CN103947686B - A kind of Traditional Chinese medicine air disinfectant and its preparation method and application - Google Patents
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Abstract
Description
技术领域 technical field
本发明属于中药提取物应用领域，涉及一种中药空气消毒剂及其制备方法和应用。 The invention belongs to the application field of traditional Chinese medicine extracts, and relates to a traditional Chinese medicine air disinfectant and its preparation method and application.
背景技术 Background technique
空气是致病微生物传播的重要途径之一，空气消毒是预防和控制空气传播疾病，防止交叉感染的重要措施，是防止公共场所如医院、出入境口岸、公共交通工具中致病微生物传播的有效工具。近年来空气传播疾病的发病率居高不下，如2003年的非典，2004年的H5N1，2009年爆发的甲型H1N1流感以及2013年H7N9禽流感的爆发严重危害我国居民的身体健康，促使人们越来越关注室内微生物污染的问题。我国人口多，密度大，如何发展高效低毒的空气消毒剂是一个急需解决的问题。 Air is one of the important ways for the transmission of pathogenic microorganisms. Air disinfection is an important measure to prevent and control airborne diseases and prevent cross-infection. tool. In recent years, the incidence of airborne diseases has remained high, such as SARS in 2003, H5N1 in 2004, H1N1 influenza in 2009 and H7N9 avian influenza in 2013. More and more attention is paid to the problem of indoor microbial contamination. my country has a large population and a high density. How to develop an air disinfectant with high efficiency and low toxicity is an urgent problem to be solved.
目前常用的空气消毒方法是化学法和物理法，化学制剂有过氧乙酸、乳酸、过氧化氢、甲醛、84消毒液、戊二醛等，物理方法有紫外线照射、臭氧发生器等。这些方法能够快速杀灭致病微生物，有效浓度低，但也存在不少缺点，如有毒性、刺激性、金属腐蚀性、持续效果差，许多本身又构成污染物，不同程度地影响人体健康，而且消毒时所有人员必须离场，操作系统要有防护措施；消毒后异味不能很快消除，长期使用会对人体有蓄积中毒的危险等。如84消毒液空气浓度较高时，对人体的黏膜、皮肤有较强的刺激或损伤。乳酸杀菌率为70%，有刺激性气味，腐蚀性强，对呼吸道有刺激，对家具、电器、金属物品有损害。过氧乙酸对粘膜有刺激性，对金属有腐蚀性。过氧化氢长期作用对大鼠特别是雄鼠肝、肾功能及肺、睾丸可能有损害。甲醛薰蒸辛辣味太浓，有致癌性。臭氧发生器释放的臭氧时间过长可使皮肤产生红斑以及臭氧中毒、脱发、掉眉毛等副作用，紫外线照射消毒效果维持不长，停止照射后细菌数很快回升，且受灯管强度、洁净度、环境温湿度、遮挡物和距离影响。目前，各国正在逐渐淘汰毒副作用较强的空气消毒方法，采用空气过滤器消毒、层流等方法净化空气，但此法投资大，对许多公共场所难以实施。 At present, the commonly used air disinfection methods are chemical and physical methods. Chemical preparations include peracetic acid, lactic acid, hydrogen peroxide, formaldehyde, 84 disinfectant, glutaraldehyde, etc. Physical methods include ultraviolet radiation and ozone generators. These methods can quickly kill pathogenic microorganisms, and the effective concentration is low, but there are also many disadvantages, such as toxicity, irritation, metal corrosion, and poor continuous effect. Many of them constitute pollutants and affect human health to varying degrees. Moreover, all personnel must leave the site during disinfection, and protective measures must be taken for the operating system; after disinfection, the peculiar smell cannot be eliminated quickly, and long-term use will cause the danger of accumulated poisoning to the human body. For example, when the air concentration of 84 disinfectant is high, it will strongly stimulate or damage the mucous membranes and skin of the human body. The bactericidal rate of lactic acid is 70%. It has a pungent smell, is highly corrosive, irritates the respiratory tract, and is harmful to furniture, electrical appliances, and metal objects. Peracetic acid is irritating to mucous membranes and corrosive to metals. The long-term effect of hydrogen peroxide may damage the liver, kidney function, lung and testis of rats, especially male rats. Formaldehyde fumigation is too spicy and carcinogenic. The ozone released by the ozone generator for too long can cause erythema on the skin and side effects such as ozone poisoning, hair loss, and eyebrow loss. The disinfection effect of ultraviolet radiation does not last long. , ambient temperature and humidity, shelter and distance. At present, countries are gradually eliminating air disinfection methods with strong toxic and side effects, and adopt air filter disinfection, laminar flow and other methods to purify the air, but this method requires a large investment and is difficult to implement in many public places.
