基于HPLC指纹图谱结合化学模式识别的山茱萸酒炙前后对比研究

Comparative Study of Cornus Officinalis before and after Wine Processing Based on HPLC Fingerprints Combined with Chemical Pattern Recognition

目的:建立山茱萸生品和酒炙品HPLC指纹图谱,结合化学模式识别方法比较酒炙前后化学成分指纹峰的差异,寻找差异标志物,为山茱萸的酒炙工艺研究提供科学依据。方法:采用HPLC法,Phenomenex Luna C_(18)色谱柱(250 mm×4.6 mm,5μm),乙腈-0.2%磷酸水溶液为流动相进行梯度洗脱,体积流量为0.9 ml·min^(-1),检测波长为240 nm(莫诺苷、獐牙菜苷、马钱苷和山茱萸新苷Ⅰ)、360 nm(齐墩果酸和熊果酸)。采用中药色谱指纹图谱相似度评价系统(2012版)计算相似度,运用聚类分析(HCA)和正交偏最小二乘法-判别分析(OPLS-DA)对山茱萸生品与酒炙品进行化学模式识别研究。结果:建立了山萸肉和酒萸肉的指纹图谱,共标定19个共有峰,通过与对照品比对,指认了6个成分,分别为莫诺苷(8号峰为),獐牙菜苷(11号峰),马钱苷(13号峰),山茱萸新苷I(15号峰),齐墩果酸(16号峰),熊果酸(19号峰);通过HCA和OPLS-DA可明显将山茱萸生品与酒炙品分为2类,熊果酸、山茱萸新苷I、齐墩果酸等7个成分可能是影响两者质量的差异标志物。结论:建立的指纹图谱方法精密度、重复性、稳定性良好,结合化学模式识别可用于山茱萸酒炙前后整体质量控制。

Objective:To establish the HPLC fingerprint of cornus officinalis raw herb and wine processed products,and compare the differences in chemical fingerprint peaks before and after wine processing with chemical pattern recognition method,so as to find the differentical markers,and provide scientific basis for the wine processing technology of cornus officinalis.Methods:An HPLC method was established on a Phenomenex Luna C_(18)column(250 mm×4.6 mm,5μm)with acetonitrile-0.2%phosphoric acid as the mobile phase with gradient elution.The flow rate was 0.9 ml·min^(-1),the detection wavelength were 240 nm(morroniside,sweroside,loganin and cornuside I)and 360 nm(oleanolic acid and ursolic acid).The fingerprints of raw and processed cornus officinalis were determined respectively and analyzed by the software of Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine.The discoiminant analysis of hierarchical cluster analysis(HCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA)were employed to distinguish raw herb and wine processed cornus officinalis.Results:The fingerprints of raw and processed cornus officinalis at different processing degrees were established,and a total of 19 common peaks were marked.By comparison with the standard,six ingredients were identified,they were morroniside(peak 8),sweroside(peak 11),loganin(peak 13),cornuside I(peak 15),oleanolic acid(peak 16)and ursolic acid(peak 19),respectively.Through HCA and OPLA-DA,cornus officinalis raw products and wine processed products could be obviously divided into two categories.Ursolic acid,cornuside I,oleanolic acid and other seven components may be the differential markers affecting the quality of the two products.Conclusion:The established fingerprint method has good precision,repeatability and stability,and combined with chemical pattern recognition,it can be used for the overall quality control of cornus officinalis before and after wine processing.