血人参水溶性化学成分的UHPLC-DAD-Q-TOF-MS/MS分析

Analysis of Water-soluble Chemical Constituents of Indigoferae Stachyoidis Radix by UHPLC-DAD-Q-TOF-MS/MS

摘要:

目的: 采用超高压液相色谱-二级管阵列检测器-四级杆-飞行时间质谱(UHPLC-DAD-Q-TOF-MS/MS)联用技术,在负离子模式下开展血人参水煎煮提取物中化学成分的一级及二级质谱研究,分析血人参的水溶性化学成分. 方法: 液相色谱条件为Synergi Hydro-RP色谱柱(2.0 mm×100 mm,2.5 μm),0.1%甲酸乙腈-0.1%甲酸水溶液梯度洗脱.质谱条件为Bruker Daltonics micro TOF-Q质谱仪,电喷雾离子源,负离子模式,毛细管电压2 800 V,二级质谱碰撞能量11～25 eV;源内碰撞诱导裂解(CID)及其二级质谱条件:源内CID能量25～35 eV,碰撞能量12～17 eV. 结果: 共推测出15个化合物的化学结构,分别为没食子酸、原儿茶酸-3-O-葡萄糖苷、原儿茶酸、原儿茶醛、丁香酸葡萄糖苷、龙胆酸、儿茶素、表儿茶素、原花青素B类同分异构体4个、棕儿茶素A类同分异构体3个. 结论: 除化合物表儿茶素外,其他14个化合物均为该植物首次报道.研究结果为血人参及其制剂的开发和质量标准制定提供了依据.

Abstract:

Objective: An ultra-high-pressure liquid chromatography-diode-array detection-quadrupole time-of-flight mass spectrometric (UHPLC-DAD-Q-TOF-MS/MS) method was established to analyze the chemical constituents of the decoction of Indigoferae Stachyoidis Radix. Method: The chromatographic separation was performed on a Synergi Hydro-RP (2.00 mm×100 mm, 2.5 μm) column using a mobile phase consisting of 0.1% formic acid in water(A)and 0.1% formic acid in acetonitrile (B). The flow rate was 0.3 g· L^-1 and the mass spectrometer was Bruker Daltonics micro TOF-Q equipped with an electrospray ionization (ESI) source. The MS and MS/MS spectra were recorded in the negative ion mode.The capillary voltage is 2 800 V. The range of collision energy of MS² is 11 eV from 25 eV. And a condition of Endogenous collision induced dissociation (CID) is 25 eV from 35 eV, the corresponded MS² energy is 12 eV from 17 eV. Result: 15 compounds in the decoction of Indigoferae Stachyoidis Radix were well isolated and identified as gallic acid, protocatechuic acid-3-O-glucoside, protocatechuic aldehyde, protocatechuic acid, glucosyringic acid, gentisic acid, catechin, epicatechin, four procyanidines and three gambiriins. Conclusion: In addition to the compound epicatechin, the other 14 compounds were first reported in this plant. The results have formed the basis for the exploitation of Indigoferae Stachyoidis Radix and the establishment of a quality control method for Indigoferae Stachyoidis Radix.