大豆胚芽中异黄酮提取研究
|
摘要
本文研究了从大豆胚芽中提取和纯化总异黄酮的方法与工艺，包括总异黄酮的初步提取和异黄酮提取物的固体酸水解两个主要过程。
在大豆胚芽总异黄酮初步提取条件优化实验中，以单因素实验方法优化了四个影响因素：提取温度、提取时间、溶剂用量、溶剂浓度。以胚芽中总异黄酮提取率和提取的选择性为指标，确定60％乙醇为提取溶剂、提取温度50℃、提取次数三次、每次提取的液固比为6:1、三次的提取时间分别为60min、10min、10min，三次总提取率为92％。
在异黄酮固体酸水解实验中，选择南开大学化工厂的D72强酸性阳离子交换树脂作为固体酸催化剂。经过两次固定床水解反应，染料木素、大豆黄素和黄豆黄素等三种异黄酮转化率可达90％。实验研究了温度、甲醇水配比、进料配比以及接触时间对游离型异黄酮转化率的影响。结果表明，在实验温度范围内(60～70℃)，温度变化对genistein和glycitein影响较明显，温度升高转化率增大；温度变化对daidzein的转化率影响不大；接触时间越长，反应的转化率越高，并逐渐趋向平衡值；甲醇水配比为1:2有利于水解反应；溶剂比例增大，三种异黄酮转化率都增大，溶剂比例增大到一定程度转化率变化不明显。
异黄酮精制过程中，探索了柱层析、溶剂萃取、溶剂沉淀等方法，最终异黄酮含量可达40％，对胚芽异黄酮收率50％左右。
建立了总异黄酮的分析方法和游离态异黄酮的HPLC分析方法。
A process for extracting and purifying isoflavones from soybean germ was studied in this paper. The process includes extraction of total isoflavones and hydrolyzation of soybean isoflavone by solid acid catalyst.
In the experiments of extraction of total isoflavones, five factors were studied: temperature, extracting time, concentration of solvent and amount of solvent. With the extraction efficiency of total isoflavones and the selectivity of extraction as study targets, the optium extraction conditions were as followed: solvent 60% ethanol, temperature 50℃, solvent volume to sobean germ weight 6:1, extracting for 3times, 60min 10min , 10min respectively. Extraction efficiency of total isoflavones was about 92%.
In the experiments of fixed bed process, soybean isoflavone glucoside can be hydrolyzed into isoflavone aglucone with methanol in a fixed bed filled with solid acid catalyst(Nankai Chemical Factory, D72). Conversion of isoflavones is about 90%. The effect of temperature, contact time, concentration of methanol and proportion of solvent to material on conversion was studied. The results show that enhancing temperature and prolonging the contact time are favor of the reaction, and the ratio of methanol to water is appropriately 1:2.
During the process of refining of isoflavones from the extraction, methods such as chromatogram, extraction, deposition were researched. Content of isoflavone was as high as 40%, and recovery percent of isoflavone based on soybean germ was about 50%. Analytical method for total isoflavones and isoflavone aglucone was established.
在大豆胚芽总异黄酮初步提取条件优化实验中，以单因素实验方法优化了四个影响因素：提取温度、提取时间、溶剂用量、溶剂浓度。以胚芽中总异黄酮提取率和提取的选择性为指标，确定60％乙醇为提取溶剂、提取温度50℃、提取次数三次、每次提取的液固比为6:1、三次的提取时间分别为60min、10min、10min，三次总提取率为92％。
在异黄酮固体酸水解实验中，选择南开大学化工厂的D72强酸性阳离子交换树脂作为固体酸催化剂。经过两次固定床水解反应，染料木素、大豆黄素和黄豆黄素等三种异黄酮转化率可达90％。实验研究了温度、甲醇水配比、进料配比以及接触时间对游离型异黄酮转化率的影响。结果表明，在实验温度范围内(60～70℃)，温度变化对genistein和glycitein影响较明显，温度升高转化率增大；温度变化对daidzein的转化率影响不大；接触时间越长，反应的转化率越高，并逐渐趋向平衡值；甲醇水配比为1:2有利于水解反应；溶剂比例增大，三种异黄酮转化率都增大，溶剂比例增大到一定程度转化率变化不明显。
异黄酮精制过程中，探索了柱层析、溶剂萃取、溶剂沉淀等方法，最终异黄酮含量可达40％，对胚芽异黄酮收率50％左右。
建立了总异黄酮的分析方法和游离态异黄酮的HPLC分析方法。
A process for extracting and purifying isoflavones from soybean germ was studied in this paper. The process includes extraction of total isoflavones and hydrolyzation of soybean isoflavone by solid acid catalyst.
