小豆皂苷提取和液质联用检测方法的优化  

Optimization on extraction of azukisaponins and assay by ultra performance liquid chromatography mass spectrometer

作　　者：侯亚楠 王思尧 魏朝俊[2] 杨凯 万平 HOU Yanan;WANG Siyao;WEI Chaojun;YANG Kai;WAN Ping(College of Plant Science and Technology/Beijing Key Laboratory of New Technology in Agricultural Application,Beijing University of Agriculture,Beijing 102206,China;College of Bioscience and Resource Environment/Key Laboratory of Urban Agriculture(North China),Ministry of Agriculture and Rural Affairs,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院植物科学技术学院/农业应用新技术北京市重点实验室 [2]北京农学院生物与资源环境学院/农业农村部华北都市农业重点实验室,北京102206

出　　处：《北京农学院学报》2020年第3期17-21,共5页Journal of Beijing University of Agriculture

基　　金：国家重点研发计划(2018YFD1000705,2018YFD1000700);北京农学院2018年本科生科研训练项目。

摘　　要：【目的】优化小豆皂苷提取方法和适用于超高效液相色谱-电喷雾质谱联用的小豆皂苷检测方法。【方法】以小豆品种‘京农6号’为材料,采用索氏提取法和超声波辅助浸泡法两种提取方法提取小豆皂苷,利用超高效液相色谱-电喷雾质谱联用检测小豆籽粒中6种小豆皂苷的相对含量。【结果】使用80%乙醇作为提取液、物料比1∶20、40℃、40 KHz超声提取90 min,提取效率较索氏提取法平均效率约高10%。采用C18柱(2.1 mm×50 mm,1.8μm),流动相0.1%甲酸/水(A)和乙腈(B),简化洗脱条件为10%乙腈至100%乙腈5 min线性梯度洗脱,柱温25℃,流速0.4 mL/min;电离电压3500 V、源温度350℃电喷雾电离、氮气35 psi、气体流速10 L/min,在5 min中内可快速完成6种小豆皂苷检测。【结论】超声波辅助浸泡法可提高小豆皂苷提取率,缩短提取时间,减少溶剂用量;对超高效液相色谱-电喷雾质谱联用检测小豆皂苷的洗脱梯度进行优化,快速检测小豆皂苷。【Objective】 Optimization of extraction and assay method for adzuki bean saponins by Ultra Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometric. 【Methods】 Azukisaponins were extracted from Jingnong 6 variety of adzuki bean by Soxhlet extraction and Ultrasonic-assisted extraction. The relative content of six azukisaponins in adzuki bean seed were assayed by Ultra Performance Liquid Chromatography-Electrospray Ionization Mass Spectrometric method. 【Results】 Using 80% aqueous ethanol as the extracting solution, 20∶1 as the efficiency ratio of solvent to the seed powder, 40 ℃ and 40 KHz ultrasonic extraction for 90 min, the total azukisaponins was 10% higher than that of Soxhlet extraction. The mobile phase of Agilent C18 column(2.1 mm×50 mm,1.8 μm) was 0.1% aqueous formic acid(A) and acetonitrile(B). The simplified elution conditions were linear gradient elution from 10% acetonitrile to 100% acetonitrile for 5 min. The column temperature was 25 ℃, and the flow rate was 0.4 mL/min. The instrument was operated at an ionization voltage of +3500 V and source temperature of 350 ℃. Nitrogen was used as nebulizer gas at 35 psi and drying gas at a flow rate of 10 L/min, within 5 min to complete the separation and detection of six azukisaponins. 【Conclusion】 Ultrasonic-assisted extraction method can increase the extraction rate, reduce time and solvent. The gradient elution of azukisaponins by Ultra Performance Liquid Chromatography-electrospray Ionization Mass Spectrometric method were optimized, and azukisaponins are rapidly assayed.

关 键 词：小豆皂苷 提取方法 超高效液相色谱-电喷雾质谱 

分 类 号：O657.33[理学—分析化学]