橘皮的化学成分与黄酮类化合物的提取、纯化研究
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摘要
本文主要是关于柑橘皮中黄酮类化合物的含量分析、提取条件优化、分离制备单一天然产物的方法研究。实验结果表明:
1、不同品种的橘皮中黄酮的种类、含量不同,并且相差很大,需要针对不同的目标产物选择原料。
2、比较常压有机溶剂、高压有机溶剂、超临界二氧化碳和常压石灰水提取柑橘皮中黄酮类化合物的条件、提取物的纯度、提取效率,结果显示:常压超声提取最经济,高压有机溶剂提取效率最高,超临界二氧化碳的提取物纯度高但效率低,饱和石灰水是提取柑橘皮中橙皮苷的理想溶剂,甲醇、乙醇、正丁醇对黄酮类化合物的提取效率依次增大,提取物的纯度也提高,高压乙酸乙酯是提取多甲氧基黄酮的理想方法。
3、比较不同极性有机溶剂萃取分离、高速逆流色谱分离、D101大孔树脂吸附、强碱性阴离子交换树脂吸附等方法对提取物的纯化效果。结果显示:不同极性的有机溶剂和高速逆流色谱并不能实现复杂混合物的完全分离;D101大孔吸附树脂对黄酮类化合物进行浓缩和纯化可作为前处理的方法。
4、研究认为:以甲醇-0.5%HAc作流动相,C_(18)硅胶色谱柱作固定相,283nm的检测波长可以对橘皮提取物的D101大孔树脂纯化物进行选择地分离,得到4种高纯度的物质。
Contents of this paper consist of optimized extracting factors of flavonoids from citrus peels and methods of separation and purification of single natural compound and stability conditions of extracting matters. The results are showed as follows:
1. They have different species and contents of flavonoids in six citrus peels. So it's essencial to select given breeds in order to obtain the most yield.
2. The optimized conditions of extracting flavonoids are studied by different ways such as organic solvents under one atm. As well as under high presure , by supercritical fluid carbon dioxide,limewater under one atm. The purity and rate of production are detected as main parameters. Results showed that extraction by ultrasound under one atm. is cheaper,by organic solvents under high presure is with high rate,by upercutical fluid carbon dioxide is with high purity but low rate. Limewater is the perfecter solvent to extract flavonoid hesperidin. Methanol or ethanol or n-butyl alcohol worked as solvents are without different,of them n-butyl alcohol is the best,ethanol is better .Acetic ether is fit to extract PDF under high pressure.
3. The laminar analyses are studied by D101 macroporous resin, strong basicity anion polymeric adsorbent. Seperation by different organic solvent and high speed countercurrent chromtography are studied as well .The rusults showed macroporous resinis is better matter to condense and purify flavonoids except seperation, strong basicity anion polymeric adsorbent adsorbs selectively certein flavonoids . Extracting flavonoids by different polar solvents and testing by HPLC afterwards, it is difficult to separate compounds by simple extraction.
4. Separation of flavonoids from citrus peel by HPLC was studied , which worked by methanol-0.5%HAc as solution,C_(18) resin column as stationary phase. The results showed that whether resolutions or retention time is ideal under chosed conditions. Now it is limited by the weight of flavonoids added to the column.
1、不同品种的橘皮中黄酮的种类、含量不同,并且相差很大,需要针对不同的目标产物选择原料。
2、比较常压有机溶剂、高压有机溶剂、超临界二氧化碳和常压石灰水提取柑橘皮中黄酮类化合物的条件、提取物的纯度、提取效率,结果显示:常压超声提取最经济,高压有机溶剂提取效率最高,超临界二氧化碳的提取物纯度高但效率低,饱和石灰水是提取柑橘皮中橙皮苷的理想溶剂,甲醇、乙醇、正丁醇对黄酮类化合物的提取效率依次增大,提取物的纯度也提高,高压乙酸乙酯是提取多甲氧基黄酮的理想方法。
3、比较不同极性有机溶剂萃取分离、高速逆流色谱分离、D101大孔树脂吸附、强碱性阴离子交换树脂吸附等方法对提取物的纯化效果。结果显示:不同极性的有机溶剂和高速逆流色谱并不能实现复杂混合物的完全分离;D101大孔吸附树脂对黄酮类化合物进行浓缩和纯化可作为前处理的方法。
4、研究认为:以甲醇-0.5%HAc作流动相,C_(18)硅胶色谱柱作固定相,283nm的检测波长可以对橘皮提取物的D101大孔树脂纯化物进行选择地分离,得到4种高纯度的物质。
Contents of this paper consist of optimized extracting factors of flavonoids from citrus peels and methods of separation and purification of single natural compound and stability conditions of extracting matters. The results are showed as follows:
1. They have different species and contents of flavonoids in six citrus peels. So it's essencial to select given breeds in order to obtain the most yield.