中草药是我国的宝贵遗产，许多中草药体外具有较强的抗菌、抗病毒作用，并具有刺激性小、毒性低的优点，越来越受到研究者的重视和市场的青睐。目前常用的中药空气消毒法有中药烟熏法（如苍术、艾叶）、中药液喷雾法、中药空气消毒片以及中药气雾剂法。其中中药烟熏法容易燃烧不充分，产生的烟雾微粒在空气中漂浮污染空气，对呼吸道黏膜有一定刺激；中药气雾剂使用方便，喷出的雾粒微小轻细，分布均匀，利于扩散，但容器成本较高，制备较复杂，特别是要以如丙烷、丁烷、二甲醚等多种挥发性有机物作抛射剂，可造成空气二次污染，只适于小范围使用；中药液喷雾法不仅具有消毒作用，还能使尘埃沉降，湿化、清新空气等多种效果，已越来越受到关注。 Chinese herbal medicine is a precious heritage of our country. Many Chinese herbal medicines have strong antibacterial and antiviral effects in vitro, and have the advantages of small irritation and low toxicity, and are increasingly valued by researchers and favored by the market. At present, the commonly used traditional Chinese medicine air disinfection methods include traditional Chinese medicine smoke method (such as atractylodes atractylodes and Artemisia argyi), traditional Chinese medicine liquid spray method, traditional Chinese medicine air disinfection tablet and traditional Chinese medicine aerosol method. Among them, the traditional Chinese medicine smoke method is easy to burn insufficiently, and the smoke particles produced float in the air and pollute the air, which has certain irritation to the respiratory mucosa; the traditional Chinese medicine aerosol is easy to use, and the sprayed mist particles are small and fine, evenly distributed, and conducive to diffusion. However, the cost of the container is relatively high, and the preparation is relatively complicated. Especially, multiple volatile organic compounds such as propane, butane, and dimethyl ether should be used as propellants, which can cause secondary air pollution and are only suitable for small-scale use; The spray method not only has disinfection effect, but also can make dust settle, humidify, freshen air and other effects, which has attracted more and more attention.
近年来中药喷雾型空气消毒剂备受关注，对其组方、剂型等方面都取得了一定进展，开发出了不少消毒效果不错的单味和复方消毒剂。但是仍有许多不足之处。由于受中药原材料质量、提取工艺等的影响，中药空气消毒剂效果不够稳定，对其质量的控制尚缺乏研究。有的原料价格昂贵，提取工艺复杂，限制了其使用，有的由于雾化效果不好利用率不高，使用效果仅与紫外线法相近，有的虽然效果较好，但是药液颜色较深，容易污染墙壁、衣物等。 In recent years, traditional Chinese medicine spray-type air disinfectants have attracted much attention, and certain progress has been made in their formulations and dosage forms. Many single-flavor and compound disinfectants with good disinfection effects have been developed. But there are still many shortcomings. Due to the influence of raw material quality and extraction process of traditional Chinese medicine, the effect of traditional Chinese medicine air disinfectant is not stable enough, and there is still a lack of research on its quality control. Some raw materials are expensive and the extraction process is complicated, which limits their use. Some have low utilization rate due to poor atomization effect, and the use effect is only similar to that of ultraviolet method. Although some have better effect, the liquid color is darker. It is easy to contaminate walls, clothing, etc.
因此，积极寻找新的适用于作为中药消毒剂的中药组合物、开发出具有天然清香、安全环保、杀菌效果突出，持续时间长的新型消毒剂具有十分重要的临床意义和社会效益。现有技术中，有采用5~30%的肉桂精油，与70~95%的乙醇混合制备的抑菌空气清新剂（CN201310239366.X）。但这种清新剂由于采用了高浓度的乙醇，易燃，使用安全性较差，另外，肉桂精油的浓度高，对人体的刺激性较大。 Therefore, it is of great clinical significance and social benefit to actively search for new Chinese medicine compositions suitable for use as traditional Chinese medicine disinfectants, and to develop new disinfectants with natural fragrance, safety and environmental protection, outstanding bactericidal effect, and long duration. In the prior art, there is an antibacterial air freshener prepared by mixing 5-30% cinnamon essential oil with 70-95% ethanol (CN201310239366.X). However, this freshener is flammable due to the use of high-concentration ethanol, and is less safe to use. In addition, the concentration of cinnamon essential oil is high, which is more irritating to the human body.
