CN115429736A - 一种知母总皂苷提取物的制备方法及在化妆品中的应用 - Google Patents
一种知母总皂苷提取物的制备方法及在化妆品中的应用 Download PDFInfo
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61Q19/005—Preparations for sensitive skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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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
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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Abstract
本发明公开一种知母总皂苷提取物的制备方法及在化妆品中的应用,知母总皂苷提取物通过下述方法制备:取知母药材,水煎煮提取,过滤,获得提取液;提取液经大孔树脂柱纯化后,减压回收溶剂,干燥,粉碎过筛即得。制备得到的知母总皂苷提取物经验证具有良好的抗透明质酸酶活性。本发明所得的总皂苷提取物纯度高,制备方法简单、安全,便于工厂化生产,且提取物功效明确,可以应用于制备抗过敏类型化妆品的中。
Description
技术领域
本发明属于化妆品领域,具体涉及一种知母总皂苷提取物的制备方法及在化妆品中的应用。
背景技术
皮肤过敏反应又称皮肤变态反应,是皮肤对某种物质产生的免疫源性炎症反应,发生在人身上又称为变应性接触皮炎,以瘙痒、红斑、丘疹、水疱、融合水疱为特征。目前化妆品的使用越来越普及,特别是很多化妆品在研发制备过程中,除了配方中的功效成分、增稠体系外,还会添加多种化学物质,例如防腐剂、香料香精、抗菌剂、防晒剂等。这类物质很容易成为皮肤过敏的过敏源,再加上部分人群对皮肤医学认知浅薄,忽略了自身皮肤状态的把握,往往会导致护肤品使用不当等情况的发生,正是这些现象容易导致皮肤过敏,引起许许多多的不良症状,如红肿、脱屑、瘙痒等,因此皮肤敏感问题越来越受到人们的重视。近年来化妆品行业正在逐渐朝着崇尚绿色、回归自然的方向发展。“天然提取,杜绝添加,没有刺激”已经逐渐变成化妆品开发领域的主要研究方向。在常见的日用护肤品和化妆品中,添加从天然产物中提取的具有抗过敏功效的成分,降低使用者的过敏风险,己经被作为重点研究指导方向应用于化妆品行业中。因此,天然产物的提取、抗过敏功效的研究和应用途径的探索,日益被国内外研究人员和学者关注。
知母为百合科植物Anemarrhena asphodeloides Bge的干燥根茎,知母中含有甾体皂苷、双苯吡酮类、木质素类和多糖类成分,在临床中应用广泛,具有抗血小板血栓、改善阿尔茨海默症、抗肿瘤、抗炎、解热等作用,目前较被关注的是皂苷类成分。研究表明,知母皂苷具有广泛的药理活性,包括抗氧化、抗衰老、抗凝血、抗肿瘤、抗骨质疏松、抗炎、降血压、降血糖及降血脂等。但是迄今为止,国内外未见关于知母提取物或者知母皂苷的抗敏抗刺激作用方面的研究或文献报道。
发明内容
本发明的目的是提供一种知母总皂苷提取物的制备方法,该提取物其具有良好的抗透明质酸酶活性,可以应用在相关化妆品中。
本发明目的通过以下技术方案实现:
一种知母总皂苷提取物的制备方法,包括下述步骤:
步骤1:取知母药材,加水煎煮三次,第一次加10倍量水,提取时间为2小时,第二次、第三次各加8倍量水,提取时间各1.5小时,过滤,合并滤液。
