CN113520900A - 一种低致敏、抗衰老的牦牛胶原蛋白组合物及应用 - Google Patents
一种低致敏、抗衰老的牦牛胶原蛋白组合物及应用 Download PDFInfo
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Abstract
本发明属于皮肤医学领域,具体涉及一种低致敏、抗衰老的牦牛胶原蛋白复合物。本发明提供了一种低内毒素、低致敏的牦牛胶原蛋白,所述牦牛胶原蛋白细胞毒性低,能够促进细胞增殖和细胞黏附,具有良好的抗氧化和抗衰老等性能;同时本发明筛选获得了一种含有牦牛胶原蛋白与透明质酸、甘草酸二钾、乙酰基六肽‑8、鞣花酸、白柳树皮提取物、烟酰胺、维生素E、白藜芦醇的牦牛胶原蛋白组合物,所述牦牛胶原蛋白组合物具有低致敏性以及更优良的抗氧化、抗衰老性能,可广泛应用在化妆品或皮肤医学领域。
Description
技术领域
本发明属于皮肤医学领域,具体涉及一种低致敏、抗衰老的牦牛胶原蛋白组合物及应用。
背景技术
胶原蛋白是人体内含量最多的蛋白质,约占人体蛋白质总量的30%。胶原蛋白是细胞外基质的主要成分,也是皮肤的主要成分,广泛分布于人体的皮肤、血管、骨骼、肌腱、眼角膜等结缔组织。目前已经发现29种不同类型的胶原蛋白,其中人体皮肤真皮层中主要含有I型胶原蛋白,I型胶原蛋白组装形成的纤维支架为皮肤提供弹性和强度。
胶原蛋白作为皮肤的天然成分,已经在化妆品和皮肤修复敷料中广泛应用。胶原蛋白中的酪氨酸残基和皮肤中的酪氨酸进行对抗,和酪氨酸酶进行结合,可防止酪氨酸转化为多巴醌,有助于抑制黑色素产生和减少色素沉着。同时,胶原蛋白含有丰富的丙氨酸、丝氨酸、甘氨酸以及天门冬氨酸,它们是天然的亲水基团和保湿因子,具有良好的保湿功能,有助于促进皮肤血液循环和新陈代谢,加强皮肤的免疫力与抵抗力,建立皮肤的保护屏障,抑制皮肤过敏的症状。研究表明,由胶原蛋白溶液和无纺布制成的胶原蛋白贴敷料,可以为皮肤提供湿润的环境,修复皮肤屏障、促进伤口愈合。但是,目前很多市售的胶原蛋白产品存在内毒素含量较高的问题,它们具有较高的致敏性,在与人体皮肤接触后容易导致皮肤发生过敏反应,甚至诱发皮肤病。
面部衰老是人体机能变缓的直接表现,如何抗衰老是化妆品和皮肤医学的永恒主题。衰老的原因包括过度氧化、细胞寿命、蛋白质老化、内分泌功能减退、微循环障碍、荷尔蒙缺失等多个方面。其中,过度氧化是导致衰老的最重要因素之一。氧化应激是指机体在遭受各种有害刺激时,体内高活性分子如活性氧等自由基产生过多,氧化程度超出氧化物的清除,氧化系统和抗氧化系统失衡,从而导致损伤。O2-·是所有活性氧自由基前体,它与生物体炎症、衰老等症状密切相关;·OH可造成糖类、氨基酸、蛋白质、核酸和脂类等物质氧化损伤,使细胞坏死或突变,是目前所知活性氧自由基中对生物体毒性最强、危害最大的一种。因此,开发高效抗氧化的产品,是抗衰老的重要方式。
针对上述问题,本发明提供了一种低内毒素、低致敏的牦牛胶原蛋白,它不仅生物相容性好、细胞毒性低、可以有效促进细胞增殖和细胞黏附,而且具有良好的抗氧化和抗衰老等性能;同时,本发明筛选获得了一种由牦牛胶原蛋白与透明质酸、甘草酸二钾、乙酰基六肽-8、鞣花酸、白柳树皮提取物、烟酰胺、维生素E、白藜芦醇等成分组成的牦牛胶原蛋白组合物,它们具有低致敏性和更优良的抗氧化和抗衰老等性能,更适用于敏感肌肤和衰老肌肤,可广泛应用在化妆品或皮肤医学领域。
发明内容
本发明的目的在于提供一种低致敏、抗衰老的牦牛胶原蛋白组合物,具体包括以下内容:
第一方面,本发明提供了一种牦牛胶原蛋白在制备抗衰老化妆品或医学美容产品中的应用。
