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葡萄叶乙酸乙酯提取物的提取分别采用索氏提取器回流提取法和超声波提取法;抑菌活性测定采用琼脂扩散法(K-B法).索氏提取法工艺优化采用单因素试验,以金黄色葡萄球菌抑菌圈直径为考察指标,分别考察提取时间、提取温度、料液比、氧化剂、还原剂、p H值、紫外线及存放时间对葡萄叶乙酸乙酯提取物抑菌活性的影响;超声波提取法工艺优化在单因素试验的基础上采用了正交试验,以金黄色葡萄球菌抑菌圈直径为考察指标,分别考察超声功率、料液比、提取温度、提取时间对葡萄叶乙酸乙酯提取物抑菌活性的影响.结果表明:葡萄叶乙酸乙酯提取物的索氏提取法最佳提取工艺条件为:料液比为1∶30、提取温度为70℃、提取时间为3 h、p H值为8、氧化剂和还原剂浓度均为最低;超声提取法最优工艺条件为A3B3C2D1,即:料液比为1∶30、提取时间为45 min、超声功率为150 W、提取温度为50℃.在超声波提取法工艺条件下,葡萄叶乙酸乙酯提取物对金黄色葡萄球菌的抑菌圈平均直径可达1.81 cm.超声波提取法对葡萄叶中脂溶性抑菌活性成分的提取效率显著优于索氏提取法.葡萄叶乙酸乙酯提取物对金黄色葡萄球菌和白假丝酵母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.葡萄叶乙酸乙酯提取物对4种菌种抑菌活性顺序为:金黄色葡萄球菌>白假丝酵母菌>大肠杆菌>枯草芽孢杆菌.葡萄叶乙酸乙酯提取物具有很高的抑菌活性,可作为脂溶性抑菌活性组分的来源.
Extraction of ethyl acetate extract from grape leaves were respectively performed by Soxhlet extractor reflux extraction and ultrasonic extraction.The antibacterial activity was determined by agar diffusion method (KB method) Soxhlet extraction process optimization using single factor test, golden yellow Staphylococcus aureus inhibition zone diameter as an index to investigate the effects of extraction time, extraction temperature, solid-liquid ratio, oxidant, reducing agent, p H value, ultraviolet light and storage time on the antibacterial activity of ethyl acetate extract; Optimization of extraction process Based on the single-factor test, orthogonal test was adopted. Taking the diameter of bacteriostatic circle of Staphylococcus aureus as the index, the effect of ultrasonic power, solid-liquid ratio, extraction temperature and extraction time on ethyl acetate The results showed that the optimum extraction conditions of Soxhlet extraction of ethyl acetate extract were as follows: the ratio of solid to liquid was 1:30, the extraction temperature was 70 ℃, the extraction time was 3 h, p H value is 8 and the concentration of oxidant and reductant is the lowest. The optimum conditions of ultrasonic extraction are A3B3C2D1, ie, the ratio of material to liquid is 1:30, the extraction time is 45 min, the ultrasonic power is 150 W , The extraction temperature was 50 ° C. Under the ultrasonic extraction process conditions, the average diameter of the inhibition zone of ethyl acetate extract of grape leaves against Staphylococcus aureus up to 1.81 cm. Ultrasonic extraction of grape leaf fat-soluble antibacterial activity The extraction efficiency of the components was significantly better than that of Soxhlet extraction.Ethyl acetate extract of grape leaves had significant antibacterial activity against Staphylococcus aureus and Candida albicans, of which Staphylococcus aureus had the highest antibacterial activity, Bacillus subtilis bacteriostatic effect is not significant.Effects of grape leaf ethyl acetate extract on the four kinds of bacteria antibacterial activity order: Staphylococcus aureus> Candida albicans> Escherichia coli> Bacillus subtilis. Grape leaves Ethyl acetate extract has a high antibacterial activity, can be used as a source of fat-soluble antimicrobial activity components.