HPLC-MS/MS法同时测定烟草中22种Amadori化合物

Simultaneous determination of 22 Amadori compounds in tobacco leaves by HPLC-MS/MS

为同时测定不同类型烟叶中的Amadori化合物,围绕12种无标样Amadori化合物,分别进行了美拉德反应合成实验,基于高分辨的高效液相色谱-四极杆飞行时间质谱联用仪(HPLC-Q-TOF-MS),通过精确分子量匹配及二级碎裂方式验证,准确定性了各反应体系中的Amadori化合物,确定了用于多反应监测(MRM)模式的定量及定性离子对。在前期建立的10种具有标样的Amadori化合物的外标绝对定量方法的基础上,对12种无标样Amadori化合物采用保留时间及结构较为近似的Amadori化合物标准品的标准曲线进行近似定量。结果表明:(1)连续进行6 d以上测试(共进样300针以上),14个Amadori化合物的日间精密度<10%,5个Amadori化合物的日间精密度在10%~15%之间。(2)烤烟、香料烟和白肋烟样品分析结果表明,Amadori化合物总量分别占烟叶干质量的2.47%、1.70%和0.14%,且不同类型烟叶中各Amadori化合物的质量分数也存在较大差异。该方法可实现烟草中22种Amadori化合物的同时快速测定。

To simultaneously determine the Amadori compounds in different types of tobacco,the Maillard model reactions for 12 Amadori compounds without standard reagents on sale were conducted respectively.Based on high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS),the 12 Amadori compounds were qualitatively identified in the model reaction systems after being matched with accurate molecular weight and confirmed with the secondary fragmentation modes of targets.The quantitative and qualitative ion pairs for MRM mode were also selected.Based on our previously established external standard quantitative method for the 10 Amadori compounds with available standard reagents,the 12 Amadori compounds without standard reagents were approximately quantified by HPLC-MS/MS using the standard curves of other targets with available standard reagents and similar retention times and structures.The results showed that:(1)With more than 300 injections in successive six days,the inter-day precisions of 14 Amadori compounds were lower than 10%,and that of 5 Amadori compounds were in the range of 10%-15%.(2)The test results of different tobacco leaf samples indicated that the total weights of 22 Amadori compounds accounted for 2.47%in flue-cured tobacco leaves,1.70%in oriental tobacco leaves,and only 0.14%in burley tobacco leaves.The concentrations of the 22 Amadori compounds differed greatly in the tobacco leaves of different types.This method could achieve rapid and simultaneous determination of 22 Amadori compounds in tobacco.