CN106511582A - 一种天然原料萃取物为有效成分的抗氧化组合物及其制法 - Google Patents
一种天然原料萃取物为有效成分的抗氧化组合物及其制法 Download PDFInfo
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Abstract
本发明公开了一种天然原料萃取物为有效成分的抗氧化组合物，所述制法包括如下步骤：第一步，粉碎，第二步，萃取，第三步，发酵，第四步，复合发酵萃取物，上述制法制成的天然原料萃取物为有效成分的抗氧化组合物，包括如下组份重量百分比：包括马黛70~30%，鱼腥草10~40%，八角茴香10~40%和海带0.1~10%。本发明的天然原料萃取物为有效成分的抗氧化组合物及其制法，使用具备抗氧化活性的马黛、鱼腥草、八角茴香及海带的复合提取物，利用酵母发酵，提升抗氧化酶的发现可能，并提升抗氧化组成物的抗氧化活性，组成物可消除DPPH自由基及ABTS自由基的活性，提升SOD‑1,SOD‑2及过氧化氢酶的表现，呈现好的抗氧化活性，利用其而制成的新化妆原料组合物，药学性组合物及食品功效显著。
Description
技术领域
本发明涉及一种抗氧化组合物及其制法，具体涉及一种天然原料萃取物为有效成分的抗氧化组合物及其制法，属于抗氧化组合物及其制法技术领域。
背景技术
空气中一般存在的分子状态稳定的三重基础抗氧化素，依据体内酵素系、还原代谢、化学药品、污染物质、光化学反应等环境及生化反应要素，转化为反应性极大的自由基活性氧，从而对细胞的构成成分进行不可逆的破坏；活性氧可根据在体内防御组织过氧化物歧化酶,过氧化氢酶,过氧化物酶,谷胱甘肽等的抗氧化性酶及维生素C,维生素E(等抗氧化物质的作用下达最小化；但当活性氧过度暴露或生物体防御力出现异常情况，平衡遭到破坏时， 活性氧可引起生物体致命的氧毒性；体内过多的活性氧会与细胞构成成分脂质、蛋白质、糖、DNA等反应起到破坏作用，从而可引发老化甚至癌症、脑中风、帕金森病等脑部疾病和心脏疾病、贫血、动脉硬化、皮肤损伤、炎症、类风湿、自体免疫疾病等多种疾病；在证实上述氧化性压力是引起老化等多种疾病重要原因的同时，目前对可清除人体内活性氧的抗氧化剂研发也在有条不紊的进行；对抗氧化剂的研究是1969年由McCord和Fridovich两位学者发现了可消除超氧自由基的SOD酵素为契机而全面展开，近期因发现老化、成人病等疾病与活性氧有关，因而对可调节活性氧的抗氧化剂的研究广泛开展，而因合成抗氧化剂存在潜在副作用的威胁，因此利用从天然物质中提取天然抗氧化剂的课题研究被广泛关注；虽然合成抗氧化剂和BHT因其效果、经济性及安全性等因素被广泛使用，但因合成添加剂除了一般禁忌现象之外，同时还存在长期过量服用时，对胃黏膜、肺、肾脏、循环系统等引起严重的毒副作用，故而其使用量被法规所限制；此外对人体无害且抗氧化力优秀的天然抗氧化剂的研究由来已久，虽然类似生育酚的天然抗氧化剂对人体无害，也具备可单独阻止氧化连锁反应的能力，但缺点是价格非常昂贵；因此，对安全且抗氧化作用卓越的经济型新抗氧化剂的开发迫在眉睫。
发明内容
为解决上述问题，本发明提出了一种天然原料萃取物为有效成分的抗氧化组合物及其制法，以马黛、鱼腥草、八角茴香及海带的复合发酵萃取物为有效成分的抗氧化组合物，并与利用其而制成的化妆品组合物、医学性组合物及食品原料相关。
本发明的天然原料萃取物为有效成分的抗氧化组合物制法，所述制法包括如下步骤：
第一步，粉碎，将马黛，鱼腥草，八角以及海带混合粉碎或单独粉碎，得到原料粉末；
第二步，萃取，将第一步得到的原料粉末利用热水萃取，得到萃取物；
第三步，发酵，使用酵母和益生菌将第二步得到的萃取物进行发酵，得到发酵物；
第四步，复合发酵萃取物，将第三步得到的发酵物浓缩及干燥，得到复合发酵萃取物。
进一步地，所述第三步其发酵过程具体如下：
首先，将所述第二步得到的萃取物中进行酵母及益生菌接种；
接着，将上一步骤完成接种的萃取物用益生菌发酵；
最后，将上一步骤完成益生菌发酵的萃取物进行酵母发酵。
上述制法制成的天然原料萃取物为有效成分的抗氧化组合物，包括如下组份重量百分比：包括马黛70~30%，鱼腥草10~40%，八角茴香10~40%和海带0.1~10%。
上述制法制成的天然原料萃取物为有效成分的抗氧化组合物，包括如下组份重量百分比：包括马黛提取物70~30%，鱼腥草提取物10~40%，八角茴香提取物10~40%和海带提取物0.1~10%。
进一步地，所述组合物还包括组合物总量0.1%的益生菌、组合物总量1%的酵母和所述组合物其调和糖度为5~30。
