银杏叶黄酮含量对银杏叶提取物大孔树脂纯化工艺条件的影响研究
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摘要
为考察银杏叶中黄酮含量对银杏叶提取物质量、得率及黄酮提取率的影响,为银杏叶提取物的生产提供原料标准和对应的工艺参数,本文选取黄酮含量分别为1.0%、0.8%和0.6%的三批银杏叶,在保证提取物质量满足中国药典要求的前提下,探究银杏叶质量的变化对提取物大孔树脂纯化工艺条件的影响。采用统计回归方法分别建立了银杏叶提取物的黄酮含量、提取物得率及黄酮提取率与纯化乙醇浓度和体积之间的关系。研究发现银杏叶提取物中黄酮含量与乙醇浓度及洗脱液体积正相关;提取物得率与洗脱液体积负相关、与乙醇浓度的关系随银杏叶中黄酮含量而变化;低黄酮含量银杏叶的提取物质量对工艺参数最为敏感;提取物中内酯含量与银杏叶中黄酮含量呈正相关关系并与提取物中黄酮含量、洗脱液体积呈负相关关系。为保证提取物质量、工艺条件和得率的稳定,建议通过不同质量原料混合的方式使银杏叶中黄酮含量维持在1%,采用2倍量15%乙醇进行洗脱,可获得黄酮含量大于24%、内酯含量6%,得率2.6%、黄酮提取率大于66%的银杏叶提取物。
This study aims to investigate the effect of flavonoids contents in Ginkgo biloba leaves on the quality,yield and flavonoid extraction rate of extract of G.biloba leaves(EGb),and provide quality standard of raw material and corresponding processing parameters for EGb production.G.biloba leaves with flavonoid content of 1.0 %,0.8 % and 0.6 % were applied to study the effects of the quality of Ginkgo biloba leaves on the macroporous resin purification process for ensuring the quality of EGb to meet the requirement of Chinese Pharmacopoeia.Statistic regression was used to investigate the relationship between EGb properties and the concentration and volume of ethanol for purification,respectively.These properties include flavonoid content and yield of EGb,and extraction rate of flavonoids.The results showed that the concentration of ethanol and eluent volume was positively correlated with the content of flavonoids in EGb.The yield of EGb was negatively correlated with the eluent volume and changed with the contents of flavonoids in the leaves.The quality of EGb extracted from Ginkgo Leaves with low flavonoid concentration was most sensitive to the processing parameters.The content of lactone in EGb had a positive relation with flavonoid content in Ginkgo biloba leaves.It also had a negative correlation with the flavonoid content of EGb and eluent volume.To keep the quality,processing conditions and yield of EGb stable,it was suggested to mix G.biloba leaves with different flavonoid concentration to make the flavonoid content in the leaves be 1.0 %.By using twice amount of 15 % ethanol to elute the resins,EGb containing more than 24 % of flavonoids and 6 % of lactone can be prepared with yield of 2.6 % and over 66 % flavonoid extraction rate.
This study aims to investigate the effect of flavonoids contents in Ginkgo biloba leaves on the quality,yield and flavonoid extraction rate of extract of G.biloba leaves(EGb),and provide quality standard of raw material and corresponding processing parameters for EGb production.G.biloba leaves with flavonoid content of 1.0 %,0.8 % and 0.6 % were applied to study the effects of the quality of Ginkgo biloba leaves on the macroporous resin purification process for ensuring the quality of EGb to meet the requirement of Chinese Pharmacopoeia.Statistic regression was used to investigate the relationship between EGb properties and the concentration and volume of ethanol for purification,respectively.These properties include flavonoid content and yield of EGb,and extraction rate of flavonoids.The results showed that the concentration of ethanol and eluent volume was positively correlated with the content of flavonoids in EGb.The yield of EGb was negatively correlated with the eluent volume and changed with the contents of flavonoids in the leaves.The quality of EGb extracted from Ginkgo Leaves with low flavonoid concentration was most sensitive to the processing parameters.The content of lactone in EGb had a positive relation with flavonoid content in Ginkgo biloba leaves.It also had a negative correlation with the flavonoid content of EGb and eluent volume.To keep the quality,processing conditions and yield of EGb stable,it was suggested to mix G.biloba leaves with different flavonoid concentration to make the flavonoid content in the leaves be 1.0 %.By using twice amount of 15 % ethanol to elute the resins,EGb containing more than 24 % of flavonoids and 6 % of lactone can be prepared with yield of 2.6 % and over 66 % flavonoid extraction rate.
