金丝苦楝的化学成分鉴定及不同产地的6个黄酮类成分含量测定分析

Identification of Chemical Constituents of Cardiospermum halicacabum L.and Analysis its Six Flavonoid Components from Different regions Content Determination

关键词:  金丝苦楝  超高效液相色谱-串联四极杆飞行时间质谱  高效液相色谱法  含量测定  主成分分析  正交偏最小二乘法-判别分析

Key words:  Cardiospermum halicacabum L.  UPLC-Q/TOF-MS/MS  HPLC  Content determination  PCA  OPLS-DA

摘要:

基于超高效液相色谱-串联四极杆飞行时间质谱( UPLC-Q/TOF-MS/MS) 技术分析金丝苦楝药材的化学成分，并基于HPLC技术建立同时测定不同产地金丝苦楝的6个黄酮类成分含量的方法。方法 UPLC-Q/TOF-MS/MS采用Cortecs UPLC C18色谱柱（2.1 mm×100 mm, 1.6 μm），流动相甲醇（A）-0.1%甲酸水（B）梯度洗脱，流速0.2 mL/min，柱温35℃，进样量2 μL。MS条件：电喷雾正、负离子模式采集数据，质谱扫描范围m/z 50~1000。HPLC采用Shim-pack GIST C18色谱柱（4.6×250 μm，5 μm），流动相为甲醇（A）-0.1%甲酸水（B），梯度洗脱，流速1 mL/min，柱温35℃，检测波长350 nm，进样量10 μL。使用 SIMCA-P 14.1进行主成分分析（PCA），筛选得到差异性（VIP＞1）的成分。结果 基于 UPLC-Q/TOF-MS/MS 技术在正、负离子模式下检测出33个化合物，基于HPLC方法测定不同产地金丝苦楝的6个黄酮类成分，进行主成分分析，结果显示福建、广东、广西3个产地的金丝苦楝各自聚为一类，不同批次金丝苦楝的主要差异性成分为异槲皮苷，不同产地的样品存在较大差异，同时通过研究发现广西产的金丝苦楝样品组间差异小，质量较稳定。结论 基于UPLC-Q-TOF-MS/MS技术，可实现对金丝苦楝药材化学成分的快速鉴定，建立的HPLC测定金丝苦楝中6种黄酮类成分的含量方法稳定、可靠，可为其质量控制提供参考。

Abstract:

Based on UPLC-Q/TOF-MS/MS technology, analyze the chemical components of Cardiospermum halicacabum L. and establish a method for simultaneous determination of six flavonoid components in Cardiospermum halicacabum L. from different regions using HPLC technology.METHODS UPLC-Q/TOF-MS/MS using Cortec UPLC C18 column (2.1 mm×100 mm, 1.6 μm) Mobile phase methanol (A) -0.1% formic acid water (B) gradient elution, flow rate 0.2 mL·min-1, column temperature 35℃, injection volume 2 μL; electrospray ionization ( ESI) source was used to acquire mass spectrometry data in positive and negative ion modes with the scanning range of m/z m/z 50~1000. HPLC was performed on Shim-pack GIST C18 (2.1 mm×100 mm, 1.6 μm) with mobile phase consisting of methanol (A) -0.1% formic acid water (B), gradient elution at the flow rate of 1 mL·min-1, column temperature of 35℃, detection wavelength of 350 nm, and sample size of 10 μL. Principal component analysis (PCA) was performed using SIMCA-P 14.1, and the components with differences (VIP > 1) were screened. RESULTS Based on the UPLC-Q/TOF-MS/MS technique, 33 compounds were detected in positive and negative ion mode.The results of principal component analysis showed that the samples from Fujian, Guangdong and Guangxi were grouped into one class respectively. The main component of different batches was isoquercitrin, and the samples from different places had great differences. At the same time, it was found that there was little difference between the samples and the quality was stable. CONCLUSION Based on the UPLC-Q-TOF-MS/MS technique, rapid identification of chemical components of Cardiospermum halicacabum L. was realized. The established HPLC method for the determination of 6 flavonoids in Cardiospermum halicacabum L. was stable and reliable, which could provide reference for quality control.