CN110269813A - 一种复合活性多肽生物微晶冻干粉制备 - Google Patents
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Abstract
本发明提供一种复合活性多肽生物微晶冻干粉制备,应用于美容抗衰化妆品领域,其采用生物技术复合活性多肽通过冻干粉的技术处理,发挥有效的美容抗衰老活性成分。预处理包含复合活性多肽混合液,将其冻干工艺进行了详细的研究并作出改进,通过溶液预冻、一次干燥、二次干燥的过程,能够使得制备的冻干粉具有散射状微晶表面。一方面提高复合生物活性多肽冻干粉的生物活性和稳定性,且所得复合活性美容多肽生物微晶冻干粉能够在冷藏或者常温下保存较长时间。另一方面可提高了冻干粉的外观性,区别于传统冻干粉疏松的块状物或粉末外观表面。本发明工艺简单易行,适合工业化生产。
Description
技术领域
本发明属于美容抗衰老化妆品领域,具体涉及一种复合活性多肽生物微晶冻干粉制备、包含所述复合活性多肽生物微晶冻干粉的组合物及制备,提高复合活性多的生物活性和稳定性,且具有散射状微晶表面,能够在冷藏或者常温下保存较长时间。
背景技术
近年来随着我国经济的快速发展和生活水平的提高,人们对于美容和抗衰老的需求呈爆发式增长。在皮肤的自然老化和护理过程中,生物活性多肽起着重要的作用,如细胞增殖、细胞迁移、血管发生、色素形成和蛋白质合成及调控等。各种活性多肽除了具备自身的功能外,相互之间还可以产生促进作用,所以,复合活性多肽在医疗和化妆品等行业具有重要作用。对于生物活性多肽,容易在生产过程中受到温度、酸碱的影响,造成不同程度的变形,影响其纯度及活性,较好的保存方法是采用冷冻干燥方法。冷冻干燥又称升华干燥,是将需要干燥的物料溶液预先冻结成固体,然后在低温低压条件下,从冻结状态不经过液态而直接升华去除水分的一种干燥方法。通过冷冻工艺改进,不仅可提高复合活性多肽冻干粉的生物活性和稳定性。同时,还可以实现将物料的组织结构和外观形态较好地保存。
现有技术中,活性多肽冻干粉的保藏温度在-20℃及以下,保存条件非常苛刻,不利于保存和运输,因此,亟需提高生物活性多肽冻干粉的保藏活性和稳定性,使其能够在冷藏或者常温下保存较长时间。
同时,传统冻干粉多为疏松的块状物或粉末外观,较少具有新颖独特外观。如何改变冻干粉原有单调的外观效果,提高观感以更好的发挥其心理功能即引起消费者的关注,成为研究的另一个热点。
发明内容
本发明的主要目的在于,提供一种复合活性多肽生物微晶冻干粉制备方法,提高复合生物活性因子冻干粉的生物活性和稳定性,且能够在冷藏或者常温下保存较长时间。同时,所制备冻干粉具有散射状微晶表面的独特外观。
为达到上述目的,本发明通过以下技术方案实现:
本发明的第一特征在于提供一种复合活性多肽生物微晶冻干粉抗衰老化妆品,按重量配比其包含下列组分:0.01%-0.05%(m/v)活性多肽、5%-20%(m/v)冻干保护剂、0.1%-2.0%(m/v)皮肤调理剂、0.1%-0.3%(m/v)保湿剂、0.5%-1.0%生物成膜剂,余量为去离子水;
进一步地,其中所述活性多肽为酸性成纤维细胞生长因子(aFGF)、碱性成纤维细胞生长因子(bFGF)、人表皮生长因子(EGF)、血管内皮生长因子(VEGF)、转化生长因子-β1(TGF-β1)、转化生长因子-β2(TGF-β2)和角质细胞生长因子(KGF)中的一种或多种;
进一步地,其中所述冻干保护剂为海藻糖、甘露醇、甘氨酸、右旋糖酐、精氨酸中的一种或多种;
进一步地,其中所述皮肤调理剂为胶原蛋白、尿囊素中一种或两种;
进一步地,其中所述保湿剂为透明质酸和/或透明质酸盐;
进一步地,其中所述生物成膜剂为普鲁兰多糖。
本发明的第二特征在于提供上述生物微晶冻干粉的制作工艺,包含的具体步骤为:
(1)配料:预处理复合生物活性多肽溶液,经过0.22um微孔滤膜过滤除菌;
