CN108498577A - 一种肉桂挥发油用于杀菌和或抑菌、抗耐药菌的应用及其制备方法 - Google Patents
一种肉桂挥发油用于杀菌和或抑菌、抗耐药菌的应用及其制备方法 Download PDFInfo
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Abstract
本发明公开了肉桂挥发油的抑菌和抗耐药菌作用的应用。采用水蒸气蒸馏法提取肉桂挥发油，肉桂挥发油对葡萄球菌属、大肠埃希菌属、念珠菌属、假单胞菌属、肠杆菌属等细菌和真菌具有一定的杀灭和/或抑制生长的作用。肉桂挥发油对耐药型葡萄球菌属、耐药性大肠埃希菌属、耐药型肠杆菌属、耐药型克雷伯氏菌属、耐药型假单胞菌属、耐药型不动杆菌属、耐药型肠球菌属等细菌具有一定杀灭和/或抑制生长的作用。肉桂挥发油可应用于食品、药品、保健品、化妆品和其他日用产品等行业，制备出具有抑菌和抗耐药菌作用的各种形式的产品。属于肉桂挥发油的新应用领域。
Description
技术领域
本发明涉及食品医药化工技术领域，尤其涉及一种肉桂挥发油用于杀菌和或抑菌、抗耐药菌的应用及其制备方法。
背景技术
肉桂Cinnamomi Cortex为樟科植物肉桂Cinnamomum cassia Presl.的干燥树皮，是我国重点开发研究的药材之一，也是日常食品中常用的调味料之一，为药食两用中药。其性热，味辛、甘，具有补火助阳，温通经脉的功效，临床常用于腰膝冷痛，肾虚作喘，虚阳上浮，虚寒吐泻等症。肉桂的主要化学成分是挥发油，其挥发油是各种药理作用的物质基础。
文献(王波,龚伟,陈国宝,宋桂萍,金俊杰,秦昆明,蔡宝昌.肉桂挥发性成分的气相色谱/质谱分析.世界中西医结合杂志,2014,9(09):941-943.)报道肉桂挥发油中主要化学成分为芳香醛、芳香酯、脂肪酸和萜类化合物，其中桂皮醛含量最高(41.86％)，其次是棕榈酸(7.66％)。
抗菌素耐药性正在变得越来越危险。据报道全球每年出现约44万耐多药结核病(MDR-TB)新发病例，至少造成15万例死亡。在多个疟疾流行国家，对较早一代抗疟药物的耐药性十分普遍。此外，有很大比例的医院感染是由耐甲氧西林金黄色葡萄球菌(MRSA)等高度耐药细菌引起的。耐药微生物引起的感染会造成长期患病和更大的死亡风险，使得许多传染病有可能变得无法控制。2011年世界卫生日的主题是“控制抗菌素耐药性：今天不采取行动，明天就无药可用”。中医药是古老的治疗手段之一，在抗菌、抗病毒、抗炎等方面的显著疗效也逐渐得到全世界的认同。抗生素细菌耐药现状的不乐观，研究者纷纷将目光投向中医药。
目前，尚未见有关肉桂挥发油抗耐药菌活性研究的相关文献报道。
发明内容
本发明目的在于提供一种抑菌和/或杀菌、抗耐药作用的肉桂挥发油及其在制备具有抑菌和/或杀菌、抗耐药作用的产品中的应用。
本发明采取的技术方案是：
肉桂挥发油用于杀菌和/或抑菌、抗耐药菌的应用。
所述的肉桂挥发油是肉桂植物的任何一部分或是几个部分的组合或是整株植物提取物。
所述的肉桂挥发油是水蒸气蒸馏或是CO2超临界萃取或是其他萃取方法所制备。
所述应用是对耐药菌有抑制和/或杀灭作用。
所述的耐药菌为耐药型葡萄球菌属、耐药性大肠埃希菌属、耐药型肠杆菌属、耐药型克雷伯氏菌属、耐药型假单胞菌属、耐药型不动杆菌属、耐药型肠球菌属、耐药型链球菌属，优选耐药型金黄色葡萄球菌、耐药型克雷伯杆菌、耐药型铜绿假单孢杆菌、耐药型大肠埃希菌、耐药型鲍曼不动杆菌、耐药型屎肠球菌、耐药型乙性溶血性链球菌。
