HPLC法同时测定烟叶中6种多酚类物质

　　摘 要：本文建立了一種可同时测定烤烟叶片中绿原酸、咖啡酸、芦丁、莨菪亭、槲皮素和山奈酚的高效液相色谱（HPLC）检测方法，并分析了不同烟草杂种F1及其亲本中上述多酚的含量。采用甲醇：水=7∶3为提取液，超声提取30 min，色谱柱为Agilent 5 TC-C18 （250 mm × 4.6 mm，5 μm） ；2%冰乙酸水溶液-甲醇为流动相，梯度洗脱，流速1 ml/min；柱温30℃；进样体积10 μl。结果显示：6种多酚类物质分离效果良好，加样回收率范围在95.53%～100.622%（RSD为0.95%～2.285%），稳定性RSD（n=3）为0.13%～1.73%。绿原酸、咖啡酸、芦丁、莨菪亭、槲皮素和山奈酚的线性范围在10～600 μg/ml之间，各多酚类物质线性方程相关系数在0.99996～0.99999之间。各材料的绿原酸和芦丁含量较高，为多酚类物质主要成分，其他多酚物质含量则比较低。杂种F1相对其亲本材料在某种多酚含量中表现出一定优势，VA116 × GDH88绿原酸含量远远高于其两个亲本材料，VA116 × TN90芦丁含量也超越其双亲，而绿原酸含量则低于其亲本VA116，Florida301 × GDH94芦丁含量超越双亲，绿原酸含量则低于其亲本GDH94。
　　关键词：烤烟；HPLC；多酚；绿原酸；芦丁
　　中图分类号：S572
　　文献标识码：A
　　文章编号：1008-0457（2018）03-0021-06 国际DOI编码：10.15958/j.cnki.sdnyswxb.2018.03.004
　　Simultaneous Determination of Six Polyphenols in Flue-cured Tobacco Leaves by HPLC Method
　　ZHANG Qifa1，2，NIE Qiong1，2*，ZOU Xushen1，2，XIE Rui1，2，LIU Yunhe1，2
　　（1. College of Agriculture， Guizhou University， Guiyang 550025， Guizhou， China；2. Guizhou Provincial Key Laboratory of Tobacco Quality，Tobacco Research Center of Guizhou University， Guiyang 550025， Guizhou， China）
　　Abstract：A high performance liquid chromatographic （HPLC） method was developed for the simultaneous determination of chlorogenic acid， caffeic acid， rutin， palatalin， quercetin， and kaempferol in flue-cured tobacco leaves. The content of these six polyphenols were analysed in the hybrid F1 and its parents for different tobaccos. The extraction solution of pure methanol：pure water =7：3 was used. Ultrasonic extraction was performed for 30min. Column T was Agilent 5 TC-C18 （250mmx4.6mm， 5 mm）. Two per cent of aqueous glacial acetic acid-methanol was used as the mobile phase. Gradient elution with the flow rate of 1ml/min was applied. Column temperature was at 30 C and injection volume was 10 L. The results showed that the six polyphenols had good separation effect. The rate of recovery ranged from 95.53% to 100.622% （RSD was 0.95% to 2.285%）， and the stability RSD （n=3） was 0.13% ～ 1.73%. Chlorogenic acid， caffeic acid， rutin， palatalin， quercetin and kaempferol was in the range of 10～600 g/ml. The correlation coefficients of the linear equation of polyphenols varied from 0.99996 to 0.99999. The content of chlorogenic acid and rutin in each material was relatively high， which was the main component of polyphenols. The content of other polyphenols is relatively low. Hybrid F1 had some advantages relative to its parent materials in certain polyphenol contents. For example， the content of chlorogenic acid of VA116 X GDH88 in hybrid F1 was much higher than that of its two parent materials； the content of rutin of VA116 X TN90 also surpassed the parents； while the content of chlorogenic acid was lower than that of its parent VA116； the content of rutin of Florda301 X GDH94 was also beyond that of the parents， while the content of chlorogenic acid was lower than that of its parent GDH94.

相关热词搜索：烟叶 测定 物质 多酚 HPLC