杜梨叶黄酮类化合物提取、纯化及抗氧化性的研究
|
摘要
黄酮类化合物在自然界中分布广泛,主要存在于植物的叶片和果实中。黄酮类药物具有抗脑缺血、抗心肌缺血、抗心律失常、镇痛、保肝护肝、对消化性溃疡的保护、抗病毒、抗肿瘤作用等。此外,还有大量研究表明黄酮类天然产物有效成分有降压、降血脂和抑制血小板聚集等多种药理作用。为此,受到各国政府和医药、食品等行业的高度重视和关注。目前为止,对杜梨叶中黄酮类物质提取、纯化工艺研究尚未见报道。杜梨药用称棠梨,主治功用为敛肺涩肠、止咳止痢,常用于治疗久咳、久泻和久痢等。为开发利用该资源,本研究对其黄酮类物质检测、提取与分离纯化工艺进行了初步探索,建立了一套经济、安全、适用的杜梨叶黄酮提取、分离与检测方法。其主要研究如下:
1.本试验采用紫外可见分光光度法和高效液相色谱法对杜梨叶黄酮作了定性分析。建立了紫外可见分光光度法对杜梨叶总黄酮的测定方法。通过对检测波长的选择、标准品的选择、预处理方法、浓度与吸光度相关性的研究,确定了最佳检测波长为入=420nm,以芦丁为标准品,在0.06-0.54mg/mL范围内,其浓度与吸光度呈良好的线性关系,回归方程为A=1.31667C-0.01678,相关系数达r~2=0.99440,稳定性、精密度和重现性均理想,其RSD分别为3.55%(n=5)、1.36%(n=5)、4.79%(n=5)。平均回收率分别为94.41%,RSD为2.33%(n=5)。
2.在提取工艺研究中,采用乙醇水溶液为浸提剂,以杜梨叶总黄酮含量为考察指标,通过单因素实验、正交实验以及方差分析,对提取条件进行了优化,确定了最佳提取工艺为A_2B_3C_1D_2.即乙醇浓度为70%、浸提温度为70℃、浸提时间为1h,固液比为1:20,黄酮提取率为4.87%。
3.本试验采用大孔树脂对杜梨叶黄酮进行富集分离。根据大孔树脂的吸附机理,运用静态吸附与解吸实验对大孔树脂进行筛选。然后,通过单因素实验、正交实验以及方差分析确定了杜梨叶黄酮分离纯化的最佳操作条件。静态吸附与解吸中,AB-8大孔树脂表现出最好的吸附性能与良好的解吸效果,其饱和静态吸附量为12.15mg/g、解吸率为95.88%。实验表明,料液浓度和上柱速度明显影响吸附效果,而洗脱效果由乙醇浓度、洗脱速度以及洗脱剂用量来决定。实验确定的最佳吸附条件为:上柱速度为2.0 BV/h,料液浓度为0.370mg/mL,最佳洗脱条件为A_1B_2C_3,即乙醇浓度为75%,洗脱流速为2BV/h,洗脱体积为4.5BV,洗脱率为88.09%。
4.研究发现乙醇提取杜梨叶黄酮类化合物具有较强的抗氧化能力。应用清除DPPH·自由基揭示了水提取杜梨叶黄酮类化合物的抗氧化能力大于乙醇提取杜梨叶黄酮类化合物。水提取粗液的IC_(50)为0.06mg/mL,70%乙醇提取粗液的IC_(50)为0.08 mg/mL。结果表明,水提取粗液清除DPPH·自由基能力强于70%乙醇提取粗液。浓度在0.8mg/mL时,其均超过其半数清除率,水提取粗液达到84.57%,70%乙醇提取粗液为70.33%。
Flavonoids are widely distributed in nature, mainly in plant leaves and fruits. Flavonoids with anti-drug role in cerebral ischemia, the role of anti-myocardial ischemia, antiarrhythmic effects, analgesic effects, liver Hugan effect on the protective effect of peptic ulcer, anti-virus, anti-tumor effect. In addition, there are a large number of studies show that flavonoids are the active ingredients of natural products antihypertensive, hypolipidemic, inhibiting platelet aggregation and other pharmacological effects on flavonoid natural products active ingredients there are a variety of biological activity. To this end, by Governments and the pharmaceutical and food industry attaches great importance and concern. The flavonoids extract active ingredients is its reseach and application of key technologies and stages. So far, Bunge leaves flavonoids on resource extraction and purification process has not been reported. Bunge said medicinal Bunge, chief of pulmonary function for the convergence Shibuya intestines,cough, commonly used in the treatment of Chronic Cough,Disrrhea, Dysentery such as. For the development and utilization of the resources, the study of its detection, extraction and purification carried out a preliminary exploration, the establishment of a set of economic, security, application of the Bunge leaf flavonoids extraction, separation and detection methods.
