摘要
玫瑰精油具有抑菌、抗氧化、抗敏感、保湿、促进细胞再生等功效;对呼吸系统、消化系统、循环系统以及生殖系统方面都有较好的功效;在情绪方面具有镇压、减压、安眠、抗冲突、缓解紧张和抗抑郁等作用,在化妆品、食品工业、医药等行业拥有广阔的应用和开发潜力。本文主要研究了水蒸气蒸馏法提取湖州产大马士革玫瑰精油的工艺条件;对水蒸气蒸馏(共水蒸馏和水上蒸馏)和顶空固相微萃取提取的玫瑰精油进行了GC-MS化学成分分析,并对比了上述三种方法得到的玫瑰精油成分的差异;分析了玫瑰精油对大肠杆菌、金黄色葡萄球菌和伤寒鼠沙门氏菌的抑菌作用;初步探索了玫瑰精油对药物利血平诱发的小鼠抑郁模型的影响。旨在对玫瑰原料及玫瑰精油的开发应用提供参考依据。
主要研究成果如下:
1、通过单因素实验研究蒸馏时间、料液比、蒸馏液中氯化钠浓度对湖州产大马士革玫瑰精油提取率的影响。在单因素实验的基础上,利用正交优化实验,分析优化了玫瑰精油的水蒸气蒸馏提取工艺。以精油提取率为指标,得到的优化后工艺条件为整花与含有15%氯化钠的蒸馏液按1:5的比例投料,蒸馏5h,在此条件下玫瑰精油的提取率达0.0799%。
2、通过分析顶空固相微萃取和两种水蒸气蒸馏(共水蒸馏和水上蒸馏)法提取的玫瑰精油的成分,分别分离鉴定出45、55和48种化学成分,相对百分含量分别占到总提取精油含量的96.34%、96.17%和96.22%,三种方法提取的玫瑰精油中含量最高的均为D-香茅醇。
3、从顶空固相微萃取和两种水蒸气蒸馏(共水蒸馏和水上蒸馏)法提取的玫瑰精油中共鉴定出87种化学成分,其中共有19种相同的化学成分。通过对比分析发现顶空固相微萃取和两种水蒸气蒸馏(共水蒸馏和水上蒸馏)法提取的玫瑰精油的成分在种类、化合物相对含量上均有较大的差异。顶空固相微萃取获得的玫瑰精油化学成分中以萜烯类、醇类和醛类为主,分别达到精油总含量的29.20%、49.10%和7.64%;水蒸气蒸馏之共水蒸馏获得的玫瑰精油化学成分中以醇类、烷烃类和酯类为主,分别占到总精油含量的46.39%、25.81%和16.77%;水蒸气蒸馏之水上蒸馏获得的玫瑰精油化学成分中以醇类、烷烃类、酯类和萜烯类为主,分别达到精油总含量的47.93%、19.98%、13.74%和9.05%。
4、抑菌试验中,通过滤纸片法测得玫瑰精油对大肠杆菌、金黄色葡萄球菌和鼠伤寒沙门氏菌的抑菌圈直径分别17.21、16.14、17.43mm,为中度敏感,具有较强的抑菌作用。通过试管梯度稀释法测得玫瑰精油对大肠杆菌、金黄色葡萄球菌和鼠伤寒沙门氏菌的最低抑菌浓度(MIC)均为1.25μl/ml,验证了滤纸片法的结果;进一步实验测得玫瑰精油对上述3种供试菌的最低杀菌浓度(MBC)分别为1.25μl/ml,2.5μl/ml,1.25μl/ml。抑菌试验证明了玫瑰精油对大肠杆菌、金黄色葡萄球菌和鼠伤寒沙门氏菌不仅有抑制作用,在一定浓度下,还可以完全杀死细菌。可见玫瑰精油在预防和治疗由细菌引起的疾病方面有一定潜能。
5、抗抑郁试验发现玫瑰精油通过滴鼻给药和香薰给药在拮抗利血平诱发的小鼠体温下降和小鼠眼睑下垂方面有不同的作用效果,拮抗小鼠体温下降无显著性差异;而拮抗造模4h后小鼠眼睑下垂有显著性作用,可能玫瑰精油发挥抗抑郁作用在长期疗效上有更好效果;自主活动实验发现玫瑰精油通过香薰可明显抑制小鼠的自主活动,可能玫瑰精油香薰对中枢有一定的镇静作用。此外,玫瑰精油滴鼻给药高、中、低剂量组对拮抗利血平诱发的小鼠眼睑下垂无显著性效果,但是低剂量组和中剂量组对拮抗小鼠体温下降有显著性效果。通过上述利血平抑郁模型初步研究发现了玫瑰精油具有一定的抗抑郁作用,但是仍需进行进一步深入的研究。
Rose essential oil has antibacterial, antioxidant effect, antiallergic effect, moisturizing effect. It can also promote cells regeneration and have good effect on respiratory system, digestive system, circulation system and reproductive system. Apart from that, rose essential oil can help reducing pressure, easing tension, promoting sleep and prevent neurological disease such as depression, anxiety and so on. It has wide application prospects in cosmetics industry, food industry and pharmaceutical industry. This paper mainly studies the steam distillation in Rosa damascena,which planted in Huzhou prefecture. The components of Rosa damascena essential oil were extracted by HS-SPME, Steam distillation, including coeno-water distillation and water and steam distillation were analysed by GC-MS. At the same time, the differences of the volatile composition of Rosa damascena were also contrasted. In this paper, Rosa damascena essential oil's bacteriostatic action and antidepressant action were studied too, which aimed at providing a reference basis for application of Rosa damascena and its essential oil. The main research results are as follows:
1.The effect of distillation time, material to liquid ratio, concentration of sodium chloride and pretreatment of Rosa damascena on the extraction rate of Rosa damascena essential oil were researched by single factor experiment. On the basis of single factor experiment, the Rosa damascena essential oil extracted by Steam distillation process were optimized by using of orthogonal optimization experiment. Optimization of process parameters for Rosa damascena essential oil was mixing the whole flowers and 5 times distillation liquid contained 15% sodium chloride and distilling 5 hours. The finally extracted rate of Rosa damascena essential oil was 0.0799%.
2.The chemical composition of Rosa damascena essential oil extracted by HS-SPME, Steam distillation including coeno-water distillation and water and steam distillation were analysed by GC-MS.45,55and48 chemical components were respectively identified in the above three methods and the percentage content of them was respectively 96.34%,96.17% and 96.22%.6-Octen-1-ol,3,7-dimethyl-, (R)-was the highest content in three methods.
