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目的:优化马钱苷、丹皮酚和毛蕊花糖苷的HPLC方法,检测山药硫熏前后的六味地黄丸中3种成分含量的差异,研究硫熏对复方的影响。方法:采用Thermo Hypersil BDS C18色谱柱(4.6 mm×250 mm,5μm)。马钱苷,流动相乙腈-0.1%磷酸梯度洗脱,检测波长236 nm;丹皮酚,流动相乙腈-0.1%磷酸(50∶50),检测波长274 nm;毛蕊花糖苷,流动相乙腈-0.1%乙酸梯度洗脱,检测波长334 nm。柱温30℃,流速1 m L·min-1,进样量10μL。结果:随SO2量的增加,六味地黄丸中丹皮酚、马钱苷和毛蕊花糖苷均有明显的下降趋势,亚硫酸钠组的3种成分含量最低,分别为0.022 6,0.691 2,0.036 2 mg·g-1。结论:该研究优化的HPLC方法准确灵敏、重复性好,适合用于六味地黄丸的含量测定。结果证实硫熏山药对六味地黄丸会有不利影响,其根源在于SO2残留。
OBJECTIVE: To optimize the HPLC method of loganin, paeonol and verbascoside, to detect the differences of the contents of three components in Liuweidihuangwan before and after yam smoked, and to study the effect of sulfur smoked on the compound prescription. Methods: A Thermo Hypersil BDS C18 column (4.6 mm × 250 mm, 5 μm) was used. Loganin, mobile phase acetonitrile -0.1% phosphoric acid (50:50), detection wavelength 274 nm; Verbascoside, mobile phase acetonitrile-0.1 % Acetic acid gradient elution, detection wavelength 334 nm. The column temperature was 30 ℃, the flow rate was 1 m L · min-1, and the injection volume was 10μL. Results: With the increase of SO2 content, the paeonol, loganin and verbascoside in Liuweidihuang Pills decreased obviously. The content of the three components in the sodium sulfite group was the lowest, which were 0.022 6,0.691 and 2.0.036 2 mg · g-1. Conclusion: The HPLC method optimized by this study is accurate, reproducible and suitable for the determination of Liuwei Dihuang Pill. The results confirmed that sulfur smoked yam Liu Wei Di Huang Wan will have an adverse effect, the root cause is the residual SO2.