2. The optimized conditions of extracting flavonoids are studied by different ways such as organic solvents under one atm. As well as under high presure , by supercritical fluid carbon dioxide,limewater under one atm. The purity and rate of production are detected as main parameters. Results showed that extraction by ultrasound under one atm. is cheaper,by organic solvents under high presure is with high rate,by upercutical fluid carbon dioxide is with high purity but low rate. Limewater is the perfecter solvent to extract flavonoid hesperidin. Methanol or ethanol or n-butyl alcohol worked as solvents are without different,of them n-butyl alcohol is the best,ethanol is better .Acetic ether is fit to extract PDF under high pressure.
3. The laminar analyses are studied by D101 macroporous resin, strong basicity anion polymeric adsorbent. Seperation by different organic solvent and high speed countercurrent chromtography are studied as well .The rusults showed macroporous resinis is better matter to condense and purify flavonoids except seperation, strong basicity anion polymeric adsorbent adsorbs selectively certein flavonoids . Extracting flavonoids by different polar solvents and testing by HPLC afterwards, it is difficult to separate compounds by simple extraction.
4. Separation of flavonoids from citrus peel by HPLC was studied , which worked by methanol-0.5%HAc as solution,C_(18) resin column as stationary phase. The results showed that whether resolutions or retention time is ideal under chosed conditions. Now it is limited by the weight of flavonoids added to the column.
引文
[1]李嗣彪.柑橘皮功能性物质种类及绿色联产工艺[J].2003,20(3):25-27
[2]徐彭.陈皮水提取物和陈皮挥发油的药理作用比较[J].江西中医学院学报,1998,10(4):172-i73
[3]YEN WJ.Isolation of Xanthophylls from Taiwanese Orange Peels and their Effects on the Oxidation Stability of Soybean Oil[J].Food Chemistry,1995,53(4):417-425
[4]汪秋安.几类具有抗癌作用的食物成分[J].中国食物与营养,2000,6(1):42-43
[5]Schalch W,Dayhaw.Barker P,Barker F M.Nutritional and Environmental Influences on the Eye.Taylor A,ed.CRC Press,Boca Raton.1999:215-249
[6]张孟民,张新利.宝鸡文理学院学报[J].2005年第四期,第25卷,286-288.
[7]陆业高.植物化学成分[M].化工出版社,2004年4月第l版:223-240
[8]吴立军.天然药物化学[M].人民卫生出版社,1988年5月第1版
[9]高锦明.植物化学[M].科学出版社,2003年4月第1版,156-i94
[10]Benavente,Garcia O,et a1.1997.Uses and properties of citrusflavonoids.J.Agric.Food Chem.,45(2):4505-4515
[11]Ninson J A,et al.A Citrus Extract Plus Ascorbic Acid Decreases Lipids,Lipids Peroxides,Lipoprotein Oxidative Susceptibility,and Atherosclerosis in Hypercholesterolemic Hamsters[J].Food Chem,1998,46:1453-1459
[12]徐怀德.天然产物提取工艺学[M].北京:中国轻工业出版社,2..6年6月第1版:351-352
[13]谭仁祥.植物化学成分分析[M].北京:科学出版社,2002年2月第1版:492-502
[14]陈复生,左锦静等.陈皮黄酮类化合物精制条件的研究[J].食品研究与开发.2006,27(9):38-4l
[15]陈复生,左锦静等.大孔树脂吸附和分离陈皮中黄酮类化合物的研究[J].食品科技.2006,7:121-124
[16]赵雪梅.浙江大学博士论文.胡柚皮化学成分及其活性研究[M].2003,26.
