CN110139569A - 黑树莓提取物、含有黑树莓提取物的美白剂和饮食品、黑树莓提取物的制造方法和使用了黑树莓提取物的美白方法 - Google Patents
黑树莓提取物、含有黑树莓提取物的美白剂和饮食品、黑树莓提取物的制造方法和使用了黑树莓提取物的美白方法 Download PDFInfo
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Abstract
本发明的黑树莓提取物具有抑制黑素细胞中的酪氨酸酶的活性的效果。
Description
技术领域
本发明涉及黑树莓提取物、含有黑树莓提取物的美白剂和饮食品、黑树莓提取物的制造方法和使用了黑树莓提取物的美白方法。
背景技术
对于大部分生物而言,氧是不可欠缺的,被摄入到体内的氧的百分之几变为活性氧,用于防御细菌感染等。但是,为了维持体内稳态,剩余的活性氧需要通过生物体内的抗氧化酶、食品中的外源抗氧化物质除去。
另外,因紫外线、吸烟、废气、大气污染、应激等产生过量的活性氧时,给生物体组织带来氧化性损伤(例如参见非专利文献1)。其结果是,引起以生活方式疾病、老年性疾病为代表的癌症、痴呆等,可以说90%以上的疾病与过量产生的活性氧存在因果关系(例如参见非专利文献2)。
因此,体内的超氧化物歧化酶(SOD,Superoxide dismutase)的活化以及食品中的抗氧化物质受到关注,建立了由美国农务部(USDA)开发的抗氧化能力的测定方法(ORAC;Oxygen Radical Absorbance Capacity,氧自由基吸收能力),其正在普及(例如参见非专利文献3)。另外,在ORAC值高的食品、即抗氧化能力高的食品最佳100中存在12个品种的莓类食品(例如参见非专利文献4)。
成为皮肤中的色素沉着的原因的黑色素的合成因各种外源性和内源性的刺激而被促进。例如,暴露于太阳光中的紫外线、应激、大气污染等时,在皮肤的表面产生活性氧,由于该刺激使得黑素细胞被活化,酪氨酸酶的表达增强。这样表达后的酪氨酸酶将酪氨酸代谢,经过几个阶段而产生黑色素。报道了通过利用抗氧化物质捕获该活性氧来抑制黑色素的产生(例如参见非专利文献5)。另外,如果能够抑制黑素细胞中的酪氨酸酶的活性,则也可以期待肌肤的美白效果。
现有技术文献
非专利文献
非专利文献1:Oberdanner,C.ROS and Antioxidant Systems in Apoptosis.VDMVerlag,2008.ISBN 978-3836482110
非专利文献2:Slavin JL et al.Health benefits of fruits andvegetables.Adv Nutr.2012.1;3(4):506-516
非专利文献3:Cao G et al.Oxygen-radical absorbance capacity assay forantioxidants.Free Radical Biol.Med.1993.14:303-311
非专利文献4:“ORAC值高的(抗氧化作用高的)食品最佳100名单”[网上(online)](2017年6月30日检索)http://www.anteaoxidant.com/orac#top100.html
非专利文献5:Choi MY1,Song HS,Hur HS,Sim SS.Whitening activity ofluteolin related to the inhibition of cAMP pathway in alpha-MSH-stimulatedB16melanoma cells.Arch Pharm Res.2008Sep;31(9):1166-71
