CN117618316A - 一种发酵组合物、面膜及其制备方法、应用 - Google Patents
一种发酵组合物、面膜及其制备方法、应用 Download PDFInfo
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- CN117618316A CN117618316A CN202311790594.6A CN202311790594A CN117618316A CN 117618316 A CN117618316 A CN 117618316A CN 202311790594 A CN202311790594 A CN 202311790594A CN 117618316 A CN117618316 A CN 117618316A
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Abstract
本发明公开了一种发酵组合物、面膜及其制备方法、应用，属于梳妆用的配制品领域。本发明提供的发酵组合物的制备方法包括步骤有：称取小麦胚芽、玉米胚芽、银耳干、甘草根、芍药根，粉碎混合，加入水，常温浸泡、加热浸泡，分离，得滤渣和水提液；将水提液浓缩成浓缩液；得到的滤渣和得到的浓缩液混合得混合物料，将混合物料进行厌氧发酵和进行有氧发酵，最后得到发酵组合物。本发明提供的发酵组合物对皮肤具有抗炎、舒缓、抗氧化的作用。
Description
技术领域
本发明属于梳妆用的配制品领域，具体涉及一种发酵组合物、面膜及其制备方法、应用。
背景技术
发酵是微生物在一定条件下进行生理活动，借助于酶的作用对有机物进行分解、转化取得蛋白质、维生素等各种营养以生长菌体，同时产生各种次级代谢产物，如：多糖、氨基酸等的过程。微生物在自身生长代谢过程中产生丰富的胞内胞外酶，如：纤维素酶、蛋白酶、果胶酶、淀粉酶等。能够进行水解、氧化、甲基化、酯化反应等多种反应，分解转化底物而生成新的活性成分，同时产生丰富的次级代谢产物。
发酵技术是指人们利用微生物的发酵作用，运用一些技术手段控制发酵过程，大规模生产发酵产品的技术，所以与其他加工方式相比，发酵能够产生新的活性物质，降低毒性，改变功效，同时由于微生物的水解作用，使发酵产物更易被吸收。
近年来，大量的研究结果显示，发酵技术在化妆品功效原料的开发方面具有良好的应用前景。首先，绿色天然的植物原料本身含有丰富的生物质营养成分，为发酵技术的应用提供了无限可能。其次，发酵技术在增强植物原料功效、降低毒副作用等方面具有得天独厚的优势。
妆食同源的植物原料中含有多种功效成分，例如维生素、氨基酸、矿物质、多糖、黄酮、多酚等，通常具有良好的保湿、美白、舒缓及延缓衰老等功效，但是，通过传统工艺制备的植物提取物，由于提取溶剂、工艺及制备方法等方面的局限，获取的功效成分有时难以达到相应的作用效果。
因此，本发明通过将多种可食用植物原料进行粗水提，并利用发酵技术作进一步的转化，通过控制发酵条件、优选原料类别、控制原料配比以及利用原料间的互促关系，得到一种具有抗氧化、抗炎舒缓的发酵组合。
发明内容
针对现有技术的不足，本发明目的在于通过将多种可食用植物原料进行粗水提，并利用发酵技术作进一步的转化，通过控制发酵条件、优选原料类别、控制原料配比以及利用原料间的互促关系，得到一种具有抗氧化、抗炎舒缓的发酵组合。
为实现上述目的，本发明公开了以下技术方案：
第一方面，本发明提供了一种发酵组合物的制备方法，包括以下步骤：
(1)称取小麦胚芽、玉米胚芽、银耳干、甘草根、芍药根，粉碎混合，加入水，料液质量比1:(10～12)，常温浸泡1～2h，然后密封加热至95～105℃下浸泡1～2h，分离得到滤渣和水提液；
(2)将水提液在55～60℃下浓缩至所述水提液体积的30～35％，得到浓缩液；水提液中的粗提营养物在长时间沸水煎煮下会失活变性，此处将滤渣和水提液分开后对水提液进行低温浓缩能够有效避免营养物质失活变性；
