CN116726119A - 草本抑菌液及其应用 - Google Patents
草本抑菌液及其应用 Download PDFInfo
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- CN116726119A CN116726119A CN202310708114.0A CN202310708114A CN116726119A CN 116726119 A CN116726119 A CN 116726119A CN 202310708114 A CN202310708114 A CN 202310708114A CN 116726119 A CN116726119 A CN 116726119A
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Abstract
本发明公开了一种草本抑菌液及其应用,包括以下原料:百部、苦参、川穹、丹参、龙胆草、蒲公英、薄荷油、天然冰片、醋酸氯己定、稳定剂。与现有技术相比,本发明制备的草本抑菌液具有较好的抑菌效果,杀菌范围大,而且稳定性强。
Description
技术领域
本发明涉及抑菌液技术领域,尤其涉及一种草本抑菌液及其应用。
背景技术
草本抑菌液是一种利用天然植物提取物制成的抗菌剂,草本抑菌液的制备离不开植物提取技术。传统的植物提取技术包括水提法、乙醇提法、超临界流体萃取法等,现代技术则包括微波辅助提取法、超声波辅助提取法、离子交换法等。这些技术能够将植物中的有效成分提取出来,制备出高纯度的草本提取物。
虽然草本抑菌液具有广泛的应用前景,但也存在一些问题,抗菌效果不稳定:草本抑菌液的抗菌效果往往受到环境因素的影响,如PH值、温度、光照等。这些因素的变化会导致草本抑菌液的抗菌效果不稳定,难以保证其应用效果。应用场景有限:草本抑菌液的应用场景主要集中在食品、医药、化妆品等领域。在其他领域,如建筑材料、纺织品等领域的应用还需要进一步研究和探索。
发明专利申请CN113144121A公开了一种草本抑菌液及其制备方法,其制备方法步骤如下:S1:将所有的待用的中草药洗净晾干;S2:将高温下成分易破环的草药切成碎片混合均匀制成药包,放于医用酒精中进行浸泡1~2月,得到中草药的酒精溶液;S3:将耐受高温的草药切成碎片放入纯净水中进行熬煮,熬煮时间为3~4小时,得到中药液;S4:将上述中药液趁热进行过滤,除去药渣,然后冷却至室温待用;S5:在过滤冷却后的中药液加入中草药的酒精溶液,搅拌混合,然后加入蒸馏水进行稀释混合,即得到抑菌液;S6:将得到的抑菌液进行装瓶,密封存放,不会妨碍皮肤的正常呼吸代谢,能够杀灭真菌及微生物,疗效快,药物利用度高。但是该发明制备的草本抑菌液存在杀菌范围受限、储存稳定性差的问题。
发明内容
有鉴于现有技术中草本抑菌液杀菌范围小、储存稳定性差的缺点,本发明所要解决的技术问题是提供一种杀菌范围大、储存稳定性好的草本抑菌液及其应用。
为了实现上述发明目的,本发明采用了如下的技术方案:
一种草本抑菌液,包括以下组分:百部、苦参、川穹、丹参、龙胆草、蒲公英、薄荷油、天然冰片、醋酸氯己定、稳定剂。
一种草本抑菌液,包括以下重量份组分:百部2~4份、苦参6~9份、川穹3~5份、丹参3~5份、龙胆草1~3份、蒲公英3~5份、薄荷油1~3份、天然冰片0.2~0.8份、醋酸氯己定0.001~0.03份、稳定剂2~5份。
所述稳定剂的制备方法如下:
S1、将β-环状糊精加入到水中,然后加入聚乙二醇,搅拌,冷却,得到乳剂;
S2、将步骤S1制备的乳剂、促进剂和紫苏籽油加入到水中,搅拌,冷却,加入吐温-20和酪蛋白酸钠,搅拌,得到悬浮液;将悬浮液冷却至室温,搅拌;再将悬浮液重新加热,将柠檬香精滴入悬浮液中,搅拌,再均质,得到稳定剂。