(2)灌装:分装到西林瓶中,放入冻干机内;
(3)预冻:开启冷冻干燥机,在4℃±1℃温度条件下,维持1-4h;降温至-20℃以下,保持冷冻干燥室温度-20±2℃,产品预冻6-8h;
(4)一次升华:开始抽真空,当真空度达到10Pa以下,进行升温;调节冷冻干燥室温度缓慢升至0.0℃,每小时升温不超过1.0℃,真空条件下保持25-30个小时;
(5)二次升华:继续调节冷冻干燥室温度,以3-10℃/h的速率升温至30-35℃,保温3-5个小时。
(6)出料:通入氮气、氦气或氩气,调节冷冻干燥室内为常压后压内塞,取出后钆盖,即得具有独特微晶表面,且含水量≤1%的生物微晶冻干粉。
本发明实施例提供的一种复合活性多肽生物微晶冻干粉制备方法,通过对其工艺进行详细的研究和改进,通过预冷、预冻和真空干燥处理,获得复合活性多肽生物微晶冻干粉,该方法工艺简单,易于进行放大生产,可提高复合生物活性多肽冻干粉的生物活性和稳定性,能够在冷藏或者常温下保存较长时间,通过增加预冷过渡期温度参数和预冻工艺中的均匀受冷,使得制备的冻干粉具有散射状微晶表面,极大的提高了冻干粉的外观性。
本发明实例提供的复合活性多肽生物微晶冻干粉,包含生物活性多肽、冻干保护剂、保湿剂和成膜剂,可以有效的与皮肤细胞发生作用,促进真皮层胶原蛋白增生、补充皮肤水分、清除体内自由基,增强肌肤的抗氧化力,防止皮肤受到紫外线侵害。因此,对皮肤具有抚平细纹,延缓皮肤老化,进一步达到抗衰老功能。
附图说明
从下面结合附图的详细描述中,本发明上述特征、目的和优点将会更明显:
图1为实施例3复合活性多肽生物微晶冻干粉外观图片
图2为本发明实施例3所得的冻干曲线图
图3为本发明实施例中利用ELISA法检测活性多肽冻干粉在4℃保存条件下碱性成纤维细胞因子(bFGF)的活性对比图。
图4为本发明实施例中利用ELISA法检测活性多肽冻干粉在4℃保存条件下表皮生长因子(EGF)的活性对比图。
具体实施方式
下面的实施例用来说明本发明技术解决方案,并不是想限制本发明的范围。
实施例1:
含bFGF活性多肽混合液配方:
制备方法:
按配方量称取物料溶解于去离子水中,4℃过夜溶胀,通过0.22um微孔膜过滤除菌后,分装到西林瓶中,放入冻干机内。开启冷冻干燥机,在4℃±1℃温度条件下,维持2h;降温至-20℃以下,保持冷冻干燥室温度-20±2℃,产品预冻6h;开始抽真空,当真空度达到10Pa以下,进行升温;调节冷冻干燥室温度缓慢升至0.0℃,每小时升温不超过1.0℃,真空条件下保持28个小时;继续调节冷冻干燥室温度,以5℃/h的速率升温至35℃,保温3个小时。通入氮气、氦气或氩气,调节冷冻干燥室内为常压后压内塞,取出钆盖,即得bFGF活性多肽生物微晶冻干粉
对实施1所得bFGF活性多肽生物微晶冻干粉进行检测,结果见表1
表1 bFGF活性多肽生物微晶冻干粉检测结果
实施例2:
含EGF活性多肽混合液配方:
制备方法:
按配方量称取物料溶解于去离子水中,4℃过夜溶胀,通过0.22um微孔膜过滤除菌后,分装到西林瓶中,放入冻干机内。开启冷冻干燥机,在4℃±1℃温度条件下,维持3h;降温至-20℃以下,保持冷冻干燥室温度-20±2℃,产品预冻7h;开始抽真空,当真空度达到10Pa以下,进行升温;调节冷冻干燥室温度缓慢升至0.0℃,每小时升温不超过1.0℃,真空条件下保持25个小时;继续调节冷冻干燥室温度,以3℃/h的速率升温至30℃,保温5个小时。通入氮气、氦气或氩气,调节冷冻干燥室内为常压后压内塞,取出钆盖,即得EGF活性多肽生物微晶冻干粉
对实施2所得EGF活性多肽生物微晶冻干粉进行检测,结果见表2
表2 EGF活性多肽生物微晶冻干粉检测结果
实施例3:
含复合活性多肽混合液配方:
制备方法:
