CN108904587A - 提取分离青钱柳黄酮的方法 - Google Patents
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Abstract
本发明公开了一种提取分离青钱柳黄酮的方法,包括下列步骤:a.将青钱柳用清水洗涤,40‑50℃烘干干燥,样品经粉碎机粉碎,过40目筛;b.酸解;c.微生物发酵降解;d.复合酶解:e.丙酮萃取;f.粗黄酮纯化:用活性炭纯化黄酮。本发明在青钱柳黄酮的提取过程中,首先采用初步微生物发酵的方法,将青钱柳的细胞壁降解掉,同时利用微生物的作用除去一部分杂质等,比如多糖等等。然后再用酶解,将黄酮充分释放出来。本发明的整个工艺过程中无污染物排放,适合于大规模生产,有益于青钱柳资源的开发利用。具有该步骤简单,成本低,可操作性强,产量高的优点,易于工业化生产。
Description
技术领域
本发明涉及一种提取分离青钱柳黄酮的方法。属于海洋微藻养殖领域。
背景技术
青钱柳,(拉丁文名:Cyclocaryapaliurus.)别名:摇钱树、麻柳,青钱李、山麻柳、山化树。胡桃科、青钱柳属植物,我国南方多省均有发现,多以零星分布。青钱柳乃冰川四纪幸存下来的珍稀树种,仅存于中国。青钱柳被誉为植物界的大熊猫,医学界的第三棵树。人们知道,医学界的第一棵树——柳树,产生了阿司匹林,消炎杀菌、抗血栓,揭开了人类健康史的第一次变革。
现代研究证实青钱柳叶具有降血糖、降血压、降血脂作用、增强机体免疫功能等作用。其主要活性成分是黄酮类和多糖类化合物。青钱柳中含有丰富的黄酮类物质,黄铜类化合物具有心血管***活性、抗菌及抗病毒活性、抗肿瘤活性、抗氧化自由基活性、抗炎镇痛活性、保肝活性等多种生物活性。青钱柳叶含有大量的黄酮类化合物,具有降血糖、降血压、降血脂、增强肌体免疫力、抗氧化、防衰老等功能。目前,青钱柳中黄酮类化合物的提取方法较复杂、工艺成本高,不利于大规模工业化。另外,传统的原料预处理方法多为高温干燥,同时,传统的水体方法也需要采用高温加热,才可以提取出较大量的提取物,不仅会大量破坏青钱柳中的有效成分,同时增加工艺能耗。
发明内容
本发明所要解决的技术问题是:提供一种提取分离青钱柳黄酮的方法。
为解决上述技术问题,本发明采用的技术方案是:
本发明提供的一种青钱柳提取多糖的方法,包括下列步骤:
a.将青钱柳用清水洗涤,40-50℃烘干干燥,样品经粉碎机粉碎,过40目筛。
b.酸解:将青钱柳加入蒸馏水打浆,青钱柳和蒸馏水的重量比为1:1-2,加入食品级盐酸(盐酸≥31.0%),加入食品级盐酸的重量为浆料重量的5-10%,30-40℃维持1-5h。
c.微生物发酵降解:将酸解后的原料加入升温到37-40℃,调节pH为5-6,加入酵母菌、枯草芽孢杆菌菌种0.05-0.3份,充分拌匀,发酵降解0.5-1天。
d.复合酶解:发酵后的待提取物料加入复合酶,复合酶和物料的重量比1:20-40,调节pH7.5-8.0,温度保持30-45℃,同时辅助超声处理,时间1-3h,过滤得滤液。
e.丙酮萃取:滤液中加入丙酮萃取,萃取2-3次,收集有机相,蒸馏后得到粗青钱柳黄酮;
f.粗黄酮纯化:用活性炭纯化黄酮。活性炭纯化黄酮根据现有技术中的操作方法即可,具体为根据总黄酮在不同温度的水中溶解度的差别,可用热水溶解、冷水结晶。使之不断提高纯度。该工艺相对简单,操作方便。在利用重结晶纯化总黄酮,加入一定量活性炭可提高纯化速度。
优选的,复合酶包括纤维素酶和果胶酶,两者的重量比例是0.5:1-2。
本发明的有益效果:
