CN107397223B - 一种柚皮苷提取方法 - Google Patents
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Abstract
本发明提供了一种柚皮苷提取方法,其中使用酶加热水法从柚子皮中提取柚皮苷。具体地说,本发明的柚皮苷提取方法中使用250U/g的菠萝蛋白酶,50U/g的纤维素酶和100U/g的果胶酶辅助热水法从柚子皮中提取柚皮苷。
Description
一技术领域
本发明涉及植物活性物质提取领域,具体地说,本发明涉及一种柚皮苷提取方法。
二背景技术
柚子是芸香科植物柚的成熟果实,产于我国福建、江西、广东、广西等南方地区,其口味甘甜,营养丰富,是广受欢迎的水果品种。中医认为柚子有清热去火、消积食、通便秘、抗血栓、消炎、降血压的功效,是最具食疗效益的水果之一。
柚子的活性成分主要是柚皮苷,其是一种双氢黄酮类化合物。在282nm有强烈的紫外吸收峰。目前已有的柚皮苷提取方法主要包括热水法、醇提取法、碱提酸沉法、超临界CO2萃取法、超声波辅助提取法、酶加热水提取法(参见申请人在先专利申请ZL201510895047.3),其中超临界CO2萃取法需要的设备和运行成本均较为昂贵,而醇提取法和碱提酸沉法往往要使用有毒的甲醇,或者不利于后续食品加工的乙醇、氢氧化钙等成分,而且现有方法的提取率均不高,按柚皮苷重量比材料重量基本不超过7%(参见湖北农业科学2011年4月第50卷第8期柚皮苷的提取方法及其应用研究进展)。
酶加热水法提取柚皮苷条件温和、设备简单、投资小,使用的试剂均为常用的食品用酶,得到的柚皮苷提取液或提取物可以方便地直接用于多种食品的制备。在现有淀粉酶、果胶酶和纤维素酶法的基础上寻找提取率更高的酶加热水提取柚皮苷的方法,对于充分利用柚子皮这一食品加工中大量产生的废弃物和降低食品成本都有重要意义。
三发明内容
本领域一般认为柚子皮主要成分为纤维素、果胶、淀粉等多糖成分,ZL201510895047.3中也以此为基础设计了淀粉酶、果胶酶和纤维素酶的酶解体系。但申请人对柚子皮的结构进行进一步研究时发现柚子皮中存在一些连续的微小纤维状和片状的结构,经过酶解实验分析,这些结构蛋白为主要成分(具体蛋白种类尚未确定),这些纤维状和片状蛋白结构在淀粉酶、果胶酶和纤维素酶酶解或热水处理时均能保持基本完整或形变收缩,会阻止柚皮苷等成分的充分释放。
基于此发现,申请人经过筛选和组合实验,选择了菠萝蛋白酶、果胶酶和纤维素酶的酶解体系辅助热水法提取柚子皮中的柚皮苷,有效的提高了柚皮苷提取率。
基于将上述方法制备的柚皮苷提取液直接用于多种食品的制备,可以减少原料和酶用量40%左右,有效降低生产成本。上述柚皮苷提取方法也可以用于提取药用、化工用柚皮苷。
一方面,本发明提供了一种酶解热水法提取的柚皮苷提取方法,
进一步地方面,该方法包括步骤:
(1)称取柚子皮粉,加入6-8倍质量的水浸泡2-6小时;
(2)加入菠萝蛋白酶250U/g,调pH至6.5-8.0,在50-55℃下酶解反应10-60min,加热至100℃使酶灭活5-10分钟后冷却;
(3)加入纤维素酶50U/g,果胶酶100U/g,调pH至4.0-5.0,在45-60℃下酶解0.5-4h,加热至100℃使酶灭活10分钟后冷却;
(4)过滤得到柚皮苷提取液。
另一方面,本发明提供了按照上述方法制备的柚皮苷。
四具体实施方式
原料:
沙田柚柚子皮粗干粉(剥去沙田柚果肉后剩余柚子皮晒干、粉碎制成)。
试剂:
食品级菠萝蛋白酶:南宁东恒华道生物科技有限责任公司,50000U/g;
食品级纤维素酶:湖南利尔康生物有限公司,10000U/g;
食品级果胶酶:湖南利尔康生物有限公司,40000U/g;
柚皮苷标准品:购自中国药品生物制品检定所;
其余试剂均为常规国产试剂。
仪器:
色谱仪:Aglient 1100型,C18色谱柱;
其余仪器均为常规化学、食品工业用仪器。
实施例1柚皮苷的制备
(1)称取鲜柚子皮1kg,加入7kg水浸泡6小时;
(2)加入菠萝蛋白酶250000U(5g),调pH至7.0,在50℃下酶解反应30min,加热至100℃使酶灭活10分钟后冷却;
(3)加入纤维素酶50000U(5g),果胶酶100000U(2.5g),调pH至4.5,在50℃下酶解1h,加热至100℃使酶灭活10分钟后冷却;
