HPLC测定三黄汤中4种成分含量

Determination of Four Components in Sanhuang Decoction by HPLC

关键词:  三黄汤  高效液相色谱法  绿原酸  盐酸药根碱  盐酸小檗碱  盐酸巴马汀

Key words:  Sanhuang decoction  HPLC  chlorogenic acid  jatrorrhizine hydrochloride  berberine hydrochloride  palmatine hydrochloride

摘要:

目的 建立高效液相色谱法测定三黄汤中4种重要活性成分(绿原酸、盐酸药根碱、盐酸小檗碱和盐酸巴马汀)的含量。方法 采用Agilent C₁₈色谱柱(4.6 mm×200 mm，5 μm)，流动相为1%醋酸甲醇溶液-1%醋酸溶液梯度洗脱；检测波长：329 nm；流速：1.0 mL·min^-1。结果 绿原酸、盐酸药根碱、盐酸小檗碱和盐酸巴马汀的线性范围分别为0.5~4.0 μg·mL^-1(r=0.999 1)，0.5~4.0 μg·mL^-1(r=0.999 7)，0.5~12.0 μg·mL^-1(r=0.999 3)，0.3~4.0 μg·mL^-1(r=0.999 9)。平均回收率分别为99.0%(RSD=1.13%)，99.3%(RSD=1.20%)，99.0%(RSD=0.27%)，99.8%(RSD=0.88%)。日内、日间RSD均<2%。结论 本方法准确、灵敏、重复性好，可同时测定三黄汤中4种重要化学成分的含量。

Abstract:

OBJECTIVE To establish an HPLC method for simultaneous determination of four important active components in Sanhuang decoction. METHODS An Agilent C₁₈ column (4.6 mm×200 mm, 5 μm) was used with the mobile phase of 1% acetic acid methanol solution-1% acetic acid by gradient dilution, at the flow rate of 1.0 mL·min^-1 and detection wavelength of 329 nm. RESULTS The linear calibration curves of chlorogenic acid, jatrorrhizine hydrochloride, berberine hydrochloride and palmatine hydrochloride were obtained in the ranges of 0.5-4.0 μg·mL^-1(r=0.999 1), 0.5-4.0 μg·mL^-1 (r=0.999 7), 0.5-12.0 μg·mL^-1(r=0.999 3), 0.3-4.0 μg·mL^-1 (r=0.999 9), respectively. Their average recoveries (n=3) for the assay were 99.0%(RSD=1.13%)，99.3%(RSD=1.20%)，99.0%(RSD=0.27%) and 99.8%(RSD=0.88%), respectively. The intra- and inter-day RSDs were all less than 2.0%. CONCLUSION This method is accurate, sensitive and suitable for quality control of Sanhuang decoction.