CN105560306B - 一种抗皮肤过敏的马齿苋气雾剂 - Google Patents
一种抗皮肤过敏的马齿苋气雾剂 Download PDFInfo
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- CN105560306B CN105560306B CN201510975000.8A CN201510975000A CN105560306B CN 105560306 B CN105560306 B CN 105560306B CN 201510975000 A CN201510975000 A CN 201510975000A CN 105560306 B CN105560306 B CN 105560306B
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Abstract
本发明公开了一种抗皮肤过敏的马齿苋气雾剂,通过将马齿苋提取物与纯净水按照1g:500‑1000mL的配比稀释得稀释液,稀释液用金属铝罐灌装,加压充氮气保存而得。本发明原料马齿苋纯天然,提取过程不使用任何有机溶剂,产品不添加任何防腐剂,产品使用方便,抗皮肤过敏效果良好,且无副作用。
Description
技术领域
本发明涉及一种气雾剂,特别涉及一种抗皮肤过敏的马齿苋气雾剂。
背景技术
面部皮肤过敏的人群逐渐增多。面部皮肤高敏感性、长期亚健康状态,反复发作,严重影响容颜美观,造成患者极大的生理和心理压力。对于面部敏感性皮肤患者,必须提供专业性的治疗和治疗后的皮肤护理,所用产品必须具备安全性、有效性、稳定性、舒适性、可依从性及方便性。气雾剂是通过将有效成分雾化成极细微水雾,均匀地作用于面部皮肤,更容易渗透于皮肤,发挥化学作用;同时利用较低温度的物理作用,收缩面部扩张的毛细血管,减轻炎症渗出,降低感觉神经的兴奋性,抑制皮脂分泌,通过化学作用和低温物理作用的协同,达到抗过敏、抗炎消肿、舒缓止痒的目的,从而缩短皮肤恢复时间,减少皮肤色素沉着,具有有效率高、疗程短和不良反应率低等优点,是改善面部皮肤敏感状态的理想方法。
目前在化妆品和个人洗护用品中常用的抗刺激物质有激素类药物,但是,间断性或长期使用激素可使皮肤产生严重的依赖,并可使皮肤发生萎缩变薄、毛细血管扩张(红血丝)、硬化、丧失保水功能而产生红肿、过敏、瘙痒、变黑、长毛、粉刺等不可逆转的后果。到目前为止,众所周知的抗刺激剂不管是皮肤安全性方面还是化妆品原料的搭配的安全性方面大部分都存在问题所以使用受到限制。因此,找到一种实际使用效果良好,且可以抗刺激、抗过敏,且无副作用的纯天然植物原料应用于化妆品和个人洗护用品显得尤为重要。
马齿苋(拉丁学名:Portulaca oleracea L.)为马齿苋科一年生草本植物。肥厚多汁,无毛,高10~30cm,生于田野路边及庭园废墟等向阳处。国内各地均有分布。该种为药食两用植物,无毒副作用。全草供药用,有清热利湿、解毒消肿、消炎、止渴、利尿作用;民间常外用治疗湿疹、烫伤、丹毒、肿疡、扁平疣、带状疱疹、黄褐斑、痤疮、臁疮、白癜风等。马齿苋有天然抗生素之称,根据现代研究其对金黄色葡萄球菌、大肠杆菌、绿脓杆菌、病毒、真菌都有抑制作用;马齿苋含有大量植物性去甲基肾上腺素、多巴胺等儿茶酚胺类物质,具有抗过敏,收缩皮肤毛细血管的作用。现代研究,马齿苋还含有丰富的SL3脂肪酸及维生素A样物质。SL3脂肪酸是形成细胞膜,尤其是脑细胞膜与眼细胞膜所必需的物质;维生素A样物质能维持上皮组织如皮肤、角膜及结合膜的正常机能,参与视紫质的合成,增强视网膜感光性能,也参与体内许多氧化过程。
