HPLC法同時(shí)測(cè)定煙葉中6種多酚類物質(zhì)
發(fā)布時(shí)間:2019-08-26 來源: 歷史回眸 點(diǎn)擊:
摘 要:本文建立了一種可同時(shí)測(cè)定烤煙葉片中綠原酸、咖啡酸、蘆丁、莨菪亭、槲皮素和山奈酚的高效液相色譜(HPLC)檢測(cè)方法,并分析了不同煙草雜種F1及其親本中上述多酚的含量。采用甲醇:水=7∶3為提取液,超聲提取30 min,色譜柱為Agilent 5 TC-C18 (250 mm × 4.6 mm,5 μm) ;2%冰乙酸水溶液-甲醇為流動(dòng)相,梯度洗脫,流速1 ml/min;柱溫30℃;進(jìn)樣體積10 μl。結(jié)果顯示:6種多酚類物質(zhì)分離效果良好,加樣回收率范圍在95.53%~100.622%(RSD為0.95%~2.285%),穩(wěn)定性RSD(n=3)為0.13%~1.73%。綠原酸、咖啡酸、蘆丁、莨菪亭、槲皮素和山奈酚的線性范圍在10~600 μg/ml之間,各多酚類物質(zhì)線性方程相關(guān)系數(shù)在0.99996~0.99999之間。各材料的綠原酸和蘆丁含量較高,為多酚類物質(zhì)主要成分,其他多酚物質(zhì)含量則比較低。雜種F1相對(duì)其親本材料在某種多酚含量中表現(xiàn)出一定優(yōu)勢(shì),VA116 × GDH88綠原酸含量遠(yuǎn)遠(yuǎn)高于其兩個(gè)親本材料,VA116 × TN90蘆丁含量也超越其雙親,而綠原酸含量則低于其親本VA116,F(xiàn)lorida301 × GDH94蘆丁含量超越雙親,綠原酸含量則低于其親本GDH94。
關(guān)鍵詞:烤煙;HPLC;多酚;綠原酸;蘆丁
中圖分類號(hào):S572
文獻(xiàn)標(biāo)識(shí)碼:A
文章編號(hào):1008-0457(2018)03-0021-06 國(guó)際DOI編碼: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.
相關(guān)熱詞搜索:煙葉 測(cè)定 物質(zhì) 多酚 HPLC
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