【作者】 胡扬帆；

【导师】 吴楚材；

【作者基本信息】 中南林业科技大学 ， 生态学， 2012， 博士

【摘要】 灰毡毛忍冬(Lonicera macranthoides Hand.-Mazz.)是芳香类药用植物。目前,对灰毡毛忍冬的研究主要集中在化学组分的鉴定、相对含量的测算及抗炎机理上。很少有人深入研究灰毡毛忍冬花提取过程中环境因子对挥发性有机物成分的影响及生态保健功效开发应用。本研究以“提取条件”为切入点,开展了灰毡毛忍冬花挥发性有机物C02超临界萃取工艺及其在旅游生产中的应用等实验。主要研究结果如下：1.C02超临界萃取灰毡毛忍冬花挥发性有机物的最佳工艺研究。分别从萃取压力、萃取温度、萃取时间、CO2流量等因子对灰毡毛忍冬花挥发性有机物萃取率的影响进行研究,实验得出C02超临界萃取灰毡毛忍冬花挥发性有机物的最佳工艺为：萃取压力为28.5MP、萃取温度为38℃、萃取时间为2h及C02流量为18L／h,且各因子对萃取率的影响大小为：萃取温度>萃取压力>萃取时间>C02流量。2.灰毡毛忍冬花挥发性有机物经GC-MS测定,共检出45个组分：其中烃类物质有10种占挥发性有机物总量的44.03%,酸类化合物有8种占24.14%,醇类化合物有9种占6.34%,酯类化合物有9种占8.452%,酮类物质有6种占8.85%。相对含量大于1%的组分有22种：其中棕榈酸含量最高达到了12.34%,其次是正十九烷、正二十九烷、二十一烷、十五烷及正三十二烷等5种化合物的总含量为35.10%。3.灰毡毛忍冬花挥发性有机物的抗炎作用和抗炎机理研究。实验证明灰毡毛忍冬花挥发性有机物对于醋酸致小鼠腹腔毛细血管通透性升高,对二甲苯致小鼠耳廓肿胀都有明显的抑制作用；对角叉菜胶致大鼠足肿胀、致大鼠胸膜炎均显示出良好的对抗作用；对于小鼠棉球肉芽肿慢性炎症模型,虽不能显著减轻肉芽重量,但呈现一定的抑制趋势,并且大剂量组能明显降低肉芽指数；对去肾上腺大鼠角叉菜胶肿胀足中释放出的PGE2和MDA影响作用明显；能明显降低胸膜炎渗出液中NO的含量及PGE2的释放。有较强的炎症抑制作用。4.灰毡毛忍冬花挥发性有机物对金黄色葡萄球菌等10种供试菌种都表现出了一定的抑菌效果,抑菌效果为革兰氏阳性菌>革兰氏阴性菌>真菌。对金黄色葡萄球菌、藤黄八叠球菌、苏云金芽孢杆菌的最低抑菌浓度都为0.03125mg/mL,对枯草芽孢杆菌为0.0625mg/mL,对腐生葡萄球菌为0.25mg/mL,对大肠杆菌为0.5mg/mL,对黑曲霉为4mg/mL,对白色念球菌、绿脓杆菌、米根霉都为8mg/mL。苏云金芽胞杆菌、藤黄八叠球菌、金黄色葡萄球菌在实验培养温度下都被抑制不生长；腐生葡萄球菌在22℃下菌落不生长,24-28℃时轻微生长,30。C后菌落不生长,全部被抑制；黑曲霉、米根霉、白色念球菌在22-26℃下生长,28-42℃时被完全抑制；大肠杆菌和绿脓杆菌在22℃下培养菌落完全不生长,30。C时候生长最旺盛,36℃后菌落不生长,完全被抑制。所有供试菌种在培养10h前菌落完全不生长。5.汽车内灰毡毛忍冬花挥发性有机物浓度为3mg/mL、温度为25℃、时间为30min时抑菌效果最好,抑制率达到67.4%。对旅游车内细菌的抑菌率在无人状态下为63-86%,在载人状态下为48-53%；对真菌的抑菌率在无人状态下为20-42%,在载人状态下为14-39%；对放线菌的抑菌率在无人状态下在车的中、后部为50%,载人状态下的抑菌率在车的中部为33%、后部为25%。抑菌效果为细菌>真菌>放线菌。6.通过测试被试验者的心率、血压变化,SCL-90量表测试、PSQI问卷调查、STAI量表问卷分析,证实灰毡毛忍冬花挥发性有机物对缓解人的焦虑、改善和调整心情具有积极作用。并通过社会学调查,证明灰毡毛忍冬对人的健康长寿和生态保健有功效。以上研究表明,灰毡毛忍冬花挥发性有机物具有较好的抗炎抗菌功效,可有效应用于制药、旅游、生态保健等生产生活领域。更多还原