发明内容
发明所要解决的问题
本发明的目的在于,提供具有抑制黑色素的产生的作用的莓类的黑树莓提取物、含有黑树莓提取物的美白剂和饮食品、黑树莓提取物的制造方法和使用了黑树莓提取物的美白方法。
本发明人着眼于作为具有抗氧化能力的食品的莓类食品,对莓类食品与上述抑制黑色素的产生的作用的相关性进行了研究,其结果发现黑树莓的提取物达成了上述目的,从而完成了本发明。
用于解决问题的方法
即,根据本发明,可提供:
(1)一种黑树莓提取物,其具有抑制黑素细胞中的酪氨酸酶的活性的效果;
(2)一种美白剂,其含有(1)所述的黑树莓提取物;
(3)一种饮食品,其含有(1)所述的黑树莓提取物;
(4)一种黑树莓提取物的制造方法,其是(1)所述的黑树莓提取物的制造方法,其包括:对黑树莓的果实进行冷冻的冷冻工序;对上述黑树莓的果实进行解冻的解冻工序;向利用上述解冻工序解冻后的上述黑树莓的果实中添加溶剂并进行搅拌的搅拌工序;和将通过上述搅拌工序搅拌后的上述黑树莓的果实和溶剂静置、进行提取的提取工序;
(5)一种美白方法,其中,使用了(1)所述的黑树莓提取物。
发明效果
根据本发明,可以提供黑树莓提取物、含有黑树莓提取物的美白剂和饮食品、黑树莓提取物的制造方法和使用了黑树莓提取物的美白方法。
附图说明
图1是示出抑制黑素细胞中的酪氨酸酶的活性的效果的研究结果的图。
具体实施方式
以下,对本发明的黑树莓提取物进行说明。本发明的黑树莓提取物具有抑制黑素细胞中的酪氨酸酶的活性的效果。
作为黑树莓提取物的原料的黑树莓属于蔷薇科悬钩子属。另外,本发明的黑树莓提取物优选使用主要来自黑树莓的果实的提取物。
从黑树莓的果实中提取黑树莓提取物的方法没有特别限定,可以使用溶剂提取、超临界提取等。
作为进行溶剂提取时所使用的提取溶剂,没有特别限定,可以使用水、有机溶剂和它们的混合溶剂。作为有机溶剂,可以使用:甲醇、乙醇、正丙醇、异丙醇、正丁醇等醇类;乙二醇、丙二醇等二醇类;乙酸乙酯;丙酮等。这些有机溶剂可以单独使用一种,也可以组合使用两种以上。另外,可以将水等水系溶剂和有机溶剂混合使用。
其中,优选乙醇、将乙醇与水混合而成的乙醇水溶液,乙醇水溶液中的乙醇的浓度优选为1~80v/v%(EtOH),更优选为10~60v/v%(EtOH),进一步优选为20~40v/v%(EtOH)。
需要说明的是,进行超临界提取的情况下,优选使用制成超临界状态的二氧化碳。
另外,作为进行溶剂提取时的提取方法,可以使用搅拌提取、浸渍提取、振荡提取、回流提取、超声波提取等。其中,优选搅拌提取。
搅拌提取的方法没有特别限定,可以进行使用搅拌器的搅拌、使用捣碎器的搅拌等。
另外,进行搅拌提取时,一边优选以500~20000rpm、更优选以1000~10000rpm、进一步优选以6000~10000rpm的搅拌速度进行搅拌,一边进行优选为几分钟~60分钟、更优选为30分钟~60分钟的提取。需要说明的是,优选在搅拌后静置、进行静置提取。更具体而言,优选使用均质器以6000~10000rpm进行30分钟~60分钟均质化、然后静置24小时进行提取。需要说明的是,通过进行静置、提取,可以进行固液分离。
另外,进行溶剂提取时的温度可以为室温,也可以为加热条件下。另外,进行溶剂提取时的压力可以为常压,也可以为加压下。
另外,通过溶剂提取得到的提取液可以根据需要进行过滤等处理。
另外,可以使通过溶剂提取得到的提取液干燥而制成粉末。作为干燥方法,可以使用公知的方法,可以使用喷雾干燥、真空干燥、冷冻干燥等方法。
需要说明的是,作为黑树莓提取物,可以使用制成液体或粉末状的市售品或现有产品。