优选地,所述牦牛胶原蛋白的制备方法包括以下步骤:
(1)将牦牛筋腱预处理,经表面活性剂处理,和/或氧化剂处理,和/或消毒处理,和/或脱脂脱钙处理,得原料粉末;
(2)在步骤(1)所述的原料粉末中加入0.01-2.0M的氢氧化钠溶液,4-30℃,时间为2-24h;
(3)离心,弃上清,水洗沉淀至中性,采用酸溶酶解法提取获得酶解液;
(4)将步骤(3)所述酶解液灭活,盐析、透析,制得牦牛胶原蛋白。
优选地,所述化妆品包括原液、乳液、精华素、精华霜、爽肤水、调理水、面霜、面膜、隔离霜、眼霜、喷雾、防晒霜、粉底、身体乳。
优选地,所述医学美容产品包括原液、敷料、冷敷贴、凝胶、海绵、膜、水光针、乳、霜、膏。
第二方面,本发明提供了一种低致敏、抗衰老的牦牛胶原蛋白组合物,所述组合物包括以下质量百分比的组分:牦牛胶原蛋白1-5%,甘草酸二钾0.1-1.0%,白柳树皮提取物0.5-5%,鞣花酸0.5-3%,白藜芦醇0.5-3%,乙酰基六肽-8 0.1-0.3%、甘油1-3%,维生素E 0.1-1.0%,透明质酸0.5-1.0%,羟苯甲酯0.05-0.2%,羟苯丙酯0.01-0.14%。
优选地,所述牦牛胶原蛋白的制备方法包括以下步骤:
(1)将牦牛筋腱预处理,经表面活性剂处理,和/或氧化剂处理,和/或消毒处理,和/或脱脂脱钙处理,得原料粉末;
(2)在步骤(1)所述的原料粉末中加入0.01-2.0M的氢氧化钠溶液,4-30℃,时间为2-24h;
(3)离心,弃上清,水洗沉淀至中性,采用酸溶酶解法提取获得酶解液;
(4)将步骤(3)所述酶解液灭活,盐析、透析,制得牦牛胶原蛋白。
优选地,所述组合物包括以下质量百分比的组分:牦牛胶原蛋白1%,甘草酸二钾0.1%,白柳树皮提取物0.5%,鞣花酸0.5%,白藜芦醇0.5%,乙酰基六肽-8 0.1%、甘油1%,维生素E 0.1%,透明质酸0.5%,羟苯甲酯0.05%,羟苯丙酯0.01%,余量为水。
优选地,所述组合物包括以下质量百分比的组分:牦牛胶原蛋白3%,甘草酸二钾0.5%,白柳树皮提取物3%,鞣花酸1.5%,白藜芦醇1.5%,乙酰基六肽-8 0.2%、甘油2%,维生素E 0.5%,透明质酸0.8%,羟苯甲酯0.1%,羟苯丙酯0.1%。
优选地,所述组合物包括以下质量百分比的组分:牦牛胶原蛋白5%,甘草酸二钾1.0%,白柳树皮提取物5%,鞣花酸3%,白藜芦醇3%,乙酰基六肽-8 0.3%、甘油3%,维生素E1.0%,透明质酸1.0%,羟苯甲酯0.2%,羟苯丙酯0.14%。
第三方面,本发明提供了一种上述第二方面所述的组合物在制备抗衰老化妆品或医学美容产品中的应用。
优选地,所述化妆品包括原液、乳液、精华素、精华霜、爽肤水、调理水、面霜、面膜、隔离霜、眼霜、喷雾、防晒霜、粉底、身体乳。
优选地,所述医学美容产品包括原液、敷料、冷敷贴、凝胶、海绵、膜、水光针、乳、霜、膏。
本发明的有益效果是:
(1)本发明提供了一种低内毒素、低致敏的牦牛胶原蛋白,所述牦牛胶原蛋白具有生物相容性好、细胞毒性低、可促进细胞增殖和细胞黏附等优良性质,而且具有良好的抗氧化和抗衰老等性能;
(2)本发明筛选获得了一种由牦牛胶原蛋白与透明质酸、甘草酸二钾、乙酰基六肽-8、鞣花酸、白柳树皮提取物、烟酰胺、维生素E、白藜芦醇等成分组成的牦牛胶原蛋白组合物,所述牦牛胶原蛋白组合物具有低致敏性和更优良的抗氧化和抗衰老等性能;
(3)本发明所述的牦牛胶原蛋白及其组合物,可广泛应用于化妆品、皮肤医学中的抗衰老产品,尤其适用于敏感肌肤。
附图说明
图1本发明实施例1制备的牦牛胶原蛋白的SDS-PAGE图;
图2本发明实施例1制备的牦牛胶原蛋白的圆二色谱图;
