超高效液相色谱测定烟叶中的糖类化合物及在筛选评价中的应用

Determination of sugars in fresh tobacco leaf by ultra high-performance liquid chromatography for screening evaluation

利用可以快速批量前处理的新型样品萃取瓶作为前处理装置,建立了新鲜烟叶中葡萄糖、果糖、麦芽糖和蔗糖4种糖类化合物的超高效液相色谱快速检测方法。试验结果表明:(1)新开发的样品前处理装置集提取、过滤和转移为一体,可以大大提高样品前处理效率;(2)利用75%的乙腈(内含0.2%三乙胺)为流动相,以ACQUITY UPLC BEH Amide(2.1 mm×100 mm,1.7μm)为色谱柱,以流速0.2mL/min进行色谱分析。4种糖类物质在5～250μg/mL时具有较好的线性关系,线性相关系数>0.999;检出限为1.0～3.2μg/mL;不同添加水平下,平均回收率为91.0%～102.0%,精密度试验RSD<5%;(3)对于不同类型的新鲜烟叶进行检测,发现糖类化合物的含量差异明显。该方法前处理简单易操作,重复性较好,可以满足烟草及相关样品检测评价的要求。

For the fast and accurate determination of sugars in tobacco leaf,a method for the determination of sugars(fructose,glucose,sucrose,maltose)in tobacco leaf by an ultra-high performance liquid chromatography(UPLC)was established,taking a novel extraction vial as the pretreatment device.The results showed that:1)the novel pretreatment device could make the extraction,filter and transfer as one step,make the procedure of the pretreatment more efficient.2) The separation of the sample was operated on ACQUITY UPLC BEH Amide column(2.1 mm × 100 mm,1.7μm),using 75%(v/v)acetonitrile containing 0.2% triethylamine as the mobile phase at a flow-rate of 0.2mL/min.The 4analytes could be separated smoothly with good linearity in the range of5~250μg/mL,with the correlation coefficients larger than 0.999.The limits of detection(LODs)were further evaluated(1.0~3.2μg/mL).Under three different spiked level The mean recovery of the target compounds was 91.0%~102%for 4sugars and the intraday and inter-day precision were below 5%.3)Different contents of sugars in different type tobacco leaf were found with the established method.The new method,with the novel pretreatment vial for extraction and the HPLC for separation,can be used for analysis of sugars in tobacco samples.