(3)将步骤(1)得到的滤渣和步骤(2)得到的浓缩液混合，得混合物料，将混合物料升温灭菌、冷却至25～30℃，加入乳酸杆菌，厌氧发酵48～72h，然后加入酵母菌，于25～30℃、400～800rpm的条件下进行有氧发酵48～72h，发酵结束后，将发酵液过滤，得到过滤液；
(4)将步骤(3)得到的过滤液除菌过滤，收集无菌滤液，然后浓缩至所述无菌滤液原体积的30～35％，得所述发酵组合物。
小麦胚芽除了含有脂肪外，其他营养成分的含量也很高，含蛋白质约30％，包括8种人体必须氨基酸等，多种氨基酸的含量超过鸡蛋和牛肉；含钙、铁、磷、镁、锰、铜、钾、钠、锌等多种矿物质；VB1、VB2、VB6、烟酸、泛酸、肌醇等多种维生素的含量相当高；碳水化合物约占40％，因而本发明中以小麦胚芽作为微生物发酵的主要基础原料；
小麦胚芽发酵产物具有丰富的苯醌类物质，如2，6-二甲氧基对苯醌(2，6-DMBQ)和2-甲氧基对苯醌(2-MBQ)，苯醌类物质能够有效的清除自由基，减少其对皮肤的损伤。
小麦胚芽与玉米胚芽中丰富的植物蛋白通过乳酸杆菌和酵母菌分别发酵后得到的发酵产物，其含有大量的小分子量的低聚肽，可被皮肤表皮直接吸收，并且皮肤对低聚肽的吸收速度快于皮肤对氨基酸的吸收速度，有效加强了抗氧化作用。
本发明优选小麦胚芽与玉米胚芽的原因是胚芽中所含有的营养成分比起其它部位的营养成份丰富很多，该优选有利于提高发酵产物的作用效果。
甘草根发酵产物含有多种抗炎舒缓成分，如甘草次酸、甘草甜素以及多种黄酮类化合物，使其具有抗炎、抗菌和抗氧化作用。
芍药根发酵产物含有芍药苷、芍药醇、维生素、没食子酸、紫云英苷、槲皮素-3-O-葡萄糖苷等成分，具有抗炎、抗氧化、抗菌作用；通过乳酸杆菌和酵母的发酵能提高芍药根的总酚、总黄酮含量以及抗氧化、抗衰老和抗胆碱酯酶活性。
具体地，所述步骤(1)中称取以下质量份数的原料：
优选地，所述乳酸杆菌为保加利亚乳酸杆菌，按每克混合物料活菌数为(50～90)×108CFU进行接种发酵。
优选地，所述酵母菌为卡尔斯伯酵母菌，按每克混合物料活菌数为(20～47)×108CFU进行接种发酵。
优选地，所述步骤(1)的分离为利用150～200目的筛网过滤分离，得到滤渣和水提液。
优选地，所述步骤(3)中将混合物料升温至100～120℃下灭菌30～45min。
优选地，所述步骤(4)除菌过滤为利用≤0.45μm聚醚砜滤芯进行过滤除菌。
第二方面，本发明提供了一种发酵组合物，由第一方面所述的发酵组合物的制备方法制备得到。
第三方面，本发明提供了一种发酵组合物面膜，所述面膜的面膜液中包含有第二方面所述的发酵组合物；
按质量百分数计，所述发酵组合物在所述面膜液中的添加量为15～20％。
第四方面，本发明提供了第二方面所述的发酵组合物在制备具有舒缓、保湿、美白效果的化妆水、护肤精华、护肤啫喱、护肤乳液、护肤膏霜和/或洁面产品、洗护产品中的应用。
本发明的有益效果：
1.本发明提供的发酵组合物具有优异的自由基清除性能，当其浓度为18wt％自由基清楚性能达到75％以上，对改善皮肤健康，延缓皮肤衰老具有重要的影响。
2.本发明提供的发酵组合物对炎症因子TNF-α具有明显的抑制效果，具有较好的抗炎、舒缓的作用。
具体实施方式
下面将结合本发明的实施例，对本发明中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅是本发明的一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
以下结合具体实施例对本发明的技术方案做进一步详细说明，应当理解，以下实施例仅仅用以解释本发明，并不用于限定本发明。
本发明中：
保加利亚乳酸杆菌的拉丁学名为Lactobacillus bulgaricus，购自派生生物科技有限公司；
嗜酸性乳酸杆菌的拉丁学名为Lactobacillus acidophilus，购自派生生物科技有限公司；
卡尔斯伯酵母菌拉丁学名为Saccharomyces carlsbergensis，购自上海谷研实业有限公司；
