牛蒡叶活性成分快速提取及对木腐菌抑制效果

研究了牛蒡叶中抑制木腐菌活性成分的快速提取工艺及其提取物对木材腐朽菌的抑制性能。在以彩绒革盖菌为提取物抑菌效果指示菌的提取预实验以及以提取率为指标的单因素实验基础上，采用响应曲面法对牛蒡叶中活性成分进行优化提取，确定最优提取方式和对木材腐朽菌(彩绒革盖菌和密粘褶菌)的抑制能力，并通过Accela U-HPLC高速液相色谱系统分离和鉴定优化条件下提取物中的活性成分。结果表明:在预实验中，酸性水提取物的抑菌效果要优于其他方式；经过响应曲面四因素三水平优化后，在液料比30 mL/g、提取温度52 ℃、提取时间77 min和溶剂pH值2.0条件下，理论提取率达到45.81%，经验证此条件下的平均提取率为44.86%，与模型理论值差异为-2.08%；此条件下的提取物，比预实验中采用酸性水方式获得的提取物抑制白腐菌的效果提高了16.47%；在此条件下获取的提取物，经U-HPLC-MS分离并鉴定出12种具有抑菌活性的成分，分别为有机酸类(奎宁酸、原儿茶酸、异绿原酸A、异绿原酸B、异绿原酸C、二聚绿原酸、绿原酸、4, 5-二咖啡酰奎尼酸)、黄酮类(牛蒡酮-a)、苷类(芦丁、5′-Propanediolmatairesinoside)、萜类(β-桉叶醇)。经验证，响应曲面优化后得到的提取物对彩绒革盖菌和密粘褶菌均具有良好的广谱抑菌能力，其对两种菌的最低抑菌质量分数分别为1.88%和2.31%。 

Rapid extraction of active components from Arctium lappa leaves and its anti-fungal properties of wood rot fungi

The purpose of this study was to determine the rapid extraction process of the active ingredient from burdock leaves and its inhibition performance of wood-rot fungus. On the basis of the preliminary experiment with the Coriolus versicolor as indicator fungi and the single factor experiments with the extraction rate as an indicator, the response surface method ( RSM) was used to optimize the extraction of active constituents from the leaves of Arctium lappa, and the anti-fungal ability to wood rot fungi was determined. And the active constituents were separated and identified by U-HPLC-MS under the optimization condition of extraction. Experiment results showed that: the anti-fungi property with acidic water extraction was superior to other ways in the preliminary experiments. After four factors and three levels of response surface optimization, the extraction rate reached 45. 81% under the following conditions:the ratio of liquid to material was 30 mL/g, the extraction temperature was 52℃, extraction time was 77 min, and the solvent pH was 2. 0. The average extraction yield was 44. 86% verified under the optimization condition, and the theoretical value difference of the model was -2. 08%. The effect of the extract on inhibiting white rot fungi was 16. 47% higher than that of the pre-experiment with the acidic water. Twelve kinds of anti-fungi active compounds were isolated and identified by U-HPLC-MS, i. e. organic acids ( quinic acid, protocatechuic acid, isochlorogenic acid A, isochlorogenic acid B,isochlorogenic acid C, dimer chlorogenic acid, chlorogenic acid, 4,5-dicaffeoylquinic acid), flavonoids ( arctinone-a ) , glycosides ( rutin, 5′-propanediolmatairesinoside ) , terpenes (β-eudesmol ) . The minimum inhibitory concentrations of the extracts obtained by this method were 1. 88% and 2. 31% for white rot fungi ( Coriolus versicolor) and brown rot fungi ( Gloceophyllum trabeum) , respectively.