CN105585548A - 一种去苦味儿茶素的制备方法 - Google Patents
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Abstract
本发明提出了一种去苦味儿茶素的制备方法,包含如下步骤:(1)向含有绿茶儿茶素提取物的溶液中添加果胶酶,35-37℃温度下静止酶解,间歇搅拌,离心过滤,得酶解液;(2)将酶解液通入微孔陶瓷膜过滤器,并用反向渗透浓缩器浓缩,收集浓缩液,将浓缩液浓缩至16-18波美度,得浓缩液;将浓缩液通过离心喷雾干燥塔进行离心干燥,得去苦味儿茶素。本发明使用果胶酶对绿茶儿茶素进行酶解,具较好的转化效果,是一种转化率高,产品纯度高的制备方法,而且制备周期短,成本低,适宜规模化生产。
Description
技术领域
本发明属于儿茶素的制备领域,具体涉及一种去苦味儿茶素的制备方法。
背景技术
儿茶素是茶叶中最重要的生理活性物质,约占多酚类80%,属黄烷醇类,其基本结构是α-苯基苯并吡喃,分为酯型和非酯型两类,结构上的主要区别在于氢基取代的数量和位置不同。儿茶素主要由表没食子儿茶素没食子酸酯(EGCG)、表没食子儿茶素(EGC)、表儿茶素没食子酸酯(ECG)和表儿茶素(EC)等单体组成。儿茶素具有明显的抗衰老、防癌抗突变、降脂减肥、降低血糖、血脂和胆固醇、预防心血管疾病等药理功能,已广泛应用于食品加工、医药保健和日用化工等领域。卫生部已批准了茶多酚为我国食品添加剂。国内外研究成果表明,儿茶素可防心血管疾病,预防癌症,抗氧化作用,具有明显的抗衰老功能,有望开发成为一种天然的护肤品。从1990开始,由于有关茶多酚/儿茶素的保健与药理功能研究的突破,不仅扩大了绿茶在国际市场上的份额,而且极大地推进了我国茶叶深加工产业的快速发展,为茶叶功能成分的研究与开发奠定了良好的产业与市场基础,目前,我国已经形成年产5000多吨茶多酚/儿茶素的产业规模。儿茶素粗品传统的提取分离工艺主要包括离子沉淀法、有机溶剂萃取法和吸附柱色谱法。由于茶叶的功能成分复杂,各功能成分的化学结构相似,从茶叶中制取无苦味的儿茶素具有相当难度。
发明内容
本发明提供一种去苦味儿茶素的制备方法,该方法简便,只使用食用添加酶制剂,制备周期短,成本低,符合绿色和安全的要求,去苦味儿茶素的提取率及产品纯度高,适宜规模化生产。
本发明提供的技术方案是:一种去苦味儿茶素的制备方法,包括如下步骤:
(1)加入纯水溶解绿茶儿茶素提取物,配置成浓度为5%w/v的溶液,加入氢氧化钠调节pH值为4.0-5.0,制得茶汤料液,备用;
(2)按照0.025%w/v的比例,在茶汤料液中添加果胶酶,在35℃-37℃下静止酶解,间歇搅拌,得酶解液;
(3)加入氢氧化钠调节调节酶解液pH值为5,离心过滤;
(4)将步骤(3)中过滤后的上清液通过微孔陶瓷膜过滤器,滤去极微小杂质,并用反向渗透浓缩器进行浓缩,收集浓缩液,将所述洗脱液浓缩至16-18波美度,得浓缩液;
(5)将所述浓缩液在180℃下通过离心喷雾干燥塔进行离心干燥,得脱去苦味的儿茶素。
本发明步骤(2)中,加入果胶酶,静置酶解16-24小时,间歇60分钟搅拌10分钟。
本发明步骤(4)中所述反向渗透浓缩器所采用的反渗透膜材料为醋酸纤维素、芳香族聚酰胺、聚醚砜或聚偏氟乙烯其中之一。
本发明具有以下有益效果:本发明以小叶绿茶儿茶素提取物为原料,经过果胶酶酶解,微孔超滤及反向渗透浓缩,从绿茶提取物中分离去苦味儿茶素,优化了产品分离纯化工艺流程,单体提取率及产品纯度高。产品可防心血管疾病,预防癌症,抗氧化作用,具有明显的抗衰老功能,有望开发成为一种天然的护肤品。也是许多功能食品开发的重要原料,可单独或者与维生素类、天然植物成分复配应用于保健食品、化妆品等领域。本发明方法简便,制备周期短,成本低,符合环保和安全的要求,适宜规模化生产。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。本发明中,果胶酶购自诺维信(中国)生物技术有限公司。
本发明中,绿茶儿茶素提取物最好是(小叶种茶树上采摘的)小叶绿茶儿茶素提取物。
实施例1
一种去苦味儿茶素的制备方法,包括如下步骤:
1、取小叶绿茶儿茶素提取物100克,配成2000ml儿茶素溶液,加入氢氧化钠调节pH值为4.0-5.0,向所述溶液中添加0.5克果胶酶,在35-37℃温度下酶解16小时,间歇60分钟搅拌10分钟,(加入氢氧化钠)将酶解液调节pH值至5,在1200r/min条件下离心过滤;