In this study, UV-Vis spectrophotometry and HPLC of Bunge made a qualitative analysis of leaf flavonoids. Detection wavelength of choice, the choice of standard, pre-treatment method, concentration and absorbance of the relevant studies to determine the best detection wavelength of income = 420nm, for the standard to rutin in 0.06～0.54mg/ml the scope of its concentration and absorbance was a good linear relationship between the regression equation was A = 1.31667C-0.01678, correlation coefficient of r~2 = 0.99440, stability, precision and reproducibility are satisfactory, the RSD were 3.55% (n = 5), 1.36% (n = 5), 4.79% (n = 5). The average recoveries were 94.41%, RSD was 2.33% (n = 5).
Extraction process in the study, the use of ethanol solution as extraction agent, a safe, non-toxic, non-hazardous solvent residues, etc., in line with the health food and Pharmaceuticals production requirements. Bunge leaves flavonoids in content inspection targets, through the single-factor experiments and orthogonal experiment analysis of variance, considering the cost of extraction and production of the extraction conditions were optimized to determine the optimal extraction process for A_2B_3C_1D_2, that is, ethanol concentration of 70%, extraction temperature of 70°C, extraction time of 1h, the solid-liquid ratio of 1:20, flavonoids extraction rate of 4.87%.
In this study, macroporous resin leaf flavonoids of Bunge for preconcentration, separation, operational, and simple equipment, the quality of stability, good economic returns and so on. Macroporous resin in accordance with the adsorption mechanism, the study of macroporous resin itself from the physical, chemical characteristics and the nature of the adsorption media to start using static adsorption and desorption experiments on the macroporous resin filter. Then, through the single-factor experiments and orthogonal experiment analysis of variance identified Bunge Purification leaf flavonoids of the best operating conditions. Of static adsorption and desorption, AB-8 macroporous resin showed the best performance and good adsorption and desorption results, the static saturated adsorption capacity of 12.15mg /g, desorption rate of 95.88%. The experimental results show that liquid concentration and the impact on the rate of adsorption column effect, and the effect of elution by ethanol concentration, elution speed, as well as to determine the amount of eluting agent. The optimum adsorption conditions were as follows: speed of the column 2BV / h, concentration of liquid 0.370mg/mL, the best elution conditions A_1B_2C_3, that is, 75% ethanol concentration, elution flow rate of 2BV / h, washing volume from 4.5BV, elution rate of 88.09%.
Bunge study found that ethanol extract of leaf flavonoids have strong antioxidant capacity. DPPH free radical application of clear ethanol extract revealed the price of Bunge leaf flavonoids antioxidant capacity than the water extract of leaf flavonoids Bunge. 70% ethanol extract of the IC50 for refined flavonoids 0.026mg/mL, water purification flavonoid extract the IC50 for the 0.058 mg / mL. The results showed that 70% ethanol extract flavonoids refined to remove free radicals DPPH·was significantly stronger than the water extract flavonoids refined. 0.477mg/mL concentration, its clearly more than half the rate of its 70% ethanol extract flavonoids refined to reach 84.57%, water extract of 70.33 %for refined flavonoids.
Bunge study found that ethanol extract of leaf flavonoids have strong antioxidant capacity. Application to remove free radicals DPPH·Bunge revealed the water extract of leaf flavonoids antioxidant capacity than the ethanol extract of leaf flavonoids Bunge. Crude water extract of the IC_(50) for liquid 0.06mg/mL, 70% ethanol extract of the IC50 for coarse liquid 0.08 mg/mL. The results showed that the crude water extract liquid clearance ability of free radical DPPH·in 70% ethanol crude extract liquid. 0.8mg/mL concentration when more than half of its clearance rate, liquid water extract crude reached 84.57%, 70% ethanol extract of 70.33% for crude liquid
1.本试验采用紫外可见分光光度法和高效液相色谱法对杜梨叶黄酮作了定性分析。建立了紫外可见分光光度法对杜梨叶总黄酮的测定方法。通过对检测波长的选择、标准品的选择、预处理方法、浓度与吸光度相关性的研究,确定了最佳检测波长为入=420nm,以芦丁为标准品,在0.06-0.54mg/mL范围内,其浓度与吸光度呈良好的线性关系,回归方程为A=1.31667C-0.01678,相关系数达r~2=0.99440,稳定性、精密度和重现性均理想,其RSD分别为3.55%(n=5)、1.36%(n=5)、4.79%(n=5)。平均回收率分别为94.41%,RSD为2.33%(n=5)。
2.在提取工艺研究中,采用乙醇水溶液为浸提剂,以杜梨叶总黄酮含量为考察指标,通过单因素实验、正交实验以及方差分析,对提取条件进行了优化,确定了最佳提取工艺为A_2B_3C_1D_2.即乙醇浓度为70%、浸提温度为70℃、浸提时间为1h,固液比为1:20,黄酮提取率为4.87%。
