HPLC-ELSD同时测定九蒸九曝法炮制的黑参中极性和非极性皂苷

目的:为评价经九蒸九曝传统工艺炮制成的黑参质量,建立同时测定黑参中7个极性和12个非极性皂苷单体含量的方法。 方法:采用高效液相色谱结合蒸发光散射检测器(HPLC-ELSD)对九蒸九曝法炮制前后的参品中皂苷单体含量进行比较分析。色谱条件:采用Discovery C₁₈(250 mm×4.6 mm,5 μm)色谱柱,以10%乙腈水溶液(含5%醋酸)(A)-80%乙腈水溶液(B)为流动相进行梯度洗脱,流速1.2 mL·min^-1,柱温30℃;漂移管温度为60℃,载气流速1.8 L·min^-1。 结果:炮制前后参品中7个极性皂苷(人参皂苷Rg₁、Re、Rf、Rb₁、Rc、Rb₂、Rd)和12个非极性皂苷(人参皂苷F₄、Rg₆、Rk₃、Rh₄、20(S)-Rg₃、20(R)-Rg₃、20(S)-Rs₃、20(R)-Rs₃、Rk₁、Rg₅、Rs₄、Rs₅)分离良好,线性关系良好(r≥0.9997),平均回收率在97%~105%之间。与白参相比,由白参经九蒸九曝炮制而成的黑参中非极性皂苷和奥克梯隆型皂苷显著增加,而极性皂苷显著减少。 结论:HPLC-ELSD法可同时测定黑参中7个极性皂苷和12个非极性皂苷成分,较好地实现对黑参产品的质量控制,同时为黑参药理作用的物质基础研究提供科学依据。

Objective: To evaluate the quality of black ginseng prepared by nine cycles of steaming and solarization approach by establishing a method to simultaneously determine the content of 7 polar and 12 non-polar saponin monomers. Methods: Comparison of saponins contents in pre-and post-processed ginsengs was carried out by high performance liquid chromatography-evaporative light-scattering detector(HPLC-ELSD).The chromatographic separation was performed on a Discovery C₁₈ column(250 mm×4.6 mm,5 μm),the column temperature was maintained at 30℃.The binary gradient elution system consisted of 10% acetonitrile aqueous solution containing 5% acetic acid(A)and 80% acetonitrile aqueous solution water(B).The flow rate was set at 1.2 mL·min^-1.ELSD was set to a probe temperature at 60℃,and the nebulizer for nitrogen gas was adjusted to 1.8 L·min^-1. Results: The good resolution was achieved among 7 polar saponins(ginsenosides Rg₁,Re,Rf,Rb₁,Rc,Rb₂,Rd)and 12 non-polar saponins(ginsenosides F₄,Rg₆,Rk₃,Rh₄,20(S)-Rg₃,20(R)-Rg₃,20(S)-Rs₃,20(R)- Rs₃,Rk₁,Rg₅,Rs₄,Rs₅)of pre-and post-processed ginsengs,and the method was validated by linearity(r≥0.9997). The average recoveries ranged 97.0%-107.5%.Compared with white ginseng,the contents of the non-polar and ocotillol-type saponins were greatly increased in black ginseng developed from white ginseng by nine cycles of steaming and solarization,while those of polar saponins were significantly decreased. Conclusion: HPLC-ELSD can be used for simultaneous determination of 7 polar and 12 non-polar saponins in black ginseng,which can be applied for the quality control of black ginseng,and provide the scientific evidence for the basic material research on black ginseng’s pharmacological effects.