CN112842950A - 乳酸菌发酵的桦树汁及其在具有抗环境压力功效的化妆品组合物中的应用 - Google Patents
乳酸菌发酵的桦树汁及其在具有抗环境压力功效的化妆品组合物中的应用 Download PDFInfo
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- CN112842950A CN112842950A CN201911189098.9A CN201911189098A CN112842950A CN 112842950 A CN112842950 A CN 112842950A CN 201911189098 A CN201911189098 A CN 201911189098A CN 112842950 A CN112842950 A CN 112842950A
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- lactic acid
- acid bacteria
- lactobacillus
- birch juice
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Abstract
本发明涉及一种生产乳酸菌发酵的桦树汁的方法,其包括在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵的步骤。本发明还涉及通过上述方法获得的乳酸菌发酵的桦树汁及其在具有抗环境压力功效的化妆品组合物中的应用。
Description
技术领域
本发明涉及一种生产乳酸菌发酵的桦树汁的方法,其包括在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵的步骤。本发明还涉及通过上述方法获得的乳酸菌发酵的桦树汁及其在具有抗环境压力功效的化妆品组合物中的应用。
背景技术
桦树为桦木科落叶乔木,桦树汁(也称桦树液)是桦树树皮被划开或树干钻孔流出的新鲜汁液,内含糖类、氨基酸、维生素、生物素、微量的矿质元素、芳香油、桦树醇、皂角甙等化合物,具有良好的保湿、抗炎、抗皱、美白等护肤功效。
发酵技术是指通过微生物的生长繁殖及代谢活动,产生和积累人们所需要的产物的工业技术。据报道,植物提取物通过发酵工艺后可以有提升功效、降低毒性、节省能源等优势。在现有技术中,存在对乳酸菌发酵滤液或菌体应用于食品或皮肤外用品中的研究。但目前尚未见有关将乳酸菌发酵的桦树汁用于化妆品领域的研究报道。
人体皮肤暴露在地球环境中,温度、湿度、紫外线的照射量等的改变都会给皮肤带来一定的压力。工作上的出差、平日的旅游都会让皮肤处于不同的环境压力下。随着工业化的进行,人们居住环境质量却日益低下,例如臭氧层的减少带来更多的紫外线照射,工业活动等带来PM2.5的细颗粒物的增多,汽车的增加带来尾气的污染,工作环境的二手烟刺激和电脑蓝光照射等,都给在暴露在其中的皮肤带来了更大的环境压力。而上述这些环境压力会诱发皮肤出现屏障受损、敏感频发、暗沉发黄、细纹增多、痤疮加重、肤质粗糙干燥、角质更新紊乱等一系列问题。
不利生长的环境,例如过高或过的低氧环境、过高或过低的渗透压环境、过高或过低的温度、过高或过低的压强、过高或过低的pH环境等都会影响微生物的生长。但在长期演化中,微生物处于不利环境下会激发抗逆基因的表达,通过更多的抗逆机制产生平时正常环境中不会生成的热激蛋白、次级代谢产物等来适应逆环境,从而正常生长。在现有技术中,有将逆环境生长的微生物代谢产物应用在农业生产中的研究,结果发现也能提高农作物的抗逆性,增加农作物产量。
为拓宽发酵的桦树汁的应用,本发明人采用乳酸菌为菌种,以桦树汁为主要底物,任选地向其中添加促生长成分,例如乳制品,在高氧环境中进行发酵,所得乳酸菌发酵的桦树汁具有优异的护肤功效,尤其是抗环境压力的功效。
发明内容
本发明人经研究发现,采用乳酸菌作为菌种,桦树汁作为主要底物,任选地向其中添加乳制品,在高氧环境中进行发酵,可以得到一种具有改进性能的乳酸菌发酵的桦树汁,其在具有桦树汁本身活性成分的同时还富含乳酸菌细胞成分和发酵工艺产生的活性成分,例如乳酸、多糖、多酚和多肽等,从而具有优异的抗炎、镇静舒缓、修复等抗环境压力的功效,可作为皮肤外用组合物、尤其是具有抗环境压力功效的化妆品组合物中的活性原料。
一方面,本发明涉及一种生产乳酸菌发酵的桦树汁的方法,其包括在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵的步骤。
所述方法还包括过滤发酵中得到的发酵液,得到作为上清液的发酵的桦树汁滤液(即,作为产物的“乳酸菌发酵的桦树汁),和得到乳酸菌菌体副产物。
进一步地,所述方法还包括破碎所得到的乳酸菌菌体,然后进行过滤,得到作为上清液的可溶性乳酸菌溶胞物,和将所得乳酸菌溶胞物与所述发酵的桦树汁滤液混合,过滤,得到发酵的桦树汁滤液产物(即作为优选产物的“乳酸菌发酵的桦树汁”)。
在一个优选的实施方案中,所述方法包括下述步骤:
(1)在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵,获得发酵液产物;
(2)过滤所述发酵液产物,分别得到乳酸菌菌体和发酵的桦树汁滤液;
(3)破碎所述乳酸菌菌体,然后进行过滤,得到作为上清液的可溶性乳酸菌溶胞物;和
(4)将所得乳酸菌溶胞物与所述发酵的桦树汁滤液混合,过滤,得到发酵的桦树汁滤液产物(即作为优选产物的“乳酸菌发酵的桦树汁”)。