酿酒酵母拉丁学名为Saccharomyces Cerevisiae，购自上海谷研实业有限公司；
其他原料均可市售得到。
实施例1-4的制备：
原料按表1中的质量份数进行称取；
步骤(1)，称取小麦胚芽、玉米胚芽、银耳干、甘草根、芍药根，粉碎混合，加入水，料液质量比1:(10～12)，常温浸泡1～2h，然后密封加热至95～105℃下浸泡1～2h，利用150目的筛网过滤分离得到滤渣和水提液；
步骤(2)，将水提液在55～60℃下浓缩至所述水提液体积的30～35％，得到浓缩液；
步骤(3)，将步骤(1)得到的滤渣和步骤(2)得到的浓缩液加入到发酵罐中混合，得混合物料，将混合物料升温至100～120℃灭菌30～45min、冷却至25～30℃，加入保加利亚乳酸杆菌，厌氧发酵48～72h，然后加入卡尔斯伯酵母菌，于25～30℃、400～800rpm搅拌转速的条件下进行有氧发酵48～72h，发酵结束后，将发酵液过滤，得到过滤液；
步骤(4)，将步骤(3)得到的过滤液利用0.22μm聚醚砜滤芯进行过滤除菌，收集无菌滤液，然后浓缩至所述无菌滤液原体积的30～35％，得实施例发酵组合物。
表1实施例1～4原料质量份数及制备参数
对比例1-10的制备：
步骤(1)，按表2、3质量份数称取原料，粉碎混合，加入水，料液质量比1:10，常温浸泡0.5h或1.5h，然后密封加热至95℃下浸泡0.5h或1h，利用150目的筛网过滤分离得到滤渣和水提液；
步骤(2)，将水提液在55～60℃下浓缩至所述水提液体积的30％，得到浓缩液；
步骤(3)，将步骤(1)得到的滤渣和步骤(2)得到的浓缩液加入到发酵罐中混合，得混合物料，将混合物料升温至120℃灭菌30min、冷却至27℃，加入保加利亚乳酸杆菌，厌氧发酵24h或48h，然后加入卡尔斯伯酵母菌，于27℃、400rpm搅拌转速的条件下进行有氧发酵24h或48h，发酵结束后，将发酵液过滤，得到过滤液；
步骤(4)，将步骤(3)得到的过滤液利用0.22μm聚醚砜滤芯进行过滤除菌，收集无菌滤液，然后浓缩至所述无菌滤液原体积的33％，得对比例1-10发酵组合物。(上述各对比例制备具体参数按表2、3对应参数进行)
表2对比例1-5原料质量份数
注：“——”表示未添加。
性能检测：
DPPH自由基清除能力的测定
待检测液：取一定量实施例1-4和对比例1-10的发酵组合物，利用生理盐水分别稀释至12wt％、16wt％、18wt％、20wt％。
DPPH用无水乙醇配制成2×10-4mol/L的DPPH溶液，取待检测液、DPPH溶液各2mL若干份，分别将混匀后室温下放置30分钟，于波长517nm处测定吸光度得Ai，每个样品平行测定3次，取平均值。
以Vc作为阳性对照，以无水乙醇代替DPPH测定吸光度值作为对照Aj空白，以DPPH溶液和无水乙醇混合测吸光度Ac。
按下式(1)计算各待测样品对DPPH自由基的清除率，并记录至下表4。
清除率(％)＝(1-(Ai-Aj)/Ac)×100％...........(1)
式中：Ai为2mL待检测液+2mLDPPH试剂混合液的吸光度；Aj为2mL待检测液+2mL无水乙醇混合液的吸光度；Ac为2mLDPPH溶液+2mL无水乙醇混合液的吸光度。
表4DPPH自由基清除率
结果分析：
根据实验结果可知，本发明提供的发酵组合物具有优异的自由基清除性能，当其浓度为18wt％自由基清楚性能达到75％以上，对改善皮肤健康，延缓皮肤衰老具有重要的影响。
炎症因子抑制实验
待测样品：取一定量实施例1-4和对比例1-10的发酵组合物，利用生理盐水分别稀释至12wt％、16wt％、18wt％、20wt％。
实验方法：采用RAW264.7巨噬细胞作为研究对象，通过脂多糖LPS(细菌内毒素)刺激细胞建立细胞炎症模型。接种巨噬细胞至12孔板，在培养箱以37℃、5％CO2通气量条件下孵育24h，分别加入实施例1-4和对比例1-10的稀释液(12wt％、16wt％、18wt％、20wt％)，2h后加入LPS(1μg/mL)，记为T1，并设置不添加稀释液只添加LPS的组记作T总以及不添加LPS只添加稀释液的组记作T0，刺激24h，收集上清液，离心，检测。采用ELISA试剂盒分析RAW264.7的促炎性炎症因子TNF-α释放量水平。