发明内容 Contents of the invention
本发明的发明目的在于提供一种中药组合物，该组合物由肉桂精油及溶剂组成，该中药组合物具有天然清香，可较好杀灭大肠杆菌、金黄色葡萄球菌、白色葡萄球菌、白色念珠菌，对空气中自然菌落具有较好的杀灭作用，安全无毒，使用方便。 The invention object of the present invention is to provide a kind of Chinese medicine composition, and this composition is made up of cinnamon essential oil and solvent, and this Chinese medicine composition has natural delicate fragrance, can kill E. Bacteria, good killing effect on natural colonies in the air, safe and non-toxic, easy to use.
本发明的另一目的在于提供所述中药组合物的制备方法。 Another object of the present invention is to provide a preparation method of the traditional Chinese medicine composition.
本发明的另一目的在于提供所述中药组合物的应用。 Another object of the present invention is to provide the application of the traditional Chinese medicine composition.
本发明的上述目的通过如下技术方案予以实现： Above-mentioned purpose of the present invention is achieved by following technical scheme:
一种中药组合物，由以下按体积百分比计算的组分组成：中药提取物1~3%，溶剂97~99%，其中，所述中药提取物为肉桂精油，所述溶剂为水。 A traditional Chinese medicine composition, which is composed of the following components calculated by volume percentage: 1-3% of Chinese medicine extract, 97-99% of solvent, wherein, the Chinese medicine extract is cinnamon essential oil, and the solvent is water.
本申请中，采用低含量的肉桂精油与水混合，既能维持中药组合物的杀菌效果，同时减少其对人体皮肤的刺激性，还提高组合物使用时的安全性，气味清新，更便于使用。 In this application, a low content of cinnamon essential oil is used to mix with water, which can not only maintain the bactericidal effect of the traditional Chinese medicine composition, but also reduce its irritation to human skin, and improve the safety of the composition when used, with a fresh smell and easier use .
优选地，所述中药组合物由以下按体积百分比计算的组分组成：中药提取物1%，水99%。 Preferably, the traditional Chinese medicine composition is composed of the following components calculated by volume percentage: Chinese medicine extract 1%, water 99%.
优选地，所述中药组合物由以下按体积百分比计算的组分组成：中药提取物3%，水97%。 Preferably, the traditional Chinese medicine composition is composed of the following components calculated by volume percentage: Chinese medicine extract 3%, water 97%.
优选地，所述水优选为蒸馏水。 Preferably, the water is preferably distilled water.
作为一种优选方案，所述中药组合物由肉桂精油1ml，蒸馏水99ml组成。 As a preferred solution, the traditional Chinese medicine composition consists of 1ml of cinnamon essential oil and 99ml of distilled water.
作为一种优选方案，所述中药组合物由肉桂精油3ml，蒸馏水97ml组成。 As a preferred version, the traditional Chinese medicine composition consists of 3ml of cinnamon essential oil and 97ml of distilled water.
所述中药组合物的制备方法为将肉桂精油与水混合均匀即得。 The preparation method of the traditional Chinese medicine composition is obtained by uniformly mixing cinnamon essential oil and water.
所述肉桂精油为采用常规方法提取的肉桂精油或市售肉桂精油。 The cinnamon essential oil is cinnamon essential oil extracted by conventional methods or commercially available cinnamon essential oil.
所述中药组合物在制备空气消毒剂中的应用。 The application of the traditional Chinese medicine composition in the preparation of air disinfectant.
所述中药组合物在制备以喷雾形式使用的空气消毒剂中的应用。 Application of the traditional Chinese medicine composition in the preparation of an air disinfectant used in spray form.
所述中药组合物在作为空气消毒剂时，可以制备成喷雾剂，或者直接喷洒使用，或与加湿器配合超声雾化使用。 When the traditional Chinese medicine composition is used as an air disinfectant, it can be prepared as a spray, or sprayed directly, or used in conjunction with a humidifier and ultrasonic atomization.
本发明所述中药组合物，作为空气消毒剂以喷雾形式施放在空气中，可以有效杀灭空气中的大肠杆菌、金黄色葡萄球菌、白色葡萄球菌、白色念珠菌。 The traditional Chinese medicine composition of the present invention is applied in the air as an air disinfectant in spray form, and can effectively kill Escherichia coli, Staphylococcus aureus, Staphylococcus albicans and Candida albicans in the air.