步骤2:滤液徐徐注入已处理好的AB-8型大孔树脂柱,用1倍柱床体积的水洗脱,弃去水洗脱液,用4倍柱床体积的0.2%NaOH水溶液洗脱,用水洗至中性后,再用收集4倍柱体积的80%乙醇洗脱,并收集洗脱液。
步骤3:将上述步骤2的80%乙醇洗脱液真空减压浓缩至相对密度为1.10~
1.15(50~60℃),60℃真空干燥,粉碎,过60-100目筛,即得知母总皂苷提取物。
通过上述方法制备得到的知母总皂苷提取物,活性测试结果表明具有良好的抗透明质酸酶作用,可以用于制备具有抗敏效果的化妆品中。
本发明进一步提供一种知母总皂苷提取物在化妆品中的应用,所述化妆品包括采用上述方法提取得到的知母总皂苷提取物及辅料。
其中,所述的知母总皂苷提取物用作化妆品中的抗过敏成分。
与现有技术相比,本发明的有益效果在于:
1、本发明以知母为原料,通过水煎煮和大孔树脂纯化获得知母总皂苷提取物,HPLC含量测定结果显示知母总皂苷提取物中皂苷单体类活性成分含量高。本发明所述的知母总皂苷提取物的制备方法工艺合理,简便易行,生产成本低,可用于大批量的工业化生产。
2、本发明制得的知母总皂苷提取物对透明质酸酶有着良好的抑制作用,表明知母总皂苷提取物具有抗过敏功效,可以作为抗过敏的活性成分或作为添加剂用于美容化妆品。
3、本发明提供了一种植物活性提取物的新应用,可为天然绿色化妆品原料提供更多的功能成分选择。
附图说明
图1知母皂苷B2对照品HPLC-ELSD含量测定色谱图
图2知母皂苷B3对照品HPLC-ELSD图
图3知母皂苷A3对照品HPLC-ELSD图
图4知母总皂苷供试品HPLC-ELSD图
具体实施方式
下面通过具体的实施例详细介绍本发明实质性内容,但不能以此限定本发明的保护范围。
实施例1:知母总皂苷提取物的制备
取知母药材,加水煎煮三次,第一次加10倍量水,提取时间为2小时,第二次、第三次各加8倍量水,提取时间各1.5小时,滤过,合并滤液。滤液徐徐注入已处理好的AB-8型大孔树脂柱(处理过程为常规通用预处理方法),用1倍柱床体积的水洗脱,弃去水洗脱液,用4倍柱床体积0.2%的NaOH水溶液洗脱,用水洗至中性后,收集4个柱体积的80%乙醇洗脱液。80%乙醇洗脱液真空减压浓缩至相对密度为1.15,60℃真空干燥,粉碎,过80目筛,即得知母总皂苷提取物。
实施例2:知母粗提物的制备
本实施例为知母粗提物常规制备工艺。用于以下与本发明方法所制备的知母总皂苷提取物进行活性对比。
取知母药材,加水煎煮三次,第一次加10倍量水,提取时间为2小时,第二次、第三次各加8倍量水,提取时间各1.5小时,滤过,合并滤液。滤液真空减压浓缩至粘稠状,再于60℃真空干燥箱中干燥即得知母粗提物。
实施例3:知母总皂苷提取物HPLC含量测定
(1)色谱条件:采用十八烷基硅烷键合硅胶为填充剂的色谱柱;以乙腈为流动相A,以0.1%的甲酸为流动相B,梯度洗脱;采用蒸发光散射检测器。梯度洗脱程序见下表。
表1梯度洗脱表
时间(min) | 流动相A(乙腈) | 流动相B(0.1%甲酸) |
0 | 10% | 90% |
30 | 90% | 10% |
(2)对照品溶液的制备:精密称取知母皂苷B2对照品适量,加水制成每1ml含0.93mg的溶液,即浓度为0.93mg/ml。精密称取知母皂苷B3对照品适量,加甲醇制成每1ml含0.81mg的溶液,即浓度为0.81mg/ml。精密称取知母皂苷A3对照品适量,加甲醇制成每1ml含0.576mg的溶液,即浓度为0.576mg/ml。
(3)供试品溶液的制备:精密称取知母总皂苷提取物样品11.1mg,加50%甲醇溶解并定容于10ml的容量瓶中,即浓度为1.11mg/ml。
(4)分别吸取知母皂苷B2、B3、A3对照品20μl、10μl,知母总皂苷提取物样品10ul注入高效液相色谱仪,测定,即得。