2、将步骤1中过滤后的上清液缓慢通过微孔陶瓷膜过滤器,出去部分肉眼不可见杂质,再通过(截留分子量为2500的反渗透膜)反向渗透浓缩器浓缩至16-18波美度,得浓缩液;
3、将所述浓缩液在180℃温度下离心喷雾干燥,得到87.25g砖红色固体。
将步骤3得到的砖红色固体与儿茶素标准品(购自美国Sigma-Aldrich公司,C15H14O6,M.W.290.27,编号:E4018,纯度≥98%(HPLC))对照鉴定并经高效液相色谱法分析检测(鉴定、检测结果为:所述砖红色固体,分子量为290,(-)ESI-MSm/z:289[M-H]-,熔点240oC,分子式为:C15H14O6,(ECG+EGCG+GCG)含量≤1.5%表明所述砖红色固体为无苦味(去苦味)儿茶素,而且纯度≥98%。
实施例2
一种去苦味儿茶素的制备方法,包括如下步骤:
1、取小叶绿茶儿茶素提取物100克,配成2000ml儿茶素溶液,加入氢氧化钠调节pH值为4.0-5.0,向所述溶液中添加0.5克果胶酶,在35-37℃温度下酶解20小时,间歇60分钟搅拌10分钟,(加入氢氧化钠)将酶解液调节pH值至5,在1200r/min条件下离心过滤;
2、将步骤1中过滤后的上清液缓慢通过微孔陶瓷膜过滤器,出去部分肉眼不可见杂质,再通过(截留分子量为2500的反渗透膜)反向渗透浓缩器浓缩至16-18波美度,得浓缩液;
3、将所述浓缩液在180℃温度下离心喷雾干燥,得到87.25g砖红色固体。
将步骤3得到的砖红色固体与儿茶素标准品(购自美国Sigma-Aldrich公司,C15H14O6,M.W.290.27,编号:E4018,纯度≥98%(HPLC))对照鉴定并经高效液相色谱法分析检测(鉴定、检测结果为:所述砖红色固体,分子量为290,(-)ESI-MSm/z:289[M-H]-,熔点240oC,分子式为:C15H14O6,(ECG+EGCG+GCG)含量≤1.5%表明所述砖红色固体为无苦味(去苦味)儿茶素,而且纯度≥98%。
实施例3
一种去苦味儿茶素的制备方法,包括如下步骤:
1、取小叶绿茶儿茶素提取物100克,配成2000ml儿茶素溶液,加入氢氧化钠调节pH值为4.0-5.0,向所述溶液中添加0.5克果胶酶,在35-37℃温度下酶解24小时,间歇60分钟搅拌10分钟,(加入氢氧化钠)将酶解液调节pH值至5,在1200r/min条件下离心过滤;
2、将步骤1中过滤后的上清液缓慢通过微孔陶瓷膜过滤器,出去部分肉眼不可见杂质,再通过(截留分子量为2500的反渗透膜)反向渗透浓缩器浓缩至16-18波美度,得浓缩液;
3、将所述浓缩液在180℃温度下离心喷雾干燥,得到87.25g砖红色固体。
将步骤3得到的砖红色固体与儿茶素标准品(购自美国Sigma-Aldrich公司,C15H14O6,M.W.290.27,编号:E4018,纯度≥98%(HPLC))对照鉴定并经高效液相色谱法分析检测(鉴定、检测结果为:所述砖红色固体,分子量为290,(-)ESI-MSm/z:289[M-H]-,熔点240oC,分子式为:C15H14O6,(ECG+EGCG+GCG)含量≤1.5%表明所述砖红色固体为无苦味(去苦味)儿茶素,而且纯度≥98%。
Claims (3)
1.一种去苦味儿茶素的制备方法,其特征在于,包括如下步骤:
(1)加入纯水溶解绿茶儿茶素提取物,配置成浓度为5%w/v的溶液,加入氢氧化钠调节pH值为4.0-5.0,制得茶汤料液,备用;
(2)按照0.025%w/v的比例,在茶汤料液中添加果胶酶,在35℃-37℃下静止酶解,间歇搅拌,得酶解液;
(3)加入氢氧化钠调节调节酶解液pH值为5,离心过滤;
(4)将步骤(3)中过滤后的上清液通过微孔陶瓷膜过滤器,滤去极微小杂质,并用反向渗透浓缩器进行浓缩,收集浓缩液,将所述洗脱液浓缩至16-18波美度,得浓缩液;
(5)将所述浓缩液在180℃下通过离心喷雾干燥塔进行离心干燥,得脱去苦味的儿茶素。
2.根据权利要求1所述一种去苦味儿茶素的制备方法,其特征在于:步骤(2)中加入果胶酶,静置酶解16-24小时,间歇60分钟搅拌10分钟。
3.根据权利要求1所述一种去苦味儿茶素的制备方法,其特征在于:步骤(4)中所述反向渗透浓缩器所采用的反渗透膜材料为醋酸纤维素、芳香族聚酰胺、聚醚砜或聚偏氟乙烯其中之一。
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