目的 分析制附片在煎煮过程中化学成分群的变化规律,探讨制附片的科学煎煮时限。方法 利用超高效液相色谱-四极杆飞行时间质谱(UHPLC-Q-TOF/MS)技术对不同煎煮时间的制附片化学成分群信息进行采集,结合多元统计分析技术手段对采集得到的高精度数据进行处理,发现并鉴定出随煎煮时间变化显著的差异化合物,并对其变化规律、毒性及活性进行综合分析。结果 经偏最小二乘判别分析(PLS-DA)法及t检验筛选得到15个差异化学标志物;通过分析4 h内不同成分的变化规律发现,制附片煎煮过程中尼奥林等众多酯性生物碱并不受热水解,成分变化主要是单酯型生物碱向醇胺类原碱的转变,而双酯型生物碱的水解是次要的。结论 临床治疗过程中,制附片若以发挥镇痛抗炎作用为主,建议常规煎煮0.5 h即可;若以发挥强心作用为主,还需要对单酯型生物碱与醇胺类原碱的强心活性进行系统比较,才能够确定久煎的必要性与科学性。

Objective To analyze the change regulation of chemical constituent groups in processed aconite during decocting, and also to discuss the scientificity connotation of decoction of processed aconite. Methods An ultra-high performance liquid chromatography coupled with time of flight mass spectrometry(UHPLC-Q-TOF/MS) was carried out to acquire the chemical constituents information in samples with different decoction time. High precise mass data were processed by multivariate statistical analysis techniques to discover and identify the constituents with significant difference. Furthermore, the content change, toxicity, and activity of these constituents were also discussed. Results Fifteen chemical markers with significant difference were screened by partial least squares discriminant analysis(PLS-DA) and t test. By analyzing the change regulation of the different constituents during 4 h decocting process, we found that it mainly happened in the constituent dissolution and chemical transformation from monoester alkaloids to aconine alkaloids, which indicates the attenuation process is secondary. Conclusion In terms of analgesic and anti-inflammatory effect, it suggests that 0.5 h of decoction would be fine, while in terms of cardiotonic effect, it needs the further systemic comparison for cardiac activity between monoester alkaloids and aconine alkaloids to be able to investigate the necessary and scientificity for long time decoction of the processed aconite.

国家自然科学基金资助项目(81403115);成都中医药大学科技发展基金(030029037);国家级中药炮制技术传承基地(中国中医药科技[2015]86号)