3.本试验采用大孔树脂对杜梨叶黄酮进行富集分离。根据大孔树脂的吸附机理,运用静态吸附与解吸实验对大孔树脂进行筛选。然后,通过单因素实验、正交实验以及方差分析确定了杜梨叶黄酮分离纯化的最佳操作条件。静态吸附与解吸中,AB-8大孔树脂表现出最好的吸附性能与良好的解吸效果,其饱和静态吸附量为12.15mg/g、解吸率为95.88%。实验表明,料液浓度和上柱速度明显影响吸附效果,而洗脱效果由乙醇浓度、洗脱速度以及洗脱剂用量来决定。实验确定的最佳吸附条件为:上柱速度为2.0 BV/h,料液浓度为0.370mg/mL,最佳洗脱条件为A_1B_2C_3,即乙醇浓度为75%,洗脱流速为2BV/h,洗脱体积为4.5BV,洗脱率为88.09%。
4.研究发现乙醇提取杜梨叶黄酮类化合物具有较强的抗氧化能力。应用清除DPPH·自由基揭示了水提取杜梨叶黄酮类化合物的抗氧化能力大于乙醇提取杜梨叶黄酮类化合物。水提取粗液的IC_(50)为0.06mg/mL,70%乙醇提取粗液的IC_(50)为0.08 mg/mL。结果表明,水提取粗液清除DPPH·自由基能力强于70%乙醇提取粗液。浓度在0.8mg/mL时,其均超过其半数清除率,水提取粗液达到84.57%,70%乙醇提取粗液为70.33%。
Flavonoids are widely distributed in nature, mainly in plant leaves and fruits. Flavonoids with anti-drug role in cerebral ischemia, the role of anti-myocardial ischemia, antiarrhythmic effects, analgesic effects, liver Hugan effect on the protective effect of peptic ulcer, anti-virus, anti-tumor effect. In addition, there are a large number of studies show that flavonoids are the active ingredients of natural products antihypertensive, hypolipidemic, inhibiting platelet aggregation and other pharmacological effects on flavonoid natural products active ingredients there are a variety of biological activity. To this end, by Governments and the pharmaceutical and food industry attaches great importance and concern. The flavonoids extract active ingredients is its reseach and application of key technologies and stages. So far, Bunge leaves flavonoids on resource extraction and purification process has not been reported. Bunge said medicinal Bunge, chief of pulmonary function for the convergence Shibuya intestines,cough, commonly used in the treatment of Chronic Cough,Disrrhea, Dysentery such as. For the development and utilization of the resources, the study of its detection, extraction and purification carried out a preliminary exploration, the establishment of a set of economic, security, application of the Bunge leaf flavonoids extraction, separation and detection methods.
In this study, UV-Vis spectrophotometry and HPLC of Bunge made a qualitative analysis of leaf flavonoids. Detection wavelength of choice, the choice of standard, pre-treatment method, concentration and absorbance of the relevant studies to determine the best detection wavelength of income = 420nm, for the standard to rutin in 0.06～0.54mg/ml the scope of its concentration and absorbance was a good linear relationship between the regression equation was A = 1.31667C-0.01678, correlation coefficient of r~2 = 0.99440, stability, precision and reproducibility are satisfactory, the RSD were 3.55% (n = 5), 1.36% (n = 5), 4.79% (n = 5). The average recoveries were 94.41%, RSD was 2.33% (n = 5).
Extraction process in the study, the use of ethanol solution as extraction agent, a safe, non-toxic, non-hazardous solvent residues, etc., in line with the health food and Pharmaceuticals production requirements. Bunge leaves flavonoids in content inspection targets, through the single-factor experiments and orthogonal experiment analysis of variance, considering the cost of extraction and production of the extraction conditions were optimized to determine the optimal extraction process for A_2B_3C_1D_2, that is, ethanol concentration of 70%, extraction temperature of 70°C, extraction time of 1h, the solid-liquid ratio of 1:20, flavonoids extraction rate of 4.87%.
In this study, macroporous resin leaf flavonoids of Bunge for preconcentration, separation, operational, and simple equipment, the quality of stability, good economic returns and so on. Macroporous resin in accordance with the adsorption mechanism, the study of macroporous resin itself from the physical, chemical characteristics and the nature of the adsorption media to start using static adsorption and desorption experiments on the macroporous resin filter. Then, through the single-factor experiments and orthogonal experiment analysis of variance identified Bunge Purification leaf flavonoids of the best operating conditions. Of static adsorption and desorption, AB-8 macroporous resin showed the best performance and good adsorption and desorption results, the static saturated adsorption capacity of 12.15mg /g, desorption rate of 95.88%. The experimental results show that liquid concentration and the impact on the rate of adsorption column effect, and the effect of elution by ethanol concentration, elution speed, as well as to determine the amount of eluting agent. The optimum adsorption conditions were as follows: speed of the column 2BV / h, concentration of liquid 0.370mg/mL, the best elution conditions A_1B_2C_3, that is, 75% ethanol concentration, elution flow rate of 2BV / h, washing volume from 4.5BV, elution rate of 88.09%.
