摘要
植物精油具有多种抑菌活性并已被许多研究结果所证明,为了给植物精油类天然消毒杀菌产品开发利用提供理论依据,本论文采用共水蒸馏法对臭柏(Sabina vulgaris Ant.)、油松(Pinus tabulaeformis Carr.)精油的提取进行了研究,对臭柏、油松精油的体外抑菌活性及其对室内空气微生物的抑菌活性进行了比较研究。主要结果如下:
1.采用共水蒸馏法对不同季节臭柏、油松针叶精油提取率进行了对比分析,探讨影响提取率的主要因素。结果表明,不同提取条件的精油提取率存在明显差异。经单因素试验和正交试验结果确定的臭柏精油最佳最经济工艺参数为:液料比为7:1,NaCl浓度2%,浸泡时间2h,蒸馏时间为3h;油松精油的最佳最经济提取工艺条件为:液料比为7:1,NaCl浓度2%,浸泡时间2h,蒸馏时间为3h。研究了臭柏与油松针叶精油提取率月动态变化,臭柏枝叶精油以8月份的含量最高,可达2.30mL/100g干重,油松针叶精油以9月份的含量最高,可达1.00mL/100g干重。共水蒸馏法提取臭柏、油松精油对提取率的主要影响因素的大小排列顺序为:蒸馏时间>液料比>NaCl浓度。
2.体外抑菌实验表明:臭柏精油对金黄色葡萄球菌抑菌不敏感,对黑曲霉抑菌敏感,对大肠杆菌、酿酒酵母菌抑菌中度敏感,对枯草芽孢杆菌抑菌高度敏感,且不同月份间差异不显著。臭柏精油对5种菌的最低抑菌浓度(MIC)分别为:大肠杆菌0.25%(V/V),金黄色葡萄球菌2.50%(V/V),枯草芽孢杆菌0.13%(V/V),酿酒酵母菌0.50%,黑曲霉0.50%(V/V)。
3.油松精油对金黄色葡萄球菌和黑曲霉抑菌不敏感,对大肠杆菌和酵母菌抑菌敏感,对枯草芽孢杆菌抑菌中度敏感,且不同月份间差异不显著。最低抑菌浓度(MIC)分别为:大肠杆菌0.75%(V/V),金黄色葡萄球菌0.50%(V/V),枯草芽孢杆0.25%(V/V),酿酒酵母菌1.00%(V/V),黑曲霉0.50%(V/V)。
4.不同生长季节(分别采于5月、7月、8月、9月、11月)的臭柏、油松针叶精油对5种菌抑制作用的方差分析结果表明:两种植物精油的抑菌活性存在差异显著,8月份的臭柏精油对不同菌种的最低抑菌浓度为枯草芽孢杆菌最低,对金黄色葡萄球菌的抑菌浓度最高;9月份的油松精油对枯草芽孢杆菌抑菌浓度最低,对黑曲霉的抑菌浓度最高。
5.采用棋盘格法对臭柏、油松精油的联合抑菌活性进行了研究,结果表明:臭柏、油松精油对大肠杆菌、金黄色葡萄球菌、酿酒酵母菌具协同效果,而对枯草芽孢杆菌和黑曲霉的协同效果不显著。
6.自然沉降法对两种精油对室内空气微生物的抑菌活性进行了研究,结果表明:臭柏、油松精油(以8月份提取精油为例),抑菌效果具有明显规律性变化:真菌>细菌>放线菌。臭柏、油松精油的抑菌能力随抑菌时间的增加而增加,30min>20min>10min,但抑菌20min后,抑菌效果增强不明显,且臭柏精油抑菌能力显著高于油松。
Antibacterial activities of plant essential oil had been proved to have inhibition effect on kinds of bacterium by many scholars. In order to provide theoretical basis for the Development and Utilization of the disinfection and sterilization products which come from plant essential oil, we study antibacterial activity of Sabina vulgaris Ant. and Pinus tabulaeformis Carr. essential oil, including extraction of essential oil, inhibition effect on experimental bacterium and inhibition effect on air microbe. The main results were as follows:
1. According to compare extraction rate of Sabina vulgaris Ant and P.tabulaeformis Carr essential oil by steam distillation in different months, we analysis on main influence factors of extraction rate. The result showed that extraction rate was different in different extraction condition. We made sure the method of essential extraction by oil single factor experiment and orthogonal test, the results showed that the best parameters were ratio between solvents and raw materil:7:1, salt adding 2%,soaking time:2h,extracting time:3h .At this condition, the extraction ration was 2.3 ml/100g. The most economic and optimum conditions of P.tabulaeformis Carr extraction were ratio between solvents and raw materil:7:1, salt adding 2%, soakingtime:2h, extracting time:3h.At this condition, the extraction ration was 1.0ml/100g. The month dynamic of extraction rate of Sabina vulgaris Ant and P.tabulaeformis Carr essential oil showed that extraction rate of Sabina vulgaris Ant was highest(2.30mL/100g) in August; P.tabulaeformis Carr was highest(1.00mL/100g) in September. The influence factors of extraction rate was extracting time>ratio between solvents and raw materil>NaCl concentration.
