微波辅助萃取-高效液相色谱-串联质谱法测定辣椒中3种辣椒素

Microwave-assisted extraction and determination of three capsaicinoids in Capsicum annuum by HPLC-MS/MS

目的:建立微波辅助萃取-高效液相色谱-串联质谱法测定辣椒中3种辣椒素类化合物(辣椒素、二氢辣椒素、降二氢辣椒素)含量。方法:辣椒样品经微波辅助萃取甲醇稀释后分析。色谱分离采用C18反相色谱柱(150 mm×2.1 mm,3.5μm),流动相为0.1%甲酸水溶液和乙腈,梯度洗脱。串联质谱在多反应监测模式下检测目标分析物,以保留时间和特征离子对(母离子和2个碎片离子)信息比较进行定性和定量分析。结果:3种辣椒素的检出限为0.01～0.11 ng.mL-1,定量限为0.03～0.36 ng.mL-1,辣椒素、二氢辣椒素、降二氢辣椒素平均回收率分别为95.4%,94.3%,93.1%;RSD分别为2.1%,2.5%,2.7%。结论:该方法简便、准确、灵敏,可用于辣椒中3种辣椒素的分析。

Objective：To establish a microwave-assisted extraction coupled with high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） method for the determination of three capsaicinoids（capsaicin,dihydrocapsaicin,nordihydrocapsaicin） in Capsicum annuum.Methods：The sample was extracted by a microwave-assisted method and diluted with methanol for further analysis.The analysis was performed on a C18 column（150 mm×2.1 mm,3.5 μm） using a gradient elution program with mobile phases of 0.1% formic acid and acetonitrile.The analyte was determined by electrospray ionization tandem mass spectrometry in multiple reactions monitoring（MRMs） mode.The qualitative and quantitative analyses were based on the retention times and characteristic ion pairs consisting of one parent ion and two fragment ions of the analyte.Results：The limits of detection（LODs） for the 3 capsaicinoids were in the range of 0.01-0.11 ng·mL-1,and the limits of quantification（LOQs） were in the range of 0.03-0.36 ng·mL-1.The average recoveries were 95.4% with RSD of 2.1% for capsaicin,94.3% with RSD of 2.5% for dihydrocapsaicin,and 93.1% with RSD of 2.7% for nordihydrocapsaicin,respectively.Conclusion：This method is simple,reliable and sensitive,and is therefore suitable for the analysis of the three capsaicinoids in Capsicum annuum.