In the experiments of extraction of total isoflavones, five factors were studied: temperature, extracting time, concentration of solvent and amount of solvent. With the extraction efficiency of total isoflavones and the selectivity of extraction as study targets, the optium extraction conditions were as followed: solvent 60% ethanol, temperature 50℃, solvent volume to sobean germ weight 6:1, extracting for 3times, 60min 10min , 10min respectively. Extraction efficiency of total isoflavones was about 92%.
In the experiments of fixed bed process, soybean isoflavone glucoside can be hydrolyzed into isoflavone aglucone with methanol in a fixed bed filled with solid acid catalyst(Nankai Chemical Factory, D72). Conversion of isoflavones is about 90%. The effect of temperature, contact time, concentration of methanol and proportion of solvent to material on conversion was studied. The results show that enhancing temperature and prolonging the contact time are favor of the reaction, and the ratio of methanol to water is appropriately 1:2.
During the process of refining of isoflavones from the extraction, methods such as chromatogram, extraction, deposition were researched. Content of isoflavone was as high as 40%, and recovery percent of isoflavone based on soybean germ was about 50%. Analytical method for total isoflavones and isoflavone aglucone was established.
引文
1．唐传核，彭志英．大豆功能性成分的开发现状[J]．中国油脂．2000，25(4)：44-48
2．毛峻琴，宓鹤鸣．大豆异黄酮的研究进展[J]．中草药．2000，31(1)：61-64
3．张青，洪青．大豆异黄酮的产品开发与应用前景[J]．中国食品添加剂．2002，(1)：23-24
4．李小满．大豆异黄酮分子结构、生物活性及其市场现状[J]．中国食品添加．2002，2：66-71
5．陈封政，王雄清，李书华．大豆异黄酮的研究进展[J]．乐山师范学院学报．2002，17(4)：59-61
6．史宣明，岳琳，武丽荣．大豆异黄酮的提取及精制[J]．中国油脂．2001，26(2)：3-5
7．鞠兴荣，袁建，汪海峰．大豆异黄酮的提取工艺的优化[J]．中国粮油学报．2001，16(6)：17-19
8. Michael Naim,Benjamin Gestetner, Shmuel Zilkahelal.Soybean Isoflavones. Characterization, Determination, and Antifungal Activity.J.Agr.Food Chem. 1974,22(5):806～810
9．郁军，裘爱泳．利用超临界CO_2技术综合开发大豆胚芽[J]．西部粮油科技．2000，25(4)：43-44
10．孙梅君等．中国大豆制品中异黄酮含量测定和分析研究[J]．食品与发酵工业．1999，26(5)：14-19
11．赵胜利，冀亚兰，黄正云等．愈风宁心片的薄层扫描定量法[J]．
12．赵世萍，章育中．葛根素中异黄酮含量的薄层光密度法测定[J]．药学学报．1985，20(3)：203-208
