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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。
In order to optimize the extraction process of the ethanol extract of grape leaves, the grape leaves collected from Chakkale Township, Turpan City were used as the materials, and the ethanol extracts of grape leaves were extracted by reflux extraction with Soxhlet extractor. Concentration, extraction temperature, extraction time, the ratio of solid to liquid as the influencing factors, respectively, with E. coli, Staphylococcus aureus, Candida albicans, Bacillus subtilis antibacterial test, and the most obvious antibacterial effect of golden yellow Staphylococcus aureus as indicator bacteria, grape leaf alcohol extract inhibition zone diameter as an index, based on the single factor test using orthogonal test and KB agar diffusion method to determine its antibacterial activity in vitro. The results showed that the order of influence of various factors on the antibacterial substances extracted from grape leaves was ethanol concentration> extraction temperature> solid-liquid ratio> extraction time. The best extraction process, ethanol concentration 70%, extraction temperature 70 ℃, the ratio of solid to liquid 1:30, extraction time 3h. The antibacterial effect of grape leaf extract obtained under the best extraction conditions on four kinds of bacteria was: Staphylococcus aureus> Escherichia coli> Candida albicans> Bacillus subtilis. Grape leaves ethanol extract of Staphylococcus aureus and Escherichia coli has a significant inhibitory effect, of which the largest antibacterial activity of Staphylococcus aureus, Bacillus subtilis antibacterial effect is not significant. The best extraction technology of grape leaf ethanol extract established in this paper has the advantages of rapid change, simple equipment, no pollution to the environment, etc. It can be used for the extraction of antibacterial active substances in grape leaves.