The linear relationship between ursolic acid and oleanolic acid was good in the range of 0.497 6~9.952 μg and 0.126 1~2.523 μg(r=0.999 9),respectively, The average recoveries of ursolic acid and oleanolic acid were 99.93% and 98.97%,respectively.The contents of ursolic acid and oleanolic acid in Paulownia leaves in Xiangxi were 4.54~7.07 mg/g and 2.50~3.20 mg/g, respectively.HPLC method has good separation effect, high sensitivity and high accuracy, and is a scientific and effective method for the determination of ursolic acid and oleanolic acid in Paulownia leaves.