超高效液相色谱法快速测定烟草样品中的植物色素

Rapid Determination of Plant Pigments in Tobacco by Ultra Performance Liquid Chromatography

建立超高效液相色谱法快速测定烟草中植物色素(新黄质、紫黄质、叶黄素、叶绿素-a、叶绿素-b、β-胡萝卜素)检测方法。90%丙酮超声萃取30 min,色谱柱为Waters ACQUITY UPLCTM BEH C18(2.1 mm×50 mm,1.7μm),流动相为乙腈-水梯度洗脱,以体积比按(乙腈90%+10%水),4.0 min(100%乙腈)的梯度条件,流速为0.8 mL/min,用二极管阵列检测器检测。6种色素在5 min内分离,检测限15～22 ng/mL,平均回收率为93%～105%。采用自主设计的样品萃取瓶进行前处理,前处理简单快速,同时,色谱分离周期明显缩短,与目前行业标准相比,样品分析效率提高了4倍以上。

Established a ultra performance liquid chromatography(UPLC)method for the rapid determination of plant pigments(neoxanthin,violaxanthin,lutein,chlorophyll-a,chlorophyll-b andβ-carotene)in tobacco.The plant pigments were extracted from samples with 90%acetone by ultra-sonic extraction.Then,the plant pigments were separated by Waters ACQUITY UPLCTM BEH C18(2.1 mm×50 mm,1.7μm)column,with and water mixture as mobile phase at a flow-rate of 0.8 mL/min.It eluted with gradient elution(90%V(acetonitrile))+10%V(water))at the beginning,then the gradient elution replaced by acetonitrile 4 min later.These plant pigments were detected by diode array detector.Under the optimal conditions,6 kinds of plant pigments could be effectively separated within 5 min.The limits of detection(LODs)of 6 kinds of plant pigments were 15~22 ng/mL and the average recoveries were in the range of 93%to 105%.Via the self-designed sample extraction bottle using for pretreatment,and improved chromatography conditions,this method efficiency was enhanced 4 times than the current tobacco standard.