Bunge study found that ethanol extract of leaf flavonoids have strong antioxidant capacity. DPPH free radical application of clear ethanol extract revealed the price of Bunge leaf flavonoids antioxidant capacity than the water extract of leaf flavonoids Bunge. 70% ethanol extract of the IC50 for refined flavonoids 0.026mg/mL, water purification flavonoid extract the IC50 for the 0.058 mg / mL. The results showed that 70% ethanol extract flavonoids refined to remove free radicals DPPH·was significantly stronger than the water extract flavonoids refined. 0.477mg/mL concentration, its clearly more than half the rate of its 70% ethanol extract flavonoids refined to reach 84.57%, water extract of 70.33 %for refined flavonoids.
Bunge study found that ethanol extract of leaf flavonoids have strong antioxidant capacity. Application to remove free radicals DPPH·Bunge revealed the water extract of leaf flavonoids antioxidant capacity than the ethanol extract of leaf flavonoids Bunge. Crude water extract of the IC_(50) for liquid 0.06mg/mL, 70% ethanol extract of the IC50 for coarse liquid 0.08 mg/mL. The results showed that the crude water extract liquid clearance ability of free radical DPPH·in 70% ethanol crude extract liquid. 0.8mg/mL concentration when more than half of its clearance rate, liquid water extract crude reached 84.57%, 70% ethanol extract of 70.33% for crude liquid
引文
[1]http://baike.baidu.com/view/172334.htm
[2]刘孟军主编.中国野生果树[M].北京:中国农业出版社,1999.216-217.
[3]赵小亮,白红进,汪河滨,周中波,罗锋.杜梨果实多糖提取方法及含量测定研究[J].西北农业学报,2007,16(4):279-281.
[4]赵小亮,邓芳,王金磊,龙叔梅.杜梨叶片中氨基酸及矿质元素含量的测定[J].塔里木大学学报,2007,19(2):57-59.
[5]塞冬,李树青,敖拉哈,等.黄芪、天麻延缓衰老的实验及临床研究[J].锦州医学院学报,1994,15(1):9.
[6]赵艳,庄晓燕,李淑云,等.枸杞中微量元素的测定[J].微量元素与健康研究,2007,24(2):21-22.
[7]李时珍.本草纲目(校实本·下册)[M].北京:人民卫生出社,2003:1766-1767.
[8]张鞍灵,高锦明,王妹清.黄酮类化合物的分布及开发利用[J],西北林学院学报,2000,15(1):69-74.
[9]Anna Wach,Krystyna Pyrzynska,Magdalena Biesaga.Quercetin content in some food and herbal samples[J].Food Chemisty,2007,100(2):699-704.
[10]徐任生,叶阳,赵维民.天然产物化学(第二版)[M].北京:科学出版社,2004.
[11]欧阳平,张高勇,康保安.类黄酮提取的基本原理、影响因素和传统方法[J].中国食品添加剂,2003,15:54-57.
[12]高金燕,曾小花,王毅辉.2004.从西芹中提取黄酮类物质的初步研究[J].中国食品添加剂.(3):19-21.
[13]丁利君,吴振辉,蔡创海,等.菊花中黄酮类物质提取方法的研究[J].食品工业科技,2002(2):20-22
[14]曹永刚.芦丁提取工艺的研究[J].医药工业杂志,1993,:251-53.
[15]郭孝武.超声波技术在提取中药苷类成分中的应用[J].中草药,1990.(27):235.
[16]武正才,梁晓原,丁永胜.超声波提取灯盏花中总黄酮的研究[J].云南中医中药杂志,2004,25(3):35-36
[17]王延峰,李延清.超声法提取银杏叶黄酮的研究[J].食品科学,2002,23(8):166-167.
[18]张永明,张倩,杨青等.现代技术与传统方法萃取古尼虫草总黄酮的比较[J].食品研究与开发,2007,28(1):136-140.
[19]卢汝梅,蒋先明.酸催化半干微波法水解蚕蛹蛋白地研究[J].天然产物研究与开发,1996,10(2):71-74.
[20]张力学,丁金昌,谷亨杰.微波技术在有机合成中的应用[J].合成化学,1996,4(14):23-29
[21]GeogerW.Kabalka,Arkin R.Mereddy.Micorwave-assisted synthesis of function alized flavones and chromones[J].Tetrahedron Letters.2005,46:6315-6317.
[22]郭锦棠,李伶,杨俊红,等.微波提取山植中总黄酮的正交实验研究[J].中国药学杂志,2005,40(2):122-115.
[23]刘峙嵘等.2005,微波萃取银杏叶黄酮类化合物.东华理工学院学报.28(2):153-155.
[24]WangH,LiuJJ.2004.Study on extraction Proeess of flavonoids from Ginkgo biloba leaves by enzymic treatment.J Chem lnd Forest Prod.38(1):14-16.
[25]YangJX,CaiJP,Zhu L.2005.Utilization of cellulose in extraction of Chinese medicine components.J Chin Med Mater.28(1):64-67.
