CN113521189A - 一种黄精黄酮提取方法 - Google Patents
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Abstract
本发明公开了一种黄精黄酮提取方法，将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛；称取粉碎后的黄精适量，按照1:6~1:24的料液比加入提取溶剂，经过水浴提取后离心，分离上清液与底部黄精粉；对黄精粉重复水浴提取，离心获得上清液，合并上清液；将上清液用提取溶剂定容至一定体积，得到提取溶液；将提取溶液离心，得上清，即为黄精黄酮提取溶液。本发明方法利用特定参数条件下的水浴提取增加黄精细胞膜的渗透性并加速黄酮类化合物向提取溶剂扩散的速度，缩短提取时间。本发明公开的黄精黄酮提取方法具有提取得率高、工艺稳定可靠、操作性强和节能环保等优点。
Description
技术领域
本发明涉及一种提取方法，特别涉及一种黄精黄酮的提取方法，属于植物次生代谢物提取技术领域。
背景技术
黄精为百合科(Liliaceae)黄精属PoLygonatum Mill.植物的干燥根茎，味甘、性平、无毒，是我国传统的中药材，也是一种使用非常广泛的药食同源植物；具有独特的天然生物活性，黄精黄酮即为其天然生物活性成分之一。黄精具有抗氧化、抗癌、抗菌消炎、抗病毒、预防心血管疾病等功效。目前已有的总黄酮提取方法多采用热水浸提法，此法成本低、操作简单，但一般只能提取少量总黄酮，且此法耗时较长，迫切需要加以改进。CN104940609专利公开的一种黄精中多种活性成分提取分离的方法采用闪式提取～脱脂～反复萃取的工艺路线，按文献所说的黄精黄酮得率为1.97％，但是此工艺对设备要求高，工艺复杂，所用试剂存在安全隐患等缺点。
发明内容
本发明所要解决的技术问题是，针对现有技术的不足，提供一种黄精黄酮提取得率高、工艺稳定可靠、操作性强、节能环保的黄精黄酮提取方法。
本发明解决上述技术问题所采用的技术方案为：一种黄精黄酮提取方法，包括以下步骤：
(1)将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛，得到黄精粉；
(2)称取步骤(1)得到的黄精粉适量，按照1:6～1:24的料液比加入提取溶剂离心，分离上清液与底部黄精粉，对黄精粉重复水浴提取和离心，得到上清液；
(3)将步骤(2)得到的上清液置于一个容量瓶，用提取溶剂定容至50mL，得到提取溶液；
(4)将步骤(3)得到的提取溶液离心，取上清，得到黄精黄酮提取溶液。
所述步骤(2)中的水浴提取的水浴温度为50～90℃，水浴时间为15～150min，水浴次数为1～3次，离心次数为1～3次；作为优选，水浴提取的水浴温度为55℃～65℃，水浴时间为60min，水浴次数为3次，离心次数为3次。
作为优选，所述步骤(2)中的料液比为1:16～1:18。
所述步骤(2)和步骤(3)中的提取溶剂为浓度65～75％的乙醇溶液。
现有技术相比，本发明的有益效果：
本发明公开的黄精黄酮提取方法，利用特定参数条件下的水浴提取增加黄精细胞膜的渗透性并加速黄酮类化合物向提取溶剂扩散的速度，缩短提取时间。本发明公开的黄精黄酮提取方法具有黄精黄酮提取得率高、工艺稳定可靠、操作性强和节能环保等优点。
附图说明
图1为乙醇浓度对黄精黄酮得率的影响；
图2为料液比对黄精黄酮得率的影响；
图3为水浴温度对黄精黄酮得率的影响；
图4为水浴次数对黄精黄酮得率的影响；
图5为水浴时间对黄精黄酮得率的影响。
具体实施方式
以下列举的仅是本发明的具体实施例的优选。本发明不限于以下实施例，还可以有许多变形。本领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形，均应认为是本发明的保护范围。
实施例1
(1)将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛，得到黄精粉；
(2)称取步骤(1)得到的黄精粉适量，按照1:17的料液比加入浓度为70％的乙醇提取溶剂水浴并离心，温度为60℃水浴时间60min，分离上清液与底部黄精粉，对黄精粉重复水浴提取及离心3次，合并提取后，得到上清液；
(3)将步骤(2)得到的上清液置于一个容量瓶，用浓度为70％的乙醇提取溶剂定容至50mL，得到提取溶液；
(4)将步骤(3)得到的提取溶液离心，取上清，得到黄精黄酮提取溶液。
实施例2
(1)将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛，得到黄精粉；
(2)称取步骤(1)得到的黄精粉适量，按照1:6的料液比加入浓度为65％的乙醇提取溶剂水浴并离心，温度为90℃水浴时间为15min，分离上清液与底部黄精粉，对黄精粉重复水浴提取及离心1次，合并提取后，得到上清液；