按配方量称取物料溶解于去离子水中,4℃过夜溶胀,通过0.22um微孔膜过滤除菌后,分装到西林瓶中,放入冻干机内。开启冷冻干燥机,在4℃±1℃温度条件下,维持3h;降温至-20℃以下,保持冷冻干燥室温度-20±2℃,产品预冻7h;开始抽真空,当真空度达到10Pa以下,进行升温;调节冷冻干燥室温度缓慢升至0.0℃,每小时升温不超过1.0℃,真空条件下保持25个小时;继续调节冷冻干燥室温度,以10℃/h的速率升温至35℃,保温3个小时。通入氮气、氦气或氩气,调节冷冻干燥室内为常压后压内塞,并保存冻干曲线(如图2),取出钆盖,即得复合活性多肽生物微晶冻干粉
对实施3所得复合活性多肽生物微晶冻干粉进行检测,结果见表3
表3复合活性多肽生物微晶冻干粉检测结果
实施例4
分别将本发明实施例1冻干粉、实施例2冻干粉、实施例3冻干粉与市场上复合因子成品冻干粉,常规4℃保存不同期限测定细胞因子bFGF、EGF含量实验:
1)分别将所述四种干细胞试验品放置4℃冰箱中,放置时间为1个月、3个月、6个月和一年;
2)分别采用bFGF、EGF的ELISA试剂盒检测这四种产品不同保存条件下细胞因子的含量;实验结果如附图3与图4所示,表明在4℃保存条件下,本发明冻干粉相较于其他细胞因子产品其活性保持时间更持久。现有技术中,多肽因子保存起来相对困难,因此,多肽因子冻干粉的保藏温度在-20℃及以下,本发明生产冻干粉在4℃保存也可保证其具有良好的生物活性。
以上为对本发明实施例的描述,所涉及的组分和工艺参数数值在上述实施例中不可能全部体现,需要理解的是,本发明并不局限于上述特定实施方式,本领域专业技术人员可以在不脱离本发明的精神范围或范围的情况下,做出各种变形或修改,这并不影响本发明的实质内容。因此,本发明将不会被限制于本文所示的这些实施例,凡在本发明的精神和原则之内,所作的任何修改、本发明各原料的等同替换、具体方式选择等,均应落在本发明的保护范围之内。
Claims (4)
1.一种复合活性多肽生物微晶冻干粉,其特征在于,所述复合活性多肽混合液包含如下重量百分比的各组分:0.01%-0.05%(m/v)活性多肽、5%-20%(m/v)冻干保护剂、0.1%-2.0%(m/v)皮肤调理剂、0.1%-0.3%(m/v)保湿剂、0.5%-1.0%生物成膜剂,余量为去离子水;
其中所述活性多肽为酸性成纤维细胞生长因子(aFGF)、碱性成纤维细胞生长因子(bFGF)、人表皮生长因子(EGF)、血管内皮生长因子(VEGF)、转化生长因子-β1(TGF-β1)、转化生长因子-β2(TGF-β2)和角质细胞生长因子(KGF)中的一种或多种;
所述冻干保护剂为海藻糖、甘露醇、甘氨酸、右旋糖酐、精氨酸中的一种或多种;
所述皮肤调理剂为胶原蛋白、尿囊素中一种或两种
所述保湿剂为透明质酸纳和/或透明质酸盐
所述生物成膜剂为普鲁兰多糖。
2.根据权利要求1所述的一种复合活性多肽生物微晶冻干粉,其特征在于,所述生物微晶冻干粉制备方法包括如下步骤:
(1)配料:预处理复合生物活性多肽溶液,经过0.22um微孔滤膜过滤除菌;
(2)灌装:分装到西林瓶中,放入冻干机内;
(3)预冻:开启冷冻干燥机,在4℃±1℃温度条件下,维持1-4h;降温至-20℃以下,保持冷冻干燥室温度-20±2℃,产品预冻6-8h。
(4)一次升华:开始抽真空,当真空度达到10Pa以下,进行升温;调节冷冻干燥室温度缓慢升至0.0℃,每小时升温不超过1.0℃,真空条件下保持25-30个小时;(5)二次升华:继续调节冷冻干燥室温度,以3-10℃/h的速率升温至30-35℃。保温3-5个小时。
(6)出料:通入氮气、氦气或氩气,调节冷冻干燥室内为常压后压内塞,取出钆盖,即得复合活性多肽生物微晶冻干粉。
3.根据权利要求1和2所述的一种复合活性多肽生物微晶冻干粉,其特征在于,所制备的冻干粉具有散射状微晶表面。
4.根据权利要求1和2所述的一种复合活性多肽生物微晶冻干粉,其特征在于,所制备的冻干粉含水量≤1%。
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