优选的,所述稳定剂的制备方法如下,以重量份计:
S1、将1.5~2份β-环状糊精加入到60~90℃的30~50份水中,然后加入3~5份聚乙二醇,100~500rpm搅拌20~40min,20~30℃自然冷却10~30h,得到乳剂;
S2、将步骤S1制备的乳剂、0.2~0.6份促进剂和0.3~0.7份紫苏籽油加入到60~80℃的80~120份水中,100~500rpm搅拌1~5h,40~50℃冷却1~3h,加入0.04~0.06份吐温-20和0.2~0.6份酪蛋白酸钠,100~500rpm搅拌1~5h,得到悬浮液;将悬浮液冷却至室温,以50~200rpm搅拌1~5h;再将悬浮液重新加热至40~50℃,将0.4~0.6份柠檬香精以0.3~0.5L/min的速度滴入悬浮液中,100~500rpm搅拌5~15min,再以8000~15000rpm的速度均质5~10min,得到稳定剂。
所述促进剂的制备方法如下:
将脱水蒜粉和百里香精油依次加入到醋酸水溶液中,室温搅拌,得到草药溶液;然后在草药溶液中加入羧甲基壳聚糖和甘油,室温搅拌,超声,得到促进剂。
优选的,所述促进剂的制备方法如下,以重量份计:
将0.02~0.06份脱水蒜粉和0.02~0.03份百里香精油依次加入到8~12份体积分数0.5~2%的醋酸水溶液中,室温下100~800rpm搅拌0.5~2h,得到草药溶液;然后在草药溶液中加入0.02~0.03份羧甲基壳聚糖和0.004~0.008份甘油,室温下100~500rpm搅拌5~15h,超声5~15min、超声功率为100~400W、超声频率为40~60kHz,得到促进剂。
所述草本抑菌液的制备方法如下:
步骤1、按重量份称取各原料,将百部、苦参、川穹、丹参、龙胆草、蒲公英分别除去杂质,洗净后干燥;
步骤2、将步骤1干燥后的川穹和百部切碎,过筛,加入到乙醇水溶液中进行浸泡后过滤,得到浸泡溶液;
步骤3、将苦参、丹参、龙胆草、蒲公英切碎研磨,过筛,然后加入到乙醇水溶液中,加热搅拌,得到醇提液,将醇提液加入到步骤2制备的浸泡溶液中,搅拌过滤,得到混合滤液;然后加入薄荷油、天然冰片、醋酸氯己定和稳定剂,搅拌,再加入水调整含乙醇浓度,即得草本抑菌液。
优选的,所述草本抑菌液的制备方法如下:
步骤1、按重量份称取各原料,将百部、苦参、川穹、丹参、龙胆草、蒲公英分别除去杂质,洗净后干燥;
步骤2、将步骤1干燥后的川穹和百部切碎,过20~50目筛,加入到质量比为3~5倍的70~85wt%乙醇水溶液中进行浸泡20~40d后300~500目筛过滤,得到浸泡溶液;
步骤3、将苦参、丹参、龙胆草、蒲公英切碎研磨,过50~200目筛,然后加入到质量比为4~6倍的40~60wt%乙醇水溶液中,加热到60~75℃、50~200rpm搅拌3~8h,得到醇提液,将醇提液加入到步骤2制备的浸泡溶液中,100~500rpm搅拌2~6h,100~600目筛过滤,得到混合滤液;然后加入薄荷油、天然冰片、醋酸氯己定和稳定剂,100~500rpm搅拌20~40min,再加入水调整含乙醇浓度为30~50wt%,即得草本抑菌液。
所述草本抑菌液在制备中药穴位贴中的应用。
天然冰片是一种中药材,由樟科植物冰片树的蒸馏油经过冷却、凝固、结晶等过程制成,是一种无色透明的晶体。具有清热解毒、祛风除湿、止痛镇痛等作用。主要成分是萜类化合物,包括蒎烷、松油萜、桉叶萜等。其中,蒎烷是天然冰片的主要活性成分,具有镇痛、止痛、抗炎、抗菌等作用。