再进一步地，所述酵母为酿酒酵母；所述益生菌为胚牙乳杆菌。
进一步地，所述萃取物其益生菌发酵具体为：用组合物总量0.1%的益生菌接种，并30~37℃的条件下发酵3~24小时；所述萃取物其酵母发酵具体为：利用组合物总量1%的酵母接种，并在15~25℃的条件下发酵3~24小时。
进一步地，所述复合发酵萃取物为体现DPPH及ABTS自由基清除活性的复合发酵萃取物。
进一步地，所述复合发酵萃取物为促进过氧化物消除酶-1，过氧化物消除酶-及过氧化氢酶的复合发酵萃取物。
进一步地，所述复合发酵萃取物作为为有效成分的抗氧化化妆原料组成物，作为有效成分的抗氧化药学性组成物，作为有效成分的抗氧化食品。
本发明与现有技术相比较，本发明的天然原料萃取物为有效成分的抗氧化组合物及其制法，使用具备抗氧化活性的马黛、鱼腥草、八角茴香及海带的复合提取物，利用酵母发酵，提升抗氧化酶的发现可能，并提升抗氧化组成物的抗氧化活性，根据此法而发明的组成物，可消除DPPH自由基及ABTS自由基的活性，提升SOD-1,SOD-2及过氧化氢酶的表现，呈现出卓越的抗氧化活性，利用其而制成的新化妆原料组合物，药学性组合物及食品功效显著。
附图说明
图1是马黛、鱼腥草、八角茴香及海带复合发酵提取物的制作工艺图。
图2是显示实例1和对照1的DPPH自由基活性试剂图。
图3是显示实例1和对照1的DPPH自由基活性测试结果显示图。
图4是显示实例2和对照1的ATBS自由试剂图。
图5是显示实例2和对照1的ATBS自由基活性测试结果显示图。
图6是在Raw 264.7老鼠巨噬细胞中根据实例1和对照1处理结果，抗氧化酶SOD-1,SOD-2及过氧化氢酶的mRNA的表现结果图。
具体实施方式
如图1所示，本发明的天然原料萃取物为有效成分的抗氧化组合物制法，所述制法包括如下步骤：
第一步，粉碎，将马黛，鱼腥草，八角以及海带混合粉碎或单独粉碎，得到原料粉末；
第二步，萃取，将第一步得到的原料粉末利用热水萃取，得到萃取物；
第三步，发酵，使用酵母和益生菌将第二步得到的萃取物进行发酵，得到发酵物；
第四步，复合发酵萃取物，将第三步得到的发酵物浓缩及干燥，得到复合发酵萃取物。
所述第三步其发酵过程具体如下：
首先，将所述第二步得到的萃取物中进行酵母及益生菌接种；
接着，将上一步骤完成接种的萃取物用益生菌发酵；
最后，将上一步骤完成益生菌发酵的萃取物进行酵母发酵。
上述制法制成的天然原料萃取物为有效成分的抗氧化组合物，包括如下组份重量百分比：包括马黛70~30%，鱼腥草10~40%，八角茴香10~40%和海带0.1~10%。
上述制法制成的天然原料萃取物为有效成分的抗氧化组合物，包括如下组份重量百分比：包括马黛提取物70~30%，鱼腥草提取物10~40%，八角茴香提取物10~40%和海带提取物0.1~10%。
所述组合物还包括组合物总量0.1%的益生菌、组合物总量1%的酵母和所述组合物其调和糖度为5~30。
所述酵母为酿酒酵母，更为理想的酵母为酿酒酵母113-4(KCTC12092BP)；所述益生菌为胚牙乳杆菌。
所述萃取物其益生菌发酵具体为：用组合物总量0.1%的益生菌接种，并30~37℃的条件下发酵3~24小时；所述萃取物其酵母发酵具体为：利用组合物总量1%的酵母接种，并在15~25℃的条件下发酵3~24小时。
所述复合发酵萃取物为体现DPPH及ABTS自由基清除活性的复合发酵萃取物。
所述复合发酵萃取物为促进过氧化物消除酶-1，过氧化物消除酶-及过氧化氢酶的的复合发酵萃取物。
所述复合发酵萃取物作为为有效成分的抗氧化化妆原料组成物，作为有效成分的抗氧化药学性组成物，作为有效成分的抗氧化食品。
实施例1：
以100克为准，将马黛、鱼腥草、八角及海带按48：25：25：2的比例混合粉碎后，投入该混合物量10倍的纯净水，用98℃的热水熬制4小时,通过热萃取制造马黛、鱼腥草、八角茴香及海带的复合提取物；所获得的复合提取物中加入1%的酿酒酵母113-4及0.1%的胚牙乳杆菌进行接种后，在37℃温度下进行24小时的首次发酵（益生菌发酵）及25℃下24小时二次发酵（酵母发酵）；之后将试样在水浴锅中80℃加热30分钟后结束发酵；然后，50℃下进行浓缩/干燥，制得10~20白利度的复合发酵萃取物溶液；其中，利用Blois方法测定DPPH(2,2-diphenyl-2-picryl-hydrazyl)自由基清除活性；首先，将各浓度试剂用甲醇溶剂溶解，将900㎕的DPPH溶液（150μM）及各试剂100㎕进行混合搅拌；将混合后的试剂注入母牛体内，约30分钟后于517nm处进行吸光度测定，对各DPPH自由基清除活性实验反复进行3次所得平均值后, 将对照组的吸光度减少程度按下列数学公式算出, 测出试剂DPPH自由基减少50%时所需的试剂浓度(IC50)： DPPH自由基清除活性(%)= (A-B)/A x 100，代替试剂添加了甲醇的DPPH的吸光度， 添加了试剂的DPPH吸光度