与现有技术相比，本发明具有如下有益效果： Compared with the prior art, the present invention has the following beneficial effects:
肉桂精油对微生物有很强的杀菌作用，还具有抗肿瘤、抗病毒、抗溃疡、解热、镇痛解痉及杀虫等多种作用，同时具有特殊的芳香气味，然而其更多用于食品和日用品等行业用做防腐剂或香料，本发明开拓性的将肉桂精油与水在一定配比下组成空气消毒剂，以喷雾的形式喷散在空气中可以杀灭空气微生物，具有突出的杀菌效果，同时对人体刺激性小，能防病保健，天然清香，安全环保；发明还发现，把组合物中肉桂精油的含量控制在1~3%，能够在较长时间维持较好的杀菌效果。 Cinnamon essential oil has a strong bactericidal effect on microorganisms, and also has various effects such as anti-tumor, anti-virus, anti-ulcer, antipyretic, analgesic, antispasmodic and insecticidal, and has a special aroma. However, it is more used for Food and daily necessities and other industries are used as preservatives or spices. The invention pioneers the composition of cinnamon essential oil and water in a certain ratio to form an air disinfectant, which can be sprayed in the air in the form of spray to kill airborne microorganisms and has outstanding bactericidal effect. effect, at the same time, it is less irritating to the human body, can prevent diseases and protect health, has a natural fragrance, is safe and environmentally friendly; the invention also found that controlling the content of cinnamon essential oil in the composition to 1-3% can maintain a good bactericidal effect for a long time .
具体实施方式 detailed description
下面结合具体实施例对本发明作进一步的解释说明，但具体实施例并不对本发明作任何限定。除非特别说明，实施例中所涉及的试剂、方法均为本领域常用的试剂和方法。 The present invention will be further explained below in conjunction with specific examples, but the specific examples do not limit the present invention in any way. Unless otherwise specified, the reagents and methods involved in the examples are commonly used reagents and methods in the art.
实施例1 空气消毒剂制备及其杀菌效果实验 Example 1 Preparation of air disinfectant and its bactericidal effect experiment
1.消毒剂制备：将1ml肉桂精油加入99ml蒸馏水中，摇匀，即得100ml消毒剂。此消毒剂无需加压，直接装入喷雾器中即可对室内空气进行消毒杀菌，操作简单，适用范围广。 1. Disinfectant preparation: add 1ml of cinnamon essential oil to 99ml of distilled water, shake well to get 100ml of disinfectant. The disinfectant does not need to be pressurized, and can be directly loaded into the sprayer to disinfect and sterilize the indoor air. The operation is simple and the application range is wide.
2.消毒剂的杀菌效果实验 2. Bactericidal effect experiment of disinfectant
参照2008年版《消毒技术规范》规定的悬液定量杀菌试验，检测上述步骤1制备的消毒剂对大肠杆菌(8099) 、金黄色葡萄球菌(ATCC 6538) 、白色葡萄球菌（ATCC 8799）白色念珠菌(ATCC 10231)的杀菌效果，具体检测方法及结果如下。 With reference to the suspension quantitative bactericidal test stipulated in the 2008 version of "Disinfection Technical Specifications", detect the disinfectant prepared in the above step 1 against Escherichia coli (8099), Staphylococcus aureus (ATCC 6538), Staphylococcus albicans (ATCC 8799), Candida albicans (ATCC 10231), the specific detection method and results are as follows.
1）检测方法：参照2008年版《消毒技术规范》规定的悬液定量杀菌试验 1) Detection method: Refer to the suspension quantitative sterilization test stipulated in the 2008 edition of "Disinfection Technical Specifications"
取大肠杆菌(8099) 、金黄色葡萄球菌(ATCC 6538) 、白色葡萄球菌（ATCC 8799）白色念珠菌(ATCC 10231)24h新鲜斜面培养物，分别用胰蛋白胨生理盐水（TPS）洗下，混匀后稀释成含菌量为1×108cfu/ml～5×108cfu/ml的菌悬液，备用。 Take Escherichia coli (8099), Staphylococcus aureus (ATCC 6538), Staphylococcus albus (ATCC 8799) and Candida albicans (ATCC 10231) 24h fresh slant culture, wash with tryptone saline (TPS) respectively, and mix well Afterwards, it is diluted to a bacterial suspension with a bacterial content of 1×10 8 cfu/ml to 5×10 8 cfu/ml, and is set aside.
