CN101496773B - 一种含林蛙抗菌肽的抗菌洗剂及其制备方法 - Google Patents

一种含林蛙抗菌肽的抗菌洗剂及其制备方法 Download PDF

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CN101496773B
CN101496773B CN2009100665678A CN200910066567A CN101496773B CN 101496773 B CN101496773 B CN 101496773B CN 2009100665678 A CN2009100665678 A CN 2009100665678A CN 200910066567 A CN200910066567 A CN 200910066567A CN 101496773 B CN101496773 B CN 101496773B
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滕利荣
孟庆繁
逯家辉
安金双
李青山
周杰
田晓乐
高朝辉
王贞佐
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Tonghua Kang Yuan Biological Technology Co ltd
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Abstract

本发明涉及一种含林蛙抗菌肽的抗菌洗剂及其制备工艺,将林蛙抗菌肽溶于pH 5-7的磷酸盐缓冲液中,加入甘露醇保护剂制成淡黄色透明液体即得抗菌洗剂。本发明制得的洗剂具杀菌、消毒、止痒、止血收敛、清热杀虫等功能,对人体无毒无害,对皮肤粘膜无任何刺激,广泛适用于人体皮肤粘膜的卫生清洁,抗菌杀菌等领域,而且杀菌效果可以达到100%;此外,因加入了甘露醇保护剂,使其有效抑菌活性持续时间大大延长。