研究报道的马齿苋常用其抗炎功效,马齿苋采用醇提法提取有效成分,如金英子等(金英子,张红英等,马齿苋提取物的抗炎消肿及镇痛作用[J].延边大学医学学报,2008,31(4):258~260)的研究表明:马齿苋乙醇水溶液提取物可以有效抑制二甲苯及巴豆油所致小鼠耳廓肿胀,具有抗炎消肿及镇痛作用。马齿苋采用醇提法得到的提取物抗炎效果较好,但是抗过敏效果不佳。
发明内容
本发明的目的在于提供一种抗皮肤过敏的马齿苋气雾剂,使用方便,抗皮肤过敏效果良好,且无副作用,原料马齿苋纯天然。
本发明解决其技术问题所采用的技术方案是:
一种抗皮肤过敏的马齿苋气雾剂,通过将马齿苋提取物与纯净水按照1g:500-1000mL的配比稀释得稀释液,稀释液用金属铝罐灌装,加压充氮气保存而得。
本发明以抗皮肤过敏效果良好的马齿苋提取物作为主要原料,氮气作为抛射剂获得一种气雾剂,原料马齿苋纯天然,用方便,抗皮肤过敏效果良好,且无副作用。
作为优选,所述马齿苋提取物通过以下方法制备而得:
(1)压榨与过滤
新鲜马齿苋用自来水洗净,再用纯净水漂洗后,第一次冷藏处理,第一次冷藏处理的新鲜马齿苋用榨汁机榨汁,所得汁液经超声细胞破碎仪处理3-7分钟得到压榨液,压榨液经第二次冷藏处理后,过滤,滤液经第三次冷藏处理后得马齿苋粗提液;
第一次冷藏处理的目的是:减少后续榨汁过程中摩擦产生的热量破坏活性成分(植物源性去甲肾上腺素、多巴胺)。
超声细胞破碎仪处理有利于细胞内成分的溶出,而且不会破坏化学成分。去甲肾上腺素及多巴胺为水溶性,因此,提取过程尽可能不使用有机溶剂等各种溶媒。为尽可能减少提取物中掺入其他化学成分,保证提取过程绿色无污染,减少可能刺激皮肤的不良化学成分,提取过程中不使用有机溶剂等任何溶媒,保证纯粹物理提取,无菌、无杂质,无化学污染。
第二次冷藏处理、第三次冷藏处理的目的是:有助于破碎细胞内活性成分的溶出和沉渣的沉淀。全程低温条件下提取,可避免加热提取法对去甲肾上腺素、多巴胺等抗过敏活性成分的破坏。
(2)离心与冷冻干燥
马齿苋粗提液离心后,取上清液,过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机冷冻干燥,得到粉末状的马齿苋粗提物;
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解,冷藏(4℃下冷藏处理48小时)后,离心,取上清液,冷冻干燥,得到粉末状的马齿苋提取物,用真空袋封装,辐照杀菌。
马齿苋提取物中的细小渣滓可能会堵塞气雾剂喷口,含有的蛋白质容易变质导致异味,多糖容易导致皮肤绷紧感,色素颗粒容易导致使用者抗拒感,所以本发明的马齿苋粗提液提取过程经过两次离心并配合冷冻冷藏处理,最后冷冻干燥,以尽可能去除沉渣、蛋白质、脂肪、多糖等大分子物质和易腐败物质,仅保留水溶性化学成分,排除致敏因素。
现有的马齿苋提取物基本都是醇提法,醇提法所得的提取物中大量的是黄酮和多糖类物质,只有极少量的植物源性去甲肾上腺素、多巴胺。且现有的醇提法提取温度较高,工艺步骤容易使得植物源性去甲肾上腺素、多巴胺发生氧化变质,从而丧失抗皮肤过敏效用。经本发明的特定方法提取的马齿苋提取物,提取物中抗皮肤过敏成分植物源性去甲肾上腺素、多巴胺的含量较高,提取方法不破坏植物源性去甲肾上腺素、多巴胺,植物源性去甲肾上腺素、多巴胺活性活性保持好。去甲肾上腺素、多巴胺等儿茶酚胺类化合物具有抗过敏、收缩皮肤毛细血管,改善皮肤敏感的效果。但是,化学合成的重酒石酸去甲肾上腺素、多巴胺具有升高血压、促进心肌收缩等作用,多用于休克、心力衰竭等急危重症的抢救治疗,不适合用于改善皮肤过敏及皮肤毛细血管扩张。新鲜马齿苋中含有大量的植物源性去甲肾上腺素及多巴胺,相比化学合成的去甲肾上腺素、多巴胺,植物源性去甲肾上腺素及多巴胺的升高血压、收缩心肌的作用明显弱化,而且改善皮肤过敏及毛细血管扩张的效能仍然存在,因此,本发明选用新鲜马齿苋提取植物源性去甲肾上腺素及多巴胺。去甲肾上腺素和多巴胺的化学结构不稳定,遇空气、遇光、遇热容易失效,马齿苋干品中的去甲肾上腺素及多巴胺已经变性失效,因此选择使用新鲜马齿苋提取。制作过程中,尽可能全程低温提取,低温保存,常温消毒。通过金属罐填充氮气制成气雾剂使用,可以隔绝空气防止氧化;氮气作为抛射剂,对皮肤无刺激,而且具有防腐功能,可减少其他有刺激防腐剂的使用,从而减少皮肤刺激;气雾罐为马齿苋提取物提供较低的保存温度,有利于马齿苋提取物的稳定。