所述肉桂挥发油的制备方法如下：取肉桂原料适当粉碎，5-15倍量蒸馏水浸泡0.5-2h后，加热料液采用水蒸气蒸馏法提取挥发油，蒸馏提取3h以上，所得挥发油及芳香水放冷静置过夜后收集挥发油，无水硫酸钠脱水处理，得肉桂挥发油，有特殊香味，得到本发明的肉桂挥发油。
优选的技术方案：所述肉桂挥发油的制备方法如下：取肉桂原料适当粉碎，十倍量蒸馏水浸泡1h后，加热料液采用水蒸气蒸馏法提取挥发油，蒸馏提取3h以上，所得挥发油及芳香水放冷静置过夜后收集挥发油，无水硫酸钠脱水处理，得肉桂挥发油，有特殊香味，得到本发明的肉桂挥发油。
所述的肉桂挥发油用于制备有抑菌和/或杀菌、抗耐药作用的食品、保健品、化妆品、药品和其他日用产品。
本发明具体涉及肉桂挥发油在制备抑菌和/或杀菌、抗耐药作用的单方或复方药物中的新应用。
本发明以肉桂挥发油为原料，可将其作为活性成分与制药学上可接受的载体或药用辅料开发成相宜的抑菌和/或杀菌、抗耐药作用的药物制剂，如注射剂、胶囊剂(软胶囊、硬胶囊)、滴丸剂、片剂、颗粒剂、散剂、口服剂等。
具体实施方式
下面的实施例是对本发明的进一步详细描述。
实施例1肉桂挥发油抗细菌作用
1材料与方法
1.1材料：
(1)供试菌株：大肠埃希菌、金黄色葡萄球菌、绿脓杆菌。
(2)培养基：营养肉汤培养基，青岛高科园海博生物技术有限公司。
1.2实验方法
1.2.1菌种的活化：取干净的3支试管，分别加入5mL的营养肉汤培养基，硅胶塞封口，121℃灭菌30min。将灭菌的试管在超净工作台放冷后用接种环取适量的3种菌放入试管中，混匀，置于培养箱中培养18-24h后观察结果，其中大肠埃希氏菌、金黄色葡萄球菌、绿脓杆菌放置于37℃培养箱中。
1.2.2牛津杯法(管碟法)筛选中药挥发油抗菌活性
将培养12-18h的微生物液体菌种，用紫外-可见分光光度计测量菌液的OD600值，将菌悬液浓度调整至OD600为0.1-0.5左右(细胞浓度为1×108CFU)。无菌条件下用移液枪吸取配制好的200μL工作菌悬液至固体培养基中，用涂布器涂布均匀。用无菌镊子夹取牛津杯放至涂抹均匀菌液的培养基上，使牛津杯与培养基充分接触，不得留有缝隙。牛津杯放置位置要适中。每个样品3个重复。向各牛津杯中加入40μL挥发油原液后，小心将培养皿放置培养箱中，大肠埃希氏菌、金黄色葡萄球菌、绿脓杆菌置于37℃培养箱，观察其抑菌圈大小，并用游标卡尺量抑菌圈直径。
2实验结果
表1肉桂挥发油抗菌实验抑菌圈测量结果(mm)
实施例2肉桂挥发油抗真菌作用
1材料与方法
1.1材料：
(1)供试菌株：白色念珠菌。
(2)培养基：营养肉汤培养基、改良马丁培养基，青岛高科园海博生物技术有限公司。
1.2实验方法
1.2.1菌种的活化：取干净的1支试管，加入5mL的改良马丁培养基，硅胶塞封口，121℃灭菌30min。将灭菌的试管在超净工作台放冷后用接种环取适量的1种菌放入试管中，混匀，置于培养箱中培养18-24h后观察结果，白色念珠菌放置于26.5℃培养箱中。
1.2.2牛津杯法(管碟法)筛选中药挥发油抗菌活性
