[关键词]
[摘要]
目的 优选湿法超微粉碎提取水蛭的最佳工艺。方法 在单因素试验基础上,以料液比、浸泡时间、提取温度和粉碎时间为自变量,根据Box-Behnken原理采用4因素3水平的响应面分析法,以抗凝血酶活性为响应值,对湿法超微粉碎提取条件进行优化。结果 湿法超微粉碎提取水蛭的最佳工艺条件:料液比为1∶12,浸泡时间为4.9 h,提取温度为6.5 ℃,粉碎时间为12 min,其抗凝血酶活性达到20.13 U/g。结论 湿法超微粉碎提取技术适用于水蛭的提取,本实验优选的提取工艺稳定可行,适于进一步推广应用。
[Key word]
[Abstract]
Objective To prepare hydroxytyrosol (HT) by alkaline hydrolysis of oleuropein and to investigate its anti-oxidative property. Methods According to single factor experiments, four variables: temperature, sodium hydroxide concentration, time, and the ratio of solid to liquid, were selected. On the basis of single factor experiments, orthogonal tests of L9(34) were carried out. HPLC-MS method was also further applied for the qualitative analysis on HT. Under the same conditions, contrast experiments on the acid and alkaline actions were made. Finally, anti-oxidative activity of HT was evaluated. Results The results showed that the selected variables were all highly significant. Also, the descending influence orders on HT were temperature > sodium hydroxide concentration > time > the ratio of solid to liquid. Lastly, the optimal conditions on HT yield were obtained as follow: temperature 90 ℃, sodium hydroxide concentration 0.2 mol/L, time 45 min, and the ratio of solid to liquid 1∶15. The HPLC retention time of HT was about 10.3 min, and its relative molecular weight of fragment ion by MS was 134.8. By comparison with the acid action, it was proved that the alkaline action was better in HT yield. In addition, the anti-oxidative activity of HT was carried out on scavenging DPPH radical. The results showed that DPPH scavenging capability of HT was 2.45 times of BHT. Conclusion HT yield is well obtained under the alkaline condition and has a good anti-oxidation.
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[基金项目]
“十二五”国家科技支撑计划(2012BAD32B10);“863”课题项目“食品高效节能干燥装备开发与新技术研究”(2011AA100802)