红茂草中黄酮类提取工艺优化及体外抑菌研究  

Optimization of Extracting Technology of Flavonoid from Dicranostigma Leptopodum(maxim.)Fedde and Study on its Antibacterial Activity in Vitro

作　　者：赵强[1] 王廷璞[1] 

机构地区：[1]天水师范学院生物工程与技术学院,甘肃天水741000

出　　处：《畜牧兽医杂志》2016年第1期4-12,共9页Journal of Animal Science and Veterinary Medicine

基　　金：2013年国家自然科学基金项目(31360603)

摘　　要：以红茂草为研究对象,在单因素水平筛选测定的基础上,通过响应面法优化其提取工艺。提取物采用Diamonsil C18(4.6mm×150mm,1.8μm)色谱柱为固定相,甲醇-0.2%磷酸(65∶35,v/v)为流动相,流速1.0mL/min,检测波长280nm;并对提取物进行抑菌活性测定,以大肠埃希氏杆菌和金黄色葡萄球菌为供试菌,进行体外抑菌活性研究和透射电镜观察。结果显示,红茂草中黄酮类化合物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、乙醇体积分数65%,在此工艺下红茂草中黄酮类化合物含量为2.60%,HPLC检测重现性好;其提取物对大肠埃希氏杆菌和金黄色葡萄球菌的MIC为0.28g/mL和0.20g/mL,MBC为0.26g/mL和0.10g/mL,IC50为0.2162g/mL和0.1805g/mL,在透射电镜下观察抑菌效果显著。研究表明,响应面法对红茂草中黄酮类化合物提取工艺的优化比较合理。On the basis of single factor level determination of screening, the extraction technology of flavonoid from Dicranostigma leptopodum （maxim.）Fedde （DLF） was optimized by using the response surface analysis. Extractive was used by Diamonsil C18 （4.6min× 150mm,1.8μm）. The stationary phase was chromatographic column, mobile phase was methanol- 0.2% phosphoric acid（65 ：35 ,v/v）, the flow rate was 1.0 mL/min, and detection wavelength was 254 nm. Antibacterial activities of the extracts were measured. Providing E. coli and S. aureus as the tested strains, antibacterial activity in vitro was researched and transmission electron microscope images were observed. The results demonstrated that the optimum condition of extraction technology of flavonoids from DLF as follow： liquid to solid ratio was 15 mL/g, ultrasound time was 35 min, refluxing time was 2.5h, and alcohol was 65%. In these conditions, the content of flavonold from DLF was 2.60% and HPLC de tection had good reproducibility. The MIC of flavonoids on E. coli and S. aureus were 0.28 and 0.20 g/mL, respectively. The MBC were 0.26 and 0.10 g/mL, respectively. The IC50 were 0. 2162 and 0. 1805g/mL, respectively. Observed under transmission electron microscopy, the effects of antibacterial activity were significant. The results showed that the response surface analysis to optimization of extraction process of flavonoid from DLF was quite reasonable.

关 键 词：响应面法 红茂草 黄酮类化合物 

分 类 号：TS272[农业科学—茶叶生产加工]