图3本发明实施例1的牦牛胶原蛋白细胞毒性结果;
图4本发明实施例1的牦牛胶原蛋白细胞增殖情况;
图5本发明实施例1的牦牛胶原蛋白细胞黏附性能;
图6本发明实施例1的牦牛胶原蛋白的DPPH自由基清除率;
图7本发明实施例1的牦牛胶原蛋白的AAPH自由基清除率;
图8本发明实施例4的牦牛胶原蛋白组合物的DPPH自由基清除率;
图9本发明实施例4的牦牛胶原蛋白组合物的AAPH自由基清除率;
图10本发明实施例4的牦牛胶原蛋白组合物的羟自由基清除率。
具体实施方式
以下具体结合实施例进一步描述本发明的技术方案,但本发明的保护范围不局限于以下所述。
实施例1牦牛胶原蛋白的制备
1.制备方法
将牦牛筋腱进行清洗,去除异物,粉碎成小块;用1%洗必泰和5%过氧化氢对粉粹后的小块进行处理,取沉淀,水洗至中性;将得到的沉淀用10%正丁醇进行脱脂,取沉淀,水洗至中性;将前步中得到的沉淀用0.5M盐酸进行脱钙,脱钙结束,取沉淀,水洗至中性;将前步中得到的组织沉淀浸泡于0.5M的氢氧化钠溶液中,10℃搅拌处理2h,取沉淀,水洗至中性;用含胃蛋白酶的0.5M乙酸溶液提取胶原蛋白,得到粗的胶原蛋白提取液;调pH至中性,进行酶灭活;用透析袋进行透析,透析结束后,冷冻干燥,最终得到低内毒素含量的牦牛胶原蛋白。
实施例2牦牛胶原蛋白的质量评价
1.内毒素含量测定
采用ToxinSensorTM显色法LAL内毒素检测试剂盒,并按试剂盒说明书所述的方法,得到内毒素含量的标准曲线;对牦牛胶原蛋白提取过程中各操作步骤后的内毒素含量进行测定,该测定方法灵敏可靠,测定结果见下表1。
表1牦牛胶原蛋白制备各步骤内毒素含量检测结果
从表1可以看出:以本发明所述的方法提取制备的牦牛胶原蛋白,能够显著降低牦牛胶原蛋白产品中内毒素的含量,使牦牛胶原蛋白产品中内毒素的含量低于检测限,是一种生物安全性好、低致敏的牦牛胶原蛋白。
2.胶原蛋白性质鉴定
凝胶电泳实验:采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法对上述制备的牦牛胶原蛋白样品进行鉴别分析。
上述制备的牦牛胶原蛋白的凝胶电泳图如图1所示,在分子量为100kDa附近有两条α带,分别为α1和α2;在分子量为200kDa附近有两条β带,分别为β11和β12,它们是两条α链形成的二聚体肽链;在低分子量处没有多余的条带,表明本发明制备的牦牛胶原蛋白具有完整的α链,完全没有发生降解。
3.胶原蛋白结构鉴定
圆二色谱实验:圆二色谱是用于表征蛋白质结构的常用方法,其中225nm附近的正峰和205nm附近的负峰,是胶原蛋白三重螺旋结构的特征峰。
本发明制备的牦牛胶原蛋白的圆二色谱结果如图2a所示,它在225nm左右具有正峰,在205nm左右具有负峰,符合胶原蛋白三重螺旋结构的特征;同时,对牦牛胶原蛋白的热变温度进行了测定,由图2b可知,牦牛胶原蛋白的热变温度为37℃,符合天然胶原蛋白的特点。
4.细胞毒性实验
将完全贴壁生长的HeLa细胞用0.25%胰酶消化,用完全培养液(1%FBS,2%Penicillin–Streptomycin,DMEM培养基)配制成细胞密度为1×105个/mL的细胞悬液。分别移取100μL细胞悬液接种于96孔培养板中并置于37℃、5%CO2饱和湿度的培养箱中培养24h。吸出完全培养液,在实验组中加入高糖DMEM培养基稀释的不同浓度的上述制备的牦牛胶原蛋白溶液(0.001-0.1mg/ml,每个浓度设置4个复孔),对照组为DMEM培养基培养的细胞,空白组为不含细胞的DMEM培养基,继续于37℃,5%CO2饱和湿度的培养箱中培养24h。向各组加入CCK-8试剂10μL,细胞培养箱内孵育1-4h,使用酶联免疫检测仪在450nm波长下测各孔吸光度值(OD值)。根据各组的吸光度均值按照如下公式计算细胞存活率:
