芦荟药材化学成分鉴定及UPLC指纹图谱分析

Chemical profile of herbal medicine aloe by UPLC-Q-TOF/MS and its UPLC fingerprinting

目的鉴定芦荟中的主要化学成分,建立UPLC特征指纹图谱,对市售15批次芦荟药材的质量进行评价。方法应用超高效液相色谱-串联四极杆飞行时间质谱(UPLC-Q-TOF/MS)采集芦荟色谱质谱数据,并进行定性分析;UPLC色谱条件为:Waters CORTECS T3(100 mm×2.1 mm,1.6μm)色谱柱,柱温40℃,检测波长295 nm,乙腈(0.1%甲酸)-水(0.1%甲酸)为流动相,体积流量0.3 mL/min,梯度洗脱;Q-TOF/MS条件为采用ESI离子源,负离子扫描模式,毛细管电压2.3 kV,离子源温度100℃;利用《中药色谱指纹图谱相似度评价系统(2012版)》软件建立15批市售芦荟药材的UPLC特征指纹图谱。结果从芦荟药材中共鉴定34个化学成分,主要包括蒽酮类成分10个、蒽醌类成分1个、色酮类成分16个、吡喃酮类成分5个、其他类成分2个;建立的UPLC指纹图谱精密度、稳定性、重复性良好,能够满足《中国药典》2020年版规定的指纹图谱要求,15批芦荟样品的指纹图谱相似度计算结果在0.867~0.986;对18个共有峰中的14个进行了结构指认,以共有峰峰面积为变量的主成分分析和聚类分析可明确对市售芦荟药材的基原进行分类。结论所建立的UPLC-Q-TOF/MS方法可快速识别中药芦荟中的主要化学成分;建立的UPLC指纹图谱方法稳定可靠、专属性强,可为芦荟的质量评价提供参考;指纹图谱中的共有峰芦荟苷A、芦荟苷B、芦荟宁B、芦荟糖苷A、芦荟糖苷B还可用于区分芦荟药材的植物基原。

Objective To identify the major chemical constituents of Aloe raw herb and establish UPLC characteristic fingerprinting of Aloe(15 batches)for evaluation its quality.Methods An ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)was applied in this study.The UPLC method was performed on a Waters CORTECS T3(100 mm×2.1 mm,1.6μm)column with 40℃at 295 nm,and gradient elution with acetonitrile(0.1%formic acid)-water(0.1%formic acid)was used at the flow rate of 0.3 mL/min.The Q-TOF/MS method was as follows:electrospray ionization(ESI)source under negative ion scan mode;capillary voltage,2.3 kV;source temperature,100 oC.The software of similarity calculation for traditional Chinese medicines fingerprints(version 2012)was used to establish the characteristic fingerprinting of Aloe(15 batches).Results A total of 34 compounds were identified from Aloe raw herb,including 10 anthrones,1 anthraquinone,16 chromones,5 pyrones and 2 other compounds.The established UPLC fingerprinting had good precision,stability,and repeatability,which can meet the demand of the Chinese Pharmacopoeia.The similarities of Aloe raw herb from 15 batches were between 0.867 and 0.986.Meanwhile,14 of 18 common peaks were identified by UPLC-Q-TOF/MS.Principal component analysis(PCA)and cluster analysis can clearly classify the original plant of Aloe vera using common peak area as the variable.Conclusion The established method of UPLC fingerprinting is stable,reliable,and specific,and the major components of Aloe raw herb can be quickly identified by the UPLC-Q-TOF/MS method.The combined method of UPLC fingerprinting and qualitative analysis by UPLC-Q-TOF/MS can provide more information for the quality evaluation of Aloe raw herb.Among the common peaks in the fingerprinting,aloin A,aloin B,aloenin B,aloinoside A and aloinoside B can be used to differentiate the origin plants of Aloe.