在无菌试管内加入0.5ml菌悬液和 0.3%小牛血清白蛋白0.5ml，混匀，置20℃水浴中5min后，加入1%肉桂精油水溶液(阳性对照为TPS)4ml，立即混匀。作用至预定时间，分别吸取0.5ml菌药混合液加于4.5ml中和剂试管中，混匀作用10min后，取该液1.0ml接种培养，37℃培养48小时后活菌计数。试验重复 3 次，计算各组的活菌浓度（cfu/mL），并换算为对数值（N），然后按下式计算杀灭对数值：杀灭对数值（KL）＝对照组平均活菌浓度的对数值（No）－实验组活菌浓度对数值（Nx）。各次的杀灭对数值均≥5.00，判定为消毒合格。 Add 0.5ml of bacterial suspension and 0.5ml of 0.3% bovine serum albumin into a sterile test tube, mix well, put it in a water bath at 20°C for 5 minutes, add 4ml of 1% aqueous solution of cinnamon essential oil (positive control is TPS), and mix immediately . After acting for a predetermined time, draw 0.5ml of the mixed solution of bacteria and medicine and add it to a 4.5ml neutralizer test tube. After mixing for 10 minutes, take 1.0ml of the solution for inoculation and culture, and count the viable bacteria after 48 hours of incubation at 37°C. Repeat the test 3 times, calculate the concentration of viable bacteria (cfu/mL) in each group, and convert it into a logarithmic value (N), and then calculate the logarithmic value of killing according to the following formula: logarithmic killing value (KL) = average viable bacteria in the control group The logarithmic value of the concentration (No) - the logarithmic value of the concentration of viable bacteria in the experimental group (Nx). The killing logarithm value of each time is ≥5.00, which is judged as qualified for disinfection.
2）试验结果 2) Test results
1%肉桂精油水溶液对大肠杆菌、金黄色葡萄球菌、白色葡萄球菌的最短杀灭时间为5min，对白色念珠菌的最短杀灭时间为10min。 The shortest killing time of 1% cinnamon essential oil aqueous solution to Escherichia coli, Staphylococcus aureus, and Staphylococcus albicans was 5 minutes, and the shortest killing time to Candida albicans was 10 minutes.
表1 肉桂精油水溶液杀菌效果 Table 1 Bactericidal effect of cinnamon essential oil aqueous solution
3.消毒剂的空气消毒实验 3. Air disinfection experiment of disinfectant
参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验，检测上述步骤1制备的消毒剂对室内空气的消毒效果。具体检测方法及结果如下。 Refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Methods in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications" to test the disinfection effect of the disinfectant prepared in the above step 1 on indoor air. The specific detection methods and results are as follows.
1）检测方法：参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验。 1) Detection method: refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Method in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications".
采样点的设置：选择40m2的房间，设置5个采样点，即室内墙角对角线交点为一采样点，该交点与四墙角连线的中点为另外4个采样点。采样点距墙1米，采样高度1.5米，避开空调、门窗等空气流通处。 Sampling point setting: select a room of 40m 2 and set 5 sampling points, that is, the intersection point of the diagonal lines of the indoor corners is a sampling point, and the midpoint between the intersection point and the line connecting the four corners is the other 4 sampling points. The sampling point is 1 meter away from the wall, and the sampling height is 1.5 meters, avoiding air circulation places such as air conditioners, doors and windows.
消毒前将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。 Before disinfection, place an ordinary agar plate with a diameter of 9 cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn over the plate, put the plate in a 37°C incubator for 48 hours, and count the colonies.
1%肉桂精油水溶液按照10ml/m3的用量，进行室内喷雾消毒。消毒作用至预定时间（30min，60min，180min）将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。菌落计数公式为：细菌数（cfu/m3）=50000N/AT（N为菌落数，A为平板面积，A为暴露时间）。 The 1% aqueous solution of cinnamon essential oil was used for indoor spray disinfection according to the dosage of 10ml/ m3 . Disinfection until the predetermined time (30min, 60min, 180min), place a common agar plate with a diameter of 9cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn the plate over, put the plate in a 37°C incubator for 48h, Perform a colony count. The formula for colony counting is: number of bacteria (cfu/m 3 )=50000N/AT (N is the number of colonies, A is the area of the plate, and A is the exposure time).
试验重复3次或以上，计算出每次的消亡率。每次的自然菌消亡率均≥90% 者为合格。。 The experiment was repeated 3 times or more, and the death rate of each time was calculated. Those with a natural bacterial death rate of ≥90% each time are qualified. .