2. The diameter of antibacterial activities circle showed that S.vulgaris Ant. essential oil had little inhibition effect on Staphylococcus aureus, and had inhibition effect on Asperhillus niger, had moderate inhibition effect on Escherichia coli and Saccharom yces Cerevisiae, had high inhibition effect on Bacillus subtilis. The differences of antibacterial activities were not significant in different months. The minimum inhibitory concentration of essential oil of Sabina vulgaris Ant were: Escherichia coli(0.25%), Staphylococcusaureus(2.5%), Bacillus subtilis(0.13%), Asperhillus niger(0.5%) , Saccharom yces Cerevisiae(0.5%).
3. The diameter of antinicrobial activities circle showed that P.tabulaeformis Carr.essential oil had little inhibition effect on Staphylococcus aureusand, had inhibition effect on Asperhillus niger, had moderate inhibition effect on Escherichia coli and Saccharom yces Cerevisiae, had high inhibition effect on Bacillus subtilis. The differences of antibacterial activities were not significant in different months. The minimum inhibitory concentration of essential oil of Sabina vulgaris Ant , Escherichia coli(0.75%), Staphylococcu saureus(0.5%), Bacillus subtilis(0.75%), Asperhillus niger(1.0%),Saccharom yces Cerevisiae (0.5%).
4. The inhibiting effects of essential oils of Sabina vulgaris and Pines tabulaeformis to bacteria was studied by Variance Analysis in different months(May,July,August,September,November). The results suggested that there were notable different in different collecting time. The best inhibiting effects of essential oils of Sabina vulgaris to bacteria was August and Pines tabulaeformis was September. The lowest minimum inhibitory concentration of Sabina vulgaris and Pines tabulaeformis : Bacillus subtilis > Escherichia coli > Asperhillus niger > Saccharom yces Cerevisiae > Staphylococcus aureus.
5. Unite antinicrobial activities of Sabina vulgaris and Pines tabulaeformis essential oil to bacteria was studied by Chessboard method. The results suggested that Sabina vulgaris and Pines tabulaeformis essential oil had collaborative effect to Staphylococcus aureus, Escherichia coli,Asperhillus niger,Saccharom yces Cerevisiae , the collaborative effect to Bacillus subtilis and Asperhillus niger is no distinctness.
6.The inhibiting effects of essential oils of Sabina vulgaris and Pines tabulaeformis to bacteria was studied by means of natural precipitation method, the results suggested that there were different antibacterial effect of essential oils of Sabina vulgaris Ant and Pines tabulaeformis in different seasons: fungi>bacteria>actinomycetes. The inhibiting effects of essential oils of Sabina vulgaris and Pines tabulaeformis to bacteria was growing as times going on, 30min>20min>10min,When the time of the antibacterial effect was more than 20min, the antibacterial ability of Sabina vulgaris and Pines tabulaeformis had no significant increase, and the antibacterial ability of essential oils of Sabina vulgaris was stronger than essential oils of Pines tabulaeformis.
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