响应面法优化秃疮花中生物碱提取工艺及抑菌活性研究

Optimization of the extracting process for alkaloids from Dicranostigma leptopodum by response surface analysis and study on its antibacterial activity in vitro

以秃疮花为研究对象,在单因素水平测定的基础上,通过响应面法优化其提取工艺,并使用响应面法提取浓缩液进行抑菌活性测定,以大肠埃希氏杆菌为研究对象,进行透射电镜观察。结果显示,秃疮花生物碱最佳提取工艺条件为:液料比15mL/g、超声时间35min、回流时间2.5h、乙醇浓度65%,在此工艺下秃疮花生物碱含量为9.33%;其响应面法提取浓缩液对大肠埃希氏杆菌、白色念珠菌、绿脓杆菌的最小抑菌浓度分别为0.20,0.15和0.35mg/mL,最低杀菌浓度分别为0.15,0.10和0.30mg/mL;在透射电镜下观察到对大肠埃希氏杆菌抑制效果显著。表明响应面法对秃疮花生物碱提取工艺的优化比较合理。

In this paper,the Dicranostigma leptopodum was researched,on the basis of single factor level determination,the optimal extraction conditions were obtained by using the response surface analysis,and the concentrated liquid of extract response surface analysis was measured by antibacterial activity,moreover,the transmission electron microscopy images of Escherichia coli of treated with concentrated liquid of extract from D.leptopodum was discussed.The experiment results demonstrated that the optimum conditions were liquid to solid ratio was 15 mL/g,time of ultrasound was 35 min,refluxing time was 2.5 h,alcohol was 65%,and the content of alkaloid from D.leptopodum was 9.33% in the optimum extraction process condition;the optimal extraction conditions were obtained by using the response surface analysis of E.coli,Candida albicans and Pseudomonas aeruginosa of minimal inhibitory concentration and minimal bactericidal concentration were 0.20,0.15,0.35 mg/mL and 0.15,0.10,0.30 mg/mL.They were observed under transmission electron microscopy to everest to E.coli antibacterial activity effect significantly remarkable.The results also showed that the response surface analysis to D.leptopodum spend alkaloid extraction process optimization are reasonable.