丹参,又称红丹、赤丹、地丹,是一种常见的中药材,属于唇形科植物丹参的根和根茎。丹参具有活血化瘀、消肿止痛、降血脂、降血压等作用。其主要药用部位为根和根茎,常用于治疗心脑血管疾病、痛经、跌打损伤等疾病,具有显著的疗效。含有多种活性成分,包括丹参酮、丹参素、丹酚酸、黄酮类、三萜类等。其中,丹参酮和丹参素是其主要活性成分,具有活血化瘀、降血脂、抗氧化等作用。
川穹,又称天麻,是一种常见的中药材,属于兰科植物天麻的地下茎。川穹具有镇痛、镇静、安神、降压等作用。其主要药用部位为地下茎,常用于治疗头痛、失眠、焦虑症、高血压等疾病,具有显著的疗效。含有多种活性成分,包括天麻素、天麻酯、天麻醇等。其中,天麻素是其主要活性成分,具有镇痛、镇静、抗癫痫等作用。
百部是一种中药材,也称为百部根,是一种多年生草本植物,属于菊科植物。百部具有清热解毒、消肿止痛、祛痰化瘀等作用。其主要药用部位为根,常用于治疗肺热、咳嗽、咽喉肿痛、痈肿疮毒等疾病,具有显著的疗效。含有多种活性成分,包括黄酮类、苷类、挥发油等。其中,百部苷是其主要活性成分,具有清热解毒、消肿止痛等作用。
苦参是一种中药材,也称为黄苦参、苦柏、苦木、苦艽等,是一种常见的多年生草本植物,属于豆科植物。苦参具有清热解毒、消肿止痛、抗菌消炎等作用。其主要药用部位为全草、种子,常用于治疗痈肿疮毒、湿疹、痤疮等疾病,具有显著的疗效。含有多种活性成分,包括黄酮类、生物碱、苦参碱等。其中,苦参碱是其主要活性成分,具有抗菌消炎、抑制炎症反应等作用。
醋酸氯己定一般指醋酸洗必泰,是一种有机物,为白色或几乎白色的结晶粉末,无臭,味苦。在乙醇中溶解,在水中微溶。水溶液是用于医治霉菌性阴道炎及其宫颈糜烂的药品,具有除菌消毒的作用。可以用于穴位贴,是一种贴在穴位上的贴片,可以通过经皮吸收的方式缓解症状。
龙胆草是一种常见的草本植物,属于龙胆科。主要用于清热解毒、利胆退黄、消炎止痛等方面。其主要药用部位为地上部分,包括叶、茎、花等,常用制成龙胆泻肝汤、龙胆清肝丸等中药方剂。龙胆草含有许多具有药用价值的化学成分,主要包括龙胆苦苷、苦参碱、绿原酸、芦丁、黄酮等。其中,龙胆苦苷是龙胆草的主要活性成分,具有明显的清热解毒、利胆退黄、抗菌消炎等作用。
蒲公英是一种常见的草本植物,属于菊科,也称为蒲公英草。在中医药中被认为具有清热解毒、利尿消肿、降血压等作用。其主要药用部位为全草,包括根、茎、叶等,常用于治疗肝炎、黄疸、水肿等疾病,具有显著的疗效。蒲公英含有多种活性成分,包括挥发油、黄酮类、苦味素类、黄酮苷类、有机酸和维生素等。其中,蒲公英苦素和蒲公英黄酮是其主要活性成分,具有利尿、消炎、解热、降血压等作用。
本发明通过科学的原料和配比将百部、苦参、川穹、丹参、龙胆草、蒲公英进行提取,然后与薄荷油、天然冰片、醋酸氯己定和稳定剂复配制备成草本抑菌液;其中所述稳定剂是将β-环状糊精加入热水中与聚乙二醇复配,得到乳剂;将乳剂、促进剂和紫苏籽油加入到热水中,搅拌冷却,加入吐温-20和酪蛋白酸钠搅拌,再滴入柠檬香精搅拌,再均质得到;所述促进剂是脱水蒜粉和百里香精油与羧甲基壳聚糖和甘油复合得到。
在脱水蒜粉、百里香精油的成分中有相当多的O-H键和C-O键。同时,壳聚糖分子在乙酰化单体上具有质子给体原子和质子受体原子。可能在壳聚糖分子和脱水蒜粉、百里香精油之间形成分子间氢键。与纯壳聚糖膜相比,经脱水蒜粉、百里香精油处理的壳聚糖基膜的碳元素含量更高。此外,由于脱水蒜粉中含硫化合物的存在,使得壳聚糖基膜中硫元素的含量增加,可以提高壳聚糖的表面粗糙度。由于草本抑菌液中功能成分的水溶性差、不耐温、易降解和在光照下不稳定,降低了储存时间。在羧甲基壳聚糖中通过与脱水蒜粉、百里香精油和甘油引入疏水段,成功制备了两亲性羧甲基壳聚糖,可以增加疏水基团之间的疏水相互作用,从而形成更致密的疏水核,增强乳化能力。壳聚糖的抗菌活性是接触型的,抗菌活性是因为壳聚糖的游离氨基(正电荷)通过静电相互作用与细菌的细胞表面(负电荷)结合,通过破坏细胞膜和细胞内成分的渗漏导致细菌细胞死亡。经脱水蒜粉、百里香精油处理具有更高的抗菌能力,这可能是由于壳聚糖和脱水蒜粉、百里香精油的协同作用。百里香精油中的香芹酚和百里香酚,以及脱水蒜粉中的大蒜素。百里香酚和香芹酚是疏水酚类成分,它们会对细菌细胞膜产生理化变化,增加细胞膜的通透性,从而导致细菌死亡。此外,香芹酚可诱导细菌细胞中ATP的渗漏和损失。