本发明中所采用的乳酸菌包括乳杆菌属(Lactobacillus)、双歧杆菌属(Bifidobacterium)以及选自乳球菌属(Lactococcus)、链球菌属(Streptococcus)、片球菌属(Pediococcus)、明串珠菌属(Leuconostoc)其它类别乳酸菌等。所述乳杆菌属(Lactobacillus)包括但不限于嗜酸乳杆菌、干酪乳杆菌、鼠李糖乳杆菌、短乳杆菌、卷曲乳杆菌、德氏乳杆菌(保加利亚亚种、乳亚种)、发酵乳杆菌、瑞士乳杆菌、鸡乳杆菌、加氏乳杆菌、约氏乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、唾液乳杆菌、消化乳杆菌、弯曲乳杆菌、干酪乳杆菌干酪亚种和清酒乳杆菌。所述双歧杆菌属(Bifidobacterium)包括但不限于青春双歧杆菌、动物双歧杆菌、两岐双岐杆菌、短双歧杆菌、乳双歧杆菌、长双歧杆菌、婴儿双歧杆菌和假小链双歧杆菌。所述其他乳酸菌包括但不限于乳酸乳球菌(乳酸亚种、乳脂亚种、双乙酰亚种)、嗜热链球菌、乳酸链球菌、乳酸片球菌、戊糖片球菌、肠膜明串珠菌和酒明串珠菌等。优选地,所述乳酸菌包括嗜酸乳杆菌、干酪乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、动物双歧杆菌、长双歧杆菌和婴儿双歧杆菌。所述乳酸菌可以菌粉形式商购,例如商购于杜邦公司、科汉森公司、荷兰皇家帝斯曼集团、北京科拓恒通生物技术股份有限公司等。
本发明中所涉及的桦树汁得自桦木科桦树属,其可来自白桦(Betula alba)、柔毛桦(Betula pubescens)、垂枝桦(Betula Pendula)和亚洲白桦(Betula platyphylla)等品种。所述桦树汁为在解冻至早春发叶之间,人工在桦树的树干基部钻孔收集而得的无色透明、无沉淀、无杂物,具有桦树清香营养丰富的汁液。所述桦树汁可商购获得并原样采用,例如可购自大兴安岭超越野生浆果开发有限责任公司。
本发明中可采用的桦树汁为桦树汁原液或浓缩的桦树汁(也称为桦树汁浓缩液),其中浓缩的桦树汁的浓缩倍数为约1.05-10.0倍,优选约1.5-6.0倍。
所述浓缩的桦树汁是通过将商购获得的桦树汁产品进行浓缩得到的。浓缩方法是本领域已知的,例如加热浓缩、低温真空浓缩、膜浓缩等。在本发明中,优选通过低温冷冻浓缩或膜浓缩工艺进行浓缩,例如,将商购的桦树汁原液输入低温干燥设备,降温至约-40℃至-70℃,抽真空至约0.1-30Pa而进行低温真空浓缩,从而得到不同浓缩倍数的浓缩桦树汁。
上述步骤(1)发酵是本领域已知的。例如,按照约60-80%(v/v)的装液量,将桦树汁培养基在约95-140℃超高温瞬时杀菌约4-30秒后加入约100-20000升已经空消无菌状态的发酵罐中,或将桦树汁培养基加入约100-20000升发酵罐中,并以约95-121℃灭菌约10-30分钟。按发酵罐中培养基体积计将乳酸菌直投式菌剂或乳酸菌种子液在无菌条件下接入发酵罐中,在约30-45℃温度和约50-350rpm的转速下进行搅拌,同时通入空气以控制整个培养基中的相对溶解氧含量为约20-200%,优选50-120%,连续发酵约24-168小时,停罐,即完成发酵过程,得到发酵液产物。
其中,上述乳酸菌直投式菌剂可直接投入发酵罐中。上述乳酸菌种子液是通过使菌粉活化培养获得的。菌粉活化培养是本领域已知的,例如,按照最终发酵体积的约1/50-1/10制备菌种活化步骤的无菌桦树汁种子液培养基;称取约0.1-10g/L、优选约0.5-5g/L以无菌桦树汁种子液培养基计的相应质量的乳酸菌菌粉,将其加入到种子液培养基中,在约30-45℃下培养约4-12小时,即得到所述乳酸菌种子液。
上述桦树汁培养基可以采用桦树汁、尤其是浓缩的桦树汁,例如约1.5-6.0倍的浓缩桦树汁作为底物来制备。桦树汁在所述桦树汁培养基中的含量在约90%以上,优选约94%以上,基于所述桦树汁培养基的总重量。
优选地,可将作为碳源、氮源的乳制品加入桦树汁培养基中。所述乳制品包括但不限于液态乳、乳粉、乳加工产品和乳加工副产物等。所述液态乳包括例如巴氏杀菌乳、灭菌乳、调制乳和复原乳等。所述乳粉包括例如全脂乳粉、低脂乳粉、脱脂乳粉和羊奶粉等。所述乳加工产品包括例如稀奶油、淡炼乳、干酪、乳酪粉和乳扇等。所述乳加工副产物包括例如乳糖、酪蛋白、水解牛奶蛋白、浓缩乳清蛋白、分离乳清蛋白和乳清蛋白肽等。优选地,所述乳制品包括脱脂乳粉、乳糖和乳清蛋白(浓缩乳清蛋白、分离乳清蛋白、乳清蛋白肽)。所述乳制品在所述桦树汁培养基中的含量通常为约0-8%,优选约1-5%,基于所述桦树汁培养基的总重量。
进一步地,可将pH调节剂加入到桦树汁培养基中,以将桦树汁培养基的pH调节到5.0-7.0。所述pH调节剂是本领域已知的,例如,包括但不限于乳酸、柠檬酸、氨水、乳酸钠、柠檬酸钠和氢氧化钠,优选柠檬酸钠。
此外,还可以将促进乳酸菌生长的无机盐加入到培养基中,其实例包括但不限于磷酸二氢钾、磷酸二氢钠、磷酸氢二钾、磷酸氢二钠、柠檬酸氢二铵、乙酸钠、硫酸锰、硫酸镁、硝酸钾等。所述无机盐的用量是本领域已知的,通常为约0-2%,优选约0.2-1%,基于所述桦树汁培养基的总重量。
在包含乳制品、pH调节剂和/或无机盐的情况下,将这些物质加入桦树汁中即可制备桦树汁培养基。
上述步骤(2)过滤所得发酵产物是本领域已知的。通常,采用碟式离心机或管式离心机,在约6000-15000rpm下,进行约0.5-15分钟。或者,可通过陶瓷膜或有机膜等进行膜分离过滤。所述过滤步骤将发酵产物中的乳酸菌菌体和发酵的桦树汁滤液(上清液)分开。
上述步骤(3)破碎所得乳酸菌菌体是本领域已知的。可采用的方式包括且不限于高压破碎法、反复冻融法、酶解法等。例如,使用约3-5倍质量的发酵桦树汁滤液将所得乳酸菌菌体重悬,得到乳酸菌菌体的重悬液,然后采用高压细胞破碎机,在约1-2升/分钟的流速和约800-1200巴的压力下,均匀处理所述重悬液,直至乳酸菌菌体的破碎率达到约98%以上。或者,也可将菌体重悬液在约零下10-20℃冻结,在约30-50℃下复溶,反复5-10次。或者,也可采用溶菌酶、胰蛋白酶等按约0.2-0.8%添加至重悬液中,基于重悬液的体积,在酶适宜温度下处理约2-5小时。然后,将经破碎的乳酸菌菌体重悬液进行过滤,通常在约10000-20000rpm下进行约10-30分钟,所得上清液即为可溶性乳酸菌溶胞物。或者,通过有机膜或陶瓷膜进行分离,分离出的清液即为可溶性乳酸菌溶胞物。
