竹叶黄酮的酶促酰化、分离与结构鉴定  

Enzymatic acylation,isolation and identification of flavonoid glycosides from bamboo leaves

作　　者：孙元奎[1] 马祥[1] 刘柳[1] 逯与运[1] 晏日安[1] 

机构地区：[1]暨南大学食品科学与工程系,广东广州510632

出　　处：《食品工业科技》2015年第9期96-100,106,共6页Science and Technology of Food Industry

基　　金：广东省自然科学基金(S2013010012931)

摘　　要：研究了固定化脂肪酶B(Novozmy 435)在低水分含量的叔戊醇溶剂中催化竹叶黄酮与硬脂酸的酰化反应。反应条件:竹叶黄酮与硬脂酸酸摩尔比1∶5,脂肪酶用量50g/L,机械搅拌转速200r/min,反应温度50℃,反应时间72h。后经液液萃取,薄层层析(TLC,UV 254nm),硅胶柱层析分离纯化,薄层层析展开剂配比为氯仿/甲醇/冰乙酸(7∶1∶0.05,v/v/v),柱层析洗脱剂配比为氯仿/甲醇/冰乙酸(10∶1∶0.05,v/v/v);洗脱液合并相同组分,得到两种黄酮苷酰化物单体。对纯化后的组分采用红外光谱(IR)、质谱(ESI-MS)、核磁共振(1H-NMR、13C-NMR)进行结构鉴定和表征,确定产物分别为异牡荆苷-6″-硬脂酸酯和异荭草苷-6″-硬脂酸酯,说明反应表现出高度选择性。Immobilised Candida Antarctica lipase B（ Novozmy 435） was used to acylate flavonoid glycosides from bamboo leaves in tert- amyl alcohol,with stearic acid as acyl donors at low moisture content.The optimum reaction condition was as follows： a molar ratio of flavonoid glycosides from bamboo leaves to Stearic acid of 1 ∶ 5,50 g / L lipase,stirring speed was 200 r / min,50℃ and reaction time was 72 h.After Liquid- liquid extraction,the products of esterification reactions were subjected to thin- layer chromatography（ TLC,UV 254nm）,a silica gel column（ 700mm× 35 mm,i.d.）,the developing solvent ratio was chloroform / methanol / acetic acid（ 7∶1∶0.05,v / v / v）,and eluted with chloroform / methanol / acetic acid（ 10 ∶ 1 ∶ 0.05,v / v / v）. Then combine fractions had the same value of retardation factor Rf,two types of acylated flavonoids were obtained. The purified products were analyzed by IR,ESI- MS,^1H- NMR and ^13C- NMR and the two types of acylated flavonoids were isoorientin-6″- stearate and isovitexin-6″-stearate.The enzyme showed strong selectivity for acylation of glycosides.

关 键 词：固定化脂肪酶B 竹叶黄酮 酰化 薄层层析 硅胶柱层析 

分 类 号：TS202.3[轻工技术与工程—食品科学]