HPLC-Q-TOF-MS和UPLC-QqQ-MS的三叶青主要成分定性与定量研究

Qualitative and quantitative analysis of major constituents in Tetrastigma hemsleyanum by HPLC-Q-TOF-MS and UPLC-Qq Q-MS

建立高效液相色谱串联四极杆飞行时间质谱法（HPLC-Q-TOF-MS）快速鉴定三叶青化学成分,建立超高效液相色谱串联三重四级杆质谱法（UPLC-Qq Q-MS）同时测定三叶青中4种成分（芦丁、异槲皮苷、山柰酚芸香糖苷、紫云英苷）的含量。HPLC-Q-TOF-MS法采用月旭Ultimate-XB C18色谱柱（4.6 mm×150 mm,5μm）,流动相为乙腈（A）-0.1%甲酸水（B）梯度洗脱,流速0.8 m L·min^-1,柱温30℃。飞行时间质谱,采用负离子模式扫描。UPLC-Qq Q-MS法采用Waters CORTECS C18（2.1 mm×100 mm,1.6μm）,流动相为乙腈（A）-0.1%甲酸水（B）梯度洗脱,柱温45℃,采用负离子多反应监测（MRM）方式进行含量测定。结果 HPLC-Q-TOF-MS法推断鉴定了三叶青中的24种成分,UPLC-Qq Q-MS法定量分析三叶青中4种成分在考察的浓度范围内,线性关系良好（r〉0.996 6）;回收率均在98.27%-101.6%,RSD 3.2%-5.9%。通过HPLC-Q-TOF-MS联用技术,为鉴定三叶青的化学成分提供了一种快速、高效的定性分析方法。建立的UPLC-Qq Q-MS同时测定三叶青中4种成分的定量分析方法简便、快捷、准确,为综合评价三叶青的质量提供参考。

A qualitative analytical method of liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry（ HPLC-Q-TOF-MS） was developed for identification of multi-constituents and an analytical method was developed for simultaneously determining 4 major compounds（ rutin,isoquercitrin,kaempferol-3-O-rutinoside,and astragalin） in Tetrastigma hemsleyanum Diels et Gilg. The HPLC-Q-TOF-MS assay was performed on a Welch Ultimate XB-C18column（ 4. 6 mm × 150 mm,5 μm） with the mobile phase consisting of acetonitrile（ A） and water containing 0. 1% Formic acid（ B） in gradient mode at a flow rate of 0. 8 m L·min-1. The column temperature was at 30 ℃,and negative ion mode was used for TOF-MS. The UPLC-Qq Q-MS assay was performed on a Waters CORTECS C18（ 2. 1 mm × 100 mm,1. 6 μm） with the mobile phase consisting of acetonitrile（ A） and water containing0. 1% formic acid（ B） in gradient mode at a flow rate of 0. 25 m L·min-1. The column temperature was at 45 ℃,and MRM mode was used for Qq Q-MS. Based on the retention time and MS spectra,24 compounds were identified or tentatively characterized by comparing with reference substances or literatures. For quantitative the linear range of 4 detected compounds were good（ r 0. 996 6）,and the overall recoveries ranged from 98. 27% to 101. 58%,with the RSD ranging from 3. 15% to 5. 88%. The results indicated that new approach conbined HPLC-Q-TOF-MS and UPLC-Qq Q-MS was applicable in qualitative and quantitative quality control of Tetrastigma hemsleyanum.