基于HPLC-Q-TOF-MS/MS技术的藏药熏倒牛化学成分分析

Analysis of Chemical Constituents in Biebersteinia heterostemon Maxim by HPLC-Q-TOF-MS/MS

目的:利用高效液相色谱-四级杆串联飞行时间质谱(HPLC-Q-TOF-MS)在负离子模式下对藏药熏倒牛中的化学成分进行分析和鉴定。方法:采用Agilent ZORBAX Eclipse Plus C_(18)柱(3.0 mm×150 mm,1.8μm);以0.1%甲酸的乙腈(A)和0.1%甲酸水溶液(B)为流动相梯度洗脱;流速为0.250 mL·min^(-1);进样体积:2μL;柱温为35℃;以电喷雾(ESI)离子源,负离子模式下采集数据,通过分析质谱数据和元素组成,结合文献数据对照,分析推测所含化合物。结果:通过分子离子峰和二级高分辨质谱数据结合文献数据鉴定了熏倒牛全草中的25个化合物,包括1个脂环族有机酸类、2个香豆素类、2个多酚类、1个蒽醌类及19个黄酮类化合物。结论:UPLC-Q-TOF-MS/MS技术为鉴别藏药熏倒牛药材中的化学成分提供快捷、准确的方法,其主要化学成分为黄酮醇类、异黄酮类、蒽醌类,并从该植物中首次鉴定出了14个黄酮类成分,可为熏倒牛药材的药效物质基础研究和质量控制提供参考。

Objective To analyze and identify the chemical constituents in Biebersteinia heterostemon Maxim by liquid chromatography-quadrupole-time-of-flight high resolution mass spectrometry(HPLC-Q-TOF-MS/MS)in a negative ion mode.Methods The analysis was carried out on a Agilent ZORBAX Eclipse Plus C_(18) chromatographic column(3.0 mm×150 mm,1.8μm)with 0.1%formic acid acetonitrile(A)-0.1%formic acid aqueous solution(B)as mobile phase by gradient elution,at a flow rate of 0.250 mL·min^(-1),the injection volume was 2μL,the column temperature was set at 35℃.The data were collected under the negative ion mode using an ESI ion source.The compounds were identified by mass spectrometry data,elemental composition and reference data.Results Totally 25 compounds from Biebersteinia heterostemon Maxim were identified by the molecular ion peaks,secondary high resolution mass spectrometry and literature data,among which 1 was Alicyclic organic acids、2 were coumarins、2 were polyphenols、1 was anthraquinone and 19 were flavonoids.Conclusion HPLC-Q-TOF-MS detecting techniques can be used to analyze and identify the main chemical constituents of flavonols、isoflavones and anthraquinone in Biebersteinia heterostemon Maxim quickly and comprehensively,at the same time,fourteen flavonoids were found from this genus for the first time which provides reference for the basic research of pharmacodynamics and the quality control of Biebersteinia heterostemon Maxim.