【Abstract】 Lonicera macranthoides Hand.-Mazz is an aromatic medicinal plant. Studies on the Lonicera macranthoides mainly focus on the analysis of its chemical composition, calculation of relative contents and ant-inflammatory. Few scholars emphasize on the influence of environmental factors to the plant volatile organic compounds during the extraction process and the development on the effect of ecological health. The research is based on the extract condition as the entry point to study the supercritical CO2extraction methods and its application in tourism. The results come out as follows:1. Study the extraction rate while pressure, temperature, CO2flow rate, time as test samples. The optimal process to extraction macranthoides flower using supercritical CO2is:pressure,28.5MP; temperature:38℃; time:2h; CO2flow rate:18kg/h. The affect size of extraction rate is:pressure>temperature> time>CO2flow rate.2. A total of45components were detected after GC-MS analysis to the volatile organic compounds of lonicera macranthoides flower. There are10kinds of hydrocarbons(44.03%), eight kinds of acids(24.14%),nine kinds of alcohols(6.34%),nine kinds of esters(8.452%) and six kinds of ketones(8.85%).Of which22showed higher than1%content, and the maximum content compounds-palmitic acid(12.34%), the total content including nonadecane(9.56%), nonacosane(8.95%),32alkyl(6.68%), pentadecane(5.23%) are over35.10%.3. Results indicated that the macranthoides flower volatile organic compounds can significantly inhibit the enhancement of the mouse peritoneal capillary permeability induced by acetic acid, Good resist effects to the xylene-induced mouse ear swelling,the carrageenan induced rat paw edema and rat pleurisy. To the cotton ball granuloma in mice model of chronic inflammation, the macranthoides flower volatile organic compounds can not significantly reduce granulation weight, but has a certain inhibitory trend, and large dose group can obviously reduce granulation index. It has an obviously effect to reduce the PGE2and MDA from the adrenal gland of rat carrageenin paw edema foot.It can significantly reduce the volume of NO and PGE2released from the pleurisy with effusion. 4. Results indicated that the macranthoides flower volatile organic compounds have antibacterial effects to all tested strains. The antibacterial effects are:gram-positive bacteria> gram-negative bacteria>fungi. The lowest antibacterial concentrations are:staphylococcus aureus(0.03125mg/mL),sarcina lutea (0.03125mg/mL),bacillus thuringiensis(0.03125mg/mL),bacillus subtilis (0.0625mg/mL), staphylococcus saprophyticus(0.25mg/mL), e. coli (0.5mg/mL), aspergillus(4mg/mL), candida albicans,pseudomonas aeruginosa,rhizopus oryzae are8mg/mL.Results also indicated that the staphylococcus aureus,sarcina lutea,bacillus thuringiensis can’t grow at any culture temperature; The staphylococcus saprophyticus can’t grow under22℃,will grow slightly at24-28℃,can’t grow over30℃; The aspergillus,rhizopus oryzae,candida albicans will grow at22-26℃,will be inhabited absolutely under28-42℃; The e. Coli,pseudomonas aeruginosa can’t grow under22℃,will grow the most productively at30℃,will be inhabited absolutely and can’t grow over36℃.All the tested strains can’t grow before10h in cultured.5. The lonicera macranthoides flower volatile organic compounds have the best antibacterial effects when concentration is3mg/mL, temperature25℃, time30min, inhibition rate reached67.4%.The bacterial inhibition rate in unmanned condition is63-86%,In manned sate is48-53%;The fungal inhibition rate in unmanned is20-42%, In manned sate is14-39%; The actinomycetes bacteriostatic rate in unmanned condition is50%in the rear, In manned sate is33%in the middle and25%in the back. The antibacterial effects are:gram-positive bacteria> fungi> actinomycetes.6. Results indicated that the lonicera macranthoides flower volatile organic compounds have positive effects to ease the anxiety, improve and adjust the mood of the people by testing changes of blood pressure and heart rate, testing the SCL-90scale, PSQI questionnaire survey,STAI scale questionnaire analysis, verified the effects to human longevity and health care through the social investigation.Conclusion:The Lonicera macranthoides flower volatile organic compounds have good effects to anti-inflammatory and antibacterial, Can be easily used in pharmaceutical, tourism, ecological health and other production areas of life更多还原