2）实验结果 2) Experimental results
1%肉桂精油水溶液对室内空气消毒1小时后，自然菌的消亡率为93.2%，达到预定的消毒效果。 After the 1% aqueous solution of cinnamon essential oil disinfects the indoor air for 1 hour, the extinction rate of natural bacteria is 93.2%, reaching the predetermined disinfection effect.
表2 肉桂精油水溶液空气消毒效果 Table 2 Air disinfection effect of cinnamon essential oil aqueous solution
实施例2 空气消毒剂制备及其杀菌效果实验 Example 2 Preparation of air disinfectant and its bactericidal effect experiment
1.消毒剂制备：将3ml肉桂精油加入97ml蒸馏水中，摇匀，即得100ml消毒剂。此消毒剂无需加压，直接装入喷雾器中即可对室内空气进行消毒杀菌，操作简单，适用范围广。 1. Disinfectant preparation: Add 3ml of cinnamon essential oil to 97ml of distilled water and shake well to get 100ml of disinfectant. The disinfectant does not need to be pressurized, and can be directly loaded into the sprayer to disinfect and sterilize the indoor air. The operation is simple and the application range is wide.
2.消毒剂的杀菌效果实验 2. Bactericidal effect experiment of disinfectant
参照2008年版《消毒技术规范》规定的悬液定量杀菌试验，检测上述步骤1制备的消毒剂对大肠杆菌(8099) 、金黄色葡萄球菌(ATCC 6538) 、白色葡萄球菌（ATCC 8799）白色念珠菌(ATCC 10231)的杀菌效果，具体检测方法及结果如下。 With reference to the suspension quantitative bactericidal test stipulated in the 2008 version of "Disinfection Technical Specifications", detect the disinfectant prepared in the above step 1 against Escherichia coli (8099), Staphylococcus aureus (ATCC 6538), Staphylococcus albicans (ATCC 8799), Candida albicans (ATCC 10231), the specific detection method and results are as follows.
1）检测方法：参照2008年版《消毒技术规范》规定的悬液定量杀菌试验 1) Detection method: Refer to the suspension quantitative sterilization test stipulated in the 2008 edition of "Disinfection Technical Specifications"
取大肠杆菌(8099) 、金黄色葡萄球菌(ATCC 6538) 、白色葡萄球菌（ATCC 8799）白色念珠菌(ATCC 10231)24h新鲜斜面培养物，分别用胰蛋白胨生理盐水（TPS）洗下，混匀后稀释成含菌量为1×108cfu/ml～5×108cfu/ml的菌悬液，备用。 Take Escherichia coli (8099), Staphylococcus aureus (ATCC 6538), Staphylococcus albus (ATCC 8799) and Candida albicans (ATCC 10231) 24h fresh slant culture, wash with tryptone saline (TPS) respectively, and mix well Afterwards, it is diluted to a bacterial suspension with a bacterial content of 1×10 8 cfu/ml to 5×10 8 cfu/ml, and is set aside.
在无菌试管内加入0.5ml菌悬液和 0.3%小牛血清白蛋白0.5ml，混匀，置20℃水浴中5min后，加入3%肉桂精油水溶液(阳性对照为TPS)4ml，立即混匀。作用至预定时间，分别吸取0.5ml菌药混合液加于4.5ml中和剂试管中，混匀作用10min后，取该液1.0ml接种培养，37℃培养48小时后活菌计数。试验重复 3 次，计算各组的活菌浓度（cfu/mL），并换算为对数值（N），然后按下式计算杀灭对数值：杀灭对数值（KL）＝对照组平均活菌浓度的对数值（No）－实验组活菌浓度对数值（Nx）。各次的杀灭对数值均≥5.00，判定为消毒合格。 Add 0.5ml of bacterial suspension and 0.5ml of 0.3% bovine serum albumin into a sterile test tube, mix well, put it in a water bath at 20°C for 5 minutes, add 4ml of 3% aqueous solution of cinnamon essential oil (positive control is TPS), and mix immediately . After acting for a predetermined time, draw 0.5ml of the mixed solution of bacteria and medicine and add it to a 4.5ml neutralizer test tube. After mixing for 10 minutes, take 1.0ml of the solution for inoculation and culture, and count the viable bacteria after 48 hours of incubation at 37°C. Repeat the test 3 times, calculate the concentration of viable bacteria (cfu/mL) in each group, and convert it into a logarithmic value (N), and then calculate the logarithmic value of killing according to the following formula: logarithmic killing value (KL) = average viable bacteria in the control group The logarithmic value of the concentration (No) - the logarithmic value of the concentration of viable bacteria in the experimental group (Nx). The killing logarithm value of each time is ≥5.00, which is judged as qualified for disinfection.
