油菜花粉中黄酮苷类的制备分离和鉴定  

Separation and Identification of Flavonoid Glycosides from Brassica napus L. Pollen

作　　者：杨开[1,2] 叶兴乾[1] 刘东红[1] 孙培龙[2] 

机构地区：[1]浙江大学生物系统工程与食品科学学院,杭州31002 [2]浙江工业大学生物与环境工程学院,杭州310032

出　　处：《中国粮油学报》2010年第8期91-97,共7页Journal of the Chinese Cereals and Oils Association

基　　金：国家科技支撑计划(2006BAD27B06);教育部创新工程重大项目培育资金项目(707034);浙江省重大攻关项目(2008C02005-2)

摘　　要：以油菜蜂花粉80%乙醇超声提取液中的正丁醇萃取物为黄酮苷分离样品,乙酸乙酯∶正丁醇∶水=2∶1∶3(体积比)为溶剂体系,主机转速900 r/min,上相为固定相,下相为流动相,1.5 mL/min流速,30℃条件下,400 mg样品经高速逆流色谱制备分离,所得产物经HPLC检测,5 h内得到2个黄酮苷单体和1个黄酮苷混合物,经UV、MS、1H-NMR和13C-NMR结构鉴定为山奈酚-3-O-β-D-吡喃葡萄糖苷(1→2)-β-D-吡喃葡萄糖苷(89.1 mg,纯度98.2%)和山奈酚-3-O-新橙皮糖苷(41.9 mg,纯度96.3%),其中山奈酚-3-O-新橙皮糖苷属首次从油菜花粉中制备分离得到,另外对混合物中所含2个黄酮苷进行了HPLC-MS分析和结构推断。High-speed counter-current chromatography（HSCCC） was developed for preparation and purification of flavonoid glycosides from pollen of Brassica napus L.400 mg n-butanol extraction partition was prepared as sample by ultrasonic extracting Brassica napus L.with 80% ethanol,and then extracted by different polarity organic solvents and freeze-dried.When in the HSCCC separation,a two-phase solvent system composed of ethyl acetate：n-butylol：water（2∶1∶3,v/v/v）,at a flow rate of 1.5 mL/min,at 900 r/min and temperature 30 ℃ were the conditions,and the obtained fractions were analyzed by HPLC.Results：Two purified flavonoid glycosides and a mixture are obtained within 5 h,and the structure of two flavonoid glycosides are elucidated as kaempferol-3-O-β-D-Glc（1→2）-β-D-Glc（89.1 mg with a purity of 98.2%）,and kaempferol-3-O-neohesperidoside（41.9 mg with a purity of 96.3%） through spectroscopic analysis of UV,MS,1H-NMR and 13C-NMR,and the latter is discovered from Brassica napus L.pollen for the first time.Another two flavonoid glycosides in the mixture are also tentatively identified by HPLC-MS.

关 键 词：油菜花粉 黄酮苷 高速逆流色谱 制备分离 鉴定 

分 类 号：TS201.2[轻工技术与工程—食品科学]