β-环状糊精分子首先与吐温-20组装成超分子复合物,然后通过氢键进一步与功能剂组装成含有β-环状糊精的两亲性颗粒。在油水界面形成薄膜。该膜可以通过物理分割强有力地分离油相和水相,并增加位阻来阻碍聚合。紫苏籽油有促进乳化和形成交联网络的能力,聚乙二醇的长链穿过β-环状糊精的空腔,在链的两端形成纳米级结构域,进一步形成网状结构,网状结构阻碍了液滴的聚集变质,更有利于草本抑菌液的稳定性,长期储存或加热处理后都是稳定的。
与现有技术相比,本发明的有益效果:
1)本发明通过将百部、苦参、川穹、丹参、龙胆草、蒲公英进行提取,然后与薄荷油、天然冰片、醋酸氯己定和稳定剂复配制备成草本抑菌液,具有较好的抑菌效果,杀菌范围大,而且稳定性强。
2)本发明所述稳定剂是将β-环状糊精加入热水中与聚乙二醇复配,得到乳剂;将乳剂、促进剂和紫苏籽油加入到热水中,搅拌冷却,加入吐温-20和酪蛋白酸钠搅拌,再滴入柠檬香精搅拌,再均质得到;稳定剂能形成网状结构,网状结构阻碍了液滴的聚集变质,有利于草本抑菌液的稳定性。
具体实施方式
主要物质来源:
百部:Stemona japonica,产于湖南常德。
苦参:Sophora flavescens,产于山西长治。
丹参:Salvia miltiorrhiza Bunge,产于四川省德阳市中江县。
川芎:Ligusticum chuanxiong hort,产于四川都江堰市。
龙胆草:Gentiana scabra Bunge,产于辽宁省清原满族自治县。
蒲公英:Taraxacum mongolicum,产于河北邯郸磁县。
薄荷油:江西亿森源植物香料有限公司,型号:DF。
天然冰片:惠州鸿泰生物科技有限公司,货号:56-11。
脱水蒜粉:兴化市味承食品有限公司,货号:02。
百里香精油:江西泰诚天然香料有限公司,货号:TC2018。
紫苏籽油:江西纤络化妆品有限公司,型号:df。
柠檬香精:杭州百瑞香精香料有限公司,型号:BR1418。
聚乙二醇:上海荟隽化工有限公司,型号:PEG-20000。
羧甲基壳聚糖:西安超邦生物科技有限公司,型号:CB。
实施例1
一种草本抑菌液的制备方法如下:
步骤1、将3kg百部、7kg苦参、4kg川穹、4kg丹参、2kg龙胆草、4kg蒲公英分别除去杂质,洗净后干燥;
步骤2、将步骤1干燥后的川穹和百部切碎,过30目筛,加入到质量比为4倍的75wt%乙醇水溶液中进行浸泡30d后400目筛过滤,得到浸泡溶液;
步骤3、将苦参、丹参、龙胆草、蒲公英切碎研磨,过100目筛,然后加入到质量比为5倍的45wt%乙醇水溶液中,加热到70℃、100rpm搅拌5h,得到醇提液,将醇提液加入到步骤2制备的浸泡溶液中,400rpm搅拌4h,500目筛过滤,得到混合滤液;然后加入2kg薄荷油、0.5kg天然冰片、0.01kg醋酸氯己定和4kg稳定剂,200rpm搅拌30min,再加入水调整含乙醇浓度为40wt%,即得草本抑菌液。
所述稳定剂的制备方法如下:
S1、将1.8kgβ-环状糊精加入到80℃的40kg水中,然后加入4kg聚乙二醇,200rpm搅拌30min,25℃自然冷却24h,得到乳剂;