本发明的黑树莓提取物可以配合在食品、饮料等中。作为食品,可以列举面包类、面类、点心类、食用肉加工品、鱼类加工品、冷冻食品、果冻类、冰淇淋类、乳制品、各种调料等。另外,除了一般食品以外,也可以配合在特定保健用食品、准药品、健康食品、补充剂中。作为饮料,可以列举清凉饮料水、乳饮料、酒类、茶、红茶饮料、咖啡、果汁饮料、碳酸饮料、矿泉水类、果蔬饮料等。
另外,可以将配合有本发明的黑树莓提取物的食品、饮料制成与片剂、胶囊剂、糖浆等口服给药制剂同样的形态。
另外,在制造配合有本发明的黑树莓提取物的食品、饮料时,可以在不妨碍本发明效果的范围内根据需要添加甜味剂、着色剂、防腐剂、增稠剂、稳定剂、胶凝剂、抗氧化剂、显色剂、漂白剂、乳化剂、膨胀剂、酸味剂、光泽剂、香料等添加剂、溶剂、油。这些添加剂可以单独使用一种,也可以组合使用两种以上。
配合在上述食品、饮料中的黑树莓提取物的比例可以根据使用目的适当调整,配合在上述食品、饮料中的黑树莓提取物的比例优选为0.0001~80重量%,更优选为0.003~50重量%,进一步优选为0.005~30重量%。
本发明的黑树莓提取物在用于得到肌肤的美白效果方面是有用的。
实施例
以下,列举实施例对本发明进行说明,但本发明并不限于此。需要说明的是,只要没有特别说明,则本实施例中的份和%为重量基准。在实施例和比较例中,进行了基于ORAC值的抗氧化作用的评价和关于抑制黑素细胞中的酪氨酸酶的活性的效果的研究。
关于试样
(试样的准备)作为试样,使用三种莓类试样(黑树莓(蔷薇科悬钩子属)(以下有时称为“BR”)、蓝莓(杜鹃花科越橘属)(以下有时称为“BB”)和树莓(蔷薇科悬钩子属)(以下有时称为“RB”))的提取物以及作为配合有黑树莓的提取物的粉末制剂的Blabina(注册商标)。
(提取液的制备)
黑树莓、蓝莓和树莓的提取液通过以下方法得到。
首先,将黑树莓、蓝莓和树莓的果实分别在冷冻状态下保存。接着,将各果实解冻并切割。分别称量(约100g)黑树莓、蓝莓和树莓,添加5倍量的30v/v%(EtOH)乙醇水溶液(约500mL),使用捣碎器以8000rpm、30分钟的条件进行搅拌提取。进而,在室温下静置24小时,由此进行静置提取,以150目进行过滤而分别得到上述黑树莓、蓝莓和树莓的乙醇提取液。后述的ORAC的测定在乙醇溶解状态下进行,因此,使用乙醇提取液进行测定。
需要说明的是,关于Blabina,按照使Blabina达到9mg/mL的方式使其溶解于30v/v%(EtOH)中。
(提取物的制备)
使用蒸发器分别从通过上述提取液的制备得到的黑树莓、蓝莓和树莓各自的乙醇提取液、以及Blabina的乙醇溶液中蒸发除去乙醇成分,得到提取物。需要说明的是,黑树莓、蓝莓和树莓的乙醇提取液含有9mg/mL的提取物(extract)。
测定和评价
(基于ORAC值的抗氧化作用的评价)
测定上述得到的三种莓类样本(黑树莓、蓝莓和树莓的乙醇提取液)的ORAC值。将结果示于表1中。需要说明的是,对于Blabina,没有进行ORAC值的测定。
[表1]
样本的种类 | ORAC值(μMTE/g) |
BR | 62 |
BB | 66 |
RB | 49 |
与一般的抗氧化食品相比,三种莓类样本的ORAC值均显示出高的值(对于一般的抗氧化食品的ORAC值,参见上述非专利文献4)。三种莓类样本中,蓝莓显示出最高的值,但这是使用乙醇提取液测定的结果,因此,不能涉及到水溶性成分。
另外,乙醇浓度为约0.1%以上时对细胞存在影响,因此,在进行使用了细胞的测定和评价的情况下,利用使用了30体积%乙醇水溶液的乙醇提取液并不合适。因此,在以下的抑制黑素细胞中的酪氨酸酶的活性的效果的研究中,使用了通过蒸发器分别从乙醇提取液中蒸发除去乙醇成分后的样本(参见上述“提取物的制备”)。
