基于^1HNMR对泽泻药材地缘区分和质量评价的代谢组学研究

^1H NMR-based metabonomics for quality evaluation and geographical origin discrimination of Rhizoma Alismatis

目的用基于1H NMR的代谢组学方法,建立泽泻的氢核磁共振指纹图谱,确定不同产地泽泻药材的整体代谢物差异和质量评价。方法用核磁共振仪测定不同产地的泽泻样品,获得1H NMR谱图,所得自由衰减信号导入MestReNova软件进行分段积分、数据归一化后用偏最小二乘法判别分析(PLS-DA)进行模式识别。结果建立了泽泻药材的1H NMR指纹图谱,并鉴定了4个主要代谢成分:23-乙酰泽泻醇B、蔗糖、β-半乳糖和丙氨酸;PLS-DA分析结果显示4个不同产地泽泻药材能够明显分开;以23-乙酰泽泻醇B的相对含量为质量评价指标,经1H NMR定量分析,发现四川产地的泽泻药材质量最优,且与福建产地的泽泻药材有统计学差异(P<0.05)。结论用基于1H NMR的代谢组学方法能有效区分不同产地的泽泻药材和进行质量评价,该方法高效快速、操作简便、无损检测、无需标准品绘制标准曲线,比常规的HPLC反映出更加全面综合的化学信息,且谱图中信号的相对强弱还能反映样品中各组分的相对含量。

Objective To establish 1H NMR fingerprint of Alisma orientalis （Sam.） Juzep., to reveal the holistic metabolite difference of samples from different geographical origins, and to conduct quality evaluation by 1H NMR-based metabonomics. Methods 1H NMR spectra of the samples from different origins were obtained by NMR spectrometer. The obtained free induction decay （FID） signal will be imported into MestReNova software and divided into segments. The data will be normalized and processed by partial least squares discriminant analysis to perform pattern recognition. Results 1H NMR fingerprint of Alisma orientalis （Sam.） Juzep. was established. Four primary metabolites were identified as 23-O-Acetylalisol B, sucrose, β-galactose, and alanine. It was found that the sample collected from Sichuan had the best quality through quantitative 1H NMR analysis of 23-O-Acetylalisol B. Sichuan and Fujian groups had significantly difference in accuracy （P〈0.05）. Conclusion 1H NMR-based metabonomics can effectively distinguish the origins of samples and perform quality evaluation. This method is efficient, fast, and non-destructive, without drawing a standard curve. 1H NMR-based metabonomics shows more comprehensive information as compared with traditional HPLC, and the relative strength of the signal spectrum can reflect the relative contents of the components.