CN110025679A - 一种短瓣金莲花抗氧化提取物的制备方法 - Google Patents
一种短瓣金莲花抗氧化提取物的制备方法 Download PDFInfo
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Abstract
本发明公开了一种短瓣金莲花抗氧化提取物的制备方法，其特征在于，将短瓣金莲花干花浸入乙醇水溶液中进行提取，并过滤；将得到的提取液经过减压蒸馏，得到浓缩物；将浓缩物溶于水，得到混合溶液；将混合溶液进行纯化后，用乙醇水溶液进行解吸；将所得解吸溶液浓缩，并冷冻干燥，得到短瓣金莲花抗氧化提取物。本发明设备要求不高，成本低、效率高，可行性高，操作过程简单方便，适于大生产，所得的短瓣金莲花抗氧化提取物具有较强的抗氧化性质，不仅可以用于医疗、保健品等行业，也可应用于化妆品行业，进一步扩宽了短瓣金莲花的应用领域。
Description
技术领域
本发明涉及一种短瓣金莲花抗氧化提取物的制备方法，属于植物提取技术领域。
背景技术
短瓣金莲花为毛茛科金莲花属植物，系野生的多年生草本植物，资源十分丰富，主要分布于黑龙江及内蒙古东北部等地区。其药用始载于赵学敏所著《本草纲目拾遗》中，可“治口疮，喉肿，牙宣，耳疼，目痛，解岚瘴”，主要含有黄酮及其苷类化合物，挥发油及有机酸等。现代药理研究表明黄酮类化合物为金莲花属植物主要有效成分，鉴于黄酮类化合物具有保肝抗炎、镇定补脑、延缓衰老、抗抑郁等多种生理活性功效，黄酮类化合物的研究日益引起人们的关注。荭草苷和牡荆苷作为该属植物黄酮类化合物的主要抗氧化成分，以荭草苷和牡荆苷为指标研究该属植物的抗氧化活性的研究目前没有。荭草苷和牡荆苷作为作为天然抗氧化剂，不但具有安全性高、无毒副作用的危险，而且抗氧化能力优于VE，和VC接近。
发明内容
本发明所要解决的技术问题是：现有技术中的抗氧化成分活性差，制备方法复杂的问题。
为了解决上述问题，本发明提供了一种短瓣金莲花抗氧化提取物的制备方法，其特征在于，包括以下步骤：
步骤1)：将短瓣金莲花干花浸入乙醇水溶液中进行提取，并过滤；
步骤2)：将步骤1)得到的提取液经过蒸馏，得到浓缩物；
步骤3)：将步骤2)得到的浓缩物溶于水，得到混合溶液；将混合溶液进行纯化后，用乙醇水溶液进行解吸；将所得解吸溶液浓缩，并冷冻干燥，得到短瓣金莲花抗氧化提取物。
优选地，所述步骤1)中的金莲花干花由短瓣金莲花的花朵自然晾晒后制得。
优选地，所述步骤1)中的提取方法采用加热回流、超声辅助或索氏提取，提取温度为40～70℃，提取时间为0.5-2.5h。
更优选地，所述提取方法采用加热回流，提取温度为60℃，提取时间为2h；其中，乙醇的质量浓度为60％，短瓣金莲花干花与乙醇的体积比为1：30。
优选地，所述步骤1)中乙醇水溶液的质量浓度为50-90％；短瓣金莲花干花与乙醇水溶液的体积比为1：(20～40)，提取时间为2h。
优选地，所述步骤2)中减压蒸馏的真空度为0.04～0.06Mpa，温度为40～50℃；减压蒸馏至无醇味，结束减压蒸馏。
更优选地，所述减压蒸馏的真空度为0.05Mpa，温度为48℃。
优选地，所述步骤3)中的纯化采用大孔树脂，大孔树脂的规格为AB-8、D-101、HPD-600、HPD-300、HPD-100、HP-20或1300。
更优选地，所述大孔树脂采用AB-8。
优选地，所述步骤3)得到的短瓣金莲花抗氧化提取物中的抗氧化成分包括荭草苷及牡荆苷。所得短瓣金莲花抗氧化提取物的抗氧化活性与相应体积浓度的VC相当，比相应体积浓度的VE好。
与现有技术相比，本发明短瓣金莲花抗氧化提取物的制备方法设备要求不高，成本低、效率高，可行性高，简单实用，操作过程简单方便，适于大生产，制得的提取物主要抗氧化成分包括荭草苷和牡荆苷，抗氧化活性好；不仅可以用于医疗、保健品等行业，也可应用于化妆品行业，进一步扩宽了短瓣金莲花的应用领域。
附图说明
图1为实施例中荭草苷的检测结果；
图2为实施例中牡荆苷的检测结果；
图3为实施例中短瓣金莲花抗氧提取物的检测结果；
图4为实施例中维生素C和维生素E对DPPH自由基的抑制结果；