上述步骤(4)中,将步骤(3)获得的可溶性乳酸菌溶胞物与步骤(2)得到的发酵的桦树汁滤液混合均匀,然后进一步过滤,所得滤液即为乳酸菌发酵的桦树汁滤液产物。
所述方法可进一步包括将所得产物经超高温瞬时杀菌,其中杀菌温度为约95-140℃,时间为约4-30秒;然后,将杀菌过的产物转至存储罐存储,并分装灌装。
通过上述方法获得的乳酸菌发酵的桦树汁色浅、透明,含有桦树汁底物自身的营养成分(其中包括B族维生素、微量元素、氨基酸、脂肪酸等),可溶性乳酸菌溶胞物的营养成分(其中包括多糖、氨基酸、多肽、蛋白、核苷酸等),还有发酵新生成的功效成分(其中包括乳酸、多糖、水解小分子肽、氨基酸等)。通常,所得乳酸菌发酵的桦树汁包含0.2-10g/L的总酚,0.2-5g/L的多糖,0.1-10g/L的氨基酸和0.2-60g/L的多肽。因此,所得乳酸菌发酵的桦树汁含有丰富的活性营养物,可以作为原料营养物用于皮肤外用组合物中,尤其是具有抗环境压力功效的化妆品组合物中,其显示了优异的抗环境压力功效,从而减少了对皮肤的损害和延长了皮肤的衰老。
另一方面,本发明涉及一种乳酸菌发酵的桦树汁,其是通过在高氧环境中采用乳酸菌作为菌种在包含桦树汁和任选的乳制品的培养基中进行发酵而获得的。
通常,所得乳酸菌发酵的桦树汁包含0.2-10g/L的总酚,0.2-5g/L的多糖,0.1-10g/L的氨基酸和0.2-60g/L的多肽。
再一方面,本发明涉及乳酸菌发酵的桦树汁在具有抗环境压力功效的化妆品组合物中的用途。
又一方面,本发明涉及一种具有抗环境压力功效的化妆品组合物,其包含(A)乳酸菌发酵的桦树汁。
所述乳酸菌发酵的桦树汁在所述化妆品组合物中的含量可以在宽范围内变化,例如大于0至小于100%,优选约20-95%,基于所述化妆品组合物的总重量。
除了所述乳酸菌发酵的桦树汁外,所述化妆品组合物还可以任选地包含(B)化妆品组合物中常用的成分,其包括但不限于媒介物、活性成分和辅料等。组分(B)是本领域已知的,本领域技术人员可根据需要选择其类型和用量,例如,组分(B)的含量为约2-82%,基于所述化妆品组合物的总重量。
所述媒介物包括例如稀释剂、分散剂或载体等,其实例包括但不限于乙醇、双丙甘醇、丁二醇等。所述媒介物在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.5-20%。
所述活性成分包括例如润肤剂、保湿剂、抗氧化剂、美白活性成分、抗炎活性成分、抗湿疹活性成分、抗衰老活性成分、抗菌活性成分等。
所述润肤剂的实例包括但不限于橄榄油、澳洲坚果油、甜杏仁油、葡萄籽油、鳄梨油、玉米油、芝麻油、大豆油、花生油、白池花籽油、红花籽油、狗牙蔷薇果油、刺阿干树仁油、霍霍巴籽油、向日葵籽油、毛瑞榈果油、角鲨烷、棕榈酸乙基己酯、肉豆蔻酸异丙酯、氢化聚异丁烯、异十六烷、异十二烷、碳酸二乙基己酯、碳酸二辛酯、月桂酰肌氨酸异丙酯、异壬酸异壬酯、氢化聚癸烯、甘油三(乙基己酸)酯、鲸蜡醇乙基己酸酯、双-二乙氧基二甘醇环己烷1,4-二羧酸酯、辛酸/癸酸甘油三酯、油醇芥酸酯、辛基十二醇肉豆蔻酸酯、辛基十二醇、聚二甲基硅氧烷、辛基聚甲基硅氧烷、鲸蜡基聚二甲基硅氧烷、环五聚二甲基硅氧烷等的一种或多种。固体润肤剂的实例包括但不限于鲸蜡醇、硬脂醇、鲸蜡硬脂醇、山嵛醇、鲨肝醇、月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、蜂蜡、小烛树蜡、巴西棕榈蜡、羊毛脂、地蜡、霍霍巴籽蜡、石蜡、微晶蜡、氢化米糠蜡、氢化椰油甘油酯类、甘油山嵛酸酯/二十酸酯、肉豆蔻醇肉豆蔻酸酯、双-二甘油多酰基己二酸酯-2、牛油果树果脂、木鲁星果棕籽脂等中的一种或多种。所述润肤剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约1-50%。
所述保湿剂的实例包括但不限于甘油、双甘油、丁二醇、丙二醇、1,3-丙二醇、双丙甘醇、1,2-戊二醇、聚乙二醇-8、聚乙二醇-32、甲基葡糖醇聚醚-10、甲基葡糖醇聚醚-20、PEG/PPG-17/6共聚物、甘油聚醚-7、甘油聚醚-26、甘油葡糖苷、PPG-10甲基葡糖醚、PPG-20甲基葡糖醚、PEG/PPG/聚丁二醇-8/5/3甘油、蔗糖、海藻糖、鼠李糖、甘露糖、棉子糖、甜菜碱、赤藓醇、木糖醇、尿素、甘油聚醚-5乳酸酯、透明质酸钠、水解透明质酸钠、乙酰化透明质酸钠、聚谷氨酸钠、水解小核菌胶、出芽短梗酶多糖、银耳多糖、酸豆籽多糖等中的一种或多种。所述保湿剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约1-30%。
所述抗氧化剂包括但不限于白茶多酚、红茂草生物碱、山楂黄酮、辅酶Q10、葡萄籽提取物、芦笋提取物、萝卜提取物、左旋C原液、红花水提物、积雪草苷、白术多糖、维生素E及其衍生物等中的一种或多种。所述抗氧化剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的0.01-30%。
所述美白活性成分,包括但不限于曲酸、抗坏血酸葡糖苷、熊果苷、传明酸、烟酰胺、植物甾醇、植物甾醇/山嵛醇/辛基癸醇月桂酰谷氨酸酯、苯乙基间苯二酚、姜黄根提取物、白桦树皮提取物、神经酰胺2、神经酰胺3、乙酰植物鞘氨醇、白藜芦醇、花榈木树皮提取物、毛喉鞘蕊花根提取物、胡椒籽提取物、泛醌、胆甾醇、胆甾醇硬脂酸酯、抗坏血酸、抗坏血酸二棕榈酸酯、生育酚(维生素E)、生育酚乙酸酯、红没药醇、抗坏血酸四异棕榈酸酯、吡哆素二辛酸酯、吡哆素二棕榈酸酯、视黄醇棕榈酸酯、植物甾醇/辛基十二醇月桂酰谷氨酸酯、双-山嵛醇/异硬脂醇/植物甾醇二聚亚油醇二聚亚油酸酯、植物甾醇澳洲坚果油酸酯、各种肽类、各种植物提取物等中的一种或多种。所述美白成分在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.01-30%。