将培养12-18h的微生物液体菌种，用紫外-可见分光光度计测量菌液的OD600值，将菌悬液浓度调整至OD600为0.1-0.5左右(细胞浓度为1×108CFU)。无菌条件下用移液枪吸取配制好的200μL工作菌悬液至固体培养基中，用涂布器涂布均匀。用无菌镊子夹取牛津杯放至涂抹均匀菌液的培养基上，使牛津杯与培养基充分接触，不得留有缝隙。牛津杯放置位置要适中。每个样品3个重复。向各牛津杯中加入40μL挥发油原液后，小心将培养皿放置培养箱中，白色念珠菌置于26℃培养箱中，培养18-24h，观察其抑菌圈大小，并用游标卡尺量抑菌圈直径。
2实验结果
表2肉桂挥发油抗真菌实验抑菌圈测量结果
实施例3肉桂挥发油抗耐药菌作用
1.材料与方法
1.1细菌株
耐药型金黄色葡萄球菌(MRSA)，菌号为：BAA-1717；耐药型克雷伯杆菌，菌号为：BAA-1705；耐药型绿脓杆菌，菌号为：BAA-2110；耐药型屎肠球菌，菌号为：BAA-2320；耐药型鲍氏不动杆菌，菌号为：19606；均购自美国模式培养物集存库(ATCC)。耐药型肠侵袭性大肠埃希菌，菌号为：186731，购自北京北纳创联生物技术研究院。
1.2试验试剂
表3
1.3试验仪器
表4
1.4试验方法
向96孔培养板内加入胰酪大豆胨液体培养基，每孔280_μl，在第一排每孔加入原药液10_μl，做对倍系列稀释，共设8个稀释度，另设细菌对照。将37℃培养18小时的菌液，用胰酪大豆胨液体培养基稀释成每毫升含105个细菌，在每孔加入各菌液10μl，将培养板放入37℃培养箱内培养24小时，以浊度为指标检查各孔有无细菌生长，以最后一个不显示浊度，细菌无生长的药液稀释度作为最低抑菌浓度(MIC)。
2.实验结果
表5药物对各细菌的抑制作用(μg/ml)
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。
Claims (8)
1.肉桂挥发油用于杀菌和/或抑菌、抗耐药菌的应用。
2.如权利要求1所述的应用，其特征在于：所述的肉桂挥发油是肉桂植物的任何一部分或是几个部分的组合或是整株植物的提取物。
3.如权利要求2所述的应用，其特征在于：所述的提取物是水蒸气蒸馏或是CO2超临界萃取或是其他萃取方法所制备。
4.如权利要求1所述的应用，其特征在于：所述应用是对耐药菌有抑制和/或杀灭作用。
5.如权利要求4所述的应用，其特征在于：所述的耐药菌为耐药型葡萄球菌属、耐药性大肠埃希菌属、耐药型肠杆菌属、耐药型克雷伯氏菌属、耐药型假单胞菌属、耐药型不动杆菌属、耐药型肠球菌属、耐药型链球菌属，优选耐药型金黄色葡萄球菌、耐药型克雷伯杆菌、耐药型铜绿假单孢杆菌、耐药型大肠埃希菌、耐药型鲍曼不动杆菌、耐药型屎肠球菌、耐药型乙性溶血性链球菌。
6.如权利要求1所述的应用，其特征在于：所述肉桂挥发油的制备方法如下：取肉桂原料适当粉碎，5-15倍量蒸馏水浸泡0.5-2h后，加热料液采用水蒸气蒸馏法提取挥发油，蒸馏提取3 h以上，所得挥发油及芳香水放冷静置过夜后收集挥发油，无水硫酸钠脱水处理，得肉桂挥发油，有特殊香味，得到本发明的肉桂挥发油。
7.如权利要求1所述的应用，其特征在于：所述肉桂挥发油的制备方法如下：取肉桂原料适当粉碎，十倍量蒸馏水浸泡1 h后，加热料液采用水蒸气蒸馏法提取挥发油，蒸馏提取3h以上，所得挥发油及芳香水放冷静置过夜后收集挥发油，无水硫酸钠脱水处理，得肉桂挥发油，有特殊香味，得到本发明的肉桂挥发油。
8.如权利要求1所述的应用，其特征在于：所述的肉桂挥发油用于制备有抑菌和/或杀菌、抗耐药作用的食品、保健品、化妆品、药品和其他日用产品。
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