血栓心脉宁片UPLC-PDA指纹图谱研究

Study on UPLC-PDA fingerprint of Xueshuan Xinmaining Tablets

目的建立血栓心脉宁片(XXT)的UPLC-PDA指纹图谱,考察不同批次XXT质量的一致性,为XXT质量稳定性提供可靠的科学依据。方法样品经甲醇提取,采用Waters Acquity UPLC HSS T3(50 mm×2.1 mm,1.8μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,190~400 nm波长扫描,体积流量0.5 m L/min,进样量3μL,柱温40℃;对30批XXT样品进行方法学考察、相似度计算和聚类分析;单味药材进行共有峰归属,并通过对照品对共有峰的化学成分进行指认。结果建立了XXT UPLC-PDA指纹图谱,280 nm波长下标定28个共有峰,其中1、10、12~16、18、21~28号峰来自丹参,2~4、6、8、9、17号峰来自川芎,3、7、9、11、16号峰来自毛冬青、5号峰来自槐花,19、20号峰来自蟾酥,经对照品比对指认出4号峰为阿魏酸、5号峰为芦丁、13号峰为丹酚酸B、19号峰为酯蟾毒配基、20号峰为华蟾酥毒基、28号峰为丹参酮IIA;通过聚类分析30批XXT可分为2类,经中药色谱指纹图谱评价软件(2012版)计算得30批XXT相似度均大于0.960。结论建立的XXT UPLC-PDA指纹图谱具有良好的精密度、稳定性和重复性,方法简便可靠,可为进一步完善XXT的质量评价提供参考依据。

Objective To assess the consistency of 30 batches of Xueshuan Xinmaining Tablets（XXT）, we establish the UPLC-PDA fingerprint of XXT and provide reliable scientific basis for the Chinese medicine quality consistency. Methods Methanol was used to extract the XXT samples, and the extracts were measured by UPLC with the absorption wavelength at 280 nm. Waters Acquity UPLC HSS T3 reversed-phase column（50 mm × 2.1 mm, 1.8 μm） was used for determining the extracts at a flow rate of 0.5 mL/min and column temperature of 40 ℃. The mobile phase condition was acetonitrile-0.1% formic acid aqueous solution with gradient mode. The methodology, similarity calculation and cluster analysis of 30 batches of XXT samples were investigated. The common peak of XXT was belonged to single medicinal materials and common peaks were identified by comparing with the reference. Results The UPLC fingerprint of XXT was established. Twenty-seven common fingerprint peaks were indicated with wavelength 280 nm, Among them, peaks 1, 10, 12—16, 18, and 21—28 belonged to Salviae Miltiorrhizae Radix et Rhizoma（SMRR）, peaks 2—4, 6, 8, 9, and 17 belonged to Chuanxiong Rhizoma（CR）, peaks 3, 7, 9, 11, and 16 belonged to Ilicis Pubescentis Radix（IPR）, peak 5 belonged to Sophora Flos（SF）, peaks 19 and 20 belonged to Bufonis Venenum（BV）. Peaks 4, 5, 13, 19, 20, and 28 were ferulic acid, rutin, salvianolic acid B, resibufogenin, cinobufagin, tanshinone IIA, respectively, compared with reference. The analyzed samples were classified into two classes and the similarity was greater than 0.960. Conclusion The method of XXT UPLC-PDA fingerprint was established with good precision, stability and repeatability, supplying helpful information for the comprehensive evaluation of XXT.