所述抗炎活性成分的实例包括但不限于甘草酸二钾、马齿苋(PORTULACAOLERACEA)提取物、燕麦(AVENA SATIVA)仁提取物(燕麦酰基邻氨基苯甲酸)、泛醇、尿囊素、生物糖胶-1、β-葡聚糖、果聚糖、黄芩(SCUTELLARIA BAICALENSIS)根提取物、欧洲七叶树(AESCULUS HIPPOCASTANUM)提取物、红没药醇、4-叔丁基环己醇、神经酰胺3、氢化卵磷脂、光果甘草(GLYCYRRHIZA GLABRA)提取物、水解蜂王浆蛋白、谷维素、植物鞘氨醇、五羟黄酮(槲皮素)、姜根提取物迷迭香叶提取物等中的一种或多种。所述抗炎活性成分在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的0.01-10%。
所述抗湿疹活性成分的实例包括但不限于糖皮质激素(包含所有强度的糖皮质激素)、氧化锌、炉甘石、硼酸溶液、盐酸小檗碱、依沙吖啶溶液、尿素、水杨酸、钙调神经磷酸酶抑制剂、水杨酸、乳酸、维A酸、焦油、维生素C、葡萄糖酸钙、硫酸锌铜等。所述抗湿疹活性成分在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的0.01-10%。
所述抗衰老活性成分的实例包括但不限于生育酚(维生素E)、视黄醇、视黄醇棕榈酸酯、水解胶原、水解弹性蛋白、尿囊素、酵母提取物、谷维素、四氢姜黄素、鞣花酸、泛醌、乳清蛋白、乙酰基六肽-8、棕榈酰五肽-4、水杨酰植物鞘氨醇、浓缩桦树汁、水飞蓟素、蚕丝胶蛋白、生育酚磷酸酯钠、核糖核酸(RNA)、二肽二氨基丁酰苄基酰胺二乙酸盐、棕榈酰三肽-5、寡肽-1、六肽-9、棕榈酰寡肽、棕榈酰四肽-7、葡萄(VITIS VINIFERA)籽提取物、花榈木(PTEROCARPUS MARSUPIUM)树皮提取物、茶(CAMELLIA SINENSIS)多酚、葡萄酒提取物、苹果籽提取物、欧洲水青冈(FAGUS SYLVATICA)芽提取物、水解猴面包树(ADANSONIA DIGITATA)提取物、卤虫(ARTEMIA)提取物、香根鸢尾(IRIS FLORENTINA)根提取物、橙皮苷、人参皂甙、丹参(SALVIA MILTIORRHIZA)提取物、烟酰胺、熊果酸、透明质酸钠、乙酰化透明质酸钠、水解透明质酸钠、番茄红素、咖啡(COFFEA ARABICA)提取物、二肽-2、乳酸、超氧化物歧化酶(SOD)、月见草(OENOTHERA BIENNIS)油、神经酰胺、二棕榈酰羟脯氨酸、羟基硬脂酸、水杨酸、麦角硫因、溶血卵磷脂、肌肽、脱羧肌肽HCL、硫辛酸、腺苷、糖原、白藜芦醇、阿魏酸、二裂酵母发酵产物溶胞物、乳酸菌发酵溶胞产物等中的一种或多种。所述抗衰老成分在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.01-10%。
所述抗菌活性成分包括但不限于熊果酸、白花蛇舌草黄酮、忍冬花提取物、茶树精油、甲壳素、桂枝提取物、珊瑚姜挥发油、丁香提取物、蘑菇提取物、芦荟提取物、艾叶提取物、1-十五醇及其衍生物、雪松烯、石竹烯、长叶烯、白桦树叶提取物、白桦树皮提取物等中的一种或多种。所述抗菌活性成分在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的0.01-30%。
所述辅料包括例如乳化剂、增稠剂、防腐剂、香料等。
所述乳化剂的实例包括但不限于鲸蜡硬脂醇橄榄油酸酯、山梨坦橄榄油酸酯、聚山梨醇酯-60、聚山梨醇酯-80、甲基葡糖倍半硬脂酸酯、PEG-20甲基葡糖倍半硬脂酸酯、PEG-40氢化蓖麻油、PPG-26-丁醇聚醚-26、PEG-4聚甘油-2硬脂酸酯、PEG-60氢化蓖麻油、硬脂醇聚醚-2、硬脂醇聚醚-21、PPG-13-癸基十四醇聚醚-24、鲸蜡硬脂基葡糖苷、PEG-100硬脂酸酯、甘油硬脂酸酯、甘油硬脂酸酯SE、椰油基葡糖苷、鲸蜡硬脂醇聚醚-25、PEG-40硬脂酸酯、聚甘油-3甲基葡糖二硬脂酸酯、甘油硬脂酸酯柠檬酸酯、聚甘油-10硬脂酸酯、聚甘油-10肉豆蔻酸酯、聚甘油-10二油酸酯、聚甘油-10月桂酸酯、聚甘油-10异硬脂酸酯、聚甘油-10油酸酯、聚甘油-10二异硬脂酸酯、聚甘油-6月桂酸酯、聚甘油-6肉豆蔻酸酯、蔗糖硬脂酸酯、蔗糖多硬脂酸酯等中的一种或多种。所述乳化剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.5-10%。
所述增稠剂的实例包括但不限于卡波姆类、丙烯酸(酯)类及其衍生物、黄原胶、阿拉伯胶、聚乙二醇-14M、聚乙二醇-90M、琥珀酰聚糖、羟乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素等高分子聚合物中的一种或多种。所述增稠剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.1-10%。
所述防腐剂的实例包括但不限于羟苯甲酯、羟苯丙酯、苯氧乙醇、苯甲醇、苯乙醇、双(羟甲基)咪唑烷基脲、山梨酸钾、苯甲酸钠、氯苯甘醚、脱氢乙酸钠、辛酰羟肟酸、1,2-己二醇、1,2-戊二醇、对羟基苯乙酮、辛甘醇、甘油辛酸酯、十一碳烯酸甘油酯、山梨坦辛酸酯、乙基己基甘油、牡丹根提取物等中的一种或多种。所述防腐剂在所述化妆品组合物中的含量是本领域已知的,例如,其通常占组分(B)总重量的约0.01-2%。