S2、将步骤S1制备的乳剂、0.4kg促进剂和0.5kg紫苏籽油加入到70℃的100kg水中,200rpm搅拌3h,45℃冷却2h,加入0.05kg吐温-20和0.4kg酪蛋白酸钠,200rpm搅拌3h,得到悬浮液;将悬浮液冷却至室温,以200rpm搅拌3h;再将悬浮液重新加热至45℃,将0.5kg柠檬香精以0.4L/min的速度滴入悬浮液中,400rpm搅拌10min,再以10000rpm的速度均质8min,得到稳定剂。
所述促进剂的制备方法如下:
将0.04kg脱水蒜粉和0.025kg百里香精油依次加入到10kg体积分数1%的醋酸水溶液中,室温下500rpm搅拌1h,得到草药溶液;然后在草药溶液中加入0.025kg羧甲基壳聚糖和0.006kg甘油,室温下400rpm搅拌9h,超声10min、超声功率为200W、超声频率为50kHz,得到促进剂。
实施例2
一种草本抑菌液的制备方法与实施例1基本相同,唯一区别仅仅在于:所述稳定剂的制备方法中不同。
所述稳定剂的制备方法如下:
S1、将1.8kgβ-环状糊精加入到80℃的40kg水中,然后加入4kg聚乙二醇,200rpm搅拌30min,25℃自然冷却24h,得到乳剂;
S2、将步骤S1制备的乳剂和0.5kg紫苏籽油加入到70℃的100kg水中,200rpm搅拌3h,45℃冷却2h,加入0.05kg吐温-20和0.4kg酪蛋白酸钠,200rpm搅拌3h,得到悬浮液;将悬浮液冷却至室温,以200rpm搅拌3h;再将悬浮液重新加热至45℃,将0.5kg柠檬香精以0.4L/min的速度滴入悬浮液中,400rpm搅拌10min,再以10000rpm的速度均质8min,得到稳定剂。
实施例3
一种草本抑菌液的制备方法与实施例1基本相同,唯一区别仅仅在于:所述稳定剂的制备方法中不同。
所述稳定剂的制备方法如下:
S1、将1.8kgβ-环状糊精加入到80℃的40kg水中,然后加入4kg聚乙二醇,200rpm搅拌30min,25℃自然冷却24h,得到乳剂;
S2、将步骤S1制备的乳剂、0.4kg促进剂加入到70℃的100kg水中,200rpm搅拌3h,45℃冷却2h,加入0.05kg吐温-20和0.4kg酪蛋白酸钠,200rpm搅拌3h,得到悬浮液;将悬浮液冷却至室温,以200rpm搅拌3h;再将悬浮液重新加热至45℃,将0.5kg柠檬香精以0.4L/min的速度滴入悬浮液中,400rpm搅拌10min,再以10000rpm的速度均质8min,得到稳定剂。
所述促进剂的制备方法与实施例1相同。
实施例4
一种草本抑菌液的制备方法与实施例1基本相同,唯一区别仅仅在于:所述稳定剂的制备方法中不同。
所述稳定剂的制备方法如下:
S1、将1.8kgβ-环状糊精加入到80℃的40kg水中,然后加入4kg聚乙二醇,200rpm搅拌30min,25℃自然冷却24h,得到乳剂;
S2、将步骤S1制备的乳剂、0.4kg促进剂和0.5kg紫苏籽油加入到70℃的100kg水中,200rpm搅拌3h,45℃冷却2h,加入0.4kg酪蛋白酸钠,200rpm搅拌3h,得到悬浮液;将悬浮液冷却至室温,以200rpm搅拌3h;再将悬浮液重新加热至45℃,将0.5kg柠檬香精以0.4L/min的速度滴入悬浮液中,400rpm搅拌10min,再以10000rpm的速度均质8min,得到稳定剂。
所述促进剂的制备方法与实施例1相同。
对比例1
一种草本抑菌液的制备方法与实施例1基本相同,唯一区别仅仅在于:所述稳定剂的制备方法中不同。
所述稳定剂的制备方法如下:
S1、将1.8kgβ-环状糊精加入到80℃的40kg水中,然后加入4kg聚乙二醇,200rpm搅拌30min,25℃自然冷却24h,得到乳剂;
