[关键词]
[摘要]
目的 基于当归Ca2+拮抗作用的质量标志物(Q-marker),建立一种利用近红外光谱快速评价当归扩血管功效的方法。方法 采用UPLC/Q-TOF结合Ca2+双荧光报告系统对当归药材的Ca2+拮抗成分进行谱效筛选,并通过细胞和离体实验评价确立Q-marker。对多批药材中的Q-marker进行含量分析,并获取其近红外光谱,建立其近红外拟合算法。将当归提取物的整体Ca2+拮抗活性与Q-marker的含量相关联构建量效拟合函数。结果 研究发现藁本内酯(X1)和欧当归内酯A(X2)具有明显的Ca2+拮抗作用,且其含量波动与当归提取物的整体活性(Y)的变化相吻合,确认其为Q-marker。经非线性回归分析,确认其量效关系符合下列函数方程:Y=31.257 9 X1+381.352 0 X2-248.979 0 X1X2+18.482 2。所优化的近红外检测方法,其预测值与真实值展示了良好的相关性。结论 整合Q-marker研究结果,确立了当归血管舒张作用的Q-marker,以及其与当归药材整体活性之间的非线性关系,并利用近红外光谱技术实现了当归药材血管扩张功效的快速评价,为中药品质的快速分析提供新的解决方案。
[Key word]
[Abstract]
Objective By screening the quality markers of Ca2+ antagonistic ingredients, a rapid evaluation system about the vasodilatory effect of Angelica sinensis (AS) by near infrared spectroscopy (NIR) was established. Methods The Ca2+ antagonists in AS were screened by UPLC/Q-TOF combined with Ca2+double luciferase reporter gene system. To establish the quality markers, the antagonistic effects were further evaluated in cells and in vitro. The quality markers in multi batches of AS were quantitatively analyzed, and the corresponding NIR was obtained, and then the NIR fitting algorithm was established. Meanwhile, the relationship between Ca2+ antagonistic holistic activity of AS extract and the content of quality markers was investigated, and the prediction model for vasodilation efficacy of AS was constructed based on quality markers check analysis via NIR technique. Results The screening result showed that ligustilide (X1) and levistilide A (X2) in AS had significant Ca2+ antagonistic effects, and the change of the content was consistent with the capacity of Ca2+ antagonistic action of AS extracts, so they were confirmed as quality markers. According to nonlinear regression analysis, the relationship between Ca2+ antagonistic effect (Y) of AS and the content of two quality markers satisfied the following functions:Y=31.257 9 X1 + 381.352 0 X2-248.979 0 X1X2 + 18.482 2. In addition, the measured values from NIR simulation method for ligustilide and levistilide A detection showed a good correlation with the predicted values. Conclusion In this study, it was found that ligustilide and levistilide A were quality markers responsible for vasodilation efficacy in AS, and the quantitative correlation between quality markers and the function of vasodilation effect was established. With the help of NIR technology, a novel solution for the rapid monitoring of the quality of traditional Chinese medicine was demonstrated.
[中图分类号]
[基金项目]
天津市科技援助项目(18YFYZCG00060)