高效液相色谱-串联质谱法同时测定番茄中的番茄苷与番茄碱

Simultaneous Determination of Tomatine and Tomatidine in Tomatoes by High Performance Liquid Chromatography-Tandem Mass Spectrometry

利用高效液相色谱-串联质谱法,建立番茄中番茄苷和番茄碱2种主要生物碱的同时检测方法。样品经匀浆提取后,利用C18固相萃取柱净化,以5 mmol/L乙酸铵(含0.05%甲酸)-乙腈为流动相,Agilent Proshell 120 ECC18进行分离,三重四极杆检测器,多反应监测模式进行检测。结果表明,番茄苷在50~1000 ng/mL范围内线性良好,相关系数为0.9991。番茄碱在5~100 ng/mL范围内线性良好,相关系数为0.9994。对番茄苷和番茄碱进行3个水平的添加回收实验,结果表明添加回收率在85.6%~105%之间,相对标准偏差在1.0%~6.8%之间。所建方法简单、快速、灵敏度高、重复性好,可用于番茄中番茄苷与番茄碱的快速、定量检测。实际样品检测结果表明,番茄中番茄苷的含量明显高于番茄碱。

An analytical method was developed for the simultaneous determination of tomatine and tomatidine in tomatoes by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Samples were homogenized,separated on a Proshell 120 EC-C18 column,and purified by solid phase extraction(SPE)with C18 as sorbent and tomatine and tomatidine were then analyzed by triple quadrupole tandem mass spectrometry under the multiple reaction monitoring(MRM)mode.The mobile phase consisted of 5 mmol/L ammonium acetate(containing 0.05%formic acid)and acetonitrile.The results showed that good linearity was observed for tomatine and tomatidine with correlation coefficients above 0.9991 and 0.9994 in the range of 50–1000 and 5–100 ng/mL,respectively.The average recoveries at three spiked levels ranged from 85.6%to 105%,with relative standard deviations(RSDs)varying from 1.0%to 6.8%.The developed method was simple and rapid with high sensitivity and repeatability,which was suitable for rapid and simultaneous determination of tomatine and tomatidine in tomatoes.Tomatine was significantly more abundant than tomatidine in tomato samples as determined by this method.