可根据皮肤化妆品组合物工业中已知的任何方法,将(A)乳酸菌发酵的桦树汁与其它化妆品成分混合,得到化妆品组合物。例如,使用化妆品领域中常用的溶解槽、乳化锅、分散器、输送泵等设备制备。制备时先将水溶性物质投入水相溶解釜,将油溶性物质投入油相溶解釜,将两个釜的温度加热至约80℃,其中对于易结块的原料,可先用分散器将其预分散。待溶解完成后将油相和水相输送至乳化锅中,均质乳化约5-15分钟。乳化完成后将料体温度降至常温,加入任选的香精、防腐剂等,并视需要调节产物的pH。相关检测指标都合格后方可灌装出货。以上制备方法可根据剂型要求进行删减或调整。
所述化妆品组合物可根据需要制成各种剂型,例如溶液、悬浮液、膏、霜、乳液、凝胶、粉末、喷剂等。
实施例
以下结合实施例,对本发明进行进一步详细说明。但是,应当理解为,这些实施例、对比例仅仅是用于更详细地说明本发明,而不应理解为用于以任何形式限制本发明所附权利要求书的范围。
实施例1-4和对比例中采用的副干酪乳杆菌(Lactobacillus para casei)菌种商购于杜邦公司,菌株编号LPC-37;瑞士乳杆菌(Lactobacillus helveticus)菌种商购于法国拉曼集团,菌株编号R0052;德氏乳杆菌保加利亚亚种(Lactobacillus bulgaricus)商购于杜邦公司,菌株编号LB340;动物双歧杆菌(Bifidobacterium animalis)商购于科汉森公司,菌株编号BB-12。
实施例1
(1)桦树汁培养基的制备
以东北大兴安岭采集的桦树汁原液(brix 1.20)为底物,添加0.2%乙酸钠、0.2%柠檬酸氢二铵、0.05%硫酸镁和0.02%硫酸锰,使用1M柠檬酸钠水溶液调整桦树汁培养基的pH为5.8±0.2。
(2)副干酪乳杆菌种子液的制备
制备8L桦树汁培养基并在105℃灭菌20分钟,冷却至37℃后,按1g/L投入副干酪乳杆菌粉,以培养基体积计;在37℃环境中培养8小时,得到副干酪乳杆菌种子液。
(3)接种发酵
按照72%(v/v)体积装液量,将步骤(1)所配制的桦树汁培养基加入到100升发酵罐中,105℃灭菌20分钟;将所制备的副干酪乳杆菌种子液无菌操作接入发酵罐中,于37℃、搅拌转速100rpm、通入空气以控制相对溶解氧量为100%的条件下,连续发酵72小时,停罐,得到发酵液产物。
(4)陶瓷膜分离发酵滤液和菌体
使用0.2μm陶瓷膜处理发酵液产物,操作压力0.15MPa,操作温度10℃;得到作为上清液的发酵的桦树汁滤液并打入无菌储存罐;和得到作为菌泥的副干酪乳杆菌菌体,并用5倍体积上清液重悬,得到副干酪乳杆菌的重悬液。
(5)破碎,过滤,获得可溶性副干酪乳杆菌溶胞物
利用高压细胞破碎设备,在900±50巴、1.5升/分钟流速的条件下,将步骤(4)所得副干酪乳杆菌的重悬液进行高压细胞破碎。然后使用0.2μm陶瓷膜过滤,即得到可溶性的副干酪乳杆菌溶胞物。
(6)将副干酪乳杆菌溶胞物与发酵桦树汁滤液混合和过滤
将步骤(5)所得副干酪乳杆菌溶胞物与步骤(4)所得发酵的桦树汁滤液混合,然后进一步使用0.2μm陶瓷膜过滤,获得最终发酵的桦树汁滤液产物。
取发酵的桦树汁滤液产物,测定其中的总酚、多糖、氨基酸和多肽含量,结果如表1所示;测定对DPPH自由基清除能力,结果如表2所示;测试对致病菌的抑制率,结果如表3所示;测试对人原代角质形成细胞的保湿相关蛋白表达的影响,结果如表4所示;测试对H2O2活性氧刺激人角质细胞的影响,结果如表5所示。
实施例2
(1)浓缩的桦树汁培养基的制备
以来自东北小兴安岭的桦树汁的浓缩液(浓缩6倍,brix 6.10)为底物,添加0.2%柠檬酸氢二铵、0.1%磷酸二氢钾、0.05%硫酸镁和0.02%硫酸锰,使用0.5M氢氧化钠水溶液调整桦树汁培养基的pH为5.8±0.2。
(2)接种发酵
按照70%(v/v)体积装液量,将步骤(1)所配制的桦树汁培养基通过超高温瞬时杀菌设备128℃灭菌4秒;加入到1000L已经空罐杀菌后的发酵罐中并将培养基冷却至35℃;将所商购的瑞士乳杆菌菌粉按3g/L无菌操作投入发酵罐中,于35℃、搅拌转速200rpm、通入空气以控制相对溶氧量80%的条件下,连续发酵60小时,停罐,得到发酵液产物。
(3)管式离心机分离发酵滤液和菌体
使用管式离心机处理发酵液产物,其中离心速度13000rpm,操作料液温度10℃,流速160升/小时;得到作为上清液的发酵的桦树汁滤液并打入无菌储存罐;和得到作为菌泥的瑞士乳杆菌菌体,并将其用4倍体积上清液重悬,获得瑞士乳杆菌的重悬液。
(4)破碎,过滤,获得可溶性瑞士乳杆菌溶胞物
将步骤(3)所得瑞士乳杆菌的重悬液进行反复冻融破碎,其中将重悬液置于零下20℃速冷冻结后,在40℃复融,周期循环5次。然后使用管式离心机过滤,即得到可溶性的瑞士乳杆菌溶胞物。
(5)将瑞士乳杆菌溶胞物与发酵桦树汁滤液混合过滤
将步骤(4)所得瑞士乳杆菌溶胞物与步骤(3)所得发酵的桦树汁滤液混合,然后进一步使用0.2μm陶瓷膜过滤,获得最终发酵的桦树汁滤液产物。
(6)将发酵桦树汁滤液杀菌灌装
将步骤(5)所得的发酵的桦树汁滤液产物经超高温瞬时杀菌设备,杀菌温度为110℃,杀菌15秒;然后,将杀菌过的滤液产物灌装于无菌袋中。
取发酵的桦树汁滤液产物,测定其中的总酚、多糖、氨基酸和多肽含量,结果如表1所示;测定对DPPH自由基清除能力,结果如表2所示;测试对致病菌的抑制率,结果如表3所示;测试对人原代角质形成细胞的保湿相关蛋白表达的影响,结果如表4所示;测试对H2O2活性氧刺激人角质细胞的影响,结果如表5所示。
实施例3
(1)桦树汁培养基的制备
以东北小兴安岭采集的桦树汁原液(brix1.05)为底物,添加7%脱脂乳粉、1%乳糖、0.05%硫酸镁和0.02%硫酸锰,使用1M柠檬酸钠水溶液调整桦树汁培养基的pH为5.8±0.2。
(2)接种发酵