[26]王晖,刘佳佳.银杏黄酮的酶法提取工艺研究[J].中药材,2003,12:887-888。
[27]王晖,刘佳佳.银杏黄酮的酶法提取工艺研究[J].林产化工通讯,2004,38(1):14-16
[28]lee ML Analytical Supercritlcal Fluid Chrcrmtography and Extraction Charmtographic Conefernees[J].IncprovoVD,,1990:437.
[29]韩玉谦,隋晓.银可叶活性成分萃取工艺的研究[J].精细化工,2000,17(9):505-506,514.
[30]曾琦华,黄少列.银杏叶超临界CO2提取条件的研究[J].广东药学院学报,2000,16(4):304-306.
[31]曾琦华,黄少烈.夹带剂在银杏叶超临界CO2萃取中的应用研究[J].广东药学,2001,11(6):7-10.
[32]吴向阳,仰榴青,陈钧,等.超临界CO2脱除银杏叶中银杏酸的研究[J].食品科学,2003,24(10):85-88
[33]潘见,张文成,谢慧明.银杏菇内酷超临界CO2萃取结晶前处理工艺研究[J].食品科学,2003,24(12):86-89
[34]潘见,张文成,谢慧明.超临界CO2萃取结晶银杏内酷关键工艺参数研究[J].食品科学,2004,25(3):132-134
[35]Chiu,Kou-Lung:Cheng,Ya-Chuan:Chen,Jun-Hao.Supereritieal fluids Extraction of Ginkgo ginkgolides and flavonoids[J].The Journal of Supercritical Fluids,2002,24(1):77-87
[36]彭洪,郭振德.银杏叶超临界CO2萃取产物的成分研究[J].中草药,1996,27(A09):45-46
[37]刘志敏,赵锁奇,王仁安,等.黄酮类化合物的超临界流体色谱分离闭[J].分析化学,1997,3:272-275
[38]Tao Bo,Ke An Li,Huwei Liu.Fast determination of flvaonoids in olycyrrhizae ardix by Capillary zone electroresis[J].Analytiea Acta,22,458:345-354
[39]揭金阶.2006.罗布麻叶总黄酮类化合物的提取及含量测定.中国医院药学杂志.26(12)1490-1491.
[40]杨荣平,王宾豪,张小梅等.2007.紫外分光光度法测定补骨脂中总黄酮含量.中国药业.16(1):22-23.
[41]穆赫塔尔·伊米尔艾山,摩尔班·吾斯曼,,自草中总黄酮含量测定[J].中草药,2001,32(10):900.
[42]彭友元,丁祥欢,叶建农.2003.沙棘黄酮口服液中芦丁和5-羟色胺的毛细管区带电泳电化学检测.分析测试学报.22(1):76-78.
[43]陈在新,李俊凯,陈义全.2003.分光光度法测定板栗黄酮含量.果树学报.20(2):149-151.
[44]谭平华,林金清等.2003.双水相萃取技术研究进展及应用.化工.10(1):19-23
[45]张廷之,侯镜德,徐秀珠.反相高效液相色谱法测定毛竹叶中总黄酮[J],理化检验一化学分册,2001,37(3):附10.
[46]胡芳弟,封士兰等,2003.HPLC法测定黄茂中黄酮类成分和黄茂甲昔的含量.分析测试技术与仪器.9(3):173-277.
[47]刘斌等.2005.HPLC法测定苦参汤有效部位中4种黄酮类成分含量.北京中医药大学学 报,28(6):59-62.
[48]孟祥颖,李向高,刘大有.西洋参不同部位中黄酮的含量测定[J].长春中医学院.2002,18:45-46.
[49]Potapovich A.I,Kostyuk V.A.2003.Comparative Study of Antioxidant Properties and Cytoprotective Activity of Flavonoids.Biochemistry(Moscow).68(5):514-519
[50]Whysner,J.Safety Assessment of Butylated Hydroxyanisole and Butylated Hydroxytolueneas as Antioxidant Food Additives,Fd.Chem.Toxic.,1994,32:215-222.
[51]HO Jin Heo,Chang Yong Lee.Epicatechin and Catehcin in Cocoa Inhibit Amyloid,Protein Induced Apoptosis[J].J.Agric.Food Chem.2005,53:1445-1448
[52]Paola Montoro,Alessandra Braca,Cosimo Pizza,et al.Structre-antioxidant activity relationships of flavonoids isolated from different plant species[J].Food Chemistry,2005,92:349-355.
[53]胡春,丁宵霖.黄酮类化合物在不同氧化体系中的抗氧化作用研究[J].食品与发酵工业,1996,(3):46-53
[54]Gao.Getal.Antioxidant and prooxidant behavior of flavonoids structure-activiy relationships[J].Frer Redical Biology and Medicine.1997,22(5):749-760.
[55]Fedosova N.F,Alisievich S.V,Lyadov K.V,Romanova E.P,Rudko I.A.,Kubatiev A.A.2004.Mechanisms Underlying Diquertin-Mediated Regulation of Neutrophil Function in Patients With Non-Insulin-Dependent Diabetes Mellitus.Bulletin of Experimental Biology and Medicine.137(2) 143-146.
