液质联用测定蜂蜜中钩吻碱和钩吻碱子

Determination of Gelsemine and Koumine in Honey by Liquid Chromatography-tandem Mass Spectrometry

建立一种高效液相色谱-质谱串联法测定蜂蜜中的钩吻碱和钩吻碱子的方法。样品采用1%盐酸超声提取,经MCX固相萃取小柱净化,再用氨化甲醇进行洗脱,洗脱液浓缩后用50%甲醇水溶液(含0.1%甲酸)定容,进行超高效液相色谱-质谱串联仪分析。采用乙腈和0.1%甲酸水溶液(含5 mmol/L甲酸铵)作为流动相进行梯度洗脱,质谱采用电喷雾离子源(ESI)正离子模式扫描多反应监测(MRM)对钩吻碱和钩吻碱子进行测定。该方法在30 min内完成对钩吻碱子和钩吻碱的分析。钩吻碱在28,56和280μg/kg添加水平的回收率为89.3%~98.1%,相对标准偏差小于4.1%(n=5),方法定量限为5.6μg/kg,检出限为2.8μg/kg;钩吻碱子在20,40和200μg/kg添加水平的回收率为86.2%~95.1%,相对标准偏差小于4.1%(n=5),方法定量限为4.0μg/kg,检出限为2.0μg/kg。该方法能快速、准确、灵敏地测定蜂蜜中的钩吻碱和钩吻碱子。

It was to establish a method for the determination of gelsemine and koumine in honey by liquid chromatographytandem mass spectrometry.The crude sample was extracted by 1%HCl with ultrasound-assisted extraction,then purified by MCX-SPE column,and eluted with ammoria-methanol(5︰95).After concentration,the residue was dissolved by 50%methanolic water(containing 0.1%formic acid).The real sample was analyzed by HPLC/MS.Gradient elution was carried out using acetonitrile and 0.1%aqueous formic acid(containing 5 mmol/L ammonium formate)as mobile phase,and mass spectrometry(ESI+)was used to monitor gelsemine and koumine by MRM mode.This method completed the analysis of gelsemine and koumine in 30 min.The recovery of gelsemine at 28,56 and 280μg/kg was 89.3%-98.1%,and the relative standard deviation was less than 4.1%(n=5).The limit of quantification was 5.6μg/kg and the limit of detection was 2.8μg/kg;The recoveries of hooking base at 20,40 and 200μg/kg were 86.2%-95.1%,the relative standard deviation was less than 4.1%(n=5),and the method limit of quantification was 4.0μg/kg.The limit of detection was 2.0μg/kg.This method was a rapid,accurate and sensitive method for the determination of gelsemine and koumine in honey.