HPLC同时测定白花蛇舌草中熊果酸、异熊果酸和对香豆酸的含量

Simultaneous Determination of Ursolic Acid, Isoursolic Acid and P-coumaric Acid of Hedyotis Diffusa Willd. by HPLC

关键词:  高效液相色谱法  白花蛇舌草  熊果酸  异熊果酸  对香豆酸

Key words:  HPLC  Hedyotis diffusa Willd.  ursolic acid  isoursolic acid  p-coumaric acid

摘要:

目的 建立HPLC测定白花蛇舌草中熊果酸、异熊果酸和对香豆酸的含量。方法 采用 Diamonsil C₁₈ ODS (4.6 mm×250 mm，5 μm)，流动相为乙腈-15 mmol·L^-1 NH₄Ac(12∶88，pH 3.5)，流速为1.0 mL·min^-1，检测波长为308 nm，柱温为35 ℃。结果 熊果酸、异熊果酸和对香豆酸线性范围分别为0.075 7~2.271 μg(R²=0.999 6)，0.010 3~0.309 μg (R²=0.999 2)和0.020 2~0.606 μg(R²=0.999 6)；3种成分测定的平均回收率分别为99.9% (RSD=0.7%)，99.0% (RSD=1.7%)和101.1% (RSD=0.7%)。结论 本法简便、准确度高、稳定性和重复性好，可用于白花蛇舌草中指标性成分定量分析和质量控制。

Abstract:

OBJECTIVE To establish an HPLC method for the simultaneous determination of ursolic acid, isoursolic acid and p-coumaric acid in Hedyotis diffusa Willd.. METHODS Diamonsil C₁₈ ODS (4.6 mm×250 mm, 5 μm) was used; mobile phase was acetonitrile and 15 mmol·L^-1 NH₄AC(12∶88, pH3.5); flow rate: 1.0 mL·min^-1; wavelength: 308 nm; column temperature: 35 ℃. RESULTS Three ingredients showed good linear relationships, ursolic acid: 0.075 7-2.271 μg (R²=0.999 6), isoursolic acid: 0.010 3-0.309 μg (R²=0.999 2) and p-coumaric acid: 0.020 2-0.606 μg (R²=0.999 6), with the mean recoveries as 99.9% (RSD=0.7%), 99.0% (RSD=1.7%) and 101.1% (RSD=0.7%), respectively. CONCLUSION This method is simple with high precision, repeatability, and stability, so it can be used to quantitative analysis and quality control of Hedyotis diffusa Willd.