[56]Balasubashini M.S,Rukkumani R,Menon V.P.2003.Protective effects of ferulic acid on hyperlipidemic diabetic rats.Acta Diabetologica.40(3):118-122.
[57]俞灵莺,李向荣.2000.植物黄酮类抗糖尿病及其并发症的研究进展.国外医学卫生学分册.27(6):333-334.
[58]姚思宇,等.黄酮降血脂功效的动物试验[J].中国热带医学,2006,6(3):531-532.
[59]谢作权,张庆华.2007.银杏叶提取物对神经及心血管系统的调节机制.国外医学.药学分册.1:56-59.
[60]唐传核,彭志英.2002.类黄酮的最新研究进展(Ⅱ)-生理功能.中国食品添加.(1):5-11.
[61]叶怀义,龚赋岚等.2004.甘草黄酮抗衰老作用的研究.哈尔滨商业大学学报.20(1):93-95.
[62]HagiwaraA,Miyashita K.etal.2001.Pronounced inhibition by antural anthocyanin,purple corn color,of-2amino2-2methyl-6-phenylimidazo[4,5-b]pyridine(phIP)-associated colorectal Carcinogenesis in maleF344 rats pretreated with 1,2-dimethyl-hydrazine.Cancer Lett.(171):17-25.
[63]Kazuyasu Samejima,ctal.A Strong Antimutagen against Dietary Carcinogen,Trp-p-2,in Peppermint,Sage,and Thyme[J].Agric.Food Chem.1995,43(1):410-414.
[64]白凤梅,蔡同一.类黄酮生物活性及其机理的研究进展[J].食品科学.1999,(8):11-13
[65]高中洪,黄开勋,徐辉碧.黄芩中黄酮类生物活性的研究进展[J].中国药学杂志.1998,(12):705-707.
[66]赵雪梅,朱大元,叶兴乾等.柑桔属中类黄酮的研究进展[J].天然产物研究与开发,2002,14(1):89-91.
[67]Yoshikazu Sakagami,Hiroko Murata,Tsutomu Nakanishi,etal.lnhibitory effect of plant Extracts on production of verotoxni by enterohemorrhagic Escherichia coliO157:H7[J].J.Health Sci,2001,47(4):473-477.
[68]Peterson Jetal.Flavoncids:dierary occurrence and biochemical activity[J].Nutr.Res.1998,18(12):1995-2018
[69]焦晶晶,张英.植物类黄酮作为护肤因子在化妆品领域的研究进展[J].精细化工,2004,24(1):98-102.
[70]武雪芬,景小琪等.金银花叶药用成分的提取及抑菌试验[J].中药材,2001,13(3):43.
[71]肖崇厚主编.中药化学[M].上海:上海科学技术出版社,2002,276-277.
[72]中国科学院上海药物研究所植物化学研究室.黄酮体化合物鉴定手册[M].北京:科学技术出版社,1981,14.
[73]于世林编,高效液相色谱方法及应用[M],化学工业出版社,2002.1,2-10.
[74]陈战国,吕九如,中草药有效成分定量分析方法研究新进展[J],陕西师范大学学报,自然科学版.2002,30(4).-116-121.
[75]汪正范,色谱定性与定量[M],化学工业出版社,2000:11-122.
[76]陈国珍,分光光度法[M],科学出版社,1961.6,1-100.
[77]吕丽爽,黄健花,周庆,等.芦蒿中黄酮类化合物提取工艺研究[J].食品工业科技,2002,23(9):48.
[78]王冬梅.2002.大孔吸附树脂在药用植物有效成分分离中的应用.西北林学院报.17(1):60.
[79]陈会良,顾有方,王月雷.中草药化学成分与抗氧化活性的研究进展[J].中国中医药科技,2006,13(1):63-64.
[80]李秀信,王荣花,李萍.紫苏黄酮对食用油脂抗氧化作用的研究[J].中国粮油学报,2007(2):63-64.
[81]陈丛瑾,黄克瀛,李德良,等.香椿叶总黄酮的超声波辅助提取及其清除DPPH自由基能力的研究[J].食品与机械,2007(1):76-77.
[82]王丽玲,焦必林,曾凡坤.保健功能因子-生物类黄酮[J].粮食与油脂,2004(9):18-19.
[83]王婷,张金超,陈瑶,等.6种淫羊藿黄酮抗氧化和抗肿瘤活性的比较[J].中国中药杂志,2007(8):715-717.
[84]许钢.红薯中黄酮提取及抗氧化研究[J].食品与生物技术学报,2007,Vol.26,No.4:22-27
[2]刘孟军主编.中国野生果树[M].北京:中国农业出版社,1999.216-217.
[3]赵小亮,白红进,汪河滨,周中波,罗锋.杜梨果实多糖提取方法及含量测定研究[J].西北农业学报,2007,16(4):279-281.
[4]赵小亮,邓芳,王金磊,龙叔梅.杜梨叶片中氨基酸及矿质元素含量的测定[J].塔里木大学学报,2007,19(2):57-59.