作为优选,步骤(1)中的冷藏处理参数为:4℃下冷藏处理12小时。
作为优选,步骤(1)中的新鲜马齿苋选用生长30-60天、开花期前的新鲜马齿苋。马齿苋花中含有大量花粉,花粉是一种过敏原,有可能对个别人有致敏作用,为减少敏感皮肤的过敏刺激,本发明选用生长约30-60天,尚未开花的新鲜马齿苋为原材料,这时的马齿苋中植物源性去甲肾上腺素、多巴胺的含量也较高。
作为优选,步骤(2)中的离心参数为:4℃,3500-4500rpm离心8-15min。
作为优选,步骤(3)中的离心参数为:4℃,3500-4500rpm离心8-15min作为优选,步骤(3)中辐照杀菌采用钴辐照杀菌,辐射剂量为2500-3500千拉德,辐照杀菌时间10-12小时。经冷冻干燥后制得的粉末状提取物,不用加热加压法消毒杀菌或者化学法杀菌,采用钴辐照杀菌,从而保证保护去甲肾上腺素及多巴胺的稳定性。
作为优选,所述氮气为高纯氮气,氮气纯度为99.99-99.9999%。本发明利用金属铝罐灌装,加压充氮气保存。高纯氮气作为抛射剂,既可以提供8-9个标准大气压的压力,为喷雾提供压力,其优点不同于其他有机溶剂的抛射剂,不会给该产品带来化学污染;又可以作为一种防腐剂及稳定剂,防止提取物的腐败、细菌滋生,保持去甲肾上腺素及多巴胺的稳定。
作为优选,100mL稀释液用200mL金属铝罐灌装,加压充氮气100mL保存。
本发明的有益效果是:原料马齿苋纯天然,提取过程不使用任何有机溶剂,产品不添加任何防腐剂,产品使用方便,抗皮肤过敏效果良好,且无副作用。
本发明的马齿苋提取物中去甲肾上腺素和多巴胺的含量分别为0.12-0.15%和0.02-0.05%。
附图说明
图1是混合对照品HPLC图谱,1-NA,2-DA。
图2是马齿苋提取物样品的HPLC图谱,1-NA,2-DA。
具体实施方式
下面通过具体实施例,对本发明的技术方案作进一步的具体说明。
本发明中,若非特指,所采用的原料和设备等均可从市场购得或是本领域常用的。下述实施例中的方法,如无特别说明,均为本领域的常规方法。
本发明实施例使用的氮气为高纯氮气,氮气纯度为99.99-99.9999%。
本发明的原料新鲜马齿苋选用生长30-60天、开花期前的新鲜马齿苋。
实施例1:
一种抗皮肤过敏的马齿苋气雾剂,通过将马齿苋提取物与纯净水按照1g:500mL的配比稀释得稀释液,100mL稀释液用200mL金属铝罐灌装,加压充氮气100mL保存。
所述马齿苋提取物通过以下方法制备而得:
(1)压榨与过滤
新鲜马齿苋用自来水洗净,再用纯净水漂洗后,第一次冷藏处理(4℃下冷藏处理12小时),第一次冷藏处理的新鲜马齿苋用榨汁机榨汁,所得汁液经超声细胞破碎仪处理3分钟得到压榨液,压榨液经第二次冷藏处理(4℃下冷藏处理12小时)后,过滤,滤液经第三次冷藏处理(4℃下冷藏处理12小时)后得马齿苋粗提液;
(2)离心与冷冻干燥
马齿苋粗提液离心(4℃,3500rpm离心15min)后,取上清液,过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机冷冻干燥,得到粉末状的马齿苋粗提物;