实验结果如图3所示,上述制备的不同浓度牦牛胶原蛋白均无细胞毒性,安全性良好。
5.细胞增殖实验
取生长至培养皿面积70%-80%的人皮肤成纤维细胞(HFF-1),0.25%胰酶消化,用完全培养液制成细胞密度为1×105个/mL的细胞悬液。取100μL细胞悬液接种于96孔培养板中并置于37℃,5%CO2饱和湿度的培养箱中培养。细胞培养24h后,吸出完全培养液。在实验组中加入高糖DMEM培养基稀释的0.1mg/ml上述制备的牦牛胶原蛋白溶液(设置4个复孔),对照组为DMEM培养基培养的细胞,空白组为不含细胞的DMEM培养基,继续于37℃,5%CO2饱和湿度的培养箱中分别培养24h、48h、72h和96h。每隔一段时间,向各组加入CCK-8试剂10μL,细胞培养箱内孵育1-4h,使用酶联免疫检测仪在450nm波长下测各孔吸光度值(OD值)。根据各组的吸光度均值按照如下公式计算细胞的相对增殖率(RGR):
实验结果如图4所示,与对照组(100%)相比,0.1mg/ml牦牛胶原蛋白实验组在第1天、第2天、第3天、第4天的细胞相对增殖率均大于100%,且随着时间的增加而呈增加的趋势,表明上述制备的牦牛胶原蛋白可以促进人皮肤成纤维细胞的增殖。
6.细胞黏附实验
用PBS将牦牛胶原蛋白稀释至0.1mg/mL,选用未TC处理的24孔细胞培养板,分别用1%热变性BSA,0.1mg/mL上述制备的牦牛胶原蛋白处理孔板,每孔500μL样品,并于4℃冰箱中孵育24h。将生长至培养皿面积70%-80%的小鼠成纤维细胞(L929)用高糖DMEM培养基制成细胞密度为3×105个/mL的细胞悬液。将24孔板中的液体吸出,每孔加入500μL细胞悬液。5h后通过倒置显微镜观察细胞黏附效果。
实验结果如图5所示,在牛血清白蛋白(BSA)中生长的成纤维细胞呈圆形、未铺展的状态,而在上述制备的牦牛胶原蛋白中生长的成纤维细胞大多呈铺展、黏附的状态,表明上述制备的牦牛胶原蛋白具有良好的细胞黏附性能。
实施例3牦牛胶原蛋白抗氧化实验
1.DPPH自由基清除实验过程
sample blank样品:分别取2ml不同浓度的实施例1制备的牦牛胶原蛋白,加入2ml甲醇,制备Sample blank样品;
control样品:取2ml溶解的DPPH甲醇溶液加入2ml纯化水,制备control样品;
参比溶液为甲醇水溶液(甲醇:水=1:1);
将2ml DPPH甲醇溶液与2ml不同浓度的牦牛胶原蛋白溶液混合,制备sample test样品,剧烈搅拌1分钟后,将上述各反应混合物在黑暗中于28℃保持30分钟;然后使用紫外可见分光光度计在517nm下测量样品和各组对照的吸光度,并按以下公式计算DPPH自由基清除率:
DPPH自由基清除率(%)={1-(Asample test-Asample blank)/Acontrol}×100。
DPPH法于1958年被提出,已经广泛用于定量测定抗氧化能力。DPPH是一种很稳定的氮中心的自由基,它的稳定性主要来自3个苯环的π-π共轭作用及空间障碍,使夹在中间的氮原子上不成对的电子不能发挥其应有的电子成对作用,而抗氧化剂(或称自由基清除剂)能使单电子配对,从而使A517nm值降低,溶液褪色。牦牛胶原蛋白清除DPPH自由基的能力结果如图6所示,本发明制备的牦牛胶原蛋白能够显著清除DPPH自由基,表明它具有良好的抗氧化和抗衰老功效。
2.AAPH自由基清除实验过程