13．孙艳梅，张永忠．水解制备大豆异黄酮苷元研究进展[J]．食品研究与开发．2002，23(3)：11-13
14．崔东善．大豆异黄酮的功能及提取方法[J]．食品研究与开发．2001，22(3)：49-50
15．何继春，卫巍．大豆异黄酮检测及四标样快速测定法研究[J]．粮食与油脂．2000(7)：46-47
16．张炳文，宋永生等．糖苷型大豆异黄酮酸水解工艺的研究[J]．食品科学．2002，23(6)：75-78
17．王晓玲，温普红等．黄豆渣中异黄酮的提取分离研究[J]．中国药学杂质．2001，36(10)：661-663
18．李文亮，韩继福等．盐酸水解大豆异黄酮的工艺研究[J]．食品科学．2002，23(4)：73-75
19．U．S Patent No．5679806
20．U．S Patent No．6140469
21．冯国洲．提取大豆中异黄酮的方法．中华人民共和国知识产权局 CN 1069903C
22．汪海波，刘大川．大豆胚芽甲醇提取物中大豆皂甙、大豆异黄酮分离纯化工艺研究[J]．食品科学．2001；22(4)：40-44
23．U．S PatenZ No．5821361
24．朱仕房，王善利等．大豆异黄酮提取条件的研究[J]．食品科学．2001，22(3)：54-57
25．江和源等，测定大豆和葛根中异黄酮的高效液相色谱法[J]．分析测试学报．2000，19(6)：28-31
26. U.S Patent No. 5670632
27．江和源，吕飞杰，邰建祥，东惠茹．高效液相色谱法测定大豆中异黄酮的含量[J]．食品科学．2000，21(4)：56-58
28. U.S Patent No. 5702752
2．毛峻琴，宓鹤鸣．大豆异黄酮的研究进展[J]．中草药．2000，31(1)：61-64
3．张青，洪青．大豆异黄酮的产品开发与应用前景[J]．中国食品添加剂．2002，(1)：23-24
4．李小满．大豆异黄酮分子结构、生物活性及其市场现状[J]．中国食品添加．2002，2：66-71
5．陈封政，王雄清，李书华．大豆异黄酮的研究进展[J]．乐山师范学院学报．2002，17(4)：59-61
6．史宣明，岳琳，武丽荣．大豆异黄酮的提取及精制[J]．中国油脂．2001，26(2)：3-5
7．鞠兴荣，袁建，汪海峰．大豆异黄酮的提取工艺的优化[J]．中国粮油学报．2001，16(6)：17-19
8. Michael Naim,Benjamin Gestetner, Shmuel Zilkahelal.Soybean Isoflavones. Characterization, Determination, and Antifungal Activity.J.Agr.Food Chem. 1974,22(5):806～810
9．郁军，裘爱泳．利用超临界CO_2技术综合开发大豆胚芽[J]．西部粮油科技．2000，25(4)：43-44
10．孙梅君等．中国大豆制品中异黄酮含量测定和分析研究[J]．食品与发酵工业．1999，26(5)：14-19
11．赵胜利，冀亚兰，黄正云等．愈风宁心片的薄层扫描定量法[J]．
12．赵世萍，章育中．葛根素中异黄酮含量的薄层光密度法测定[J]．药学学报．1985，20(3)：203-208
13．孙艳梅，张永忠．水解制备大豆异黄酮苷元研究进展[J]．食品研究与开发．2002，23(3)：11-13
14．崔东善．大豆异黄酮的功能及提取方法[J]．食品研究与开发．2001，22(3)：49-50
15．何继春，卫巍．大豆异黄酮检测及四标样快速测定法研究[J]．粮食与油脂．2000(7)：46-47
16．张炳文，宋永生等．糖苷型大豆异黄酮酸水解工艺的研究[J]．食品科学．2002，23(6)：75-78
17．王晓玲，温普红等．黄豆渣中异黄酮的提取分离研究[J]．中国药学杂质．2001，36(10)：661-663
18．李文亮，韩继福等．盐酸水解大豆异黄酮的工艺研究[J]．食品科学．2002，23(4)：73-75
19．U．S Patent No．5679806
20．U．S Patent No．6140469
21．冯国洲．提取大豆中异黄酮的方法．中华人民共和国知识产权局 CN 1069903C
22．汪海波，刘大川．大豆胚芽甲醇提取物中大豆皂甙、大豆异黄酮分离纯化工艺研究[J]．食品科学．2001；22(4)：40-44
23．U．S PatenZ No．5821361
24．朱仕房，王善利等．大豆异黄酮提取条件的研究[J]．食品科学．2001，22(3)：54-57
25．江和源等，测定大豆和葛根中异黄酮的高效液相色谱法[J]．分析测试学报．2000，19(6)：28-31
26. U.S Patent No. 5670632
27．江和源，吕飞杰，邰建祥，东惠茹．高效液相色谱法测定大豆中异黄酮的含量[J]．食品科学．2000，21(4)：56-58
28. U.S Patent No. 5702752