RRLC-QTOF／MS研究不同比例制川乌与瓜蒌配伍前后化学成分变化

Chemical constituent changes of Aconiti Cocta Radix before and after combination with Trichosanthes Fructus in different ratios by RRLC-QTOF/MS

目的分析制川乌与瓜蒌配伍前后化学成分变化规律,为阐释制川乌与瓜蒌配伍禁忌提供物质基础。方法利用RRLC-QTOF/MS分析制川乌与瓜蒌配伍提取液,色谱柱为Agilent Zorbax Eclipse Plus C18柱(100 mm×2.1 mm,1.8μm),流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液,梯度洗脱;采用Mass Hunter Workstation软件和偏最小二乘判别法进行化合物鉴别和差异分析;以化合物与内标峰面积之比来表示化学标志物的相对质量分数。结果采用此方法共检测到制川乌-瓜蒌合煎液中65种化学成分,进一步鉴别了其中57种成分,15种成分在配伍前后有差异,其溶出受制川乌与瓜蒌配伍比例的变化影响。结论 RRLC-QTOF/MS方法灵敏、准确,能够较全面反映制川乌与瓜蒌配伍前后化学成分的变化;从体外化学物质基础角度分析,中药"十八反"之乌头与瓜蒌是否相反尚无定论,还需结合体内实验进一步深入研究。

Objective To analyze the chemical constituent changes ofAconiti Cocta Radix （ACR） before and after combination with Trichosanthes Fructus （TF） in different ratios and to provide the material bases for the incombination. Methods The rapid resolution liquid chromatography with quadrupole and time-of-flight mass spectrometry （RRLC-QTOF/MS） was used to analyze the extract solution from ACR and TF combination. The chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C18 column （100 mm × 2.1 mm, 1.8 μm） with 0.1% formic acid water solution-0.1% formic acid in acetonitrile solution as mobile phase for gradient elution. Mass Hunter Workstation and partial least squares-discriminant analysis （PLS-DA） were utilized for the identification of components and chemical biomarkers. Peak-area ratio of the analyte/internal standard was used to represent the contents of the variance analysis. The amounts of chemical markers were indicated by the ratio of compound and internal standard peak area. Results By an overall analysis ofRRLC-QTOF/MS, a total of 65 compounds were detected in the extracts from ACR in combination with TF, including 57 compounds with confirmed structures and 15 compounds with significant differences before and after combination, and the relative dissolution rates were affected by the changes of the combination ratios. Conclusion The sensitive and accurate RRLC-QTOF/MS method could fully reflect the chemical constituent changes of ACR before and after combination with TF. On the basis of in vitro chemical constituent and contents, it is still unclear whether the combination of ACR and TF should be prohibited, which requires further in vivo investiaation.