CN117281751B - 一种发酵金雀花提取物的制备方法及其应用 - Google Patents
一种发酵金雀花提取物的制备方法及其应用 Download PDFInfo
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- CN117281751B CN117281751B CN202310263557.3A CN202310263557A CN117281751B CN 117281751 B CN117281751 B CN 117281751B CN 202310263557 A CN202310263557 A CN 202310263557A CN 117281751 B CN117281751 B CN 117281751B
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Abstract
本发明提供一种发酵金雀花提取物的制备方法及其应用,其具体步骤包括粉碎、提取、发酵。本发明制备的发酵金雀花提取物含有较高的活性成分含量,温和高效,能有效抑制络氨酸酶、TRP‑1、TRP‑2的表达,具有优异的抗衰美白效果。本发明同时提供一种含有发酵金雀花提取物的美白组合物,含有不同的植物成分,并配合多种保湿、美白成分共同作用,具有优异的美白保湿、舒缓、抗氧化的效果,使用时能更深层次的穿透皮肤、增强皮肤的自然防御能力。并且本发明的组合物稳定性好、使用多种天然植物成分、刺激性低,可用于各类化妆品产品中。
Description
技术领域
本发明涉及化妆品技术领域,具体涉及一种发酵金雀花提取物的制备方法及其应用。
背景技术
由于作息不规律、空气污染、电子屏幕的长期使用带来的辐射、不健康的生活习惯等造成的皮肤黑色素异常沉着、肤色暗沉、皮肤发黄等现象,成为困扰人们的一大问题。美白护肤也就成为了当下人们的一大需求。
目前市面上的美白产品众多,其核心成分是添加的各种活性成分,活性成分在使用过程中与人体皮肤接触、吸收、并作用于黑色素细胞和色斑等。不同的成分其美白机理均有不同,部分可通过一直黑色素的形成实现美白,部分可通过阻断黑色素的传播实现美白,部分即可通过加快皮肤的新陈代谢实现美白。使用植物提取物作为美白成分,是当下人们的一种选择倾向,其可以有效减少化学试剂成分的使用,降低对皮肤的刺激性、增强使用安全性。
中国专利CN115068391A公开了一种美白组合物及含该组合物的美白精华及美白水,其美白组合物包括以下组分:白果紫草根提取物;美丽拟伊藻提取物;石榴果皮提取物;苹果根提取物;溶剂。通过使用几种美白成分的共同使用来达到美白的效果,但是其活性成分的利用率和组合物的稳定性等效果仍有待商榷。
金雀花提取物是一种中药植物,其根部含有大量的皂苷、多酚类活性成分,用于化妆品产品中,具有极好的美白护肤的效果。但是由于植物提取物中成分复杂且通常对酸碱敏感、对光照敏感,因此提高植物提取物中活性成分的利用率及提高其制得的组合物的稳定性仍是需要解决的问题。
发明内容
针对上述问题,本发明提供一种发酵金雀花提取物的制备方法,其具体步骤如下:
步骤S1:粉碎:将金雀花根进行清洗,110-130℃烘干,粉碎后过筛,得金雀花根粉末;
步骤S2:提取:将金雀花根粉末加入提取器中,加入乙醇,在50-60℃下提取3-5h,离心取上清液,灭菌后即得金雀花提取物;
步骤S3:发酵:将菌种种子液按10-15%的接种量接入装有金雀花提取物的发酵培养基中,搅拌均匀,在温度为26-29℃、湿度为70-80%的条件下发酵4-6天后,灭菌即得发酵金雀花提取物。
优选的,所述步骤S1中,金雀花根粉碎后过100目筛。
优选的,所述步骤S2中,乙醇与金雀花根的体积比为20-25:1。
优选的,所述乙醇浓度为75%。
优选的,所述步骤S2和步骤S3中,灭菌的具体条件为:110-125℃灭菌30-40min。
优选的,所述步骤S3中,菌种种子液的具体制备方法为:
将原始菌种接种至PDA培养基上,培养温度为28℃,培养时间为72-96h;挑取菌丝至新的培养基上,在28℃条件下培养48h;加入10-20ml生理盐水,使用无菌刮铲将表面的孢子刮下,将孢子悬液转入三角瓶中,即得菌种种子液。
进一步优选的,所述菌种为黑曲霉:Aspergillus niger CICC 41481。
金雀花根中含有大量的皂苷、多酚类活性成分,其具有优异的抗氧化、自由基清除能力,用于化妆品产品中,具有极好的美白护肤的效果。发明人发现,通过对提取物进行进一步的发酵处理,能进一步增强其抗氧化、美白效果。可能是由于经过发酵处理,使得提取物种含有的多酚类活性成分进一步分解,使其生理活性增加,作为化妆品成分使用时,更易于被皮肤吸收而具有根本更好的抗氧化、美白效果。发明人对其中的活性成分进行简单分析,发现发酵处理后,特别是白藜芦醇的含量有极大的增加,而白藜芦醇具有极强的清除自由基的效果。
本发明另一方面提供一种可美白的发酵金雀花提取物在化妆品中的应用。
优选的,所述发酵金雀花提取物可用于制备一种含有发酵金雀花提取物的美白组合物,按重量份计,原料包括:
发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物2-5份、积雪草(CENTELLAASIATICA)提取物0.1-1份、光果甘草(GLYCYRRHIZAGLABRA)根提取物0.1-2份、烟酰胺0.1-3份、皮肤调理剂5-10份、二元醇5-20份、EDTA-2Na 0.1-2份、水40-60份。
积雪草(CENTELLAASIATICA)提取物中含有多种三萜类成分,包括积雪草酸、积雪草苷、羟基积雪草苷等成分,具有优异的抗菌消炎作用,并能降低组织细胞内自由基的浓度,同时抑制其活性,有较强的抗氧化作用;同时可以抑制酪氨酸酶活性,淡化黑色素沉着。光果甘草(GLYCYRRHIZAGLABRA)根提取物能抑制酪氨酸酶的活性,又能抑制多巴色素互变和DHICA氧化酶的活性。烟酰胺是一种维生素B3,能够抑制黑色素的生成、有效清除自由基。本发明的组合物中通过多种成分配合使用,具有优异的美白、抗氧化的效果。
优选的,所述皮肤调理剂为甘油、透明质酸钠、壬二酸、生育酚乙酸酯、维生素e、卵磷脂中的一种或多种组合。
进一步优选的,所述皮肤调理剂为透明质酸钠和维生素E。
进一步优选的,所述透明质酸钠和维生素E的质量比为1-3:1。
优选的,所述透明质酸钠为小分子透明质酸钠和中分子透明质酸钠按质量比为2-4:1的混合。
进一步优选的,所述小分子透明质酸钠的分子量为10KDa-500 KDa,所述中分子透明质酸钠的分子量为500KDa─2000KDa。
为了进一步增强组合物的保湿、抗氧化效果等效果,本发明中同时还使用透明质酸钠和维生素E作为皮肤调理剂。中分子透明质酸钠具有更长的分子链,使用时能够在皮肤表面形成保湿膜,有效补水保湿,配合小分子透明质酸钠、和维生素E使用,能够同时有效渗透皮肤,达到深层保湿滋养的效果。但是中分子透明质酸钠若含量过多,可能会导致其在体系中分散不均,并且使用感不好。
发明人发现,当使用透明质酸钠和维生素E,特别是当两者质量比为1-3:1时,不仅具有保湿、抗氧化的效果,同时还能进一步增强组合物的稳定性。由于本发明中含有多种植物提取成分,虽然同时使用多种植物提取成分具有优异的美白、抗氧化的效果,但是由于植物提取物中成分较为复杂,含有各种微量成分,受重金属、光热、酸碱等影响较大,易造成活性成分失活等问题。通过使用EDTA-2Na,能有效去除重金属等的影响,同时含有透明质酸钠等成分,由于离子间的斥力,避免小分子成分的团聚,使体系中的各种成分能稳定存在,增强了体系的稳定性。
优选的,所述二元醇为丁二醇、丙二醇、戊二醇、己二醇中的一种或多种混合。
进一步优选的,所述二元醇为丁二醇。
优选的,所述含发酵金雀花提取物的美白组合物的制备方法如下:
(1)将发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物、积雪草(CENTELLAASIATICA)提取物、光果甘草(GLYCYRRHIZAGLABRA)根提取物、二元醇混合均匀,得到混合物A;
(2)将混合物A、烟酰胺、皮肤调理剂、EDTA-2Na依次加入水中,在35-45℃条件下混合均匀,即得所述含发酵金雀花提取物的组合物。
有益效果:
本发明制备的发酵金雀花提取物含有较高的活性成分含量,温和高效,能有效抑制络氨酸酶、TRP-1、TRP-2的表达,具有优异的抗衰美白效果。本发明同时提供一种含有发酵金雀花提取物的美白组合物,含有不同的植物成分,并配合多种保湿、美白成分共同作用,具有优异的美白保湿、抗炎、抗氧化的效果,使用时能更深层次的穿透皮肤、增强皮肤的自然防御能力。并且本发明的组合物稳定性好、使用多种天然植物成分、刺激性低,可用于各类化妆品产品中。
附图说明
图1为本发明实施例1制备的发酵金雀花提取物的样品图;
图2为本发明实施例4制备的美白组合物的样品图。
具体实施方式
以下结合具体实施例对本发明进一步详细说明。应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。需要说明的是,在不冲突的情况下,本申请中的实施例及实施例中的特征可以相互组合。
实施例
实施例1
本实施例提供了一种发酵金雀花提取物的制备方法,其具体步骤如下:
步骤S1:粉碎:将金雀花根进行清洗,120℃烘干,粉碎后过100目筛,得金雀花根粉末;
步骤S2:提取:将金雀花根粉末加入提取器中,加入20倍体积的75%乙醇,在55℃下提取4h,离心取上清液,121℃灭菌30min后即得金雀花提取物;
步骤S3:发酵:将原始菌种接种至PDA培养基上,培养温度为28℃,培养时间为72h;挑取菌丝至新的培养基上,在28℃条件下培养48h;加入15ml生理盐水,使用无菌刮铲将表面的孢子刮下,将孢子悬液转入三角瓶中,即得菌种种子液;
将菌种种子液按10%的接种量接入装有金雀花提取物的发酵培养基中,搅拌均匀,在温度为28℃、湿度为75%的条件下发酵5天后,121℃灭菌30min后即得发酵金雀花提取物。
菌种为黑曲霉:Aspergillus niger CICC 41481。购于中国工业微生物菌种保藏管理中心。
实施例2
本实施例提供一种含有发酵金雀花提取物的美白组合物,所述美白组合物按重量份计,原料包括:
发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物2份、积雪草(CENTELLA ASIATICA)提取物0.1份、光果甘草(GLYCYRRHIZA GLABRA)根提取物0.1份、烟酰胺0.1份、皮肤调理剂5份、二元醇5份、EDTA-2Na 0.1份、水40份。
发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物的制备方法同实施例1。
积雪草(CENTELLA ASIATICA)提取物购于广州花之王化工有限公司;光果甘草(GLYCYRRHIZA GLABRA)根提取物购于武汉克米克生物医药技术有限公司。
皮肤调理剂为质量比为2:1的透明质酸钠和维生素E。透明质酸钠为质量比为3:1的小分子透明质酸钠和中分子透明质酸钠;小分子透明质酸钠的分子量为200KDa,中分子透明质酸钠的分子量为1000KDa;购于成都欧恩瑞思化学试剂。二元醇为丁二醇。
本实施例的美白组合物的制备方法如下:
(1)将发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物、积雪草(CENTELLAASIATICA)提取物、光果甘草(GLYCYRRHIZAGLABRA)根提取物、二元醇混合均匀,得到混合物A;
(2)将混合物A、烟酰胺、皮肤调理剂、EDTA-2Na依次加入水中,在40℃条件下混合均匀,即得所述含发酵金雀花提取物的组合物。