[5]塞冬,李树青,敖拉哈,等.黄芪、天麻延缓衰老的实验及临床研究[J].锦州医学院学报,1994,15(1):9.
[6]赵艳,庄晓燕,李淑云,等.枸杞中微量元素的测定[J].微量元素与健康研究,2007,24(2):21-22.
[7]李时珍.本草纲目(校实本·下册)[M].北京:人民卫生出社,2003:1766-1767.
[8]张鞍灵,高锦明,王妹清.黄酮类化合物的分布及开发利用[J],西北林学院学报,2000,15(1):69-74.
[9]Anna Wach,Krystyna Pyrzynska,Magdalena Biesaga.Quercetin content in some food and herbal samples[J].Food Chemisty,2007,100(2):699-704.
[10]徐任生,叶阳,赵维民.天然产物化学(第二版)[M].北京:科学出版社,2004.
[11]欧阳平,张高勇,康保安.类黄酮提取的基本原理、影响因素和传统方法[J].中国食品添加剂,2003,15:54-57.
[12]高金燕,曾小花,王毅辉.2004.从西芹中提取黄酮类物质的初步研究[J].中国食品添加剂.(3):19-21.
[13]丁利君,吴振辉,蔡创海,等.菊花中黄酮类物质提取方法的研究[J].食品工业科技,2002(2):20-22
[14]曹永刚.芦丁提取工艺的研究[J].医药工业杂志,1993,:251-53.
[15]郭孝武.超声波技术在提取中药苷类成分中的应用[J].中草药,1990.(27):235.
[16]武正才,梁晓原,丁永胜.超声波提取灯盏花中总黄酮的研究[J].云南中医中药杂志,2004,25(3):35-36
[17]王延峰,李延清.超声法提取银杏叶黄酮的研究[J].食品科学,2002,23(8):166-167.
[18]张永明,张倩,杨青等.现代技术与传统方法萃取古尼虫草总黄酮的比较[J].食品研究与开发,2007,28(1):136-140.
[19]卢汝梅,蒋先明.酸催化半干微波法水解蚕蛹蛋白地研究[J].天然产物研究与开发,1996,10(2):71-74.
[20]张力学,丁金昌,谷亨杰.微波技术在有机合成中的应用[J].合成化学,1996,4(14):23-29
[21]GeogerW.Kabalka,Arkin R.Mereddy.Micorwave-assisted synthesis of function alized flavones and chromones[J].Tetrahedron Letters.2005,46:6315-6317.
[22]郭锦棠,李伶,杨俊红,等.微波提取山植中总黄酮的正交实验研究[J].中国药学杂志,2005,40(2):122-115.
[23]刘峙嵘等.2005,微波萃取银杏叶黄酮类化合物.东华理工学院学报.28(2):153-155.
[24]WangH,LiuJJ.2004.Study on extraction Proeess of flavonoids from Ginkgo biloba leaves by enzymic treatment.J Chem lnd Forest Prod.38(1):14-16.
[25]YangJX,CaiJP,Zhu L.2005.Utilization of cellulose in extraction of Chinese medicine components.J Chin Med Mater.28(1):64-67.
[26]王晖,刘佳佳.银杏黄酮的酶法提取工艺研究[J].中药材,2003,12:887-888。
[27]王晖,刘佳佳.银杏黄酮的酶法提取工艺研究[J].林产化工通讯,2004,38(1):14-16
[28]lee ML Analytical Supercritlcal Fluid Chrcrmtography and Extraction Charmtographic Conefernees[J].IncprovoVD,,1990:437.
[29]韩玉谦,隋晓.银可叶活性成分萃取工艺的研究[J].精细化工,2000,17(9):505-506,514.
[30]曾琦华,黄少列.银杏叶超临界CO2提取条件的研究[J].广东药学院学报,2000,16(4):304-306.
[31]曾琦华,黄少烈.夹带剂在银杏叶超临界CO2萃取中的应用研究[J].广东药学,2001,11(6):7-10.
[32]吴向阳,仰榴青,陈钧,等.超临界CO2脱除银杏叶中银杏酸的研究[J].食品科学,2003,24(10):85-88
[33]潘见,张文成,谢慧明.银杏菇内酷超临界CO2萃取结晶前处理工艺研究[J].食品科学,2003,24(12):86-89
[34]潘见,张文成,谢慧明.超临界CO2萃取结晶银杏内酷关键工艺参数研究[J].食品科学,2004,25(3):132-134
[35]Chiu,Kou-Lung:Cheng,Ya-Chuan:Chen,Jun-Hao.Supereritieal fluids Extraction of Ginkgo ginkgolides and flavonoids[J].The Journal of Supercritical Fluids,2002,24(1):77-87
[36]彭洪,郭振德.银杏叶超临界CO2萃取产物的成分研究[J].中草药,1996,27(A09):45-46
[37]刘志敏,赵锁奇,王仁安,等.黄酮类化合物的超临界流体色谱分离闭[J].分析化学,1997,3:272-275
[38]Tao Bo,Ke An Li,Huwei Liu.Fast determination of flvaonoids in olycyrrhizae ardix by Capillary zone electroresis[J].Analytiea Acta,22,458:345-354
[39]揭金阶.2006.罗布麻叶总黄酮类化合物的提取及含量测定.中国医院药学杂志.26(12)1490-1491.
