薄层生物自显影技术筛选两色金鸡菊中的活性成分

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 for 60 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 tinctoriacontained 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 inCoreopsis tinctoria,marein,chlorogenic acid,flavanokanin and okanin have a strong antioxidation.Marein and okanin have strong α-glucosidases inhibitory effect.