2）试验结果 2) Test results
3%肉桂精油水溶液对大肠杆菌、金黄色葡萄球菌、白色葡萄球菌以及白色念珠菌的最短杀灭时间均为1min。 The shortest killing time of 3% cinnamon essential oil aqueous solution to Escherichia coli, Staphylococcus aureus, Staphylococcus albicans and Candida albicans was 1 min.
表3 肉桂精油水溶液杀菌效果 Table 3 Bactericidal effect of cinnamon essential oil aqueous solution
3.消毒剂的空气消毒实验 3. Air disinfection experiment of disinfectant
参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验，检测上述步骤1制备的消毒剂对室内空气的消毒效果。具体检测方法及结果如下。 Refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Methods in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications" to test the disinfection effect of the disinfectant prepared in the above step 1 on indoor air. The specific detection methods and results are as follows.
1）检测方法：参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验。 1) Detection method: refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Method in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications".
采样点的设置：选择40m2的房间，设置5个采样点，即室内墙角对角线交点为一采样点，该交点与四墙角连线的中点为另外4个采样点。采样点距墙1米，采样高度1.5米，避开空调、门窗等空气流通处。 Sampling point setting: select a room of 40m 2 and set 5 sampling points, that is, the intersection point of the diagonal lines of the indoor corners is a sampling point, and the midpoint between the intersection point and the line connecting the four corners is the other 4 sampling points. The sampling point is 1 meter away from the wall, and the sampling height is 1.5 meters, avoiding air circulation places such as air conditioners, doors and windows.
消毒前将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。 Before disinfection, place an ordinary agar plate with a diameter of 9 cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn over the plate, put the plate in a 37°C incubator for 48 hours, and count the colonies.
3%肉桂精油水溶液按照10ml/m3的用量，进行室内喷雾消毒。消毒作用至预定时间（30min，60min，180min）将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。菌落计数公式为：细菌数（cfu/m3）=50000N/AT（N为菌落数，A为平板面积，A为暴露时间）。 The 3% aqueous solution of cinnamon essential oil was used for indoor spray disinfection according to the dosage of 10ml/ m3 . Disinfection until the predetermined time (30min, 60min, 180min), place a common agar plate with a diameter of 9cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn the plate over, put the plate in a 37°C incubator for 48h, Perform a colony count. The formula for colony counting is: number of bacteria (cfu/m 3 )=50000N/AT (N is the number of colonies, A is the area of the plate, and A is the exposure time).
试验重复3次或以上。计算出每次的消亡率。每次的自然菌消亡率均≥90% 者为合格。。 The experiment was repeated 3 times or more. Calculate the death rate for each time. Those with a natural bacterial death rate of ≥90% each time are qualified. .
2）实验结果 2) Experimental results
3%肉桂精油水溶液对室内空气消毒30min后，自然菌的消亡率为97.2%，达到预定的消毒效果。 After the 3% aqueous solution of cinnamon essential oil disinfected the indoor air for 30 minutes, the extinction rate of natural bacteria was 97.2%, which reached the predetermined disinfection effect.
表4 肉桂精油水溶液空气消毒效果 Table 4 Air disinfection effect of cinnamon essential oil aqueous solution
对比例1空气消毒剂制备及其杀菌效果实验 Comparative example 1 air disinfectant preparation and its bactericidal effect experiment
1.消毒剂制备：将5ml肉桂精油加入95ml蒸馏水中，摇匀，即得100ml消毒剂。 1. Disinfectant preparation: add 5ml cinnamon essential oil to 95ml distilled water, shake well to get 100ml disinfectant.
2.消毒剂的空气消毒实验 2. Air disinfection experiment of disinfectant
参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验，检测上述步骤1制备的消毒剂对室内空气的消毒效果。具体检测方法及结果如下。 Refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Methods in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications" to test the disinfection effect of the disinfectant prepared in the above step 1 on indoor air. The specific detection methods and results are as follows.
1）检测方法：参照GB/T18204.1-2000《公共场所空气微生物检验方法-细菌总数测定》以及2008年版《消毒技术规范》规定的空气消毒试验。 1) Detection method: refer to the air disinfection test specified in GB/T18204.1-2000 "Air Microbiological Examination Method in Public Places-Determination of Total Bacteria" and the 2008 edition of "Disinfection Technical Specifications".
