气相色谱-串联质谱检测新鲜烟叶中的萜类成分

Determination of terpenoids in fresh tobacco leaves by GC-MS/MS

为研究烟叶中的萜类成分含量(质量分数),采用选择反应监测模式,建立了同时测定新鲜烟叶中反,反-金合欢醇、α-植醇、顺冷杉醇、β-植醇、(1S,2E,4S,6R,7E,11E)-2,7,11-西柏烷三烯-4,6-二醇(α-CBD)、(1S,2E,4R,6R,7E,11E)-2,7,11-西柏烷三烯-4,6-二醇(β-CBD)、香紫苏醇、赖百当-13-烯-8,15-二醇、角鲨烯、2,3-环氧化鲨烯、双氢速甾醇、胆固醇、菜籽甾醇、麦角甾醇、菜油甾醇、豆甾醇、羊毛甾醇、β-谷甾醇、β-香树脂醇、α-香树脂醇、鹅去氧胆酸等21种萜类成分的气相色谱-串联质谱(GC-MS/MS)方法;并采用该方法对云南、河南、贵州等3个产区的成熟期新鲜烟叶样品进行了分析。结果表明:在线性范围内21种萜类成分的R^2为0.9990~0.9998;方法检测限为12.7~308.7ng·g^-1,定量限为35.8~984.6ng·g^-1;日内、日间精密度(RSD)分别为0.19%~8.28%、1.02%~9.35%。不同产区烟叶中主要萜类成分α-CBD的含量高低顺序为云南(98.83μg·g^-1)>河南(57.04μg·g^-1)>贵州(38.61μg·g^-1),且3个产区之间存在显著性差异。贵州产区烟叶中萜类成分含量的分布较云南和河南产区烟叶差异更大,遵义种植的南江3号品种烟叶中萜类含量与其他3个品种差异明显。该方法前处理简单,分析的萜类物质覆盖面广,可用于批量烟叶样品的检测。

In order to quantify tobacco terpenoids, a method for simultaneous determination of 21 terpenoids, including t,t-farnesol,α-phytol, cis-abienol,β-phytol,(1S,2E,4S,6R,7E,11E)-2,7,11-cembratriene-4,6-diol (α-CBD),(1S,2E,4R,6R,7E,11E)-2,7,11-cembratriene-4,6-diol (β-CBD), sclareol, labd-13-ene-8,15-diol, squalene, 2,3-oxosqualene, dihydrotachysterol, cholesterol, brassicasterol, ergosterol, campesterol, stigmasterol, lanosterol,β-sitosterol,β-amyrin,α-amyrin and chenodeoxycholic acid in tobacco was developed using gas chromatography triple quadruples mass spectrometry (GC-MS/MS). The characteristic fragment ion peaks of the 21 terpenoids were obtained under selective reaction monitoring (SRM) mode in positive channels. The results noted that under the optimized conditions, the determination coefficients (R^2) of the 21 terpenoids were 0.999 0-0.999 8 in the linear ranges. The limits of detection (LOD) for analytes ranged from 12.7 to 308.7 ng·g^-1, and the limits of quantification (LOQ) were 35.8-984.6 ng·g^-1. The intra- and inter-day relative standard deviations (RSDs) were 0.19%-8.28% and 1.02%-9.35% respectively. Mature fresh tobacco leaves from Yunnan, Henan and Guizhou Provinces were analyzed using this method. The results showed that the total amount of α-CBD was Yunnan (98.83 μg·g^-1)> Henan (57.04μg·g^-1)> Guizhou (38.61 μg·g^-1), and there was significant difference among them. In particular, the terpenoid contents in leaves from Guizhou differed substantially from Yunnan and Henan, and that in leaves of cv. Nanjiang No.3 from Zunyi was significantly different from the other three varieties. This method uses only a simple pretreatment and is reproducible, can cover a wide range of terpenoids for batch analysis of tobacco leaves.