[17]唐传核.植物功能性食品[M].北京:化学工业出版社,2004年10月第1版:197
[18]Ortuno AM,et al.1999.Evolution of polymethoxy flavones during development of tangelo Nova fruits.Food Chenistry,66:217-220
[19]胡燕飞,王长海.微波法提取陈皮中3,5,6,7,8,3',4'-七甲氧基黄酮及其含量的测定[J].烟台大学学报,2005,18(1):45-49
[20]Bear W L,TelI R W.Anticancer Res[J].2000,20(5A):3323-3329
[21]罗水忠.柑橘籽中柠檬苦素的提取与抑菌性研究[J].2006,10:105-107
[22]齐海鸥等.柑橘皮的综合利用[J].浙江柑橘,2003,20(3):31-35
[23]孙崇德.柑橘果实中天然柠檬苦素和诺米林的提取、鉴别与检测[J].2004,4(1):6-11
[24]祝渊等.柑橘果实纤维的研究[J].果树学报,2003,20(4):256-260
[25]Woodman O L,Meeker W F.Vasovelaxant and antioxidant activity of flavonoids and flavones:structure-activity relationships[J].Cardiovasc Pharmacol,2005,46:302-309
[26]Ma Y L,Li Q M,Vandenheuvel H,Claaeys M.Rapid commun.Mass Spectrom.,1997,(11):1357-1364.
[27]J.B.哈本.黄酮类化合物[M].科学出版社,1983年4月第1版
[28]周大勇,徐青,薛兴亚等.高效液相色谱质谱一电喷雾质谱法测定枳壳中黄酮苷类化合物.分析化学研究报告[J].2006年第34卷:31-35.
[29]蓝航莲.柑橘类黄酮的生理活性和研究进展[J].2001
[30]苏东林.柑橘皮中功能性成分的作用概述[J].现代食品科技,2006,22(2):260-263
[31].食品卫生检验方法(理化部分),GB/T,5009-2003
[32]岳红.超临界二氧化碳萃取柿叶黄酮的工艺研究[J].2005,25(3):55-58
[33]刘文娟,耿秋菊等.枳实药材中总黄酮提取工艺的研究[J].时珍国医国药.2006,17(11):2233-2234
[34]Zhang Chongwei,Wei Dongzhi 等。Citrus flavonoids simultaneous HPLC anylysis.中成药[J]2007年第29卷第一期,99-104.
[35]李行利,郭秋月.陈皮提取工艺研究[J].中成药.1997,19(1):6-8
[36]廖传华,黄振仁.超临界C02流体萃取技术[M].北京:化学工业出版社,2004年7月第1版:118-212
[37]何扩等.超临界流体萃取银杏叶中黄酮类化合物的研究[J].山西食品工业, 2005,24(4):31-35
[38]Chen C,Shen C,Chow M,et al.Flavone inhibition of tumor growth via apoptosis in vitro and in vivo[J].Int J Oncol,2004,25(3):661-670.
[39]董岩,崔庆新,魏兴国等.陈皮挥发油化学成分气相色谱一质谱分析[J].山东中医杂志,2004,23(6):370-372
[40]刘成梅,游海.天然产物有效成分的分离和应用[M].中国轻工业出版社,2002年8月第1版
[41]刘均玉,许晨等.不同品种橘皮中的黄酮分析.中国食品添加剂[J],2008,vol.90,NO.5 2008:160
[42]Wang X,Li F W,Zhang H X,et al.Preparative isolation and purification of polymethoxylated flavones from Tangerine peel using high-speed counter-current chromatography[J].Journal of Chromatography A,2005,1090(1-2):188-192
[43]严赞开,胡春菊.橙皮苷的抑菌效果研究[J].西北农业学报,2004,13(2):87-89
[44]易美华.生物资源开发利用[M].中国轻工业出版社,2003年6月第1版:159-259
[45]曹学丽.高速逆流色谱分离技术及应用[M].化学工业出版社,2005年3月第1版:30-252
[46]J.A.Vinson,X.Liang,J.Proch,B.A.Hontz,J.Dancel and N.Sandone,Adv.Exp.Med.Biol.[J],2002(505):113
[47]S.C.Choe,H.S.Kim,T.S.Jeong,S.H.Bok and Y.B.Park,J.Cardiovasc.Pharmacol.[J],2001(38):947
[48]Ninson J A,et al.A Citrus Extract Plus Ascorbic Acid Decreases Lipids,Lipids Peroxides,Lipoprotein Oxidative Susceptibility,and Atherosclerosis in Hypercholesterolemic Hamsters[J].Food Chem,1998,46:1453-1459
[49]张孟民,张新利.宝鸡文理学院学报[J].2005年第四期,第25卷,286-288.