按照70%(v/v)体积装液量,将步骤(1)所配制的桦树汁培养基通过超高温瞬时杀菌设备135℃灭菌3秒后无菌转移到500升无菌发酵罐中并将培养基冷却至38℃;将所商购的动物双歧杆菌菌粉按5g/L无菌操作投入发酵罐中,于38℃、搅拌转速500rpm、通入空气以控制相对溶解氧量20%的条件下,连续发酵54小时,停罐,得到发酵液产物。
(3)离心机分离发酵滤液和菌体
使用碟式离心机处理步骤(2)得到的发酵液,其中离心速度9000rpm,操作温度20℃,分离时间20分钟;得到作为上清液的发酵的桦树汁滤液并打入无菌储存罐;和得到作为菌泥的动物双歧杆菌菌体,并将其用4倍体积上清液重悬,获得动物双歧杆菌的重悬液。
(4)破碎,过滤,获得可溶性动物双歧杆菌溶胞物
将步骤(3)所得瑞士乳杆菌的重悬液进行反复冻融破碎,其中将重悬液置于零下20℃速冷冻结后,在40℃复融,周期循环5次。然后使用管式离心机过滤,即得到可溶性的动物双歧杆菌溶胞物。
(5)将动物双歧杆菌溶胞物与发酵桦树汁滤液混合和过滤
将步骤(4)所得动物双歧杆菌溶胞物与步骤(3)所得发酵的桦树汁滤液混合,然后进一步使用0.2μm陶瓷膜过滤,获得最终发酵的桦树汁滤液产物。
取发酵的桦树汁滤液产物,测定其中的总酚、多糖、氨基酸和多肽含量,结果如表1所示;测定对DPPH自由基清除能力,结果如表2所示;测试对致病菌的抑制率结果,如表3所示;测试对人原代角质形成细胞的保湿相关蛋白表达的影响,结果如表4所示;测试对H2O2活性氧刺激人角质细胞的影响,结果如表5所示。
实施例4
(1)桦树汁培养基的制备
以来自东北小兴安岭的桦树汁的浓缩液(浓缩3倍,brix2.95)为底物,添加4%水解乳清蛋白和2%的乳糖,使用0.5M氢氧化钠水溶液调整桦树汁培养基的pH为5.8±0.2。
(2)接种发酵
按照60%(v/v)体积装液量,将步骤(1)所配制的桦树汁培养基通过超高温瞬时杀菌设备115℃灭菌20秒后无菌转移到20000升无菌发酵罐中;将德氏乳杆菌保加利亚亚种菌粉无菌操作接入发酵罐中,于42℃、搅拌转速200rpm、通入空气控制相对溶解氧量60%的条件下,连续发酵84小时,停罐,得到发酵液产物。
(3)离心机分离发酵滤液和菌体
使用管式离心机处理发酵液,其中离心速度14000rpm,操作温度10℃,分离流速150升/小时;得到作为上清液的发酵的桦树汁滤液并打入无菌储存罐;和得到作为菌泥的德氏乳杆菌保加利亚亚种菌菌体,并将其用5倍体积上清液重悬,获得德氏乳杆菌保加利亚亚种菌的重悬液。
(4)破碎,过滤,获得可溶性德氏乳杆菌保加利亚亚种菌溶胞物
将步骤(3)所得德氏乳杆菌保加利亚亚种菌的重悬液进行酶解细胞破碎,其中添加800微克/毫升溶菌酶在35℃酶解处理60分钟,在200巴、1.3升/分钟流速的条件下进行均质破碎。然后使用0.2μm陶瓷膜过滤,即得到可溶性的德氏乳杆菌保加利亚亚种菌溶胞物。
(5)将德氏乳杆菌保加利亚亚种菌溶胞物与发酵桦树汁滤液混合和过滤
将步骤(4)所得德氏乳杆菌保加利亚亚种菌溶胞物与步骤(3)所得发酵的桦树汁滤液混合,然后进一步使用离心机过滤,获得最终发酵的桦树汁滤液产物。
取发酵的桦树汁滤液产物,测定其中的总酚、多糖、氨基酸和多肽含量,结果如表1所示;测定对DPPH自由基清除能力,结果如表2所示;测试对致病菌的抑制率,结果如表3所示;测试对人原代角质形成细胞的保湿相关蛋白表达的影响,结果如表4所示;测试对H2O2活性氧刺激人角质细胞的影响,结果如表5所示。
对比例1
(1)桦树汁培养基的制备
以与实施例1相同的工序,制备桦树汁培养基。
(2)副干酪乳杆菌种子液的制备
以与实施例1相同的工序,制备副干酪乳杆菌种子液。
(3)接种发酵
按照72%(v/v)体积装液量,将步骤(1)所配制的桦树汁培养基加入到100升发酵罐中,105℃灭菌20分钟;将所制备的副干酪乳杆菌种子液无菌操作接入发酵罐中,于37℃、搅拌转速100rpm、不通入空气的条件下,连续发酵72小时,停罐,得到发酵液产物。
(4)陶瓷膜分离发酵液产物
以与实施例1相同的工序,采用陶瓷膜分离发酵液产物,得到发酵的桦树汁滤液,和副干酪乳杆菌菌体悬浮液。
(5)破碎,过滤,获得可溶性副干酪乳杆菌溶胞物
以与实施例1相同的工序,获得可溶性副干酪乳杆菌菌体溶胞物。
(6)将副干酪乳杆菌菌体溶胞物与发酵的桦树汁滤液混合和过滤
以与实施例1相同的工序,获得最终的发酵的桦树汁滤液产物。
取发酵的桦树汁滤液产物,测定其中的总酚、多糖、氨基酸和多肽含量,结果如表1所示;测定对DPPH自由基清除能力,结果如表2所示;测试对致病菌的抑制率结果,如表3所示;测试对人原代角质形成细胞的保湿相关蛋白表达的影响,结果如表4所示;测试对H2O2活性氧刺激人角质细胞的影响,结果如表5所示。
表1:发酵的桦树汁滤液产物的总酚、多糖、氨基酸、多肽含量
从结果表明,与桦树汁原液相比,通过乳酸菌发酵桦树汁获得了营养物质明显增加的发酵产物(对比例);进一步地,与常规发酵环境(对比例)相比,在高氧环境中进行发酵,所得发酵产物中的多项营养物质显著进一步增加,尤其是其中的多糖含量和氨基酸含量。
表2:发酵的桦树汁滤液产物对DPPH的清除率
样品 | DPPH清除率(%) |
桦树汁原液(未发酵) | 27.3 |
对比例 | 36.4 |
实施例1 | 87.0 |
实施例2 | 90.5 |
实施例3 | 93.9 |
实施例4 | 98.1 |
DPPH自由基通常用来评价各种抗氧化剂对一般自由基的清除能力,用于抗氧化成分的体外抗氧化性评价;皮肤中的自由基的增多,自由基会争夺各个正常生理功能的电子而造成皮肤损伤。DPPH自由基清除能力的结果表明,与桦树汁原液和对比例相比,在高氧环境下发酵获得的发酵产物能够显著清除DPPH自由基,具有优异的抗氧化能力。
表3:发酵的桦树汁滤液产物对致病菌的抑菌能力