引文
1 Laws KR,Sweetnam H,Kondel TK.Is Ginkgo biloba a cognitive enhancer in healthy individuals?A meta-analysis.Hum Psychopharm Clin,2012,27:527-533.
2 Zhiyan Consulting Group(智研咨询集团).Report on market and investment opportunities of China Ginkgo biloba extract industry during year 2015-2020.http://www.chyxx.com/research/201507/329542.html#catalogue,2015,7.
3 Chinese Pharmacopoeia Commission(国家药典委员会).Pharmacopoeia of the People's Republic of China(中华人民共和国药典).Beijing:China Medical Science Press,2015.Vol I,316-416.
4 United States Pharmacopoeia,USP36-NF31.Rockville,MD:The United States Pharmacopoeia Convention,2013.1476.
5 Zhou XH(周小华),Zeng H(曾晖),Li DH(李端华).Purification of total flavones of Ginkgo biloba L.leaves by HZ-841 adsorption resin.Nat Prod Res Dev(天然产物研究与开发),2005,17:76-79.
6 Liu XG,Wu SQ,Li P,et al.Advancement in the chemical analysis and quality control of flavonoid in Ginkgo biloba.J Pharm Biomed Anal,2015,113:212-225.
7 Chen XP,Zhang YD.Rapid microwave-assisted hydrolysis for determination of Ginkgo flavonol glycosides in extracts of Ginkgo bilobaleaves.J Chromatogr Sci,2008,46:117-121.
8 Yang RH(杨荣华),Ren JD(任京东),Qu YX(屈云霞).Study on extracting flavonoids from the leaves of Ginkgo biloba by ultrasonic.China Med Herald(中国医药导报),2010,7(20):25-27.
9 Wu ML(吴梅林),Zhou CS(周春山),Chen LS(陈龙胜).Study on the extraction of total flavonoids from Ginkgo leaves by enzyme hydrolysis.Nat Prod Res Dev(天然产物研究与开发),2004,16:557-560.
10 Miao SF,Yu JP,Du Z,et al.Supercritical fluid extraction and micronization of ginkgo flavonoids from Ginkgo biloba Leaves.Ind Eng Chem Res,2010,49:5461-5466.
11 Zhang Q,Chen LJ,Ye HY,et al.Isolation and purification of ginkgo flavonol glycosidesfrom Ginkgo biloba leaves by highspeed counter-current chromatography.J Sep Sci,2007,30:2153-2159.
12 Li YJ(李艳杰).Study on biological characteristics and pharmacological constituents of Ginkgo biloba leaves in Changchun and Nanjing.Changchun:Northeast Normal University(东北师范大学),MSc.2008.
13 Liu FY(刘芳瑞),Hu JH(胡晶红),Li J(李佳),et al.Analysis of influencing factors on the synthesis and accumulation of active substance in Ginkgo plants.J Shandong Univ TCM(山东中医药大学学报),2011,35:199-202.
14 Qa'dan F,Mansoor K,AL-Adham I,et al.Proanthocyanidins from Ginkgo biloba leaf extract and their radical scavenging activity.Pharm Biol,2011,49:471-476.
15 Xie HY,Wang JR,Yau LF,et al.Catechins and procyanidins of Ginkgo bilobashow potent activities towards the inhibition ofβ-amyloid peptideaggregation and destabilization of preformed fibrils.Cell Biochem Funct,2013,31:344.
16 Park YG,Kim SJ,Kang HYM,et al.Variation of ginkgolides and bilobalide contents in leaves and cell culture of Ginkgo biloba L.Biotechnol Bioproc Eng,2004,9:35-40.
17 van Beeka TA,Montorob P.Chemical analysis and quality control of Ginkgo biloba leaves,extracts,and phytopharmaceuticals.J Chromatogr A,2009,1216:2002-2032.
18 Qin B(秦兵),Zhang GB(张根葆),Chen DY(陈冬云),et al.Protective effects of ginkgolide B on neuron injury induced by cerebral ischemia/reperfusion.Chin J TCM WM Crit Care(中国中西医结合急救杂志),2005,12(1):17-20.