图5为实施例制得短瓣金莲花抗氧提取物对DPPH自由基的抑制结果。
具体实施方式
为使本发明更明显易懂，兹以优选实施例，并配合附图作详细说明如下。
实施例所用到的仪器：Agilent 1200型高效液相色谱(配备四元梯度泵，在线真空脱气机，DAD检测器等)；ShangHai Ultra Sonics SY-800Type超声波清洗器；A604A电子天平(上海科天电子仪器有限公司)；色谱柱：Shim-pack VP-ODS，4.6mm×250mm，5μm(岛津中国(上海)有限公司)；多功能酶标仪(美国伯腾仪器有限公司)；冷冻干燥机(上海比朗仪器制造有限公司)。
实施例
一种短瓣金莲花抗氧化提取物的制备方法，其步骤如下：
(1)将5.00g短瓣金莲花干花，在提取温度为60℃，以乙醇体积浓度为60％，液料比为1:30的条件，加热回流提取2h，过滤，合并滤液得到提取液；
(2)提取液在48℃、真空度0.05Mpa条件下减压蒸馏浓缩至无醇味；
(3)将得到的浓缩物加入300mL的去离子水溶解，得到混合溶液；采用大孔树脂AB-8进行纯化，用体积浓度60％的乙醇进行洗脱，将所得溶液在48℃、真空度0.05Mpa条件下减压蒸馏浓缩至无醇味；冷冻干燥，得到短瓣金莲花抗氧化提取物粉末。
为了探讨短瓣金莲花抗氧化提取物中荭草苷和牡荆苷的含量，以及对短瓣金莲花抗氧化提取物的体外抗氧化能力进行评价，作了如下实验，其步骤如下：
一、短瓣金莲花抗氧化提取物中荭草苷和牡荆苷的含量测定
对照品溶液制备：分别精确称取荭草苷、牡荆苷对照品适量，置于20.00mL的容量瓶中，加乙腈-0.1wt％磷酸水溶液溶解稀释定容至刻度，摇匀，作为对照品溶液。
供试品溶液：取适量的短瓣金莲花抗氧化提取物粉末，置于20.00mL的容量瓶中，加乙腈-0.1wt％磷酸水溶液溶解稀释定容至刻度，摇匀，作为供试品溶液。
检测条件：采用高效液相色谱仪，以十八烷基硅烷键合硅胶为键合相的色谱柱，以乙腈、0.1wt％磷酸水溶液为流动相，在流速为1.0mL/min；柱温为35℃；紫外检测器波长为350nm，进样体积为5μL的条件，按照梯度洗脱的条件为：第0-8min：乙腈为10-20wt％，0.1wt％的冰醋酸水溶液为90-80wt％；第8-23min：乙腈为20-45wt％，0.1wt％的冰醋酸水溶液为80-55wt％；第23-27min：乙腈为45-85wt％，0.1wt％的冰醋酸水溶液为65-15wt％；第27-32min到最后：乙腈为10wt％，0.1wt％的冰醋酸水溶液为90wt％。
检测方法：吸取对照品、供试品溶液，进样检测。
荭草苷标品色谱分离结果见图1所示，从图1中可以看出，荭草苷的保留时间为12.557min，牡荆苷标品色谱分离结果见图2所示，从图2中可以看出，牡荆苷的保留时间为13.948min，荭草苷和牡荆苷能够实现完全分离，分离度良好，满足中国药典的要求。
供试品色谱分离结果见图3所示，从图2中可以看出，短瓣金莲花抗氧化提取物中荭草苷和牡荆苷能够实现完全分离，分离度良好。本人通过大量实验研究表明短瓣金莲花的抗氧化活性与荭草苷和牡荆苷的含量呈正相关。
二、短瓣金莲花抗氧化提取物的体外抗氧化性测定如下：
(1)精确取30.00mg DPPH粉末，用无水乙醇定容至25mL的容量瓶中，超声10min，0℃保存；
(2)将VC、VE标准品分别配制成浓度为0.1000mg/mL、0.0500mg/mL、0.0250mg/mL、0.0125mg/mL、0.0063mg/mL的样品液；
(3)将样品(短瓣金莲花抗氧提取物)分别配制成浓度为0.1800mg/mL、0.0900mg/mL、0.0450mg/mL、0.0225mg/mL、0.0113mg/mL的样品液；
(4)分别取各浓度的样品液0.5mL，加入0.5mL的DPPH储备液，置于暗室反应10min，用酶标仪于517nm处测定吸光度。
每个反应体系对DPPH·的抑制率采用下式计算：
抑制率(％)＝(1-A/A0)×100％，其中，A0表示未加样的DPPH溶液的吸光度，A表示样品与DPPH·反应后的吸光度。
实验结果如表1所示。
