摘要
本文建立了一种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,5μm);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10μl。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10~600μg/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。
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 30 min. Column T was Agilent 5 TC-C18（ 250 mmx4. 6 mm,5 mm）. Two per cent of aqueous glacial acetic acid-methanol was used as the mobile phase. Gradient elution with the flow rate of 1 ml/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.
作者
张启发
聂琼
邹序生
谢瑞
刘云鹤
ZHANG Qi-fa;NIE Qiong;ZOU Xu-shen;XIE Rui;LIU Yun-he(College of Agriculture,Guizhou University,Guiyang 550025,Guizhou,China;Guizhou Provincial Key Laboratory of Tobacco Quality,Tobacco Research Center of Guizhou University,Guiyang 550025,Guizhou,China)
出处
《山地农业生物学报》
2018年第3期21-26,共6页
Journal of Mountain Agriculture and Biology
基金
中国烟草总公司贵州省公司项目“烟草杂种优势分子基础及优势组合筛选”(黔烟科合201302)
关键词
烤烟
HPLC
多酚
绿原酸
芦丁
Tobacco polyphenols
HPLC
chlorogenic acid
rutin
method validation