TNF-α抑制率由如下公式计算得到：
TNF-α抑制率(％)＝[(T总-T1)/(T总-T0)]×100％。
TNF-α抑制率越高，说明待测样品的抗炎效果越好。TNF-α抑制测试结果如表5所示。
表5TNF-α抑制率
结果分析：
从实验结果可以看出，本发明提供的发酵组合物对炎症因子TNF-α具有明显的抑制效果。实施例1-4的发酵组合物的浓度12％(wt)、16％(wt)、18％(wt)、20％(wt)对炎症因子TNF-α的抑制率分别达到了12％、18％、20％和15％，具有较好的抗炎、舒缓的作用。
最后应说明的是：以上所述仅为本发明的优选实施例而已，并不用于限制本发明，尽管参照前述实施例对本发明进行了详细的说明，对于本领域的技术人员来说，其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分技术特征进行等同替换。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (10)
1.一种发酵组合物的制备方法，其特征在于，包括以下步骤：
(1)称取小麦胚芽、玉米胚芽、银耳干、甘草根、芍药根，粉碎混合，加入水，料液质量比1:(10～12)，浸泡1～2h，然后密封加热至95～105℃下浸泡1～2h，分离得到滤渣和水提液；
(2)将水提液在55～60℃下浓缩至所述水提液体积的30～35％，得到浓缩液；
(3)将步骤(1)得到的滤渣和步骤(2)得到的浓缩液混合，得混合物料，将混合物料升温灭菌、冷却至25～30℃，加入乳酸杆菌，厌氧发酵48～72h，然后加入酵母菌，于25～30℃、400～800rpm的条件下进行有氧发酵48～72h，发酵结束后，将发酵液过滤，得到过滤液；
(4)将步骤(3)得到的过滤液除菌过滤，收集无菌滤液，然后浓缩至所述无菌滤液原体积的30～35％，得所述发酵组合物。
2.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述步骤(1)中称取以下质量份数的原料：
3.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述乳酸杆菌为保加利亚乳酸杆菌，按每克混合物料活菌数为(50～90)×108CFU进行接种发酵。
4.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述酵母菌为卡尔斯伯酵母菌，按每克混合物料活菌数为(20～47)×108CFU进行接种发酵。
5.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述步骤(1)的分离为利用150～200目的筛网过滤分离，得到滤渣和水提液。
6.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述步骤(3)中将混合物料升温至100～120℃下灭菌30～45min。
7.根据权利要求1所述的发酵组合物的制备方法，其特征在于，所述步骤(4)除菌过滤为利用≤0.45μm聚醚砜滤芯进行过滤除菌。
8.一种发酵组合物，其特征在于，由权利要求1～7任一所述的发酵组合物的制备方法制备得到。
9.一种发酵组合物面膜，其特征在于，所述面膜的面膜液中包含有权利要求8所述的发酵组合物；
按质量百分数计，所述发酵组合物在所述面膜液中的添加量为15～20％。
10.权利要求8所述的发酵组合物在制备具有舒缓、保湿、美白效果的化妆水、护肤精华、护肤啫喱、护肤乳液、护肤膏霜和/或洁面产品、洗护产品中的应用。
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