采取酮提取茶树鲜叶中的叶黄素循环组分,分别在15℃、22℃和30℃下,用Spherisorb C18(5μm,250βmm×4.6βmm)色谱柱,乙腈︰甲醇(85︰15)和甲醇︰乙酸乙酯(68︰32)为流动相,线性洗脱,在445βnm下检测。结果表明,不同温度条件下,高效液相色谱法都能够从茶树鲜叶提取物中检测到紫黄素、环氧玉米黄素和玉米黄素,而且分离效果好,重现性和精密度也很高。强光下,叶黄素循环库和紫黄素脱环氧化程度皆明显增加,表明其在茶树中有重要的光保护作用。
The xanthophyll cycle components in fresh leaves of tea plant (Camellia sinensis cv. Fudingdabai)were extracted by acetone. The pigments were analyzed with a Spherisorb column (C18 , 5μm,250㎜×4.6βmm),and were eluted using acetonitrile: methanol(85:15) and methanol:ethyl acetate(68:32) by a linear gradient. The pigments were detected by their absorbance at 445βnm. It was showed that violaxanthin, antheraxanthin and zeaxanthin from fresh leaves of tea plant were detected by HPLC, respectively, under temperatures of 15℃, 22℃ and 30℃, meanwhile, the pigments were separated well, and it is a sensitive, well-reappeared method. Tea plant had a larger xanthophyll cycle pool and a larger de-epoxidation state in higher sun-light, which indicated xanthophyll cycle plays an important role in photo-protection in tea plant.
[1] Demmig-Adams B, Adams ww Ⅲ. The role of xanthophyll cycle Carotenois in the protection of photosynthesis[J]. Trends in Plant Science,1996,1:21~26.
[2] Pfündel E, Bilger W.Regulation and possible function of violaxanthin cycle[J]. Photosynthsis Research 1994, 42: 89-109.
[3] Glimore AM.Mechanistic aspects of xanthophyll cyce-dependent photoprotection on higher chloroplasts and leaves[J]. Physiology Plant, 1997, 99:197-209.
[4] Eskling M, Arvidsson PO, Åkerlund H E.The xanthophyll cycle, its regulation and components[J]. Physiology Plant, 1997, 100:806-816.
[5] Müller P, Xiao-Ping Li, Niyogi KK.Non-Photochemical quenching. A response to excess light energy[J]. Plant Physiology, 2001,125:1558~1566.
[6] Thayer S S, Bjorkman O.Leaf xanthophylls content and composition in sun and shade determined by HPLC[J]. Photosynthesis Research, 1990, 23:331-343.
[7] Glimore A M, Yamamoto H.Y. Resolution of lutein and zeaxanthin using a non-endcapped, lightly carbon-loaded C18 high-performance liquid chromatographic column[J]. Journal of Chromatography, 1991, 543:137-145.
[8] 赵世杰, 孟庆伟, 等. 植物组织中叶黄素循环组分的高效液相色谱分析法[J]. 植物生理通讯,1995,31(6):438-442.