响应面法优化航天杂交构树叶总黄酮提取工艺及体外抗弧菌活性研究

Optimization of Extraction of Total Flavonoids from Spaceflight Hybrid Broussonetia Papyrifera Leaves by Box-Behnken Response Surface Methodology and Anti-Vibrio Activity in Vitro

【目的】优化航天杂交构树叶总黄酮提取工艺,测定其体外抗弧菌活性。【方法】以总黄酮得率为指标,采用加热回流提取法,通过单因素试验结合Box-Behnken响应面法优化航天杂交构树叶总黄酮的最佳提取工艺。同时采用纸片扩散法和微量稀释法测定航天杂交构树叶总黄酮提取物及不同极性有机溶剂萃取物对哈维氏弧菌、副溶血弧菌、溶藻弧菌、创伤弧菌、非01霍乱弧菌、嗜水气单胞菌的抑菌圈直径和最低抑菌浓度(MIC)。【结果】(1)最佳提取工艺:乙醇浓度70%、料液比1∶20[(m/g)∶(V/mL)]、2 h/次、100℃加热回流提取3次,航天杂交构树叶总黄酮得率为(6.02±0.14)%。(2)抑菌环直径:哈维氏弧菌、副溶血弧菌、创伤弧菌、嗜水气单胞菌,>17 mm;溶藻弧菌、非01霍乱弧菌,>12 mm。较不同极性有机溶剂萃取物敏感。(3)MIC值:航天杂交构树叶总黄酮提取物对不同弧菌的MIC值由低至高依次为嗜水气单胞菌<副溶血弧菌<哈维弧菌=溶藻弧菌=创伤弧菌<非01霍乱弧菌,较不同极性有机溶剂萃取物的抗菌活性强。【结论】该工艺条件适用于航天杂交构树叶总黄酮的制备,得率较高。航天杂交构树叶总黄酮提取物具有较强的抗弧菌活性,是不同极性成分间协同作用的结果。

Objective To optimize the exaction technology of total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves,and to determine the anti-vibrio activity in vitro.Methods Taking the yield rate of total flavonoids as evaluation indicator,Box-Behnken response surface methodology was used to optimize the extraction technology of total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves by heating reflux extraction method based on single factor tests.And the diameter of inhibition zone and minimal inhibitory concentration of total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves and their different organic solventextractants for V.harveyi,V.prarhaemolyticus,V.alginolyticus,V.vulnificus,V.cholerae and Aeromonas hydrophila were determined by the slip method and micro-dilution methods.Results(1)The best extraction conditions were as follows:ethanol concentration was 70%,solid-liquid ratio was 1∶20[(m/g)∶(V/mL)],2 h per time,and 100℃ heating reflux extraction for 3 times.Under this condition,the yield rate of total flavonoids was(6.02±0.14)%.(2)The stagnation of inhibition zone:V.harveyi,V.prarhaemolyticus,V.vulnificus and Aeromonas hydrophila,>17 mm;V.alginolyticus and V.cholerae,>12 mm.The effects were more sensitive than those of their different polar organic solvent-extractants.(3)The minimal inhibitory concentration of total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves from low to high was Aeromonas hydrophila<V.parahaemolyticus<V.harveyi<V.alginolyticus=V.vulnificus<V.cholerae,and its antibacterial activity being stronger than that of their different polar organic solvent-extractants.Conclusion This extraction technology is suitable for the preparation of total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves,its yield rate being higher.The total flavonoids from spaceflight hybrid Broussonetia papyrifera leaves displayed obvious antivibrio activity resulting in the synergistic effect of their different polar components.