(抑制黑素细胞中的酪氨酸酶的活性的效果)使用黑素细胞作为实验中使用的细胞。作为黑素细胞,使用正常人表皮黑素细胞(Normal Human Epidermal Melanocytes)、幼体包皮(juvenile foreskin)(C-12400,购自PromoCell公司),使用专用培养基(黑素细胞生长培养基补充包(Melanocyte Growth Medium Supplement Pack);C-39410,购自PromoCell公司),在37℃、5%CO2的条件下进行培养。在即将实验前利用0.25%胰蛋白酶(Trypsin)/EDTA溶液回收细胞,进行台盼蓝染色处理后,利用自动细胞计数仪Countess(Invitrogen)算出细胞数。然后,接种于24孔板或96孔板,进行24小时预培养后用于实验。
另外,将在上述提取物的制备中得到的三种莓类(黑树莓、蓝莓和树莓)提取物、Blabina的除去乙醇后的提取物分别溶解于超纯水中而制成约11.5mg/mL,得到样本。接着,将各样本进行连续稀释(从稀释100000倍至稀释10倍),分别添加至黑素细胞中。
接着,向0.1M磷酸缓冲液(pH6.5)60μL中添加33.3μL的2.5mM酪氨酸(Tyrosine)水溶液(25℃)。向其中添加3.3μL的L-酪氨酸底物溶液并进行搅拌,添加3.3μL的酪氨酸酶水溶液(约1300单位(unit)/mL),使酶反应开始。将酶标仪内设定为25℃,进行酶反应。
酶活性的评价以利用酪氨酸酶合成的DOPA铬的吸光度(475nm)为指标。需要说明的是,使用维生素C 20mg/mL作为阳性对照(Positive control)(例如参见上述非专利文献5和Matsuki M,Watanabe T,Ogasawara A,Mikami T,Matsumoto T.Inhibitory mechanismof melanin synthesis by glutathione.[Article in Japanese]Yakugaku Zasshi.2008Aug;128(8):1203-7 12.)。
将结果示于图1中。关于DOPA铬的50%抑制(Inhibition)(IC50),以概算计,对于阳性对照(维生素C)而言为稀释100000倍,对于Blabina而言为稀释50000倍,对于BR而言为稀释10000倍,对于BB而言为稀释200倍,对于RB而言为稀释100倍。这显示出:与BB和RB相比,Blabina和BR具有几百倍以上的抑制活性。
Claims (5)
1.一种黑树莓提取物,其具有抑制黑素细胞中的酪氨酸酶的活性的效果。
2.一种美白剂,其含有权利要求1所述的黑树莓提取物。
3.一种饮食品,其含有权利要求1所述的黑树莓提取物。
4.一种黑树莓提取物的制造方法,其是权利要求1所述的黑树莓提取物的制造方法,其包括:
对黑树莓的果实进行冷冻的冷冻工序;
对所述黑树莓的果实进行解冻的解冻工序;
向利用所述解冻工序解冻后的所述黑树莓的果实中添加溶剂并进行搅拌的搅拌工序;和
将通过所述搅拌工序搅拌后的所述黑树莓的果实和溶剂静置、进行提取的提取工序。
5.一种美白方法,其中,使用了权利要求1所述的黑树莓提取物。
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CN116370382A (zh) | 2023-07-04 |
WO2019030956A1 (ja) | 2019-02-14 |
JP2019030267A (ja) | 2019-02-28 |
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