表1
| 样品名称 | IC50(mg/mL) |
| VC | 0.034 |
| VE | 0.077 |
| 短瓣金莲花抗氧提取物 | 0.050 |
由表1可见，随着短瓣金莲花抗氧化提取物浓度的增加，对DPPH自由基的抑制率也相应呈上升趋势。当短瓣金莲花抗氧化提取物浓度为50μg/ml及以上时，对DPPH自由基的清除率可以达到50％以上，且与相应体积浓度的VC相当，比相应体积浓度的VE抑制效果要好。
Claims (10)
1.一种短瓣金莲花抗氧化提取物的制备方法，其特征在于，包括以下步骤：
步骤1)：将短瓣金莲花干花浸入乙醇水溶液中进行提取，并过滤；
步骤2)：将步骤1)得到的提取液经过蒸馏，得到浓缩物；
步骤3)：将步骤2)得到的浓缩物溶于水，得到混合溶液；将混合溶液进行纯化后，用乙醇水溶液进行解吸；将所得解吸溶液浓缩，并冷冻干燥，得到短瓣金莲花抗氧化提取物。
2.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤1)中的金莲花干花由短瓣金莲花的花朵自然晾晒后制得。
3.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤1)中乙醇水溶液的质量浓度为50-90％；短瓣金莲花干花与乙醇水溶液的体积比为1：(20～40)，提取时间为0.5-2.5h。
4.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤1)中的提取方法采用加热回流、超声辅助或索氏提取，提取温度为40～70℃。
5.如权利要求4所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述提取方法采用加热回流，提取温度为60℃，提取时间为2h；其中，乙醇的质量浓度为60％，短瓣金莲花干花与乙醇的体积比为1：30，提取时间为2h。
6.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤2)中减压蒸馏的真空度为0.04～0.06Mpa，温度为40～50℃；减压蒸馏至无醇味，结束减压蒸馏。
7.如权利要求6所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述减压蒸馏的真空度为0.05Mpa，温度为48℃。
8.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤3)中的纯化采用大孔树脂，大孔树脂的规格为AB-8、D-101、HPD-600、HPD-300、HPD-100、HP-20或1300。
9.如权利要求8所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述大孔树脂采用AB-8。
10.如权利要求1所述的短瓣金莲花抗氧化提取物的制备方法，其特征在于，所述步骤3)得到的短瓣金莲花抗氧化提取物中的抗氧化成分包括荭草苷及牡荆苷。
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| CN113208961A (zh) * | 2021-05-08 | 2021-08-06 | 广州市络捷生物科技有限公司 | 一种金莲花提取物的制备方法和应用 |
| CN114904016A (zh) * | 2022-04-14 | 2022-08-16 | 华南理工大学 | 一种含金莲花提取物的包合物及其制备方法与应用 |
| CN114904016B (zh) * | 2022-04-14 | 2023-12-19 | 华南理工大学 | 一种含金莲花提取物的包合物及其制备方法与应用 |
| CN114948815A (zh) * | 2022-06-09 | 2022-08-30 | 内蒙古科技大学 | 一种以天然草本植物制备的防晒定妆散粉及其制备方法 |
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