(5)知母总皂苷提取物的HPLC含量测定色谱图见图1-图4,测定结果显示知母总皂苷提取物中知母皂苷B2含量为40.6%,知母皂苷B3含量为7.7%,知母皂苷A3含量为2.7%。
实施例4:知母总皂苷提取物的活性测定
(1)实验原理本方法采用透明质酸酶体外抑制实验Elson-Morgan测定抗过敏功效。透明质酸酶是过敏反应的参与者,研究表明透明质酸酶与炎症、过敏有强相关性,许多抗过敏药物有强抑制透明质酸酶活性的作用。实验过程中,透明质酸酶能够不加选择地水解透明质酸中的β-N-乙酰葡糖胺与D-葡糖醛酸盐之间的1,4-键,得到β-N-乙酰葡糖胺,β-N-乙酰葡糖胺在碱性条件下可以与乙酰丙酮缩合生成生色原2-甲基-3-二乙酰吡咯衍生物,生色原与对-二甲氨基苯甲醛在浓盐酸乙醇中生色。
(2)实验样品制备方法将实施例1制得的知母总皂苷提取物样品用去离子水配制成0.25和2mg/ml的溶液,另将实施例2制得的知母粗提物样品用去离子水配制成2mg/ml的溶液。
(3)实验方法A、B号试管(0.5mL样品溶液和0.5mL透明质酸酶500U/mL),C、D号试管(0.5mL蒸馏水和0.5mL pH=5.4醋酸缓冲溶剂),37℃保温20min;加入0.1mL(2.5mol/L)氯化钙溶液,37℃保温20min;A、C号试管中加入0.5mL透明质酸钠(0.5mg/ml),B、D号试管中加入0.5mL pH=5.4醋酸缓冲溶剂,37℃保温40min;室温放置10min,加入0.5mL乙酰丙酮溶液(3.5mL乙酰丙酮溶于50mL 0.1mol/L碳酸钠溶液中)、0.1mL NaOH溶液(5mol/L)和0.5mL蒸馏水,放入沸水浴中15min;冷却5min,放入冰水浴10min;室温放置10min,后加入蒸馏水0.5mL、Ehrlich试剂0.5mL、浓HCl 2mL,补加无水乙醇至10mL,振摇混匀,静置30min后在于530nm处测定各管吸光度值。按下式计算:透明质酸酶抑制率/%=[(C-D)-(A-B)]/(C-D)×100%。式中:A为A管(试样溶液)的OD值;B为B管(试样空白溶液)的OD值;C为C管(对照溶液)的OD值;D为D管(对照空白)的OD值。
(4)实验结果结果见表2。
表2对透明质酸酶的抑制率(%)
由表2中数据可知,知母粗提物在2mg/ml浓度下,对透明质酸酶无抑制活性。而知母总皂苷提取物在浓度为2mg/ml时,对透明质酸酶的体外抑制率达到62.29%,显示出较好的抗敏效果,可作为天然抗敏剂应用于抗过敏效果的化妆品中。
以上所述为本发明的较佳实施例而已,但本领域技术人员应当知道,不应将本发明的保护范围局限于该具体实施例。
Claims (4)
1.一种知母总皂苷提取物的制备方法,其特征在于:该方法包括下述步骤:
步骤1:取知母药材,加水煎煮三次,第一次加10倍量水,提取时间为2小时,第二次、第三次各加8倍量水,提取时间各1.5小时,过滤,合并滤液;
步骤2:滤液徐徐注入已处理好的AB-8型大孔树脂柱,用1倍柱床体积的水洗脱,弃去水洗脱液,用4倍柱床体积的0.2%NaOH水溶液洗脱,用水洗至中性后,再用收集4倍柱体积的80%乙醇洗脱,并收集洗脱液;
步骤3:将上述步骤2的80%乙醇洗脱液真空减压浓缩至相对密度为1.10~1.15,60℃真空干燥,粉碎,过60-100目筛,即得知母总皂苷提取物。
2.根据权利要求1所述的一种知母总皂苷提取物的制备方法,其特征在于:所述步骤2中真空减压浓缩时的温度为50~60℃。
3.一种知母总皂苷提取物在化妆品中的应用,其特征在于:所述化妆品包括采用上述方法提取得到的知母总皂苷提取物及辅料。
4.根据权利要求3所述的一种知母总皂苷提取物在化妆品中的应用,其特征在于:所述的知母总皂苷提取物用作化妆品中的抗过敏成分。
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