高效液相色谱-紫外/质谱检测法联合测定新鲜烟叶中的25种酚类物质

建立了烟叶中25种酚类物质的高效液相色谱-紫外/质谱(HPLC-UV/MS)测定方法,其中11种主要酚类物质采用HPLC-UV法进行绝对定量分析,其余采用HPLC-MS法进行半定量分析。方法验证结果表明:11种酚类标准物质在0.90~99.00 mg/L(绿原酸及芸香苷为0.95~380.00 mg/L)范围内呈现很好的线性关系,相关系数为0.999 3~0.999 9。在22.5~24.8、45.0~49.5、67.5~74.3 mg/L 3个水平的加标回收率为91.0%~112.4%。方法重复性的相对标准偏差(RSD)为1.48%~13.40%,日内日间精密度的RSD为0.35%~15.54%。采用本方法对云南、河南、贵州的成熟期新鲜烟叶样品进行了分析,主要酚类物质总量为贵州云南河南,贵州与河南两地间存在显著性差异;芸香苷含量为云南贵州河南,且相互间有显著性差异。本方法预处理简单,酚类物质的分析覆盖面广,可用于批量烟叶样品的检测。

Determination of phenolic compounds in fresh tobacco leaves by high performance liquid chromatography-ultraviolet/mass spectrometry

Twenty-five phenolic compounds in tobacco were analyzed using high performance liquid chromatography-ultraviolet/mass spectrometry (HPLC-UV/MS), including absolute quantification of 11 main phenols by HPLC-UV and relative quantification of other 14 phenols by HPLC-MS. The validation results of this method were satisfactory. Eleven phenol standards showed good linearity with correlation coefficients (r²) of 0.9993-0.9999 over the mass concentration range from 0.90 mg/L to 99.00 mg/L (the range of chlorogenic acid and rutin was from 0.95 mg/L to 380.00 mg/L). The recoveries of the 11 phenol standards were 91.0%-112.4% with the RSDs of 0.33%-8.11% at the three spiked levels of 22.5-24.8, 45.0-49.5 and 67.5-74.3 mg/L. The reproducibility of the method was good with the RSDs of 1.48%-13.40%. In addition, the intra-day and inter-day precisions were also satisfactory with the RSDs of 0.35%-15.54%. Mature fresh tobacco leaves from Yunnan, Henan and Guizhou Provinces, China were analyzed using this method. The results showed that the total amount of main phenols was Guizhou>Yunnan>Henan and there was significant difference between Guizhou and Henan. The amount of rutin was Yunnan>Guizhou>Henan, and there were significant differences among them each other. The method is reproducible with a simple pretreatment covering a wide range of phenols, and can be applied in batch analysis of tobacco leaves.