目的 采用核磁共振的代谢组学技术研究“杀青”对连翘Forsythia suspense化学成分的影响。方法 对连翘生品与杀青品的¹H-NMR图谱进行分析;将核磁数据导入SMICA-P软件进行多元统计分析,寻找二者的差异代谢物;并通过生物合成途径分析主要差异代谢物的降解途径。结果 共指认24个代谢物,主成分分析(PCA)显示连翘生品与杀青品能明显区分,说明它们的代谢物组成有显著差异。正交偏最小二乘判别分析(OPLS-DA)的S-plot结合变异权重参数(VIP)显示生品与杀青品的化学差异主要涉及10个代谢物,杀青品中连翘酯苷A、连翘酯苷C、连翘苷、芦丁、蔗糖的量较高,而生品中连翘环己醇、连翘环己醇苷、连翘环己醇酮、α-葡萄糖、β-葡萄糖的量较高。并推测生品中连翘酯苷A大幅降低的原因在于降解为连翘环己醇。结论 从整体代谢物轮廓分析“杀青”对连翘化学组成的影响,证实了传统连翘产地加工“杀青”环节的合理性和必要性,也为连翘的活性成分群研究和多指标质量标准建立奠定了基础。

Objective To investigate the effect of deactivation of enzymes on chemical composition of Forsythiae Fructus using NMR based metabolomics approach. Methods ¹H-NMR-based metabolomics approach combined with multivariate statistical analysis was used to investigate the differential metabolites between raw Forsythiae Fructus (RF) and Forsythiae Fructus with deactivation of enzymes (DF). And degradation pathway of significant metabolites was inferred by biosynthetic pathway. Results Twenty-four metabolites were identified in the ¹H-NMR spectrum of Forsythiae Fructus, and principal component analysis (PCA) showed clear separation between RF and DF. The S-plot of orthogonal partial least square discriminate analysis (OPLS-DA) revealed that ten compounds contributed to the separation of RF and DF, and the levels of forsythiaside A, forsythiaside C, phillyrin, rutin, and surcose were higher in DF than those in RF, while the levels of rengyol, rengyoside, rengyolone, α-glucose, and β-glucose were higher in RF than those in DF. Moreover, the degradation of forsythiaside A into rengyol was proposed. Conclusion This study reveals the chemical effect of deactivation of enzymes on Forsythiae Fructus in a holistic manner, which confirms rationality and necessity of deactivation of enzymes during the processing of Forsythiae Fructus. This study also serves as a basis for the bioactive compounds and quality standards research of Forsythiae Fructus.

科技部科技支撑计划项目(2011BAI07B06);山西省科学技术发展计划项目(20110313001-1);山西省科技创新重点团队(2013131015)