基于UPLC指纹图谱结合化学模式识别的冠心丹参胶囊质量控制研究

Quality control research of Guanxin Danshen Capsule based on UPLC fingerprint combined with chemical pattern recognition

目的采用UPLC法建立冠心丹参胶囊的指纹图谱,并结合化学模式识别技术对其进行系统、全面和科学的质量评价。方法采用Waters UPLC超高效液相色谱仪,Acquity UPLC?HSS T3色谱柱,以乙腈-0.1%甲酸水为流动相进行梯度洗脱,检测波长为256 nm,建立10批次冠心丹参胶囊的指纹图谱。通过相似度分析并结合聚类分析(CA)、主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA)等模式识别技术对冠心丹参胶囊的总体质量进行分析评价。结果建立的指纹图谱共标定75个共有峰,经对照品进行化学指认共鉴定了其中的13个色谱峰,分别是甜菜碱、琥珀酸、丹参素、原儿茶醛、咖啡酸、芦丁、木犀草苷、迷迭香酸、丹酚酸B、丹酚酸A、二氢丹参酮I、隐丹参酮、丹参酮IIA。10批供试品的相似度均大于0.97,表明该药物总体质量较为稳定;但通过CA及PCA均发现不同批次药物质量之间存在微小差异,且主要分为2类,最后进一步采用OPLS-DA筛选出了导致批次药物质量差异的3种主要成分,分别为甜菜碱、芦丁和丹酚酸B。结论本研究建立的分析方法科学、准确、可靠且简便,指纹图谱结合化学模式识别技术可更加系统、全面地评价冠心丹参胶囊的药物质量,同时为今后中药制剂更进一步的质量控制研究奠定了理论基础。

Objective To establish the UPLC fingerprint of Guanxin Danshen Capsule（GDC） and conduct a systemic, comprehensive, and scientific quality evaluation of GDC using a chemical pattern recognition method. Methods Ultra-high performance liquid chromatography instrument and Acquity UPLC？ HSS T3 chromatographic column was employed, the separation was performed with the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution, and the detection wavelength was set at 256 nm to establish the UPLC fingerprint of ten batches of GDC. Then, the further quality assessment of the drug was carried out by similarity evaluation, cluster analysis（CA）, principal component analysis（PCA）, and orthogonal partial least squares discriminant analysis（OPLS-DA）. Results In this research, 75 peaks were recognised as common peaks in the fingerprint, 13 peaks were identified using standard references, they were betaine, succinic acid, tanshinol, protocatechualdehyde, caffeic acid, rutin, galuteolin, rosmarinic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone Ⅰ, cryptotanshinone, and tanshinone ⅡA. The similarity values of the drugs were all above 0.97, indicating a relatively stable quality of the drugs. Little difference was then discovered between the batches of the drug by CA and PCA. Finally, glycinebetaine, rutin, and salvia acid B were recognised as the quality makers using a OPLS-DA method. Conclusion The analysis method established in this study was scientific, accurate, reliable, and simple; The drug quality of GDC could be evaluated systematically and comprehensively using a fingerprint combined with chemical pattern recognition technique. Moreover, it will also lay a solid theoretical basis for the further quality control of traditional Chinese medicine and its preparations at the same time.