Description

一种含林蛙抗菌肽的抗菌洗剂及其制备方法
技术领域:
本发明涉及一种从两栖动物林蛙中得到的抗菌肽制成的产品,尤其涉及一种含林蛙抗菌肽的抗菌洗剂及其制备方法,属于洗涤用品制备技术领域。
背景技术;
目前市场上销售的抗菌洗剂产品,多为化学合成制品,因其含有化学成分,对人体易造成不同程度的刺激伤害等毒副作用。如醋酸氯已定,苯甲溴铵为主成分的抗菌洗剂,其抑菌效果在10分钟以上可达90%,无明显杀菌效果,而且具有极强的刺激性酸味,对皮肤粘膜一有刺激性。相比之下,市售的中药洗剂,100%  为混悬液,液体浑浊,瓶底明显有沉淀物;刺激味亦很明显。且以上几类产品的适用范围,适用于女性阴部卫生清洁,而适合用在全身各部位又能达到满意效果的产品目前还为之甚少。
从最初人们研究北美天蚕的免疫机制时发现了其血淋巴产生具有抑菌作用的多肽物质以来,相继从其他昆虫以及两栖类动物、哺乳动物中,也分离到结构相似的抗菌多肽。迄今为止,在不同动物组织中已发现了很多具有抗菌作用的蛋白质和多肽,已有70多种抗菌多肽的结构被测定,抗菌肽的概念得到了极大的扩展。然而,作为一种多肽类物质,其生物活性也易随时间的延长而丧失,因此,如何在较长时间使其维持在较高的活性水平是近年来研究多肽类物质的热点。
发明内容:
本发明的目的在于提供一种含林蛙抗菌肽的抗菌洗剂,具有杀菌、抗病毒、止痒、止血收敛、清热等功能,解决了现有抗菌剂刺激性强,杀菌效果不明显的问题。
本发明还一种含抗菌肽抗菌洗剂的制备方法,适用工业化生产。
本发明提供的含林蛙抗菌肽的抗菌洗剂,是由以下原料按重量份数比制成的:
林蛙抗菌肽0.0005-0.05wt%  甘露醇5-20wt%
磷酸盐缓冲液(0.25M pH5-7)75-95wt%、余量为水。
本发明林蛙抗菌洗剂的制备方法,包括如下步骤得到的:
(1)将林蛙干蛙皮,用4%的醋酸溶液以料液比为1∶10-15(m/v)的比例浸泡24h(室温)。
(2)将所得浸提液8000r.p.m离心15min,弃沉淀,留上清。
(3)将上清液煮沸5min,冷却,8000r.p.m离心15min,取上清过30KDa膜。
(4)将过30KDa膜溶液冻干,将冻干粉溶于0.25M pH 5-7的75-95wt%磷酸盐缓冲液中,加入5-20wt%多肽保护剂甘露醇制成淡黄色或棕黄色透明液体即得抗菌洗剂。
以下实验表明本发明的含林蛙抗菌肽的抗菌洗剂的抑菌活性:
(1)细菌培养使用LB肉汤(Luria broth)培养基。
(2)在无菌96孔板中每孔加入100μl液体,其中50μl为对数生长期的菌稀释至106 CFU ml-1,另外50μl为抗菌洗剂水溶液的系列2倍稀释液。
(3)37℃培养18-20h后,用酶标仪(Elx 800 universal microplate reader)读630nm的吸光度值。
(4)同一菌株的OD630 PB/OD630 B(PB:抗菌洗剂+菌液;B:菌液)表征抗菌洗剂对细菌增殖活性的影响。
表1本发明的抗菌洗剂的抑菌活性
Figure G2009100665678D00021
表2与其它消毒产品的比较
名称   pH值 杀菌时间/min 杀灭效果/% 透明度 稀释倍数 刺激性
实施例1   4.5  3 100 淡黄色透明 50
实施例2   5.4  3 100 淡黄色透明 50
实施例3   6.4   3   100   淡黄色透明   50   无
  妇康洁   5   未定   未定   灰黑色明显沉淀   5   轻度
  舒洁   7.0   未定   未定   黑灰色明显沉淀   5-10   轻度
  洁尔阴   4.5   未定   未定   黑灰色明显沉淀   10-20   轻度
表3添加了多肽保护剂的抗菌洗剂与未加多肽保护剂的抗菌洗剂抑菌时间比较
Figure G2009100665678D00031
注:I:添加了多肽保护剂的抗菌洗剂;II:未加多肽保护剂的抗菌洗剂
本发明的抗菌洗剂pH在4.5-6.5之间,对人体无毒无害,对皮肤粘膜无任何刺激。对人体皮肤及皮肤粘膜的抗菌杀菌效果显著,可以预防各种感染和传染。长期使用既可以达到杀菌清洁的作用,又可以使皮肤柔嫩而光滑。且婴幼儿、孕妇使用安全,对环境无污染。
本发明的抗菌洗剂制备过程中,加入了多肽保护剂,使此抗菌洗剂的抑菌活性时间大大延长;此外采用的冻干工艺,既可以消除醋酸的刺激味,又使制成洗剂的操作工艺大大简化,免去了除去其他水溶性杂质的步骤,适用于工业化生产。
具体实施方案:
通过以下实施例进一步举例描述本发明,并不以任何方式限制本发明,在不背离本发明的技术解决方案的前提下,对本发明所作的本领域普通技术人员容易实现的任何改动或改变都将落入本发明的权利要求范围之内。
实施例1
(1)将林蛙干蛙皮120g,用4%的醋酸溶液1300ml以料液比为1∶10-15(m/v)的比例浸泡24h(室温)。
(2)将所得浸提液8000r.p.m离心15min,弃沉淀,留上清。
(3)将上清液煮沸5min,冷却,8000r.p.m离心15min,取上清过30KDa膜。
(4)将过30KDa膜溶液冻干,将0.0005wt%冻干粉溶于0.25M pH 5.5的磷酸盐缓冲液1800mL中,加入5wt%甘露醇90mL制成淡黄色或棕黄色透明液体即得抗菌洗剂。
实施例2
(1)将林蛙干蛙皮200g,用4%的醋酸溶液2500ml以料液比为1∶10-15(m/v)的比例浸泡24h(室温)。
(2)将所得浸提液8000r.p.m离心15min,弃沉淀,留上清。
(3)将上清液煮沸5min,冷却,8000r.p.m离心15min,取上清过30KDa膜。
(4)将过30KDa膜溶液冻干,将0.005wt%冻干粉溶于0.25M pH6.0的磷酸盐缓冲液4200mL中,加入10wt%甘露醇600mL制成淡黄色或棕黄色透明液体即得抗菌洗剂。
实施例3
(1)将林蛙干蛙皮320g,用4%的醋酸溶液4500ml以料液比为1∶10-15(m/v)的比例浸泡24h(室温)。
(2)将所得浸提液8000r.p.m离心15min,弃沉淀,留上清。
(3)将上清液煮沸5min,冷却,8000r.p.m离心15min,取上清过30KDa膜。
(4)将过30KDa膜溶液冻干,将0.05wt%冻干粉溶于0.25M pH6.5的磷酸盐缓冲液7500mL中,加入20wt%甘露醇1500mL制成淡黄色或棕黄色透明液体即得抗菌洗剂。

Claims (2)

1.一种含林蛙抗菌肽的抗菌洗剂,是由以下原料按重量份数比制成的:
林蛙抗菌肽0.0005-0.05wt%、甘露醇5-20wt%、
0.25M pH 5-7的磷酸盐缓冲液75-95wt%,余量为水。
2.根据权利要求1所述的抗菌洗剂的制备工艺,如下步骤制得的:
将林蛙抗菌肽溶于0.25M pH 5-7的磷酸盐缓冲液中,加入甘露醇保护剂制成淡黄色透明液体即得。
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Denomination of invention: An Antibacterial Lotion Containing Forest Frog Antibacterial Peptide and Its Preparation Method

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