UPLC-MS^n法分析烟叶中蔗糖四酯类化合物

Analysis of sucrose tetraesters in tobacco leaves by UPLC-MS^n method

建立了烟叶样品中蔗糖四酯类化合物的超高效液相色谱-多级质谱(UPLC-MS^n)分析方法。采用超声辅助萃取技术,用甲醇萃取烟叶样品中的蔗糖四酯,采用正交试验对超声萃取条件进行优化;萃取液经分离除杂后,在选择反应监测(SRM)模式下进行UPLC-MS^n定性和定量分析,通过与标准品对照及对目标物的多级质谱数据进行分析,定性鉴定样品中的5组蔗糖四酯;并采用内标标准曲线法同时定量测定5组蔗糖四酯化合物的质量分数。结果表明:(1)优化后的最佳萃取条件为料液比1∶30[质量(g)∶体积(mL)],超声功率120 W,萃取时间40 min;(2)定量分析方法的回收率在80.67%~89.35%之间,相对标准偏差均小于5%;相关系数r均大于或等于0.999 0;方法的检出限(LOD)和定量限(LOQ)分别为14.71~38.42、49.02~128.05μg/kg。该方法前处理简单,定量分析灵敏度和准确度均较高,适合烟叶中蔗糖四酯类化合物的定量分析。

A method for analyzing sucrose tetraesters（STEs） in tobacco leaves by ultra-performance liquid chromatography-multi-stage mass spectrometry（UPLC-MS^n） was developed. The STEs in tobacco samples were extracted by methanol with ultrasonic-assisted extraction under conditions optimized by orthogonal experimental design. The extracts from tobacco samples were cleaned up and then analyzed qualitatively and quantitatively by UPLC-MS^nunder selected reaction monitoring（SRM） mode. Five groups of STEs were separated and identified by comparing with standard compounds and analyzing the MS^ndata of the target objects, and the contents of the five groups of STEs were simultaneously and quantitatively determined by using internal standard curves. The results showed that： 1） The optimum conditions were solid/liquid ratio 1∶30 [mass（g）∶volume（m L）], ultrasonic power 120 W and extraction time 40 min. 2） The standard recoveries of the quantitative analysis method was in the range of 80.67%-89.35%. The relative standard deviations were less than 5% and the correlation coefficients were no less than 0.999 0. The limits of detection（LODs） and quantitation（LOQs） were 14.71-38.42, 49.02-128.05 μg/kg, respectively. This method is suitable for the quantitative analysis of STEs in tobacco leaves with the advantages of simple pretreatment, higher sensitivity and accuracy.