3.87 chemical components were identified in Rosa damascena essential oil extracted by HS-SPME, Steam distillation including coeno-water distillation and water and steam distillation,19 chemical components could find in all three methods. According to a contrast and analysis, the author found that the chemical composition and percentage of content was quite different. The major chemical components of Rosa damascena essential oil extracted by HS-SPME were tepenes, alcohols and aldehydes, respectively accounting for 29.20%,49.10% and 7.64% of the total amount of rose essential oil. The major chemical components of Rosa damascena essential oil extracted by coeno-water distillation were alcohols, esters and alkanes, respectively accounting for 46.39%、25.81% and 16.77% of the total amount of rose essential oil. The major chemical components of Rosa damascena essential oil extracted by water and steam distillation were alcohols, esters, alkanes and terpenes, respectively accounting for 47.93%、19.98%、13.74% and 9.05% of the total respectively accounting for rose essential oil.
4.The in vitro antibacterial effect of Rosa damascena essential oil was studied. The bacteriostasis function of rose essential oil on Eschirichia coli, Staphylococcus aureus and Salmonella typhimrium were tested by filtering paper method and tube double dilution method. The determination of diameter of inhibiting Eschirichia coli, Staphylococcus aureus and Salmonella typhimurium were 17.21,16.14,17.43mm, which belonged to moderate sensitive. The method of doubling deletion in test tube was used to detect the minimal inhibitory concentration (MIC) and minimal bactericidal concentration. The MICs of Rosa damascena essential oil on Eschirichia coli, Staphylococcus aureus and Salmonella typhimurium were all 1.25μl/ml, and the MBCs on three tested bacteria were 1.25μl/ml,2.5μl/ml,1.25μl/ml. Bacteriostatic experiment proved that Rosa damascena essential oil did not only have the bacteriostatic effect, but also have sterilization effect at certain concentration. These results show that Rosa damascena essential oil have great potential application in the prevention and treatment of disease caused by bacteria.
5.Experimental model mice of depression were induced by reserpine for observing the correlation of the drug with Rosa damascena essential oil through olfactory system, including the degree of ptosis and body temperature. The results showed that there were different antagonisms in the degree of ptosis and body temperature. In reserpine model, there was no significant difference in antagonist mice hypothermia, however, there was significant difference in antagonist mice the degree of ptosis after building model 4 hours. Rose essential oil may play a better role in the long-term antidepressant effect. Independent activity test showed that rose essential oil could significantly inhibit the mice aromatherapy independent activities, which suggested that rose essential oil might restrain central nervous system. Furthermore, there were no statistical differences in antagonist mice the degree of ptosis among the high dosage group, middle dose group and the low dosage group. Whereas, the middle dose group and the low dosage group exhibited an excellent antagonism to mice hypothermia which induced by reserpine. In preliminary, it can predict that rose essential oil has the potential of preventing depression.
引文
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