本发明在青钱柳黄酮的提取过程中,首先采用初步微生物发酵的方法,将青钱柳的细胞壁降解掉,同时利用微生物的作用除去一部分杂质等,比如多糖等等。然后再用酶解,将黄酮充分释放出来。
本发明的整个工艺过程中无污染物排放,适合于大规模生产,有益于青钱柳资源的开发利用。具有该步骤简单,成本低,可操作性强,产量高的优点,易于工业化生产。
具体实施方式
实施例1
本实施例提供的一种青钱柳提取多糖的方法,包括下列步骤:
a.将青钱柳用清水洗涤,50℃烘干干燥,样品经粉碎机粉碎,过40目筛。
b.酸解:将青钱柳加入蒸馏水打浆,青钱柳和蒸馏水的重量比为1:2,加入食品级盐酸(盐酸≥31.0%),加入食品级盐酸的重量为浆料重量的10%,40℃维持2h。
c.微生物发酵降解:将酸解后的原料加入升温到40℃,调节pH为6,加入酵母菌、枯草芽孢杆菌菌种0.3份,充分拌匀,发酵降解1天。
d.复合酶解:发酵后的待提取物料加入复合酶,复合酶和物料的重量比1:40,调节pH.0,温度保持45℃,同时辅助超声处理,时间3h,过滤得滤液。
e.丙酮萃取:滤液中加入丙酮萃取,萃取3次,收集有机相,蒸馏后得到粗青钱柳黄酮;
f.粗黄酮纯化:用活性炭纯化黄酮。
青钱柳黄酮得率:
将1千克的青钱柳黄酮按照本实施例的方法进行制备提取,得纯化黄酮13.4g,将纯化黄酮以芦丁为对照品经UV检测,测到其中黄酮含量为84.6%。
实施例2
本实施例提供的一种青钱柳提取多糖的方法,包括下列步骤:
a.将青钱柳用清水洗涤,40℃烘干干燥,样品经粉碎机粉碎,过40目筛。
b.酸解:将青钱柳加入蒸馏水打浆,青钱柳和蒸馏水的重量比为1:1,加入食品级盐酸(盐酸≥31.0%),加入食品级盐酸的重量为浆料重量的5%,30℃维持1h。
c.微生物发酵降解:将酸解后的原料加入升温到37℃,调节pH为5,加入酵母菌、枯草芽孢杆菌菌种0.05份,充分拌匀,发酵降解0.5天。
d.复合酶解:发酵后的待提取物料加入复合酶,复合酶和物料的重量比1:20,调节pH7.5,温度保持30℃,同时辅助超声处理,时间1h,过滤得滤液。
e.丙酮萃取:滤液中加入丙酮萃取,萃取2次,收集有机相,蒸馏后得到粗青钱柳黄酮;
f.粗黄酮纯化:用活性炭纯化黄酮。
青钱柳黄酮得率:
将1千克的青钱柳黄酮按照本实施例的方法进行制备提取,得纯化黄酮11.2g,以芦丁为对照品经UV检测,黄酮含量为86.2%。
Claims (2)
1.一种提取分离青钱柳黄酮的方法,其特征在于,包括下列步骤:
a.将青钱柳用清水洗涤,40-50℃烘干干燥,样品经粉碎机粉碎,过40目筛;
b.酸解:将青钱柳加入蒸馏水打浆,青钱柳和蒸馏水的重量比为1:1-2,加入食品级盐酸(盐酸≥31.0%),加入食品级盐酸的重量为浆料重量的5-10%,30-40℃维持1-5h;
c.微生物发酵降解:将酸解后的原料加入升温到37-40℃,调节pH为5-6,加入酵母菌、枯草芽孢杆菌菌种0.05-0.3份,充分拌匀,发酵降解0.5-1天;
d.复合酶解:发酵后的待提取物料加入复合酶,复合酶和物料的重量比1:20-40,调节pH7.5-8.0,温度保持30-45℃,同时辅助超声处理,时间1-3h,过滤得滤液;
e.丙酮萃取:滤液中加入丙酮萃取,萃取2-3次,收集有机相,蒸馏后得到粗青钱柳黄酮;
f.粗黄酮纯化:用活性炭纯化黄酮。
2.根据权利要求1所述的方法,其特征在于,复合酶包括纤维素酶和果胶酶,两者的重量比例是0.5:1-2。
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