(3)将步骤(2)得到的上清液置于一个容量瓶，用浓度为65％的乙醇提取溶剂定容至50mL，得到提取溶液；
(4)将步骤(3)得到的提取溶液离心，取上清，得到黄精黄酮提取溶液。
实施例3
(1)将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛，得到黄精粉；
(2)称取步骤(1)得到的黄精粉适量，按照1:24的料液比加入浓度为75％的乙醇提取溶剂水浴并离心，温度为50℃水浴时间为150min，分离上清液与底部黄精粉，对黄精粉重复水浴提取及离心2次，合并提取后，得到上清液；
(3)将步骤(2)得到的上清液置于一个容量瓶，用浓度为75％的乙醇提取溶剂定容至50mL，得到提取溶液；
(4)将步骤(3)得到的提取溶液离心，取上清，得到黄精黄酮提取溶液。
以上实施例使用的多花黄精药材由浙江淳安县临岐镇黄精基地提供，经浙江理工大学生命与医药学院浙江省植物次生代谢调控重点实验室鉴定为多花黄精的干燥根茎；主要试剂：无水乙醇、亚硝酸钠、氢氧化钠、硝酸铝等试剂均为AR；主要仪器:QGJ-A中药切片机(吉首市中诚制药机械厂)；LG-01一两装高速中药粉碎机(瑞安市百信制药机械有限公司)；GZX-9023MBE电热鼓风干燥箱(上海博迅实业有限公司医疗设备厂)；TP-213电子天平(赛多利斯科学仪器有限公司)；DK-8D电热恒温水槽(上海一恒科技有限公司)；UV-5500紫外可见分光光度计(上海元析仪器有限公司)。
总黄酮含量测定方法：吸取样品液2.0mL，置于25mL容量瓶中，加入5％亚硝酸钠溶液1mL，摇匀，放置6min，再加入10％硝酸铝溶液1mL，摇匀，放置6min，再加入4％氢氧化钠溶液10mL，用70％乙醇定容至刻度，摇匀，放置15min，以70％乙醇为参比溶液，于分光光度计波长510nm处测定吸光度值。按照标准曲线所得回归方程计算浓度。
采用Surfer8.0软件对试验数据进行可视化分析，通过将获得的数据与考察变量绘成多幅二维图的方法，同时考察2种不同的影响因素与考察指标之间的关系，可直接从数幅图找出最佳工艺的优化区域。
见附图1，在乙醇浓度＞85％和＜65％的区域内，总黄酮提取效果影响浅，提取率低；在乙醇浓度65％～75％的区域内，总黄酮提取效果影响度最深，提取率高。故乙醇浓度影响总黄酮提取效果的最佳范围为65％～75％。见附图2，在料液比1:6～1:8和1:16～1:18的区域内，总黄酮提取影响较深，提取率较高，尤其是在1:16～1:18的区域内，总黄酮提取影响最深，故选择料液比的最佳范围为1:16～1:18。见附图3，在水浴温度＞65℃时，总黄酮提取效果影响浅，提取率低。在水浴温度55℃～65℃的区域内，总黄酮提取效果影响度最深，提取率高。故乙醇浓度影响总黄酮提取效果的最佳范围为55℃～65℃。见附图4，从图可以看出不同的水浴次数在整个二维图中都有分布表明水浴次数对总黄酮提取率影响并不高，且水浴次数为3次时，总黄酮提取的影响较深，故选择水浴次数为3次。见附图5，在水浴时间45～75min的区域内，总黄酮提取效果影响度最深，提取率高，尤其是在60min效果最佳，故选择水浴时间60min。
Claims (4)
1.一种黄精黄酮提取方法，其特征在于，包括以下步骤：
(1)将黄精切片，在45℃烘干至恒重，然后经粉碎机粉碎，过60目筛，得到黄精粉；
(2)称取步骤(1)得到的黄精粉适量，按照1:6～1:24的料液比加入提取溶剂离心，分离上清液与底部黄精粉，对黄精粉重复水浴提取和离心，得到上清液；
(3)将步骤(2)得到的上清液置于一个容量瓶，用提取溶剂定容至50mL，得到提取溶液；
(4)将步骤(3)得到的提取溶液离心，取上清，得到黄精黄酮提取溶液。
2.根据权利要求1所述的一种黄精黄酮提取方法，其特征在于，所述步骤(2)中的水浴提取的水浴温度为50～90℃，水浴时间为15～150min，水浴次数为1～3次，离心次数为1～3次；优化的所述的水浴提取的水浴温度为55℃～65℃，水浴时间为60min，水浴次数为3次，离心次数为3次。
3.根据权利要求1所述的一种黄精黄酮提取方法，其特征在于，优选的所述步骤(2)中的料液比为1:16～1:18。
4.根据权利要求1所述的一种黄精黄酮提取方法，其特征在于，所述步骤(2)和步骤(3)中的提取溶剂为浓度65～75％的乙醇溶液。
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| Title |
|---|
| 刘清华等: ""多花黄精中总黄酮提取工艺优化及可视化分析"", 《中国中医药信息杂志》 * |
| 钱森和等: ""响应面法优化黄精黄酮提取工艺"", 《安徽工程大学学报》 * |
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