(4)过滤得到柚皮苷提取液;
实施例2柚皮苷提取率测定
色谱条件:
流动相为甲醇-水-冰醋酸(35:61:4),流速1.0ml/min,检测波长283nm,柱温度25℃。标准曲线的制备:
精确称取柚皮苷标准品20mg,甲醇定容至20mL,得1mg/mL的标准溶液,吸取标准溶液1mL、2mL、4mL、6mL、9mL定容于10ml容量瓶中,取样20μL,按上述色谱条件测定峰面积,以峰面积为纵坐标,浓度为横坐标,进行线性回归。回归方程为:Y=28.753X+680.2(r=0.9993)。
取实施例1步骤(4)中得到的柚皮苷提取液和ZL201510895047.3中2015年12月7日提交的权利要求第5项步骤(1)的方法得到的柚皮苷提取液(淀粉酶、果胶酶和纤维素酶的酶解体系)各三份,3000r/min离心5min后,取上清以上述色谱条件和标准曲线测定柚皮柑浓度,计算三份的平均值,换算成柚皮苷含量并计算提取率。
各种方法柚皮苷提取率如下表所示(文献中的多个提取率取最高值):
*该方法中柚皮苷提取率记载为26.04%,按照柚皮苷重量比材料重量的计算方法明显不符合常理,参照类似结果的文献,此处提取率单位应为mg柚皮苷/g材料,换算成柚皮苷重量比材料重量,提取率应为2.64%。
上述数据表明,本发明方法的柚皮苷提取率是现有最高的乙醇提取法的约1.5倍,而且没有醇提法中所用的甲醇或乙醇影响后续直接使用的问题。是之前的酶加热水法的2倍。
上述实施例证明了菠萝蛋白酶、果胶酶和纤维素酶的酶解体系辅助热水法提取柚子皮中的柚皮苷提取率明显高于现有方法,按照需要,本发明的菠萝蛋白酶、果胶酶和纤维素酶的酶解体系也可以与醇提取法、碱提酸沉法、超临界CO2萃取法、超声波辅助提取法等方法联合使用,原理上也可以提高这些方法的柚皮苷提取率。
Claims (1)
1.一种柚皮苷提取方法,具体步骤包括:
(1)称取柚子皮粉,加入6-8倍质量的水浸泡2-6小时;
(2)加入菠萝蛋白酶250U/g,调pH至6.5-8.0,在50-55℃下酶解反应10-60min,加热至100℃使酶灭活5-10分钟后冷却;
(3)加入纤维素酶50U/g,果胶酶100U/g,调pH至4.0-5.0,在45-60℃下酶解0.5-4h,加热至100℃使酶灭活10分钟后冷却;
(4)过滤得到柚皮苷提取液。
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CN102783692A (zh) * | 2012-05-12 | 2012-11-21 | 林钧 | 橄榄汁的创新除涩技术、柑橙汁的创新除苦技术 |
CN104957328A (zh) * | 2015-06-17 | 2015-10-07 | 符洁梅 | 保健柚子皮袋泡茶及其制备方法 |
CN105533579A (zh) * | 2015-12-07 | 2016-05-04 | 广东生和堂健康食品股份有限公司 | 一种含有柚皮苷的龟苓膏及其制备方法 |
CN106083940A (zh) * | 2016-07-11 | 2016-11-09 | 湖南华诚生物资源股份有限公司 | 一种从枳实中提取高纯度新橙皮苷的方法 |
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CN102783692A (zh) * | 2012-05-12 | 2012-11-21 | 林钧 | 橄榄汁的创新除涩技术、柑橙汁的创新除苦技术 |
CN104957328A (zh) * | 2015-06-17 | 2015-10-07 | 符洁梅 | 保健柚子皮袋泡茶及其制备方法 |
CN105533579A (zh) * | 2015-12-07 | 2016-05-04 | 广东生和堂健康食品股份有限公司 | 一种含有柚皮苷的龟苓膏及其制备方法 |
CN106083940A (zh) * | 2016-07-11 | 2016-11-09 | 湖南华诚生物资源股份有限公司 | 一种从枳实中提取高纯度新橙皮苷的方法 |
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