[50]唐传核.植物功能性食品[M].北京:化学工业出版社,2004年10月第1版:197
[51]J.A.Manthey,Microcirculation,7(2000):947
[52]Cristina Garcia-Viguera.Determination of citrus jams genuineness by flavonoid analysis.Zlebensm Unters Forsch(1993)197:255-259
[53] E. Alissandrakis,D.Daferera,et.al.Ultrasound-assisted extraction of volatile compounds from citrus flowers and citrus honey.Food Chem.82(2003)575-482.
[54] E.Belajova,M.Suhaj.Determination of phenolic constituents in citrus juices:Method of HPLC . Food Chem.86(2004)339-343.
[55] Jun Yu,Deepak V. Dandekar.Supercritical fluid extraction of limomoids and naringin from grapefruit seeds. Food Chem.105(2007)1026-1031.
[2]徐彭.陈皮水提取物和陈皮挥发油的药理作用比较[J].江西中医学院学报,1998,10(4):172-i73
[3]YEN WJ.Isolation of Xanthophylls from Taiwanese Orange Peels and their Effects on the Oxidation Stability of Soybean Oil[J].Food Chemistry,1995,53(4):417-425
[4]汪秋安.几类具有抗癌作用的食物成分[J].中国食物与营养,2000,6(1):42-43
[5]Schalch W,Dayhaw.Barker P,Barker F M.Nutritional and Environmental Influences on the Eye.Taylor A,ed.CRC Press,Boca Raton.1999:215-249
[6]张孟民,张新利.宝鸡文理学院学报[J].2005年第四期,第25卷,286-288.
[7]陆业高.植物化学成分[M].化工出版社,2004年4月第l版:223-240
[8]吴立军.天然药物化学[M].人民卫生出版社,1988年5月第1版
[9]高锦明.植物化学[M].科学出版社,2003年4月第1版,156-i94
[10]Benavente,Garcia O,et a1.1997.Uses and properties of citrusflavonoids.J.Agric.Food Chem.,45(2):4505-4515
[11]Ninson J A,et al.A Citrus Extract Plus Ascorbic Acid Decreases Lipids,Lipids Peroxides,Lipoprotein Oxidative Susceptibility,and Atherosclerosis in Hypercholesterolemic Hamsters[J].Food Chem,1998,46:1453-1459
[12]徐怀德.天然产物提取工艺学[M].北京:中国轻工业出版社,2..6年6月第1版:351-352
[13]谭仁祥.植物化学成分分析[M].北京:科学出版社,2002年2月第1版:492-502
[14]陈复生,左锦静等.陈皮黄酮类化合物精制条件的研究[J].食品研究与开发.2006,27(9):38-4l
[15]陈复生,左锦静等.大孔树脂吸附和分离陈皮中黄酮类化合物的研究[J].食品科技.2006,7:121-124
[16]赵雪梅.浙江大学博士论文.胡柚皮化学成分及其活性研究[M].2003,26.
[17]唐传核.植物功能性食品[M].北京:化学工业出版社,2004年10月第1版:197
[18]Ortuno AM,et al.1999.Evolution of polymethoxy flavones during development of tangelo Nova fruits.Food Chenistry,66:217-220
[19]胡燕飞,王长海.微波法提取陈皮中3,5,6,7,8,3',4'-七甲氧基黄酮及其含量的测定[J].烟台大学学报,2005,18(1):45-49
[20]Bear W L,TelI R W.Anticancer Res[J].2000,20(5A):3323-3329
[21]罗水忠.柑橘籽中柠檬苦素的提取与抑菌性研究[J].2006,10:105-107
[22]齐海鸥等.柑橘皮的综合利用[J].浙江柑橘,2003,20(3):31-35
[23]孙崇德.柑橘果实中天然柠檬苦素和诺米林的提取、鉴别与检测[J].2004,4(1):6-11
[24]祝渊等.柑橘果实纤维的研究[J].果树学报,2003,20(4):256-260
[25]Woodman O L,Meeker W F.Vasovelaxant and antioxidant activity of flavonoids and flavones:structure-activity relationships[J].Cardiovasc Pharmacol,2005,46:302-309
[26]Ma Y L,Li Q M,Vandenheuvel H,Claaeys M.Rapid commun.Mass Spectrom.,1997,(11):1357-1364.
