葡萄叶乙醇提取物的提取工艺及体外抑菌活性研究

Extraction Process and Vitro Antibacterial Activity of Grape Leaves' Ethanol Extract

为优选出葡萄叶乙醇提取物的最佳提取工艺条件,以采自吐鲁番市恰特喀勒乡的葡萄叶为材料,采用索氏提取器回流提取法提取葡萄叶乙醇提取物;工艺优化以乙醇浓度、提取温度、提取时间、料液比为影响因素,分别用大肠杆菌、金黄色葡萄球菌、白假丝酵母菌、枯草芽孢杆菌做抑菌试验,并以其中抑菌效果最明显的金黄色葡萄球菌为指示菌,以葡萄叶乙醇提取物的抑菌圈直径为指标,在单因素试验的基础上采用了正交试验,并用K-B琼脂扩散法测定其体外抑菌活性。结果表明,各因素对提取葡萄叶抑菌活性物质的影响顺序为,乙醇浓度>浸提温度>料液比>浸提时间。最佳提取工艺为,乙醇浓度70%、浸提温度70℃、料液比1︰30、浸提时间3h。在最佳提取条件下获得的葡萄叶提取物对4种菌的抑菌作用大小为:金黄色葡萄球菌﹥大肠杆菌﹥白假丝酵母菌﹥枯草芽孢杆菌。葡萄叶中乙醇提取物对金黄色葡萄球菌和大肠杆菌具有较显著的抑菌作用,其中对金黄色葡萄球菌抑菌活性最大,而对枯草芽孢杆菌的抑菌作用不太显著。本文所建立的葡萄叶乙醇提取物的最佳提取工艺具有渐变快速、仪器设备简单、不污染环境等优点,可用于葡萄叶中具有抗菌活性物质的提取。

To identify the optimum extraction process of ethanol extract from grape leaves, taking grape leaves collected Chatqal Township of Turpan city as research materials, Soxhlet extractor reflux extraction were used to extract ethanol. The optimum process conditions were studied based on antibacterial tests of Escherichia coli, Staphylococcus aureus, Canidia albicans and Bacillus subtilis with consideration of several factors such as ethanol concentration, extraction temperature, extraction time and solid-liquid ratio. Orthogonal test was conducted based on single-factor test, and vitro antibacterial activity was measured through K-B agar diffusion method. In this test, the indicator bacteria was Staphylococcus aureus, and the indictor was the diameter of bacterial inhibition zone. The results showed that the order of the factors that influence the antibacterial activity was ethanol concentration〉extraction temperature〉solid-liquid ratio〉extraction time, and the optimum extraction process was 70% ethanol concentration, extraction temperature 70℃, solid-liquid ratio 1 ： 30, extraction time 3h. The influence order of ethanol extract on the antibacterial activity of the 4 selected bacteria was Staphyloccocus aureus 〉Escherichia coli〉 Canidia albicans〉Bacillus subtilis, and among them, the ethanol extract had significant effect on Staphylococcus aureus and Escherichia coli, but not significant for Bacillus subtilis.