S2、将步骤S1制备的乳剂加入到70℃的100kg水中,200rpm搅拌3h,45℃冷却2h,加入0.4kg酪蛋白酸钠,200rpm搅拌3h,得到悬浮液;将悬浮液冷却至室温,以200rpm搅拌3h;再将悬浮液重新加热至45℃,将0.5kg柠檬香精以0.4L/min的速度滴入悬浮液中,400rpm搅拌10min,再以10000rpm的速度均质8min,得到稳定剂。
对比例2
一种草本抑菌液的制备方法与实施例1基本相同,唯一区别仅仅在于:草本抑菌液的制备方法中不添加稳定剂。
测试例1
抑菌活性测试
将金黄色葡萄球菌(Staphylococcus aureus,菌株编号:CICC 21600)、枯草芽孢杆菌(Bacillus subtilis,菌株编号:CICC 25064)、产黄青霉(Penicillium chrysogenum,菌株编号:CICC 4017)、酿酒酵母(Saccharomyces cerevisiae,菌株编号:CICC 31362)菌种接入经灭菌的试管斜面培养基上。将金黄色葡萄球菌、枯草芽孢杆菌放在35℃±l℃的恒温培养箱内培养24h;将产黄青霉、酿酒酵母在28℃±l℃下培养48h;将供试菌株活化之后,备用。
培养基的制备:按照每45g营养琼脂粉配lL培养基的比例配制,搅拌加热煮沸至完全溶解,分装试管或三角瓶,高压蒸汽灭菌(121℃,30min),备用。
孟加拉红培养基的制作:按照每34.5g孟加拉红粉配lL培养基的比例配制,搅拌加热煮沸至完全溶解,分装试管或三角瓶,高压蒸汽灭菌(121℃,30min)备用。
菌悬液的制备:取活化培养好的菌株,用接种环分别挑取少量金黄色葡萄球菌、枯草芽孢杆菌、产黄青霉、酿酒酵母,分别接入10mL的无菌生理盐水中,摇匀,制成菌体、孢子悬浮液,然后分别从第一管取lmL加入9mL的无菌生理盐水中,摇匀,依次类推,采用平板菌落计数法将上述各菌液配成终浓度为2×107个/mL的细菌悬液和104个/mL的孢子悬液,备用。
抑菌活性测试:将配制好的各菌种培养基冷却至55℃,在无菌洁净工作台上,将培养基倒入无菌培养皿内,每个皿20mL,放平至培养基凝固。在无菌操作下,吸取已校正浓度的200μL细菌悬液涂布于培养基表面,吸取已校正浓度的200μL孢子悬液涂布于孟加拉红培养基表面,制成含菌平板。室温2~5min后,等距离放入4个灭菌的牛津杯,其中3个逐一加入本发明的草本抑菌液200μL,剩下一个加入无菌生理盐水,作为空白对照,培养基在35℃±1℃下恒温培养24h,孟加拉红培养基在28℃±1℃下恒温培养48h后,观察抑菌情况,测量抑菌圈直径,结果以mm表示,各菌平行实验3次,取平均值,以此评价抑菌效果。测试结果见表1。
表1:抑菌活性测试结果
-表示无抑菌圈。
测试例2
稳定性测试
pH对草本抑菌液抑菌效果的影响:用0.2mol/L的碳酸钠水溶液和12mol/L盐酸将本发明的草本抑菌液调成pH为3、9的系列溶液,参照测试例1的方法,测定金黄色葡萄球菌抑菌圈直径,试验重复3次,比较pH对石榴皮提取液抑菌效果的影响。测试结果见表2。
热处理对草本抑菌液抑菌活性的影响:将草本抑菌液分别置于70℃、120℃下热处理15min,参照测试例1的方法,测定金黄色葡萄球菌抑菌圈直径,试验重复3次,比较温度对石榴皮提取液抑菌效果的影响。测试结果见表3。
紫外光对草本抑菌液抑菌活性的影响:将草本抑菌液置于紫外光下分别照射10、25min后,紫外光照射功率为200W,参照测试例1的方法,测定金黄色葡萄球菌抑菌圈直径,试验重复3次,比较紫外线对草本抑菌液抑菌效果的影响。测试结果见表4。
保存时间对草本抑菌液活性的影响:将草本抑菌液在室温下保存7d、14d,以金黄色葡萄球菌为供试菌进行一次抑菌活性(抑菌圈)的测定,比较保存时间对草本抑菌液抑菌效果的影响。测试结果见表5。