19 Maclennan KM,Darlington CL,Smith PF.The CNS effects of Ginkgo biloba extracts and ginkgolide B.Progr Neurobiol,2002,67:235-257.
2 Zhiyan Consulting Group(智研咨询集团).Report on market and investment opportunities of China Ginkgo biloba extract industry during year 2015-2020.http://www.chyxx.com/research/201507/329542.html#catalogue,2015,7.
3 Chinese Pharmacopoeia Commission(国家药典委员会).Pharmacopoeia of the People's Republic of China(中华人民共和国药典).Beijing:China Medical Science Press,2015.Vol I,316-416.
4 United States Pharmacopoeia,USP36-NF31.Rockville,MD:The United States Pharmacopoeia Convention,2013.1476.
5 Zhou XH(周小华),Zeng H(曾晖),Li DH(李端华).Purification of total flavones of Ginkgo biloba L.leaves by HZ-841 adsorption resin.Nat Prod Res Dev(天然产物研究与开发),2005,17:76-79.
6 Liu XG,Wu SQ,Li P,et al.Advancement in the chemical analysis and quality control of flavonoid in Ginkgo biloba.J Pharm Biomed Anal,2015,113:212-225.
7 Chen XP,Zhang YD.Rapid microwave-assisted hydrolysis for determination of Ginkgo flavonol glycosides in extracts of Ginkgo bilobaleaves.J Chromatogr Sci,2008,46:117-121.
8 Yang RH(杨荣华),Ren JD(任京东),Qu YX(屈云霞).Study on extracting flavonoids from the leaves of Ginkgo biloba by ultrasonic.China Med Herald(中国医药导报),2010,7(20):25-27.
9 Wu ML(吴梅林),Zhou CS(周春山),Chen LS(陈龙胜).Study on the extraction of total flavonoids from Ginkgo leaves by enzyme hydrolysis.Nat Prod Res Dev(天然产物研究与开发),2004,16:557-560.
10 Miao SF,Yu JP,Du Z,et al.Supercritical fluid extraction and micronization of ginkgo flavonoids from Ginkgo biloba Leaves.Ind Eng Chem Res,2010,49:5461-5466.
11 Zhang Q,Chen LJ,Ye HY,et al.Isolation and purification of ginkgo flavonol glycosidesfrom Ginkgo biloba leaves by highspeed counter-current chromatography.J Sep Sci,2007,30:2153-2159.
12 Li YJ(李艳杰).Study on biological characteristics and pharmacological constituents of Ginkgo biloba leaves in Changchun and Nanjing.Changchun:Northeast Normal University(东北师范大学),MSc.2008.
13 Liu FY(刘芳瑞),Hu JH(胡晶红),Li J(李佳),et al.Analysis of influencing factors on the synthesis and accumulation of active substance in Ginkgo plants.J Shandong Univ TCM(山东中医药大学学报),2011,35:199-202.
14 Qa'dan F,Mansoor K,AL-Adham I,et al.Proanthocyanidins from Ginkgo biloba leaf extract and their radical scavenging activity.Pharm Biol,2011,49:471-476.
15 Xie HY,Wang JR,Yau LF,et al.Catechins and procyanidins of Ginkgo bilobashow potent activities towards the inhibition ofβ-amyloid peptideaggregation and destabilization of preformed fibrils.Cell Biochem Funct,2013,31:344.
16 Park YG,Kim SJ,Kang HYM,et al.Variation of ginkgolides and bilobalide contents in leaves and cell culture of Ginkgo biloba L.Biotechnol Bioproc Eng,2004,9:35-40.
17 van Beeka TA,Montorob P.Chemical analysis and quality control of Ginkgo biloba leaves,extracts,and phytopharmaceuticals.J Chromatogr A,2009,1216:2002-2032.
18 Qin B(秦兵),Zhang GB(张根葆),Chen DY(陈冬云),et al.Protective effects of ginkgolide B on neuron injury induced by cerebral ischemia/reperfusion.Chin J TCM WM Crit Care(中国中西医结合急救杂志),2005,12(1):17-20.
19 Maclennan KM,Darlington CL,Smith PF.The CNS effects of Ginkgo biloba extracts and ginkgolide B.Progr Neurobiol,2002,67:235-257.