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解,冷藏后(4℃下冷藏处理48小时),离心(4℃,3500rpm离心15min),取上清液,冷冻干燥,得到粉末状的马齿苋提取物,用真空袋封装,辐照杀菌,辐照杀菌采用钴辐照杀菌,辐射剂量为2500千拉德,辐照杀菌时间12小时。
实施例2:
一种抗皮肤过敏的马齿苋气雾剂,通过将马齿苋提取物与纯净水按照1g:1000mL的配比稀释得稀释液,100mL稀释液用200mL金属铝罐灌装,加压充氮气100mL保存。
所述马齿苋提取物通过以下方法制备而得:
(1)压榨与过滤
新鲜马齿苋用自来水洗净,再用纯净水漂洗后,第一次冷藏处理(4℃下冷藏处理12小时),第一次冷藏处理的新鲜马齿苋用榨汁机榨汁,所得汁液经超声细胞破碎仪处理7分钟得到压榨液,压榨液经第二次冷藏处理(4℃下冷藏处理12小时)后,过滤,滤液经第三次冷藏处理(4℃下冷藏处理12小时)后得马齿苋粗提液;
(2)离心与冷冻干燥
马齿苋粗提液离心(4℃,4500rpm离心8min)后,取上清液,过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机冷冻干燥,得到粉末状的马齿苋粗提物;
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解,冷藏后(4℃下冷藏处理48小时),离心(4℃,4500rpm离心8min),取上清液,冷冻干燥,得到粉末状的马齿苋提取物,用真空袋封装,辐照杀菌,辐照杀菌采用钴辐照杀菌,辐射剂量为3500千拉德,辐照杀菌时间10小时。
实施例3:
一种抗皮肤过敏的马齿苋气雾剂,通过将马齿苋提取物与纯净水按照1g:800mL的配比稀释得稀释液,100mL稀释液用200mL金属铝罐灌装,加压充氮气100mL保存。
所述马齿苋提取物通过以下方法制备而得:
(1)压榨与过滤
新鲜马齿苋用自来水洗净,再用纯净水漂洗后,第一次冷藏处理(4℃下冷藏处理12小时),第一次冷藏处理的新鲜马齿苋用榨汁机榨汁,所得汁液经超声细胞破碎仪处理5分钟得到压榨液,压榨液经第二次冷藏处理(4℃下冷藏处理12小时)后,过滤,滤液经第三次冷藏处理(4℃下冷藏处理12小时)后得马齿苋粗提液;
(2)离心与冷冻干燥
马齿苋粗提液离心(4℃,4000rpm离心10min)后,取上清液,过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机冷冻干燥,得到粉末状的马齿苋粗提物;
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解,冷藏(4℃下冷藏处理48小时)后,离心(4℃,4000rpm离心10min),取上清液,冷冻干燥,得到粉末状的马齿苋提取物,用真空袋封装,辐照杀菌,辐照杀菌采用钴辐照杀菌,辐射剂量为3000千拉德,辐照杀菌时间12小时。
马齿苋提取物具体提取方案:
(1)压榨与过滤
新鲜马齿苋15.29kg用自来水洗净,再用纯净水漂洗后,第一次冷藏处理(4℃下冷藏处理12小时),第一次冷藏处理的新鲜马齿苋用SKG-19911榨汁机榨汁,所得汁液经超声细胞破碎仪处理5分钟得到压榨液,压榨液经第二次冷藏处理(4℃下冷藏处理12小时)后,4层纱布过滤3次,滤液经第三次冷藏处理(4℃下冷藏处理12小时)后得马齿苋粗提液。
(2)离心与冷冻干燥
马齿苋粗提液离心(4℃,3500rpm离心10min)后,取上清液,滤纸过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机(-80℃)冷冻干燥,得到粉末状的马齿苋粗提物117g。