采样点的设置：选择40m2的房间，设置5个采样点，即室内墙角对角线交点为一采样点，该交点与四墙角连线的中点为另外4个采样点。采样点距墙1米，采样高度1.5米，避开空调、门窗等空气流通处。 Sampling point setting: select a room of 40m 2 and set 5 sampling points, that is, the intersection point of the diagonal lines of the indoor corners is a sampling point, and the midpoint between the intersection point and the line connecting the four corners is the other 4 sampling points. The sampling point is 1 meter away from the wall, and the sampling height is 1.5 meters, avoiding air circulation places such as air conditioners, doors and windows.
消毒前将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。 Before disinfection, place an ordinary agar plate with a diameter of 9 cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn over the plate, put the plate in a 37°C incubator for 48 hours, and count the colonies.
5%肉桂精油水溶液按照10ml/m3的用量，进行室内喷雾消毒。消毒作用至预定时间（30min，60min，180min）将直径9cm的普通琼脂平板置于采样点处，打开平皿盖暴露5min后，盖上皿盖，翻转平板，将平板至37℃培养箱内48h，进行菌落计数。菌落计数公式为：细菌数（cfu/m3）=50000N/AT（N为菌落数，A为平板面积，A为暴露时间）。 The 5% aqueous solution of cinnamon essential oil was used for indoor spray disinfection according to the dosage of 10ml/ m3 . Disinfection until the predetermined time (30min, 60min, 180min), place a common agar plate with a diameter of 9cm at the sampling point, open the plate cover and expose it for 5 minutes, cover the plate, turn the plate over, put the plate in a 37°C incubator for 48h, Perform a colony count. The formula for colony counting is: number of bacteria (cfu/m 3 )=50000N/AT (N is the number of colonies, A is the area of the plate, and A is the exposure time).
试验重复3次或以上。计算出每次的消亡率。每次的自然菌消亡率均≥90% 者为合格。。 The experiment was repeated 3 times or more. Calculate the death rate for each time. Those with a natural bacterial death rate of ≥90% each time are qualified. .
2）实验结果 2) Experimental results
5%肉桂精油水溶液对室内空气消毒30min后，自然菌的消亡率为95.9%，达到预定的消毒效果。 After the 5% aqueous solution of cinnamon essential oil disinfected the indoor air for 30 minutes, the extinction rate of natural bacteria was 95.9%, reaching the predetermined disinfection effect.
表5 肉桂精油水溶液空气消毒效果 Table 5 Air disinfection effect of cinnamon essential oil aqueous solution
一般认为，肉桂精油的浓度越高杀菌性能越好，但是从表2、表4和表5的结果可以看出，肉桂精油在1%、3%、5%浓度下，均能达到90%以上的消亡率，但是随着时间的延长，5%浓度的组合物细菌消亡率明显下降，在180min以后，已经下降至70%，表明肉桂精油的浓度过高时，空气中的持续消毒性能受到明显限制。发明人推断，肉桂精油的浓度与喷雾形成的液滴大小相关，当肉桂精油浓度过高时，形成的液滴尺寸相对较大，更容易发生沉降，从而导致其在空气中的持续消毒效果不佳。肉桂精油浓度在1~3%时，形成的液滴尺寸更小，更能持续地分散于空气中，在180min内仍能保持明显的消毒效果。 It is generally believed that the higher the concentration of cinnamon essential oil, the better the bactericidal performance, but from the results of Table 2, Table 4 and Table 5, it can be seen that cinnamon essential oil can reach more than 90% at the concentration of 1%, 3%, and 5%. However, as time goes on, the bacterium extinction rate of the 5% concentration composition drops significantly, and after 180 minutes, it has dropped to 70%, indicating that when the concentration of cinnamon essential oil is too high, the continuous disinfection performance in the air is significantly affected. limit. The inventor infers that the concentration of cinnamon essential oil is related to the size of the droplets formed by spraying. When the concentration of cinnamon essential oil is too high, the size of the formed droplets is relatively large, and sedimentation is more likely to occur, resulting in poor continuous disinfection effect in the air. good. When the concentration of cinnamon essential oil is 1-3%, the formed droplets are smaller in size, can be more continuously dispersed in the air, and can still maintain obvious disinfection effect within 180 minutes.
以上所述仅为本发明的较佳实施例，并不用于限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。 The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection of the present invention. within range.
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