通过测定秦岭中蜂蜂蜜理化指标,采用超高效液相色谱串联四极杆串联轨道离子阱质谱(ultra high performance lliquid chromatography Q exactive HFX mass spectrometer,UHPLC-QE-MS)对其酚类提取物进行定性分析,结合牛津杯实验、最小抑菌浓度实验,生长曲线测定及冷场发射扫描电镜分析,探究秦岭中蜂蜂蜜酚类提取物组分及其对大肠杆菌及金黄色葡萄球菌抑菌活性。结果表明:秦岭中蜂蜂蜜pH为5.61,水分含量为(17.86±0.06)%,总糖质量分数为69.78%,其中果糖36.33%,葡萄糖含量32.04%,蔗糖1.41%,淀粉酶活性为26.4 °Gothe,总酚酸和总黄酮含量分别为(37.85±1.33)mg GAE/kg和(109.38±0.05)mg RUE/kg;UHPLC-QE-MS拟鉴定出40种酚类组分,其中酚酸类11种,黄酮类29种,山奈素及山奈酚为代表性组分;提取物对金黄色葡萄球菌及大肠杆菌的最低抑菌浓度分别为2.5、5.0 mg/mL,生长曲线抑制活性呈现浓度依赖性,整体抑菌效果优于枣花蜜提取物;提取物可使致病菌细胞表面皱缩及裂解并抑制其生长。综上所述,秦岭中蜂蜂蜜具有水分含量低,淀粉酶活性高、总酚含量高等特点,其酚类提取物可有效抑制金黄色葡萄球菌及大肠杆菌生长,具有较高的天然抑菌资源开发潜能。
To investigate the phenolic components of Qinling Apis cerana honey, the physiochemical indexes, and ultra high performance liquid chromatography Q exactive HFX mass spectrometer(UHPLC-QE-MS) were tested in the honey sample. The Oxford cup tests, minimum inhibitory concentration (MIC), growth curves and field emission scanning electron microscopy (FESEM) were used to analysis its phenolic extract antibacterial properties against Staphylococcus aureus ATCC 29213 and Escherichia coli ATCC 25922. Physicochemical indexes of Qinling A. cerana honey were pH 5.61, moisture content (17.86±0.06)%, total sugar concentration 69.78%,fructose content 36.33%, glucose content 32.04%, sucrose content 1.41%, and amylase activity 26.4 °Gothe. The total phenolic contents and total flavonoids in the extract were (377.85±1.33) mg GAE/kg and (109.38±0.05) mg RUE/kg, respectively. A total of 40 phenolic compounds were qualitative based on UHPLC-QE-MS, which 11 were phenolic compounds, and 29 were flavonoid compounds. Kaempferol and kaempferide were two typical components of our sample. The MIC against S. aureus and E. coli were 2.5 and 5.0 mg/mL, respectively. The growth curves of S. aureus and E. coli were inhibited with the increase of Qinling A. cerana honey phenolic extract. The cell surface of S. aureus and E. coli were significantly collapsed and shrunk, and two pathogens were inhibited after treated with the phenolic extracts. Overall, the Qinling A. cerana honey exhibited low moisture content, high sugar concentration, high amylase activity and high TPC and TFC properties. And its phenolic extracts could effectively inhibit the growth of S. aureus, E. coli, and exhibit great potential as a natural antibacterial resource.
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