采用酶-无溶剂微波法对东北野生鲜薤白进行精油提取、化学成分鉴定和抑菌活性的研究。利用GC-MS技术对水蒸气蒸馏法(hydrodistillation,HD)、无溶剂微波法(solvent-free microwave extraction,SFME)和酶-无溶剂微波法(enzymatic pretreatment combined with solvent-free microwave extraction,EP-SFME)提取精油的化学成分进行分析。运用滤纸片琼脂板扩散法和二倍稀释法,测定了精油对5种细菌(大肠杆菌、铜绿假单胞菌、枯草芽孢杆菌、金黄色葡萄球菌和单核增生李斯特菌)的抑制效果,最后探究了对大肠杆菌的抑制机理。结果表明,EP-SFME的精油提取率[(0.115±0.005)%]比HD和SFME分别高出76.92%和21.10%。GC-MS分析表明,EP-SFME提取的精油中草酸-4-氯苯基壬酯(+19.53%)、二硫代乙酸烯丙酯(+19.48%)、1-十六炔(+11.86%)、3, 4-二甲基噻吩(+8.86%)、3, 4-二甲基噻吩-2-硫醇(+5.94%)、邻苯二甲酸环丁基十三烷基酯(+3.34%)、1, 2, 4-苯三甲酸-1, 2-二甲酯(+3.07%)、反1, 3-二恶烷-2, 2, 4, 6-四甲基(+2.84%)和2-巯基-3, 4-二甲基-2, 3-二氢噻吩(+2.04%)等成分含量显著增加2%以上,增加成分以含硫化合物和含氧化合物为主。EP-SFME所获得精油对5种细菌的抑菌活性均强于HD和SFME。其中对大肠杆菌的生长抑制作用最强,最小抑菌浓度和最低杀菌浓度分别为3.13 mg/mL和6.25 mg/mL。精油主要通过破坏大肠杆菌的细胞膜和细胞壁,诱导蛋白和核酸泄漏,从而抑制大肠杆菌的生长。研究表明,EP-SFME是从东北野生鲜薤白中提取高品质活性精油的1种有效方法。
The extraction, chemical composition, and antibacterial activity of volatile oil of wild fresh Allium macrostemon Bunge in northeast China were studied by enzymatic pretreatment combined with a solvent-free microwave method.GC-MS was used to analyze the chemical components of volatile oils extracted by hydrodistillation (HD), solvent-free microwave extraction (SFME), and enzymatic pretreatment combined with solvent-free microwave extraction (EP-SFME).The inhibitory effect of volatile oil on five bacteria (Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, and Listeria monocytogenes) was determined by diffusion method and twofold dilution method of filter paper agar plate, and finally, the inhibition mechanism of E.coli was explored.The results showed that the essential oil yield extracted by the EP-SFME [(0.115±0.025)%] was 76.92% and 21.10% higher than that by the HD and SFME.GC-MS analysis showed that the main compound contents in the essential oil extracted by the EP-SFME were significantly increased, and the main compounds with an increase of more than 2%, including oxalic acid-4-chlorophenyl nonyl ester (+19.53%), allyl dithioacetate (+19.48%), 1-hexadecyne (+11.86%), 3, 4-dimethylthiophene (+8.02%), 3, 4-dimethylthiophene-2-thiol (+5.94%), phthalic acid-cyclobutyl tridecyl ester (+3.34%), 1, 2, 4-benzenetricarboxylic acid-1, 2-dimethyl ester (+3.07%), 1, 3-dioxane-2, 2, 4, 6-tetramethyl-, trans-1, 3-dioxane-2, 2, 4, 6-tetramethyl (+2.84%) and 2-mercapto-3, 4-dimethyl-2, 3-dihydrothiophene (+2.04%), etc.Among them, sulfur-containing and oxygen-containing compounds were the main components.The antibacterial activity of the volatile oil obtained by the EP-SFME against the five bacteria was stronger than that of the HD and SFME.Among them, the growth inhibition effect on E.coli was the strongest, with the minimum inhibitory concentration and minimum bactericidal concentration of 3.13 mg/mL and 6.25 mg/mL, respectively.Volatile oils inhibited the growth of E.coli mainly by destroying the cell membrane and cell wall of E.coli, inducing protein and nucleic acid leakage.The EP-SFME could effectively extract high-quality essential oil with high antibacterial activity from northeast wild fresh Allium macrostemon Bunge.
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