实施例3
本实施例提供一种含有发酵金雀花提取物的美白组合物,所述美白组合物按重量份计,原料包括:
发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物5份、积雪草(CENTELLAASIATICA)提取物1份、光果甘草(GLYCYRRHIZA GLABRA)根提取物2份、烟酰胺3份、皮肤调理剂10份、二元醇20份、EDTA-2Na 2份、水60份。
本实施例具体实施方式同实施例2。
实施例4
本实施例提供一种含有发酵金雀花提取物的美白组合物,所述美白组合物按重量份计,原料包括:
发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物3.5份、积雪草(CENTELLAASIATICA)提取物0.5份、光果甘草(GLYCYRRHIZA GLABRA)根提取物1份、烟酰胺1.5份、皮肤调理剂8份、二元醇15份、EDTA-2Na 1份、水50份。
本实施例具体实施方式同实施例2。
对比例1
本实施例提供一种发酵金雀花提取物的制备方法,其具体步骤如下:
步骤S1:粉碎:将金雀花根进行清洗,120℃烘干,粉碎后过100目筛,得金雀花根粉末;
步骤S2:提取:将金雀花根粉末加入提取器中,加入28倍体积的80%乙醇,在55℃下提取4h,离心取上清液,121℃灭菌30min后即得金雀花提取物;
步骤S3:发酵:将原始菌种接种至PDA培养基上,培养温度为28℃,培养时间为72h;挑取菌丝至新的培养基上,在28℃条件下培养48h;加入15ml生理盐水,使用无菌刮铲将表面的孢子刮下,将孢子悬液转入三角瓶中,即得菌种种子液;
将菌种种子液按10%的接种量接入装有金雀花提取物的发酵培养基中,搅拌均匀,在温度为28℃、湿度为75%的条件下发酵5天后,121℃灭菌30min后即得发酵金雀花提取物。
菌种为黑曲霉:Aspergillus niger CICC 41481。购于中国工业微生物菌种保藏管理中心。
对比例2
本实施例提供一种发酵金雀花提取物的制备方法,其具体步骤同实施例1,与实施例1的区别在于未进行发酵步骤。
对比例3
本实施例提供一种含有发酵金雀花提取物的美白组合物,具体实施例方式同实施例4,与实施例4的区别在于,所述发酵金雀花(SAROTHAMNUS SCOPARIUS)提取物的制备方法同对比例1。
对比例4
本实施例提供一种含有发酵金雀花提取物的美白组合物,具体实施例方式同实施例4,与实施例4的区别在于,所述皮肤调理剂为维生素E。
对比例5
本实施例提供一种含有发酵金雀花提取物的美白组合物,具体实施例方式同实施例4,与实施例4的区别在于,美白组合物的制备方法为:将发酵金雀花(SAROTHAMNUSSCOPARIUS)提取物、积雪草(CENTELLAASIATICA)提取物、光果甘草(GLYCYRRHIZAGLABRA)根提取物、烟酰胺、皮肤调理剂、二元醇、EDTA-2Na依次加入水中,在40℃条件下混合均匀,即得所述含发酵金雀花提取物的组合物。
性能测试
1、白藜芦醇含量测试:使用高效液相色谱仪进行白藜芦醇含量测试。
称取白藜芦醇标准品5mg,甲醇溶解定容至25m L,配制成0.2mg/mL白藜芦醇储备液,备用。分别取储备液0.25、0.5、1.0、2.0、4.0m L,甲醇定容至10mL,0.45μm滤膜过滤,分别得到5μg/m L、10μg/mL、20μg/mL、40μg/mL、80μg/mL标准品溶液。
色谱条件:色谱柱Kromasil 100-5-C18(250mm×4.6mm),流动相为甲醇:水(40:60,v/v),流速1.0mL/min,波长306nm,柱温25℃,进样量10μL。
称取样品溶液,按色谱条件进样,平行3次,根据峰面积计算样品溶液中白藜芦醇含量。
2、DPPH自由基清除试验
