CN108633882B - 一种结核分枝杆菌消毒剂 - Google Patents
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
本发明公开了一种结核分枝杆菌消毒剂。本发明探讨LVA‑SDS对结核分枝杆菌标准株H37Rv和耐多药结核分枝杆菌临床分离株的杀灭作用。并通过相关动物实验对LVA‑SDS进行安全性毒理学试验,从而评价其在实际应用时的安全性。LVA‑SDS是一种安全的,有效的结核分枝杆菌消毒剂,能够有效的杀灭结核分枝杆菌,能够应用于切断结核分枝杆菌的空气传播,从而降低肺结核,这种严重危害人类健康的呼吸道传染性疾病的传播,同时还具有很好的安全性、无腐蚀性、无害无污染、无刺激性、无副作用等优点。
Description
技术领域
本发明涉及一种安全的,有效的结核分枝杆菌消毒剂。
背景技术
结核分枝杆菌是引起结核病的病原菌,严重威胁着人类的健康,切断结核分枝杆菌传播途径是控制结核病传播的重要措施之一。目前常用的结核分枝杆菌消毒剂有:含碘类、含氯类、含醛类、过氧化物类等。这些消毒剂对结核分枝杆菌都有一定的杀灭作用但也均存在不同的副作用。如含碘类消毒剂会导致过敏,甚至会引起甲状腺疾病等严重后果。含氯类消毒剂对人体皮肤黏膜造成损害,如高浓度的84消毒液(次氯酸钠)可致使皮肤发白、水肿发炎、水疱,致使眼部疼痛、流泪、溃疡等;84消毒液对操作台面和金属器械存在腐蚀性。含醛类有强刺激性、毒性和腐蚀性,甚至能引起过敏和哮喘。过氧化物类消毒剂易分解、刺激性强,可引起DNA损伤、基因突变、致癌等。因此,迫切需要更为安全有效的消毒剂切断结核分枝杆菌的扩散传播。
乙酰丙酸-十二烷基硫酸钠(levulinic acid,sodium dodecyl sulfate,LVA-SDS)是一种广谱的,杀灭作用较强的新型消毒剂。LVA主要生产原料是含纤维素的可再生原料(比如高粱等),因此不仅成本低,产量高,而且无害无污染。经过美国食品和药物管理局(U.S.Food and Drug Administration,FDA)认证和批准,LVA可以加入到化妆品、调味剂、烟草等食品或者日常用品中,具有很好的安全性。SDS也是FDA认证的多功能添加剂,是一种活性添加剂,已经广泛用于日常生活中,比如添加到蛋清、果汁、植物油和明胶中等,其安全性也得到了认可。
发明内容
本发明的目的是提供一种安全的,有效的结核分枝杆菌消毒剂。
本发明提供一种消毒剂,其有效成分为乙酰丙酸和十二烷基硫酸钠。
所述消毒剂中含有体积百分含量为10-30%的乙酰丙酸和质量百分含量为1.0-3.0%的十二烷基硫酸钠。
所述消毒剂中具体可含有体积百分含量为15%的乙酰丙酸和质量百分含量为1.5%的十二烷基硫酸钠。
所述消毒剂中具体可含有体积百分含量为10%的乙酰丙酸和质量百分含量为1.0%的十二烷基硫酸钠。
所述消毒剂由体积百分含量为10-30%的乙酰丙酸、质量百分含量为1.0-3.0%的十二烷基硫酸钠和溶剂组成。
所述消毒剂具体可由体积百分含量为20%的乙酰丙酸、质量百分含量为2.0%的十二烷基硫酸钠和溶剂组成。
所述消毒剂具体可由体积百分含量为15%的乙酰丙酸、质量百分含量为1.5%的十二烷基硫酸钠和溶剂组成。
所述消毒剂具体可由体积百分含量为10%的乙酰丙酸、质量百分含量为1.0%的十二烷基硫酸钠和溶剂组成。
以上任一所述溶剂具体可为生理盐水。
以上任一所述消毒剂的用途为杀灭细菌。
本发明保护以上任一所述的消毒剂在杀灭细菌中的应用。
以上任一所述消毒剂可以用于医疗卫生用品、皮肤、粘膜、餐饮具、水、空气、化妆品、调味剂、烟草等食品或者日常用品的消毒、灭菌。
本发明还保护一种产品,其活性成分为以上任一所述的消毒剂;所述产品的用途为杀灭细菌。
以上任一所述细菌为结核分枝杆菌。
以上任一所述结核分枝杆菌具体可为结核分枝杆菌标准株H37Rv(ATCC,编号:27294)或耐多药结核分枝杆菌临床分离株。
本发明探讨LVA-SDS对结核分枝杆菌标准株H37Rv和耐多药结核分枝杆菌临床分离株的杀灭作用。并通过相关动物实验对LVA-SDS进行安全性毒理学试验,从而评价其在实际应用时的安全性。LVA-SDS是一种安全的,有效的结核分枝杆菌消毒剂,能够有效的杀灭结核分枝杆菌,能够应用于切断结核分枝杆菌的空气传播,从而降低肺结核,这种严重危害人类健康的呼吸道传染性疾病的传播,同时还具有很好的安全性、无腐蚀性、无害无污染、无刺激性、无副作用等优点。
具体实施方式