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解(约1L),呈混悬液,冷藏后(放置在4℃冰箱冷藏48h),离心(4℃,3500rpm离心10min),取上清液,冷冻干燥,得到粉末状的马齿苋提取物64g,用真空袋封装,辐照杀菌,辐照杀菌采用钴辐照杀菌(放射同位素钴60产生的γ射线),辐射剂量为2500千拉德,辐照杀菌时间12小时。取少量马齿苋提取物用水溶解,呈澄清溶液。
将马齿苋提取物按照1g:500-1000mL浓度,用纯净水稀释,(1g提取物溶解于1000ml水;按0.13%去甲肾上腺素含量计算,相当于1000ml水内含有去甲肾上腺素1.3mg;去甲肾上腺素的静脉用药浓度是1mg/100ml,即10mg/1000ml;因此皮肤用去甲肾上腺素的浓度约相当于静脉用药浓度1/5--1/10);200ml金属铝罐,填装稀释液100ml,填充氮气100ml,压强8-9千克力/平方厘米(8-9个标准大气压),气雾量保证喷射3秒钟即可达到面部全覆盖。
马齿苋提取物中去甲肾上腺素、多巴胺的含量检测
[1]仪器与试药
仪器:Agilent 1260高效液相色谱仪(美国Agilent公司),色谱柱为ZORBAX SB-C18(4.6×150mm,5μm),DAD检测器。BP 211D型分析电子天平(德国赛多利斯公司);Simplicity纯水系统(美国密理博公司)。
对照品:本实验含量测定用的重酒石酸去甲肾上腺素对照品(批号:100169-201404,按C8H11NO3.C4H6O6计,含量为94.3%)和盐酸多巴胺对照品(批号:100070-201507,按C8H11NO2.HCL计,含量为99.8%)均系中国食品药品检定研究院的供含量测定用标准品。
试剂:超纯水;甲醇为色谱纯;其他试剂均为分析纯。
[2]色谱条件的确定
①色谱柱:色谱柱为ZORBAX SB-C18(4.6×150mm,5μm)
②流动相:甲醇-水[57.5:47.5,含30mmol·L-1十二烷基硫酸钠和0.02M磷酸二氢钾(PH=3.0)]
③检测波长280nm
④流速:1.0mL/min
⑤进样量:10μL
⑥柱温:40℃
⑧数据处理定量方法:外标峰面积法
[3]混合对照品溶液和供试品溶液的制备
混合对照品溶液的制备:
(1)取重酒石酸去甲肾上腺素对照品适量,精密称定,加0.1M盐酸水溶液制成每1mL含0.3mg的溶液。
(2)取盐酸多巴胺对照品适量,精密称定,加0.1M盐酸水溶液制成每1mL含1mg的溶液。
将(1)和(2)精密吸取1ml和0.1ml置10ml容量瓶中,用0.1M盐酸水溶液定容至刻度,摇匀。则每1mL混合对照品溶液含30μg去甲肾上腺素和10μg多巴胺。
供试品溶液的制备:取本品约0.5g,精密称定,置25ml量瓶中,用0.1M盐酸水溶液溶解并定容,摇匀,0.45μm微孔滤膜过滤,取续滤液。
将混合对照品溶液和供试品溶液各10μL注入液相色谱仪检测,混合对照品HPLC图谱和马齿苋提取物样品的HPLC图谱见图1和图2。
[4]线性关系的考察:
精密称取重酒石酸去甲肾上腺素对照品(供含量测定用,含量以94.3%计,中国食品药品检定研究院)20.35mg,盐酸多巴胺对照品(供含量测定用,含量以99.8%计,中国食品药品检定研究院)19.11mg同置10mL量瓶中,加0.1M盐酸水溶液溶解并稀释至刻度,则混合对照品溶液中去甲肾上腺素(NA)和多巴胺(DA)的浓度分别为962.6μg/mL和1540.5μg/mL;将上述混合对照品溶液用0.1M盐酸水溶液分别精密稀释2倍(1→2)、5倍(2→10)、10倍(1→10)、20倍(0.5→10)、100倍(0.1→10),则混合对照品溶液中NA和DA的浓度分别为481.29μg/mL和770.25μg/mL、192.52μg/mL和308.10μg/mL、96.26μg/mL和154.05μg/mL、48.13μg/mL和77.03μg/mL、9.63μg/mL和15.41μg/mL;再将混合对照品溶液(NA和DA的浓度分别为9.62μg/mL和15.41μg/mL)精密稀释10倍(1→10)、50倍(0.2→10)、100倍(0.1→10),则混合对照品溶液中NA和DA的浓度分别为0.9626μg/mL和1.5405μg/mL、0.1925μg/mL和0.3081μg/mL、0.0963μg/mL和0.1541μg/mL。