以上结果表明,与桦树汁原液相比,对比例中获得的发酵产物没有显著提高对致病菌的抑制能力。进一步地,与桦树汁原液和对比例相比,在高氧环境下发酵获得发酵产物具有显著抑制痤疮丙酸杆菌和金黄色葡萄球菌的能力。
表4:发酵的桦树汁滤液产物对人原代角质形成细胞的保湿相关蛋白表达的影响
注:*表示与原汁相比为显著,P值小于0.05;
**表示与原汁对照相比为极显著,P值小于0.01。
表4中给出了二次发酵的桦树汁对人原代角质形成细胞的角质转谷酰胺酶TGM1、表皮紧密连接蛋白(ZO-1和CLDN1)、丝聚蛋白FLG和水通道蛋白AQP3的影响的结果。结果表明,对比桦树汁原液,对比例中的发酵产物中有个别指标显著增强了保湿和屏障修复相关蛋白的表达;而在高氧环境下发酵获得的发酵产物对于保湿和屏障修复相关蛋白的表达有极显著的提升,这表明在高氧环境下进行发酵获得的发酵产物具有更好的保湿和修复功效。
表5:发酵的桦树汁滤液产物对人角质细胞中活性氧ROS的影响
样品 | ROS降低率(%) |
桦树汁原液(未发酵) | 12.1 |
对比例 | 13.6 |
实施例1 | 33.8 |
实施例2 | 36.5 |
实施例3 | 37.9 |
实施例4 | 39.1 |
表5中的试验采用人角质细胞,通过加入过氧化氢,模拟细胞内氧化应激过程,,使用荧光酶标仪检测无荧光的2',7'二氯荧光素二乙酸被氧化后荧光值的变化,检测物质对细胞内的抗氧化能力的提升。结果表明,桦树汁原液和对比例的发酵产物能够对角质细胞中的活性氧水平有一定的降低,而在高氧环境下进行发酵获得的发酵产物能够极显著的降低角质细胞中的活性氧的水平,具有优异的抗细胞氧化损伤的能力。
实施例5:具有抗环境压力功效的爽肤水
采用实施例1中制备的发酵的桦树汁滤液产物来制备所述爽肤水,其配方如下:
成分 | 含量(质量%) |
发酵的桦树汁滤液产物 | 94.25 |
苯甲酸钠 | 0.25 |
1,2-戊二醇 | 5 |
焦亚硫酸钠 | 0.5 |
上述爽肤水如下制备:
将所述发酵的桦树汁滤液产物、苯甲酸钠和戊二醇、焦亚硫酸钠混合,然后过滤,即得所述爽肤水。
选择生活城市的月度空气质量优良天数不足50%的20名受试者使用上述爽肤水4周后,对使用情况进行主观评估。结果有17人反映皮肤含水量显著增加,16人反映面部刺痛、发热或红血丝情况得到改善;且有16人反映皮肤较之前出现粉刺、发痒、皲裂等意外状况减少。
实施例6:具有抗环境压力功效的面霜
采用实施例2中制备的发酵的桦树汁滤液产物来制备所述面霜,其配方如下:
上述面霜如下制备:
1)油相:将4、6、7、8、9、11、12、13、15、21号原料加入油相锅,并加热至80℃,溶解,混合均匀;
2)将2、16、18号原料在常温下混合均匀;
3)将10、14、17、20号原料等原料在常温下混合均匀;
4)水相:将1、3、5号原料加热到80℃,加入步骤2的混合物,溶解,混合均匀;
5)乳化:将水相和油相加入乳化罐中,保温在80℃,在3000rpm速度下均质乳化5分钟,乳化完成后加入19号原料;
6)搅拌降温至40℃时加入步骤3的混合物,搅拌均匀后出料,得到所述面霜。
选择生活城市的月度空气质量优良天数不足50%的20名受试者使用上述面霜4周后,对使用情况进行主观评估。结果有18人反映皮肤含水量显著增加,16人反映面部刺痛、发热或红血丝情况得到改善;且有17人反映皮肤较之前出现粉刺、发痒、皲裂等意外状况减少。
实施例7:具有抗环境压力功效的乳液
采用实施例3中制备的发酵的桦树汁滤液产物来制备所述乳液,其配方如下:
上述乳液如下制备:
1)油相:将4、5、6、7、12、13号原料加入油相锅,并加热至80℃,溶解,混合均匀;
2)将2、8、11号原料在常温下混合均匀;
3)将9、10、16号原料等原料在常温下混合均匀;
4)水相:将1、3、14号原料加热到80℃,加入步骤2的混合物。溶解,混合均匀;
5)乳化:将水相和油相加入乳化罐中,保温在80℃,在3000rpm速度下均质乳化5分钟,乳化完成后加入15号原料;
6)搅拌降温至40℃时加入步骤3的混合物,搅拌均匀后出料,得到所述乳液。
选择生活城市的月度空气质量优良天数不足50%的20名受试者使用上述乳液4周后,对使用情况进行主观评估。结果有17人反映皮肤含水量显著增加,16人反映面部刺痛、发热或红血丝情况得到改善;且有16人反映皮肤较之前出现粉刺、发痒、皲裂等意外状况减少,皮肤更加光滑细嫩。
实施例8:具有抗环境压力功效的眼霜
采用实施例3中制备的发酵的桦树汁滤液产物来制备所述眼霜,其配方如下:
序号 | 原料 | 重量% |
1 | 发酵的桦树汁滤液产物 | 70.75 |
2 | 甘油 | 6.00 |
3 | 甘油三(乙基己酸)酯 | 3.00 |
4 | 丁二醇 | 5.00 |
5 | 聚二甲基硅氧烷 | 2.00 |
6 | C12-20烷基葡糖苷、C14-22醇 | 2.00 |
7 | 鲸蜡醇 | 2.00 |
8 | 肌酸 | 2.00 |
9 | 苯基聚三甲基硅氧烷 | 1.00 |
10 | 六肽-9 | 3.00 |
11 | 甘油硬脂酸酯类、PEG-100硬脂酸酯 | 0.50 |
12 | 苯氧乙醇 | 0.50 |
13 | 泛醇 | 0.30 |
14 | 二肽二氨基丁酰苄基酰胺二乙酸盐 | 1.00 |
15 | 精氨酸 | 0.15 |
16 | 卡波姆 | 0.25 |
17 | 乙酰化透明质酸钠 | 0.10 |
18 | 羟苯甲酯 | 0.10 |
19 | 尿囊素 | 0.15 |
20 | 聚谷氨酸钠 | 0.20 |
上述眼霜如下制备:
1)油相:将3、5、6、7、9、11号原料加入油相锅,并加热至80℃,溶解,混合均匀;
2)将2、16号原料在常温下混合均匀;
3)将10、14、17号原料等原料在常温下混合均匀;
4)水相:将1、4、8、13、18、19、20号原料加热到80℃,加入步骤2的混合物,溶解,混合均匀;