[27]J.B.哈本.黄酮类化合物[M].科学出版社,1983年4月第1版
[28]周大勇,徐青,薛兴亚等.高效液相色谱质谱一电喷雾质谱法测定枳壳中黄酮苷类化合物.分析化学研究报告[J].2006年第34卷:31-35.
[29]蓝航莲.柑橘类黄酮的生理活性和研究进展[J].2001
[30]苏东林.柑橘皮中功能性成分的作用概述[J].现代食品科技,2006,22(2):260-263
[31].食品卫生检验方法(理化部分),GB/T,5009-2003
[32]岳红.超临界二氧化碳萃取柿叶黄酮的工艺研究[J].2005,25(3):55-58
[33]刘文娟,耿秋菊等.枳实药材中总黄酮提取工艺的研究[J].时珍国医国药.2006,17(11):2233-2234
[34]Zhang Chongwei,Wei Dongzhi 等。Citrus flavonoids simultaneous HPLC anylysis.中成药[J]2007年第29卷第一期,99-104.
[35]李行利,郭秋月.陈皮提取工艺研究[J].中成药.1997,19(1):6-8
[36]廖传华,黄振仁.超临界C02流体萃取技术[M].北京:化学工业出版社,2004年7月第1版:118-212
[37]何扩等.超临界流体萃取银杏叶中黄酮类化合物的研究[J].山西食品工业, 2005,24(4):31-35
[38]Chen C,Shen C,Chow M,et al.Flavone inhibition of tumor growth via apoptosis in vitro and in vivo[J].Int J Oncol,2004,25(3):661-670.
[39]董岩,崔庆新,魏兴国等.陈皮挥发油化学成分气相色谱一质谱分析[J].山东中医杂志,2004,23(6):370-372
[40]刘成梅,游海.天然产物有效成分的分离和应用[M].中国轻工业出版社,2002年8月第1版
[41]刘均玉,许晨等.不同品种橘皮中的黄酮分析.中国食品添加剂[J],2008,vol.90,NO.5 2008:160
[42]Wang X,Li F W,Zhang H X,et al.Preparative isolation and purification of polymethoxylated flavones from Tangerine peel using high-speed counter-current chromatography[J].Journal of Chromatography A,2005,1090(1-2):188-192
[43]严赞开,胡春菊.橙皮苷的抑菌效果研究[J].西北农业学报,2004,13(2):87-89
[44]易美华.生物资源开发利用[M].中国轻工业出版社,2003年6月第1版:159-259
[45]曹学丽.高速逆流色谱分离技术及应用[M].化学工业出版社,2005年3月第1版:30-252
[46]J.A.Vinson,X.Liang,J.Proch,B.A.Hontz,J.Dancel and N.Sandone,Adv.Exp.Med.Biol.[J],2002(505):113
[47]S.C.Choe,H.S.Kim,T.S.Jeong,S.H.Bok and Y.B.Park,J.Cardiovasc.Pharmacol.[J],2001(38):947
[48]Ninson J A,et al.A Citrus Extract Plus Ascorbic Acid Decreases Lipids,Lipids Peroxides,Lipoprotein Oxidative Susceptibility,and Atherosclerosis in Hypercholesterolemic Hamsters[J].Food Chem,1998,46:1453-1459
[49]张孟民,张新利.宝鸡文理学院学报[J].2005年第四期,第25卷,286-288.
[50]唐传核.植物功能性食品[M].北京:化学工业出版社,2004年10月第1版:197
[51]J.A.Manthey,Microcirculation,7(2000):947
[52]Cristina Garcia-Viguera.Determination of citrus jams genuineness by flavonoid analysis.Zlebensm Unters Forsch(1993)197:255-259
[53] E. Alissandrakis,D.Daferera,et.al.Ultrasound-assisted extraction of volatile compounds from citrus flowers and citrus honey.Food Chem.82(2003)575-482.
[54] E.Belajova,M.Suhaj.Determination of phenolic constituents in citrus juices:Method of HPLC . Food Chem.86(2004)339-343.
[55] Jun Yu,Deepak V. Dandekar.Supercritical fluid extraction of limomoids and naringin from grapefruit seeds. Food Chem.105(2007)1026-1031.