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目的:利用响应面法优化樟叶精油的亚临界萃取技术并考察其抑菌活性。方法:在单因素试验的基础上,以樟叶精油得率为响应值,利用Box-Behnken中心组合试验优选亚临界萃取条件。采用固相微萃取与气相色谱-质谱法(GC-MS)结合对樟叶精油的成分进行分析,通过峰面积归一法计算各成分的相对百分含量。利用滤纸片琼脂扩散法和肉汤稀释法测定樟叶精油对4种供试菌的抑菌活性。结果:在最优的萃取条件(萃取时间30 min,萃取温度40℃,解析温度65℃)下樟叶精油得率为3.54%,这与模型预测值3.56%相匹配。GC-MS鉴定出47个化合物,其中相对百分含量超过1%的有12种成分,主要为桉油精(24.74%)、4-亚甲基-1-(1-甲基乙基)-[3.1.0]二环乙烷(7.05%)、芳樟醇(5.82%)、石竹烯(4.75%)。樟叶精油对大肠杆菌、金黄色葡萄球菌、绿脓杆菌、枯草杆菌均有不同程度的抑制作用。结论:该萃取工艺安全可靠,精油得率较高且具有抑菌性。
Objective: To optimize the subcritical extraction technology of camphor leaf essential oil by response surface methodology and investigate its antibacterial activity. Methods: On the basis of single factor test, the yield of camphor leaf oil was taken as the response value, and the subcritical extraction condition was optimized by the Box-Behnken central combination test. Solid phase microextraction coupled with gas chromatography-mass spectrometry (GC-MS) was used to analyze the composition of camphor leaf essential oil, and the relative percentage of each component was calculated by peak area normalization. The antibacterial activities of camphor leaf oil on four tested bacteria were determined by filter paper agar diffusion method and broth dilution method. Results: Under the optimal extraction conditions (extraction time 30 min, extraction temperature 40 ℃, analytical temperature 65 ℃), the yield of camphor leaf oil was 3.54%, which was consistent with the predicted value of 3.56%. Forty-seven compounds were identified by GC-MS. There were 12 components in the relative percentage of more than 1%, mainly eucalyptol (24.74%), 4-methylene-1- (1-methylethyl) [3.1.0] bicycloethane (7.05%), linalool (5.82%), and caryophyllene (4.75%). Camphor leaf oil on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis have different degrees of inhibition. Conclusion: The extraction process is safe and reliable, with high yield of essential oil and antibacterial activity.