以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为自常规生化试剂商店购买得到的。以下实施例中的定量试验,均设置三次重复实验,结果取平均值。
结核分枝杆菌标准株H37Rv:ATCC,编号:27294。
耐多药结核分枝杆菌临床分离株:同时对利福平、异烟肼耐药,临床药敏编号:2202,来自首都医科大学附属北京胸科医院国家结核病临床实验室菌株库。
LVA(乙酰丙酸):Acros Organics,Fair Lawn,NJ,纯度98%,密度1.134g/ml。
SDS(十二烷基硫酸钠):北京鼎国生物技术有限责任公司。
实施例1、LVA-SDS消毒剂及其效果评价
供试菌株:结核分枝杆菌标准株H37Rv和耐多药结核分枝杆菌临床分离株。
一、结核分枝杆菌菌悬液制备
在无菌磨菌瓶中加入1~2滴10%(体积百分含量)吐温-80水溶液,用无菌吸管尖端或无菌接种环刮取2~3周改良罗氏培养基(珠海市银科医学工程股份有限公司)处于对数生长期的新鲜菌落,置于磨菌瓶中,之后旋紧瓶盖在涡旋震荡器上震荡30~60s。静置10min后,打开瓶盖,加入约2ml无菌生理盐水,轻轻晃动后静置片刻,使菌液中的大块物质沉淀。用无菌吸管吸取中上部的菌液约1ml,转移至另一无菌试管中,与麦氏比浊管比浊。逐渐滴加无菌生理盐水,直至菌液浊度与1号标准麦氏比浊管一致,即得到菌含量为1mg/ml的菌悬液,备用。
二、LVA-SDS消毒剂及其效果评价
在无菌试管内加入0.5ml步骤一制备的菌悬液和0.5ml含有3%(体积百分含量)牛血清白蛋白(BSA)有机干扰物的水溶液),置于20℃±1℃水浴中,加入4ml待测消毒剂(生理盐水作为阳性对照组),作用不同时间(5min、15min或30min)后,取0.5ml混合液,加入4.5ml中和剂,混合中和作用10min。然后取0.1ml混合液涂布于7H10固体培养基(19g 7H10粉末(BD公司),5ml甘油,去离子水定容至1L),37℃恒温箱中培养1个月,观察结果并记录菌落总数(CFU/ml),计算平均杀灭对数值(N)。
所有试验均重复3次。按照下列公式计算平均杀灭对数值:
平均杀灭对数值(N)=对照组平均活菌浓度的对数值(No)-试验组活菌浓度的对数值(Nx)。
待测消毒剂A:含有20%(体积百分含量)LVA的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂B:含有15%(体积百分含量)LVA的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂C:含有10%(体积百分含量)LVA的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂D:含有5%(体积百分含量)LVA的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂E:含有2%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂F:含有1.5%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂G:含有1.0%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂H:含有0.5%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1MPBS溶液。
待测消毒剂I:含有20%(体积百分含量)LVA和2%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1M PBS溶液。
待测消毒剂J:含有15%(体积百分含量)LVA和1.5%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1M PBS溶液。
待测消毒剂K:含有10%(体积百分含量)LVA和1.0%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1M PBS溶液。
待测消毒剂L:含有5%(体积百分含量)LVA和0.5%(质量百分含量)SDS的生理盐水,其对应的中和剂为0.1M PBS溶液。
待测消毒剂M:有效氯含量为1000mg/L的次氯酸钠(北京洗得宝消毒制品有限公司),商用名“84消毒液”。其对应的中和剂为含有5g/L硫代硫酸钠、3g/L卵磷脂、10g/L吐温-80的PBS溶液。
结果如表1和表2所示。