sample blank样品:将2.0ml 60mM邻苯三酚红(PGR)与1.6ml样品(实施例1制备的不同浓度的牦牛胶原蛋白)和0.4ml去离子水混合;
sample test样品:将2.0ml 60mM邻苯三酚红(PGR)与1.6ml样品(实施例1制备的不同浓度的牦牛胶原蛋白)和0.4ml 600mM的AAPH混合;
control test样品:将2.0ml 60mM邻苯三酚红(PGR)与1.6ml样品缓冲液和0.4ml600mM的AAPH混合;
空白对照即control样品:将2.0ml 60mM邻苯三酚红(PGR)与1.6ml样品缓冲液和0.4ml去离子水混合;
将上述各组样品在37℃的水浴中孵育2小时。温育后立即将反应混合物冷却,并在540nm处测量吸光度。清除率的计算如下:
AAPH(偶氮二异丁脒盐酸盐)是水溶液中最常用的自由基源,牦牛胶原蛋白对于AAPH自由基的清除结果如图7所示,本发明制备的牦牛胶原蛋白能够显著清除AAPH自由基,表明它具有良好的抗氧化和抗衰老功效。
实施例4牦牛胶原蛋白组合物的制备
牦牛胶原蛋白组合物1,按质量比由以下组分制成:牦牛胶原蛋白1%,甘草酸二钾0.1%,白柳树皮提取物0.5%,鞣花酸0.5%,白藜芦醇0.5%,乙酰基六肽-8 0.1%、甘油1%,维生素E 0.1%,透明质酸0.5%,羟苯甲酯0.05%,羟苯丙酯0.01%,余量为水。
牦牛胶原蛋白组合物2,按质量比由以下组分制成:牦牛胶原蛋白3%,甘草酸二钾0.5%,白柳树皮提取物3%,鞣花酸1.5%,白藜芦醇1.5%,乙酰基六肽-8 0.2%、甘油2%,维生素E 0.5%,透明质酸0.8%,羟苯甲酯0.1%,羟苯丙酯0.1%,余量为水。
牦牛胶原蛋白组合物3,按质量比由以下组分制成:牦牛胶原蛋白5%,甘草酸二钾1.0%,白柳树皮提取物5%,鞣花酸3%,白藜芦醇3%,乙酰基六肽-8 0.3%、甘油3%,维生素E1.0%,透明质酸1.0%,羟苯甲酯0.2%,羟苯丙酯0.14%,余量为水。
制备过程:将A相物质(成分:水、甘油、透明质酸)加热至85度,搅拌至完全溶解后,均质3min,保温10-15min;预溶B相物质(成分:白藜芦醇、鞣花酸、维生素E)、C相物质(成分:甘草酸二钾、牦牛胶原蛋白、乙酰基六肽-8、白柳树皮提取物、烟酰胺)、D相物质(成分:羟苯甲酯、羟苯丙酯);降温至40度加入B相物质,再降温至35度加入C相物质和D相物质,搅拌至完全溶解透明。降温至30度,用洁净滤网过滤,出料静置。分别得到牦牛胶原蛋白液体组合物1、组合物2、组合物3。
实施例5牦牛胶原蛋白组合物抗氧化实验
1.DPPH自由基清除实验
sample样品:分别取2ml不同浓度的实施例4制备的牦牛胶原蛋白组合物1-3,加入2ml DPPH甲醇;
其他操作同实施例3中1所述。
牦牛胶原蛋白组合物清除DPPH自由基的能力结果如图8所示,本发明制备的牦牛胶原蛋白组合物能够显著清除DPPH自由基,组合物1清除率高于90%,组合物2和组合物3清除率均高于95%,表明本发明制备的牦牛胶原蛋白组合物具有优异的抗氧化和抗衰老功效。
2.AAPH自由基清除实验
sample test样品:将2.0ml 60mM邻苯三酚红(PGR)与1.6ml实施例4制备的牦牛胶原蛋白组合物1-3和0.4ml 600mM的AAPH混合;
其他操作同实施例3中2所述。
牦牛胶原蛋白组合物对于AAPH自由基的清除结果如图9所示,本发明制备的牦牛胶原蛋白组合物能够显著清除AAPH自由基,其清除率均高于95%,表明它具有优异的抗氧化和抗衰老功效。
3.羟基自由基清除实验
