薄层生物自显影技术筛选两色金鸡菊中的活性成分  

TLC bioautographic method for screening of active constituents of Coreopsis tinctoria

作　　者：梁乐[1] 李琳琳[1] 古扎力努尔.艾尔肯 李新霞[2] 毛新民[3] 

机构地区：[1]新疆医科大学基础医学院药理教研室,乌鲁木齐830011 [2]新疆医科大学分析测试中心,乌鲁木齐830011 [3]新疆医科大学中医学院,乌鲁木齐830011

出　　处：《药物分析杂志》2015年第3期425-429,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：新疆维吾尔自治区科技计划项目自治区科技支撑计划(201333119)

摘　　要：目的:建立薄层生物自显影技术的方法筛选具有抗氧化作用和α-葡萄糖苷酶抑制作用的活性物质。方法:将两色金鸡菊水提物、醇提物和乙酸乙酯提取物在硅胶板上以甲苯-乙酸乙酯-甲酸(9∶7∶3)为展开剂展开,以1,1-二苯基-2-苦肼基自由基甲醇溶液进行显色,获得具有抗氧化作用物质;将两色金鸡菊3种提取物以上述同样的方法展开后在硅胶板上先喷α-葡萄糖苷酶溶液孵育60 min,再以2-萘基-α-D-葡萄糖苷溶液和固蓝B盐溶液(1∶1)为显色剂,显色后在白光下对比获得具有α-葡糖糖苷酶抑制作用的物质。结果:薄层板上斑点周围可以观察到白色或类白色抑制区,说明两色金鸡菊水提物、醇提物和乙酸乙酯3种提取物中均有抗氧化活性成分和α-葡糖糖苷酶抑制成分。结论:建立的薄层生物自显影技术可以快速筛选具有抗氧化作用和α-葡糖糖苷酶抑制作用的化合物,经指认两色金鸡菊中花色苷-3-葡萄糖苷、马里苷、绿原酸、黄酮奥卡宁、奥卡宁有较强的抗氧化作用,马里苷和奥卡宁具有较强的α-葡糖糖苷酶抑制成分。Objective： To develop a TLC bioautographic method for screening active ingredients with functions of antioxidant activity and α- glucosidase inhibition. Methods： The aqueous extract,alcohol extract and ethyl acetate extract from Coreopsis tinctoria were eluted with methylbenzene- acetic ether- formic acid（ 9 ∶ 7 ∶ 3） on the silica gel plate,and the antioxidant components were identified after the coloration with the 1,1- diphenyl- 2- picrylhydrazyl（ DPPH） methanol solution. The three Coreopsis extracts were eluted with the same method,incubated for60 minutes in α- glucosidase solution which was sprayed on the silica gel plate and were colored with the solution of 2- naphthyl- α- D- glucopyranoside and fast blue B salt（ 1∶ 1） as the color developing agent,thus α- glucosidase inhibitors were obtained and identified by resolution in visible light. Results： The white spots on the TLC plate showed effect of antioxidant potency and α- glucosidases inhibition,indicating that the aqueous extract,alcohol extract and ethyl acetate extract from Coreopsis tinctoria contained active ingredients with functions of antioxidant activity and α- glucosidase inhibition. Conclusion： The established rapid TLC bioautography method for screening the compounds of antioxidant and inhibitory effect of α- glucosidases was provided. Anthocyanin- 3- glucoside in Coreopsis tinctoria,marein,chlorogenic acid,flavanokanin and okanin have a strong antioxidation. Marein and okanin have strong α- glucosidases inhibitory effect.

关 键 词：生物活性测定 活性物质筛选方法 薄层生物自显影 两色金鸡菊 α-葡糖糖苷酶抑制作用 花色苷 马里苷 抗氧化作用 

分 类 号：R917[医药卫生—药物分析学]