照高效液相色谱法(中国药典2010年版一部附录VI D)测定,以质量浓度为横坐标,峰面积值为纵坐标,得NA和DA的回归方程分别为y=10.122x-0.4503(R2=1)和y=10.366x-2.415(R2=1),说明本品NA和DA进样量分别在0.0963-481.29μg/mL和0.1541-770.25μg/mL范围内,均与各自峰面积之间呈良好的线性关系。结果见表1和表2。
表1去甲肾上腺素线性关系考察结果
表2多巴胺线性关系考察结果
[5]仪器精密度:取混合对照品溶液重复测定6次,NA峰面积的RSD为0.03%;DA峰面积的RSD为1.89%。
[6]重复性试验
取同一批样品6份,每份约0.5g,精密称定,置25mL量瓶中,用0.1M盐酸水溶液溶解并定容,摇匀,0.45μm微孔滤膜过滤,取续滤液,注入液相色谱仪检测。结果见表3。
表3重复性试验结果
[7]回收率:配制混合对照品溶液,含NA为715.2μg/ml和DA为404.7μg/ml。采用加样回收,取同一批样品6份,每份约0.25g,精密称定,置25mL量瓶中,用平头注射器精密加入混合对照品溶液500ul,用0.1M盐酸水溶液溶解并定容,摇匀,0.45μm微孔滤膜过滤,取续滤液,注入液相色谱仪检测。结果如表4、表5。
表4回收率试验结果
表5回收率试验结果
结论:该批马齿苋提取物中去甲肾上腺素和多巴胺的含量分别为0.13%和0.03%。
安全性验证试验
1.急性经口毒性试验:
试验动物:昆明小鼠10只,体重18-22g,雌雄各半。
供试液:马齿苋提取物加纯净水溶解成10%浓度(100mg/ml)
饲养条件:实验室温度21-23℃,相对湿度65-75%,给药前禁食12小时。
试验方法:参照《化妆品卫生规范》2007版,采用一次限量法,给予小鼠口服5000mg/kg(1ml/20g体重)的马齿苋提取物水溶液1ml。
试验结果:观察10天,小鼠活动正常,未见明显毒性反应,无死亡,LD50﹥5000mg/kg。
2.急性皮肤刺激性试验:
试验动物:家兔4只,体重2-3Kg,雌雄各半。
供试液:马齿苋气雾剂1:500浓度。
饲养条件:实验室温度21-23℃,相对湿度65-75%,左右两侧各备皮3×3cm。
试验方法:参照《化妆品卫生规范》2007版,马齿苋气雾剂液体3ml涂搽左侧,右侧涂搽纯净水作为对照。
试验结果:涂搽马齿苋气雾剂溶液后24、48、72小时观察激发部位皮肤反应,马齿苋气雾剂溶液未致家兔皮肤出现过敏反应。
3.多次皮肤刺激性试验:
试验动物:家兔4只,体重2-3Kg,雌雄各半。
供试液:马齿苋气雾剂1:500浓度。
饲养条件:实验室温度21-23℃,相对湿度65-75%,左右两侧各备皮3×3cm。
试验方法:参照《化妆品卫生规范》2007版,马齿苋气雾剂液体3ml涂搽左侧,右侧涂搽纯净水作为对照,每日1次,连续涂搽14天。
试验结果:连续14天观察激发部位皮肤反应,马齿苋气雾剂溶液未致家兔皮肤出现刺激反应。
4.急性眼刺激性试验:
试验动物:家兔4只,体重2-3Kg,雌雄各半。
供试液:马齿苋气雾剂1:500浓度。
饲养条件:实验室温度21-23℃,相对湿度65-75%。
试验方法:参照《化妆品卫生规范》2007版,马齿苋气雾剂液体0.1ml滴于左侧眼结膜囊,右侧眼睛作为对照。