表1待测消毒剂在不同浓度、时间条件下对结核分枝杆菌标准株H37Rv的杀灭作用
表2待测消毒剂在不同浓度、时间条件下对临床分离株的杀灭作用
上述结果表明,20%LVA,20%LVA+2%SDS作用30min,对菌悬液内结核分枝杆菌标准株H37Rv、耐多药结核分枝杆菌临床分离株的杀灭对数值分别为5.03、5.53、5.03和5.12(表1、2)。20%LVA+2%SDS的杀灭作用略强于20%LVA。含有效氯1000mg/L的“84消毒液”作用30min,对菌悬液内结核分枝杆菌标准株H37Rv、耐多药结核分枝杆菌临床分离株的杀灭对数值为6.20和6.17。含有效氯1000mg/L的“84消毒液”对结核分枝杆菌的杀灭作用虽然略强于20%LVA,20%LVA+2%SDS,但LVA+2%SDS也可以杀大部分结核分枝杆菌,如果提高消毒剂中LVA和SDS的浓度或延长作用时间,可达到完全杀灭结核分枝杆菌的效果。
实施例2、LVA-SDS消毒剂安全性评价
受试液:含有20%(体积百分含量)LVA和2%(质量百分含量)SDS的生理盐水。
实验动物:SPF级BALB/c小鼠(北京维通利华实验动物技术有限公司),体重18~20g。
一、急性经口毒性试验
将实验动物随机分为两组(每组6只,雌雄各半)。
试验前小鼠隔夜禁食12h,不禁水。
采用一次最大限度试验法。参照2002年版《消毒技术规范》,采用灌胃方式,按照0.4ml/20(g·bw)给予受试液,用灌胃针头将受试液经口一次给予动物,连续观察14天,观察并记录动物中毒的表现和死亡情况,计算LD50。试验结束后,处死存活动物,进行解剖,肉眼观察是否出现组织和脏器异常。
结果为LD50>5000mg/(kg·bw),参照《消毒技术规范》中的毒性评价可知,该消毒剂属实际无毒级。试验期间实验动物未出现异常,无死亡。实验结束后,解剖处死的实验动物,肉眼观察未见任何组织或脏器异常。
二、蓄积毒性试验
将实验动物随机分为两组(每组6只,雌雄各半)。
试验前小鼠隔夜禁食12h,不禁水。
采用灌胃方式,每天灌胃一次,按照0.4ml/20(g·bw)计算染毒量,每4天为一期。第一期剂量为0.1LD50,以后每期的剂量按照《消毒技术规范》中定期递增染毒剂量用表中所示递增,直至半数动物死亡为止,计算累积总剂量即为LD50(n)。如果给药28天,死亡动物仍不足半数,即停止实验。蓄积系数(K)公式:K=LD50(n)/LD50,计算并确定蓄积毒性分级。
试验连续进行28天,小鼠均无死亡,蓄积系数K>5,参照《消毒技术规范》中的蓄积毒性分级可知,该消毒剂为轻度蓄积毒性。
三、皮肤刺激试验
随机选择6只实验动物,雌雄各半。
试验前24h观察小鼠耳缘皮肤是否完好,无破损,次日将受试液直接涂抹于一侧完整的皮肤上,另一侧作为空白对照。分别于1h、24h、48h后观察皮肤的局部反应,根据有无红斑和水肿形成,进行皮肤反应刺激评分。
观察1h、24h、48h后,结果显示实验动物皮肤刺激反应无红斑和水肿形成,皮肤刺激积分均为0,皮肤刺激指数<0.5,参照《消毒技术规范》中的皮肤刺激评价标准可知,对小鼠皮肤无刺激性。
上述结果表明,LVA-SDS消毒剂对结核分枝杆菌除了有很好的杀灭作用,还具有很好的安全性、无腐蚀性、无害无污染、无刺激性、无副作用等优点。LVA-SDS混合液在常温下保存60天性质稳定且杀灭活性不变。
现有消毒剂例如84消毒剂对人体皮肤黏膜造成损害,如高浓度的84消毒液(次氯酸钠)可致使皮肤发白、水肿发炎、水疱,致使眼部疼痛、流泪、溃疡等;84消毒液对操作台面和金属器械存在腐蚀性。此外,84消毒液的稳定性较差,储存期一般只有3~6月。次氯酸钠的含氯消毒剂易产生致畸、致变、致癌的三氯甲烷,其安全性被世界卫生组织(WHO)列为A1级。
因此,LVA-SDS消毒剂作为一种安全的,有效的结核分枝杆菌消毒剂,能够有效的杀灭结核分枝杆菌,能够应用于切断结核分枝杆菌的空气传播,从而降低肺结核,这种严重危害人类健康的呼吸道传染性疾病的传播。
Claims (3)
1.一种消毒剂在杀灭结核分枝杆菌中的应用,所述消毒剂有效成分为乙酰丙酸和十二烷基硫酸钠;所述消毒剂中含有体积百分含量为20%的乙酰丙酸和质量百分含量为2.0%的十二烷基硫酸钠。
2.如权利要求1所述的应用,其特征在于:所述消毒剂由体积百分含量为20%的乙酰丙酸、质量百分含量为2.0%的十二烷基硫酸钠和溶剂组成。
3.如权利要求2所述的应用,其特征在于:所述溶剂为生理盐水。
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