5)乳化:将水相和油相加入乳化罐中,保温在80℃,在3000rpm速度下均质乳化5分钟,乳化完成后加入12、15号原料;
6)搅拌降温至40℃时加入步骤3的混合物,搅拌均匀后出料,得到所述眼霜。
采用半脸对照的测试方法,在使用所述眼霜前和4周后分别对来自生活环境月度空气质量优良天数不足50%的20名志愿者进行如下测试:
1)用primos对志愿者左右两侧外眼角拍照,用软件计算皱纹参数,其中包括皱纹数量、皱纹面积、皱纹深度等;
2)用Corneometer测试左右两侧眼角的皮肤水分含量。
结果显示,20名受试者中有18人眼角皮肤含水量显著增加,其中有15人眼角皱纹明显变浅变淡,皱纹面积减小,皱纹数量减少。且有16人反映眼部皮肤更加细嫩光泽。
实施例8:具有抗环境压力功效的精华液
采用实施例4中制备的发酵的桦树汁滤液产物来制备所述精华液,其配方如下:
上述精华液如下制备:
1)将1、2、3、4、6、12、13号原料加入乳化罐中,保温在80℃;
2)将5、10、11号原料在常温下混合均匀;
3)将7、8、9号原料在常温下混合均匀;
4)加入14号原料,调节pH值;
5)搅拌降温至40℃时加入15号、16号和步骤3原料,搅拌均匀后出料,得到所述精华液。
选择生活城市的月度空气质量优良天数不足50%的20名受试者使用上述精华液4周后,对使用情况进行主观评估。结果有17人反映皮肤含水量显著增加,16人反映面部刺痛、发热或红血丝情况得到改善;且有18人反映皮肤较之前出现粉刺、发痒、皲裂等意外状况减少。
以上所述实施例的技术方案是本发明优选实施方式,在不脱离本发明原理的前提下还可以进行若干改进和变换,这些改进和变化也应视为在本发明的保护范围内。
Claims (22)
1.一种生产乳酸菌发酵的桦树汁的方法,其包括在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵的步骤。
2.权利要求1所述的方法,其包括下述步骤:
(1)在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵,获得发酵液产物;
(2)过滤所述发酵液产物,分别得到乳酸菌菌体和发酵的桦树汁滤液;
(3)破碎所述乳酸菌菌体,然后进行过滤,得到可溶性乳酸菌溶胞物;和
(4)将所得乳酸菌溶胞物与所述发酵的桦树汁滤液混合,过滤,得到发酵的桦树汁滤液产物,即,乳酸菌发酵的桦树汁。
3.权利要求1或2所述的方法,其中所述桦树汁是浓缩倍数为1.05-10.0倍、优选1.5-6.0倍的浓缩的桦树汁。
4.权利要求1-3任一项所述的方法,其中所述乳酸菌选自乳杆菌属,双歧杆菌属,以及选自乳球菌属、链球菌属、片球菌属和明串珠菌属的其它类别乳酸菌。
5.权利要求1-4任一项所述的方法,其中所述乳酸菌选自嗜酸乳杆菌、干酪乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、动物双歧杆菌、长双歧杆菌和婴儿双歧杆菌。
6.权利要求1-5任一项所述的方法,其中所述桦树汁在所述桦树汁培养基中的含量在90%以上,优选94%以上,基于所述桦树汁培养基的总重量。
7.权利要求1-6任一项所述的方法,其中所述乳制品选自液态乳、乳粉、乳加工产品和乳加工副产物。
8.权利要求1-7任一项所述的方法,所述乳制品在所述桦树汁培养基中的含量为0-8%,优选1-5%,基于所述桦树汁培养基的总重量。
9.权利要求1-8任一项所述的方法,其中所述高氧环境是指在培养基中的相对溶解氧含量为20-200%,优选50-120%。
10.可通过权利要求1-9任一项所述的方法获得的乳酸菌发酵的桦树汁。
11.一种乳酸菌发酵的桦树汁,其是通过在高氧环境中,采用乳酸菌作为菌种,在包含桦树汁和任选的乳制品的培养基中进行发酵而获得的。
12.权利要求10或11所述的乳酸菌发酵的桦树汁,其包含0.2-10g/L的总酚,0.2-5g/L的多糖,0.1-10g/L的氨基酸和0.2-60g/L的多肽。
13.权利要求11或12所述的乳酸菌发酵的桦树汁,其中所述桦树汁是浓缩倍数为1.05-10.0倍、优选1.5-6.0倍的浓缩的桦树汁。
14.权利要求11-13任一项所述的乳酸菌发酵的桦树汁,其中所述乳酸菌选自乳杆菌属,双歧杆菌属,以及选自乳球菌属、链球菌属、片球菌属和明串珠菌属的其它类别乳酸菌。
15.权利要求11-14任一项所述的乳酸菌发酵的桦树汁,其中所述乳酸菌选自嗜酸乳杆菌、干酪乳杆菌、德氏乳杆菌保加利亚亚种、瑞士乳杆菌、副干酪乳杆菌、植物乳杆菌、罗伊氏乳杆菌、动物双歧杆菌、长双歧杆菌和婴儿双歧杆菌。
16.权利要求11-15任一项所述的乳酸菌发酵的桦树汁,其中所述桦树汁在所述桦树汁培养基中的含量在90%以上,优选94%以上,基于所述桦树汁培养基的总重量。
17.权利要求11-16任一项所述的乳酸菌发酵的桦树汁,其中所述乳制品选自液态乳、乳粉、乳加工产品和乳加工副产物。
18.权利要求11-17任一项所述的乳酸菌发酵的桦树汁,所述乳制品在所述桦树汁培养基中的含量为0-8%,优选1-5%,基于所述桦树汁培养基的总重量。
19.权利要求11-18任一项所述的乳酸菌发酵的桦树汁,其中所述高氧环境是指在培养基中的相对溶解氧含量为20-200%,优选50-120%。
20.权利要求10-19任一项所述的乳酸菌发酵的桦树汁在具有抗环境压力功效的化妆品组合物中的用途。
21.一种具有抗环境压力功效的化妆品组合物,其包含权利要求10-19任一项所述的乳酸菌发酵的桦树汁。
22.权利要求21所述的具有抗环境压力功效的化妆品组合物,其中乳酸菌发酵的桦树汁在所述化妆品组合物中的含量为大于0至小于100%,优选为20-95%,基于所述化妆品组合物的总重量。
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