试验结果:马齿苋气雾剂溶液后24、48、72小时观察,马齿苋气雾剂溶液未致家兔眼睛出现刺激反应。
功效性验证试验
1.抗过敏试验
目的:采用瑞敏过敏原斑贴(瑞典化学诊断公司生产),作为过敏原,评价本发明中的马齿苋气雾剂的抗过敏能力。瑞敏过敏原斑贴试剂有20种过敏原,包括苯唑卡因、硫氢混合物、咪唑烷基尿素、N-环己基硫酞内醋、对苯二胺、重铬酸钾、乙二胺、松香、甲醛、环氧树醋、溴硝丙二醇、秋兰姆混合物、对苯类、硫酸镍、香内酷类、芳香混合物、皮质类固醇组分、黑橡胶混合物、卡巴混合物和硫柳汞。
试剂:马齿苋气雾剂1:500,纯净水喷雾剂、瑞敏过敏原斑贴试剂。
方法:将滴加有20种过敏原的斑贴测试片贴敷在脊柱左右两侧,两侧过敏原完全相同,贴敷后48h将斑贴测试片移除,移除后24h判读结果。根据国际接触性皮炎研究组(IGDRG)推荐的标准判读,具体标准:①“-”阴性反应;②“±”可疑阳性(仅有轻度红斑);③“+”弱阳性(红斑、浸润、可有少量丘疹);④“++”强阳性(红斑、浸润、丘疹、水疤);⑤“+++”极强反应(红斑、浸润、丘疹、出现水疱大疱)。选择10名(5男,5女)极强阳性反应者,左侧外用马齿苋气雾剂,右侧外用纯净水气雾剂,双侧对照,观测其皮疹消退至可疑阳性程度的时间,皮疹提前好转超过24小时的认定为有效,统计有效率。马齿苋气雾剂、纯净水喷雾剂的使用剂量为每次喷雾1分钟,保留5分钟,每日3次。
结果:
结论:本发明的马齿苋气雾剂能够有效拮抗瑞敏过敏原所导致的皮肤过敏,马齿苋气雾剂的制作过程能够有效保持马齿苋鲜草中的抗过敏成分及作用。
以上所述的实施例只是本发明的一种较佳的方案,并非对本发明作任何形式上的限制,在不超出权利要求所记载的技术方案的前提下还有其它的变体及改型。
Claims (8)
1.一种抗皮肤过敏的马齿苋气雾剂,其特征在于:通过将马齿苋提取物与纯净水按照1g:500-1000mL的配比稀释得稀释液,稀释液用金属铝罐灌装,加压充氮气保存而得;
马齿苋提取物通过以下方法制备而得:
(1)压榨与过滤
新鲜马齿苋用自来水洗净,再用纯净水漂洗后,第一次冷藏处理,第一次冷藏处理的新鲜马齿苋用榨汁机榨汁,所得汁液经超声细胞破碎仪处理3-7分钟得到压榨液,压榨液经第二次冷藏处理后,过滤,滤液经第三次冷藏处理后得马齿苋粗提液;
(2)离心与冷冻干燥
马齿苋粗提液离心后,取上清液,过滤,得到澄清溶液,于-80℃冰箱内冷冻12小时,再用冷冻干燥机冷冻干燥,得到粉末状的马齿苋粗提物;
(3)二次离心与冷冻干燥
将马齿苋粗提物用超纯水溶解,冷藏后,离心,取上清液,冷冻干燥,得到粉末状的马齿苋提取物,用真空袋封装,辐照杀菌。
2.根据权利要求1所述的马齿苋气雾剂,其特征在于:步骤(1)中的冷藏处理参数为:4℃下冷藏处理12小时。
3.根据权利要求1所述的马齿苋气雾剂,其特征在于:步骤(1)中的新鲜马齿苋选用生长30-60天、开花期前的新鲜马齿苋。
4.根据权利要求1所述的马齿苋气雾剂,其特征在于:步骤(2)中的离心参数为:4℃,3500-4500rpm离心8-15min。
5.根据权利要求1所述的马齿苋气雾剂,其特征在于:步骤(3)中的离心参数为:4℃,3500-4500rpm离心8-15min。
6.根据权利要求1所述的马齿苋气雾剂,其特征在于:步骤(3)中辐照杀菌采用钴辐照杀菌,辐射剂量为2500-3500千拉德,辐照杀菌时间10-12小时。
7.根据权利要求1所述的马齿苋气雾剂,其特征在于:所述氮气为高纯氮气,氮气纯度为99.99-99.9999%。
8.根据权利要求1所述的马齿苋气雾剂,其特征在于:100mL稀释液用200mL金属铝罐灌装,加压充氮气100mL保存。
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