将4mL浓度为6×10-5mol/L的DPPH溶液分别加入4mL测试样品中,充分混合均匀,立即测量在518nm下的吸光度值,记为Ai,然后置于室温避光静置30分钟后,测定吸光度值,记为Aj,最后测定只添加DPPH溶液的吸光值,记为Ac,按以下公式计算DPPH自由基的清除率:清除率=[1-(Ai-Aj)/Ac]×100%。
3、络氨酸酶抑制率测试
在96孔板中加入50uL的5mg/mL的样品稀释液及40uL酪氨酸酶溶液,振荡混匀后于37℃孵育10min。随后分别加入110uLL-酪氨酸溶液,振荡混匀后于37℃孵育,15min后检测OD450nm,得出样品对酪氨酸酶活性的影响。
将实施例1和对比例1-2进行白藜芦醇含量测试、络氨酸酶抑制率测试,将实施例2-4、对比例3-5的美白组合物进行络氨酸酶抑制率、抗氧化DPPH自由基清除试验,测得结果如下表1。
表1
实施例 | 白藜芦醇含量 | 络氨酸酶抑制率(%) | 自由基清除率(%) |
实施例1 | 1289ppm | 86.10 | / |
实施例2 | / | 87.31 | 68.12 |
实施例3 | / | 89.32 | 70.01 |
实施例4 | / | 90.21 | 71.31 |
对比例1 | 1014ppm | 79.32 | / |
对比例2 | 853ppm | 56.43 | / |
对比例3 | / | 71.35 | 46.67 |
对比例4 | / | 80.63 | 60.35 |
对比例5 | / | 69.24 | 50.14 |
4、稳定性测试:
试验条件:将样品使用白色聚乙烯塑料瓶包装,分别放置在40℃恒温恒湿箱、室温(25℃)、冷藏(-8~-4℃)。分别于第1、2个月观察组合物是否有变色、分层、析出固体或其他不良现象。
将实施例2-4、对比例3-5的美白组合物进行稳定性测试,测试结果如下表2。
表2
Claims (3)
1.一种发酵金雀花提取物的制备方法,其特征在于,其具体步骤如下:
步骤S1:粉碎:将金雀花根进行清洗,110-130℃烘干,粉碎后过筛,得金雀花根粉末;
步骤S2:提取:将金雀花根粉末加入提取器中,加入75%乙醇,在50-60℃下提取3-5h,离心取上清液,灭菌后即得金雀花提取物;
步骤S3:发酵:将菌种种子液按10-15%的接种量接入装有金雀花提取物的发酵培养基中,搅拌均匀,在温度为26-29℃、湿度为70-80%的条件下发酵4-6天后,灭菌即得发酵金雀花提取物;
所述步骤S2中,乙醇与金雀花根的体积比为20-25:1;
所述步骤S3中,菌种为黑曲霉:Aspergillus niger CICC 41481;所述步骤S3中,菌种种子液的具体制备方法为:
将原始菌种接种至PDA培养基上,培养温度为28℃,培养时间为72-96h;挑取菌丝至新的培养基上,在28℃条件下培养48h;加入10-20ml生理盐水,使用无菌刮铲将表面的孢子刮下,将孢子悬液转入三角瓶中,即得菌种种子液。
2.一种根据权利要求1所述的制备方法制备的发酵金雀花提取物在化妆品中的应用。
3.一种含有权利要求1所述的制备方法制备的发酵金雀花提取物的美白组合物,其特征在于,所述美白组合物按重量份计,原料由以下成分组成:
发酵金雀花提取物 2-5份、积雪草提取物0.1-1份、光果甘草提取物 0.1-2份、烟酰胺0.1-3份、皮肤调理剂 5-10份、二元醇 5-20份、EDTA-2Na 0.1-2份、水 40-60份;
所述皮肤调理剂为透明质酸钠和维生素E,所述透明质酸钠和维生素E的质量比为1-3:1;所述二元醇为丁二醇、丙二醇、戊二醇、己二醇中的一种或多种混合,所述美白组合物的制备方法如下:
将发酵金雀花提取物、积雪草提取物、光果甘草提取物、二元醇混合均匀,得到混合物A;
将混合物A、烟酰胺、皮肤调理剂、EDTA-2Na依次加入水中,在35-45℃条件下混合均匀,即得所述含发酵金雀花提取物的美白组合物。
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