上述实验结果如图2和图3所示，实例1和比较示例1的DPPH自由基清除活性受浓度变化影响，比较示例1的IC50为306.6 ㎍/mL，实例1的IC50为266.1 ㎍/mL，实例1的IC50值较于比较示例1低26.2%。
实施例2：
ABTS自由基清除活性采用Roberta等方式确认ABTS的自由基清除活性；蒸馏水中加入7mM ABTS和2.45 mM过硫酸钾常温反应16小时后，ABTS阳离子生成后，保持734 nm处吸光度值为0.7以下进行稀释，制造出ABTS自由基溶液；之后制造多种浓度的试剂，将900㎕的ABTS自由基溶液和100㎕的各试剂混合搅拌；将此混合试剂30℃下反应6分钟后测定吸光度数值，各实验反复进行三次后获得ABTS自由基清除活性平均值，对照组的吸光度减少程度根据下列数学公式算出, 测出试剂ABTS自由基减少50%时所需试剂浓度(IC50)：
ABTS自由基清除活性(%)= (A-B)/A x100
A：代替试剂添加了甲醇的ABTS吸光度， B：添加试剂的ABTS吸光度。
上述结果如图4和图5所示，实例1和比较示例1的ABTS自由基清除活性受浓度增加影响，比较示例1的IC50为612.6 ㎍/mL，实例1的IC50为504.3 ㎍/mL，实例1的IC50值低于比较示例117.7%；此结果显示，实例1相较于比较示例1的ABTS自由基清除活性明显更为优秀。
实施例3：
帮助促进发现抗氧化酵素，为确认促进过氧化物消除酶-1，过氧化物消除酶-2及过氧化氢酶等抗氧化酶的发现，进行以下实验：
首先，把小鼠的巨噬细胞 RAW 264.7(American Type Culture Collection®(ATCC), Manassas, VA, USA;ATCC TIB-71™)培养于含10%胎牛血清(FBS)的DMEM(Dulbecco's modified eagle's medium)培养基培养后，以1x10 cells/well放入微量滴定板(6-well microtiter plate)中，在37℃，5%的二氧化碳(CO2)条件下培养24小时；将50μM过氧化氢(hydrogen peroxide)，实例1（10ppm）及比较示例1（25ppm）的试剂一起处理24小时后，从小鼠巨噬细胞RAW 264.7开始使用TRIzol试剂(Invitrogen, Carlsbad, CA,USA)得到总RNA后用逆转录酶进行逆转后实施以下RT-PCR分析；首先对上述RNA利用逆转录酶(reverse transcriptase)进行逆转合成cDNA；RT-PCR则为利用以下特定引物(Primerpairs)进行；
β-Actin，
Forward primer: 5′- CAGCTCAGTAACAGTCCGCC - 3′，
Reverse primer: 5′- TCACTATTGGCAACGAGCGG - 3′，
SOD-1，
Forward primer: 5′- AAGGCCGTGTGCGTGCTGAA - 3′，
Reverse primer: 5′- CAGGTCTCCAACATGCCTCT - 3′，
SOD-2，
Forward primer: 5′- GCACATTAACGCGCAGATCA - 3′，
Reverse primer: 5′- AGCCTCCAGCAACTCTCCTT - 3′，
Catalase，
Forward primer: 5′- GCAGATACCTGTGAACTGTC - 3′，
Reverse primer: 5′- GTAGAATGTCCGCACCTGAG - 3′。
上述结果如图6所示，比较示例1和实例1表明依据过氧化氢的处理而减少的SOD-1, SOD-2及Catalase的mRNA发现率随浓度变化而增加，相较于比较示例1来看，实例1中抗氧化酶的mRNA发现率的增加更有效；此结果表明经过处理的本发明发酵物的实例1与比无处理的比较示例1相比，因氧化压力下减少的抗氧化酶增长更有效，保护巨噬细胞的能力更为优秀。
以上述依据本发明的理想实施例为启示，通过上述的说明内容，相关工作人员完全可以在不偏离本项发明技术思想的范围内，进行多样的变更以及修改。本项发明的技术性范围并不局限于说明书上的内容，必须要根据权利要求范围来确定其技术性范围。