A0的测定:在比色管中依次加入1ml 9mmol/L FeSO4,1ml 9mmol/L乙醇-水杨酸,接着加入12ml去离子水,最后加入1ml 8.8mmol/L H2O2后摇匀,37℃加热15min后取出,测其吸光度即为A0。其中参比溶液为不加双氧水的体系。
Ax的测定:在比色管中依次加入1ml 9mmol/L FeSO4,1ml 9mmol/L乙醇-水杨酸,接着加入6ml待测样品,6ml去离子水,最后加入1ml 8.8mmol/L H2O2后摇匀,37℃加热15min后取出,测其吸光度即为Ax。其中参比溶液为去离子水。
Ax0的测定:在比色管中依次加入1ml 9mmol/L FeSO4,1ml 9mmol/L乙醇-水杨酸,接着加入6ml待测样品,再加入7ml去离子水摇匀,37℃加热15min后取出,测其吸光度即为Ax0。其中参比溶液为去离子水。
羟自由基的清除率由以下公式计算:
羟自由基清除率(%)=(A0+Ax0-Ax)/A0×100。
羟基自由基是一种重要的活性氧,从分子式上看是由氢氧根(OH-)失去一个电子形成,它具有极强的得电子能力和氧化能力。牦牛胶原蛋白组合物对于羟基自由基的清除结果如图10所示,本发明制备的牦牛胶原蛋白组合物能够显著清除羟基自由基,其清除率均高于80%,表明它具有优异的抗氧化和抗衰老功效。
Claims (9)
1.牦牛胶原蛋白在制备抗衰老化妆品或医学美容产品中的应用。
2.如权利要求1所述的应用,其特征在于,所述牦牛胶原蛋白的制备方法包括以下步骤:
(1)将牦牛筋腱预处理,经表面活性剂处理,和/或氧化剂处理,和/或消毒处理,和/或脱脂脱钙处理,得原料粉末;
(2)在步骤(1)所述的原料粉末中加入0.01-2.0M的氢氧化钠溶液,4-30℃,时间为2-24h;
(3)离心,弃上清,水洗沉淀至中性,采用酸溶酶解法提取获得酶解液;
(4)将步骤(3)所述酶解液灭活,盐析、透析,制得牦牛胶原蛋白。
3.如权利要求1或2所述的应用,其特征在于,所述化妆品包括原液、乳液、精华素、精华霜、爽肤水、调理水、面霜、面膜、隔离霜、眼霜、喷雾、防晒霜、粉底、身体乳。
4.如权利要求1或2所述的应用,其特征在于,所述医学美容产品包括原液、敷料、冷敷贴、凝胶、海绵、膜、水光针、乳、霜、膏。
5.一种低致敏、抗衰老的牦牛胶原蛋白组合物,其特征在于,所述组合物包括以下质量百分比的组分:牦牛胶原蛋白1-5%,甘草酸二钾0.1-1.0%,白柳树皮提取物0.5-5%,鞣花酸0.5-3%,白藜芦醇0.5-3%,乙酰基六肽-8 0.1-0.3%、甘油1-3%,维生素E 0.1-1.0%,透明质酸0.5-1.0%,羟苯甲酯0.05-0.2%,羟苯丙酯0.01-0.14%。
6.如权利要求5所述的组合物,其特征在于,所述牦牛胶原蛋白的制备方法包括以下步骤:
(1)将牦牛筋腱预处理,经表面活性剂处理,和/或氧化剂处理,和/或消毒处理,和/或脱脂脱钙处理,得原料粉末;
(2)在步骤(1)所述的原料粉末中加入0.01-2.0M的氢氧化钠溶液,4-30℃,时间为2-24h;
(3)除去碱溶液,水洗至中性,采用酸溶酶解法提取得酶解液;
(4)将步骤(3)所述酶解液灭活,盐析、透析、冷冻干燥,制得牦牛胶原蛋白。
7.如权利要求书5或6所述的组合物在制备抗衰老化妆品或医学美容产品中的应用。
8.如权利要求7所述的应用,其特征在于,所述化妆品包括原液、乳液、精华素、精华霜、爽肤水、调理水、面霜、面膜、隔离霜、眼霜、喷雾、防晒霜、粉底、身体乳。
9.如权利要求7所述的应用,其特征在于,所述医学美容产品包括原液、敷料、冷敷贴、凝胶、海绵、膜、水光针、乳、霜、膏。
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