基于UPLC-DAD/ESI-Q-TOF_MS技术的丹红注射液指纹图谱的构建

目的:建立丹红注射液的UPLC指纹图谱,并对丹红注射液的指纹图谱共有峰进行鉴定。方法:采用UPLC法,ACQUITY UPLC^® HSS T3色谱柱(2.1 mm×100 mm,1.8μm),柱温40℃,乙腈-0.1%甲酸水二元梯度洗脱,体积流量0.4 mL·min^-1,检测波长为286 nm,进样量2μL;采用飞行时间质谱仪获得化合物准分子离子和碎片离子的精确质量数,正、负离子模式扫描。结果:建立了丹红注射液的UPLC指纹图谱,共标定了24个共有峰;鉴定了丹参素、原儿茶醛、原儿茶酸、咖啡酸、羟基红花黄色素A(HSYA)、丹酚酸A、丹酚酸B等21个成分;采用《中药色谱指纹图谱相似度评价系统》(2004A版)评价了30批丹红注射液指纹图谱相似度均大于0.95。采用UPLC-DAD/ESI-Q-TOF MS技术鉴定了丹红注射液的指纹图谱中的21个共有峰的化学结构。结论:该方法构建的指纹图谱灵敏度高,稳定可靠,为丹红注射液质量控制研究提供了科学依据。

Objective:To establish chromatographic fingerprint of Danhong injection with UPLC-DAD technology. UPLC-DAD/ESI-Q-TOF MS was adopted to characterize the common peaks of chromatographic fingerprints of Danhong injection.Methods: The UPLC method was established by employing ACQUITY UPLC^® HSS T3 column(2.1 mm×100 mm, 1.8μm) with acetonitrile-0.1% formic acid aqueous solution as mobile phase in a gradient elution.The detection wavelength was 286 nm. The flow rate was 0.4 mL·min^-1, the injection volume was 2μL and the column temperature was 40℃. Time of flight mass spectrometer was employed to obtain exact mass weight of quasi-molecular ion and fragment ion in positive and negative modes, respectively.Results: By choosing rosmarinic acid as reference peak, the UPLC chromatographic fingerprint of Danhong injection was established, 24 common peaks were obtained. The similarity of 30 batches of Danhong injection was over 0.95, which was evaluated by similarity evaluation system for chromatographic fingerprint of TCM(Version 2004A).21 of 24 common peaks of Danhong injection were identified by UPLC-DAD/ESI-Q-TOF MS, such as tanshinol, protocatechuic aldehyde, protocatechuic acid, caffeic acid, hydroxy safflower yellow A(HSYA), salvianolic acid A, salvianolic acid B, etc.Conclusion: This method is stable and reliable, and can be used for the quality control of Danhong injection.