Claims (10)
1.一种天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于，所述制法包括如下步骤：
第一步，粉碎，将马黛，鱼腥草，八角以及海带混合粉碎或单独粉碎，得到原料粉末；
第二步，萃取，将第一步得到的原料粉末利用热水萃取，得到萃取物；
第三步，发酵，使用酵母和益生菌将第二步得到的萃取物进行发酵，得到发酵物；
第四步，复合发酵萃取物，将第三步得到的发酵物浓缩及干燥，得到复合发酵萃取物。
2.根据权利要求1所述的以天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于：所述第三步其发酵过程具体如下：
首先，将所述第二步得到的萃取物中进行酵母及益生菌接种；
接着，将上一步骤完成的接种萃取物用益生菌发酵；
最后，将上一步骤完成益生菌发酵萃取物进行酵母发酵。
3.一种以天然原料萃取物为有效成分的抗氧化组合物，其特征在于，所述组合物包括如下组份重量百分比：包括马黛70~30%，鱼腥草10~40%，八角茴香10~40%和海带0.1~10%。
4.一种以天然原料萃取物为有效成分的抗氧化组合物，其特征在于，所述组合物包括如下组份重量百分比：包括马黛提取物70~30%，鱼腥草提取物10~40%，八角茴香提取物10~40%和海带提取物0.1~10%。
5.根据权利要求3或4所述的以天然原料萃取物为有效成分的抗氧化组合物，其特征在于：所述组合物还包括组合物总量0.1%的益生菌、组合物总量1%的酵母和所述组合物其调和糖度为5~30。
6.根据权利要求5所述的以天然原料萃取物为有效成分的抗氧化组合物，其特征在于：所述酵母为酿酒酵母；所述益生菌为胚牙乳杆菌。
7.根据权利要求1所述的以天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于：所述萃取物其益生菌发酵具体为：用组合物总量0.1%的益生菌接种，并30~37℃的条件下发酵3~24小时；所述萃取物其酵母发酵具体为：利用组合物总量1%的酵母接种，并在15~25℃的条件下发酵3~24小时。
8.根据权利要求1所述的以天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于：所述复合发酵萃取物为体现DPPH及ABTS自由基清除活性的复合发酵萃取物。
9.根据权利要求1所述的以天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于：所述复合发酵萃取物为促进过氧化物消除酶-1，过氧化物消除酶-及过氧化氢酶的复合发酵萃取物。
10.根据权利要求1所述的以天然原料萃取物为有效成分的抗氧化组合物制法，其特征在于：所述以复合发酵萃取物作为有效成分的抗氧化化妆原料组成物，以其作为有效成分的抗氧化药学性组成物，及以其作为有效成分的抗氧化食品。
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| KR102206166B1 (ko) | 2019-04-03 | 2021-01-21 | 동의대학교 산학협력단 | 흑임자 전처리용 조성물로 전처리된 흑임자 오일의 제조방법 |
| KR102251306B1 (ko) * | 2019-11-01 | 2021-05-12 | (주)정코스메틱 | 락토바실러스 플란타룸 배양 추출물을 포함하는 화장료 조성물 |
| KR102255580B1 (ko) | 2020-07-14 | 2021-05-25 | (주)지에프씨생명과학 | 마테 추출물 및 트라넥삼산의 혼합물을 유효성분으로 포함하는 화장료 조성물 |
| KR102310691B1 (ko) * | 2020-09-02 | 2021-10-12 | 주식회사 네오캠코리아 | 산야초 발효 추출물, 이의 제조방법 및 이의 용도 |
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| CN110368337A (zh) * | 2019-07-19 | 2019-10-25 | 广州无添加主义化妆品有限公司 | 一种含玫瑰果提取物的面膜及制备方法 |
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