按下列公式确定其保留抑菌率随pH、热处理、紫外光、保存时间的变化。
保留抑菌率(%)=D1/D0
其中:D1-处理后测得抑菌圈直径;D0-处理前测得抑菌圈直径。
表2:pH稳定性测试结果
试验方案 | pH=3保留抑菌率(%) | pH=9保留抑菌率(%) |
实施例1 | 86.3 | 88.2 |
实施例2 | 74.4 | 76.8 |
实施例3 | 75.1 | 76.9 |
实施例4 | 73.6 | 75.3 |
对比例1 | 53.2 | 57.2 |
对比例2 | 51.4 | 52.5 |
表3:热处理稳定性测试结果
表4:紫外光稳定性测试结果
表5:保存时间稳定性测试结果
试验方案 | 7d保留抑菌率(%) | 14d保留抑菌率(%) |
实施例1 | 98.8 | 98.3 |
实施例2 | 97.3 | 96.3 |
实施例3 | 97.1 | 96.4 |
实施例4 | 96.1 | 95.8 |
对比例1 | 94.5 | 91.5 |
对比例2 | 92.4 | 89.8 |
从测试例1和2的测试结果可以看出,实施例和对比例中实施例1的抑菌活性和稳定性最好,可能原因在于本发明脱水蒜粉、百里香精油的成分中有相当多的O-H键和C-O键。同时,壳聚糖分子在乙酰化单体上具有质子给体原子和质子受体原子。可能在壳聚糖分子和脱水蒜粉、百里香精油之间形成分子间氢键。与纯壳聚糖膜相比,经脱水蒜粉、百里香精油处理的壳聚糖基膜的碳元素含量更高。此外,由于脱水蒜粉中含硫化合物的存在,使得壳聚糖基膜中硫元素的含量增加,可以提高壳聚糖的表面粗糙度。由于草本抑菌液中功能成分的水溶性差、不耐温、易降解和在光照下不稳定,降低了储存时间。在羧甲基壳聚糖中通过与脱水蒜粉、百里香精油和甘油引入疏水段,成功制备了两亲性羧甲基壳聚糖,可以增加疏水基团之间的疏水相互作用,从而形成更致密的疏水核,增强乳化能力。壳聚糖的抗菌活性是接触型的,抗菌活性是因为壳聚糖的游离氨基(正电荷)通过静电相互作用与细菌的细胞表面(负电荷)结合,通过破坏细胞膜和细胞内成分的渗漏导致细菌细胞死亡。经脱水蒜粉、百里香精油处理具有更高的抗菌能力,这可能是由于壳聚糖和脱水蒜粉、百里香精油的协同作用。百里香精油中的香芹酚和百里香酚,以及脱水蒜粉中的大蒜素。百里香酚和香芹酚是疏水酚类成分,它们会对细菌细胞膜产生理化变化,增加细胞膜的通透性,从而导致细菌死亡。此外,香芹酚可诱导细菌细胞中ATP的渗漏和损失。然后,β-环状糊精分子首先与吐温-20组装成超分子复合物,然后通过氢键进一步与功能剂组装成含有β-环状糊精的两亲性颗粒。在油水界面形成薄膜。该膜可以通过物理分割强有力地分离油相和水相,并增加位阻来阻碍聚合。紫苏籽油有促进乳化和形成交联网络的能力,聚乙二醇的长链穿过β-环状糊精的空腔,在链的两端形成纳米级结构域,进一步形成网状结构,网状结构阻碍了液滴的聚集变质,更有利于草本抑菌液的稳定性,长期储存或加热处理后都是稳定的。
Claims (9)
1.一种草本抑菌液,其特征在于,包括以下组分:百部、苦参、川穹、丹参、龙胆草、蒲公英、薄荷油、天然冰片、醋酸氯己定、稳定剂。
2.如权利要求1所述的一种草本抑菌液,其特征在于,包括以下重量份组分:百部2~4份、苦参6~9份、川穹3~5份、丹参3~5份、龙胆草1~3份、蒲公英3~5份、薄荷油1~3份、天然冰片0.2~0.8份、醋酸氯己定0.001~0.03份、稳定剂2~5份。
3.如权利要求1或2所述的一种草本抑菌液,其特征在于,所述稳定剂的制备方法如下:
S1、将β-环状糊精加入到水中,然后加入聚乙二醇,搅拌,冷却,得到乳剂;
S2、将步骤S1制备的乳剂、促进剂和紫苏籽油加入到水中,搅拌,冷却,加入吐温-20和酪蛋白酸钠,搅拌,得到悬浮液;将悬浮液冷却至室温,搅拌;再将悬浮液重新加热,将柠檬香精滴入悬浮液中,搅拌,再均质,得到稳定剂。
4.如权利要求3所述的一种草本抑菌液,其特征在于,所述稳定剂的制备方法如下,以重量份计:
S1、将1.5~2份β-环状糊精加入到60~90℃的30~50份水中,然后加入3~5份聚乙二醇,100~500rpm搅拌20~40min,20~30℃自然冷却10~30h,得到乳剂;
S2、将步骤S1制备的乳剂、0.2~0.6份促进剂和0.3~0.7份紫苏籽油加入到60~80℃的80~120份水中,100~500rpm搅拌1~5h,40~50℃冷却1~3h,加入0.04~0.06份吐温-20和0.2~0.6份酪蛋白酸钠,100~500rpm搅拌1~5h,得到悬浮液;将悬浮液冷却至室温,以50~200rpm搅拌1~5h;再将悬浮液重新加热至40~50℃,将0.4~0.6份柠檬香精以0.3~0.5L/min的速度滴入悬浮液中,100~500rpm搅拌5~15min,再以8000~15000rpm的速度均质5~10min,得到稳定剂。
5.如权利要求3或4所述的一种草本抑菌液,其特征在于,所述促进剂的制备方法如下:
将脱水蒜粉和百里香精油依次加入到醋酸水溶液中,室温搅拌,得到草药溶液;然后在草药溶液中加入羧甲基壳聚糖和甘油,室温搅拌,超声,得到促进剂。
6.如权利要求5所述的一种草本抑菌液,其特征在于,所述促进剂的制备方法如下,以重量份计:
将0.02~0.06份脱水蒜粉和0.02~0.03份百里香精油依次加入到8~12份体积分数0.5~2%的醋酸水溶液中,室温下100~800rpm搅拌0.5~2h,得到草药溶液;然后在草药溶液中加入0.02~0.03份羧甲基壳聚糖和0.004~0.008份甘油,室温下100~500rpm搅拌5~15h,超声5~15min、超声功率为100~400W、超声频率为40~60kHz,得到促进剂。
7.一种制备如权利要求1~6任一项所述草本抑菌液的方法,其特征在于,步骤如下:
步骤1、按重量份称取各原料,将百部、苦参、川穹、丹参、龙胆草、蒲公英分别除去杂质,洗净后干燥;
步骤2、将步骤1干燥后的川穹和百部切碎,过筛,加入到乙醇水溶液中进行浸泡后过滤,得到浸泡溶液;
步骤3、将苦参、丹参、龙胆草、蒲公英切碎研磨,过筛,然后加入到乙醇水溶液中,加热搅拌,得到醇提液,将醇提液加入到步骤2制备的浸泡溶液中,搅拌过滤,得到混合滤液;然后加入薄荷油、天然冰片、醋酸氯己定和稳定剂,搅拌,再加入水调整含乙醇浓度,即得草本抑菌液。
8.如权利要求7所述的方法,其特征在于,步骤如下:
步骤1、按重量份称取各原料,将百部、苦参、川穹、丹参、龙胆草、蒲公英分别除去杂质,洗净后干燥;
步骤2、将步骤1干燥后的川穹和百部切碎,过20~50目筛,加入到质量比为3~5倍的70~85wt%乙醇水溶液中进行浸泡20~40d后300~500目筛过滤,得到浸泡溶液;
步骤3、将苦参、丹参、龙胆草、蒲公英切碎研磨,过50~200目筛,然后加入到质量比为4~6倍的40~60wt%乙醇水溶液中,加热到60~75℃、50~200rpm搅拌3~8h,得到醇提液,将醇提液加入到步骤2制备的浸泡溶液中,100~500rpm搅拌2~6h,100~600目筛过滤,得到混合滤液;然后加入薄荷油、天然冰片、醋酸氯己定和稳定剂,100~500rpm搅拌20~40min,再加入水调整含乙醇浓度为30~50wt%,即得草本抑菌液。
9.一种如权利要求1~6任一项所述草本抑菌液在制备穴位贴中的应用。
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