[40]杨荣平,王宾豪,张小梅等.2007.紫外分光光度法测定补骨脂中总黄酮含量.中国药业.16(1):22-23.
[41]穆赫塔尔·伊米尔艾山,摩尔班·吾斯曼,,自草中总黄酮含量测定[J].中草药,2001,32(10):900.
[42]彭友元,丁祥欢,叶建农.2003.沙棘黄酮口服液中芦丁和5-羟色胺的毛细管区带电泳电化学检测.分析测试学报.22(1):76-78.
[43]陈在新,李俊凯,陈义全.2003.分光光度法测定板栗黄酮含量.果树学报.20(2):149-151.
[44]谭平华,林金清等.2003.双水相萃取技术研究进展及应用.化工.10(1):19-23
[45]张廷之,侯镜德,徐秀珠.反相高效液相色谱法测定毛竹叶中总黄酮[J],理化检验一化学分册,2001,37(3):附10.
[46]胡芳弟,封士兰等,2003.HPLC法测定黄茂中黄酮类成分和黄茂甲昔的含量.分析测试技术与仪器.9(3):173-277.
[47]刘斌等.2005.HPLC法测定苦参汤有效部位中4种黄酮类成分含量.北京中医药大学学 报,28(6):59-62.
[48]孟祥颖,李向高,刘大有.西洋参不同部位中黄酮的含量测定[J].长春中医学院.2002,18:45-46.
[49]Potapovich A.I,Kostyuk V.A.2003.Comparative Study of Antioxidant Properties and Cytoprotective Activity of Flavonoids.Biochemistry(Moscow).68(5):514-519
[50]Whysner,J.Safety Assessment of Butylated Hydroxyanisole and Butylated Hydroxytolueneas as Antioxidant Food Additives,Fd.Chem.Toxic.,1994,32:215-222.
[51]HO Jin Heo,Chang Yong Lee.Epicatechin and Catehcin in Cocoa Inhibit Amyloid,Protein Induced Apoptosis[J].J.Agric.Food Chem.2005,53:1445-1448
[52]Paola Montoro,Alessandra Braca,Cosimo Pizza,et al.Structre-antioxidant activity relationships of flavonoids isolated from different plant species[J].Food Chemistry,2005,92:349-355.
[53]胡春,丁宵霖.黄酮类化合物在不同氧化体系中的抗氧化作用研究[J].食品与发酵工业,1996,(3):46-53
[54]Gao.Getal.Antioxidant and prooxidant behavior of flavonoids structure-activiy relationships[J].Frer Redical Biology and Medicine.1997,22(5):749-760.
[55]Fedosova N.F,Alisievich S.V,Lyadov K.V,Romanova E.P,Rudko I.A.,Kubatiev A.A.2004.Mechanisms Underlying Diquertin-Mediated Regulation of Neutrophil Function in Patients With Non-Insulin-Dependent Diabetes Mellitus.Bulletin of Experimental Biology and Medicine.137(2) 143-146.
[56]Balasubashini M.S,Rukkumani R,Menon V.P.2003.Protective effects of ferulic acid on hyperlipidemic diabetic rats.Acta Diabetologica.40(3):118-122.
[57]俞灵莺,李向荣.2000.植物黄酮类抗糖尿病及其并发症的研究进展.国外医学卫生学分册.27(6):333-334.
[58]姚思宇,等.黄酮降血脂功效的动物试验[J].中国热带医学,2006,6(3):531-532.
[59]谢作权,张庆华.2007.银杏叶提取物对神经及心血管系统的调节机制.国外医学.药学分册.1:56-59.
[60]唐传核,彭志英.2002.类黄酮的最新研究进展(Ⅱ)-生理功能.中国食品添加.(1):5-11.
[61]叶怀义,龚赋岚等.2004.甘草黄酮抗衰老作用的研究.哈尔滨商业大学学报.20(1):93-95.
[62]HagiwaraA,Miyashita K.etal.2001.Pronounced inhibition by antural anthocyanin,purple corn color,of-2amino2-2methyl-6-phenylimidazo[4,5-b]pyridine(phIP)-associated colorectal Carcinogenesis in maleF344 rats pretreated with 1,2-dimethyl-hydrazine.Cancer Lett.(171):17-25.
[63]Kazuyasu Samejima,ctal.A Strong Antimutagen against Dietary Carcinogen,Trp-p-2,in Peppermint,Sage,and Thyme[J].Agric.Food Chem.1995,43(1):410-414.
[64]白凤梅,蔡同一.类黄酮生物活性及其机理的研究进展[J].食品科学.1999,(8):11-13
[65]高中洪,黄开勋,徐辉碧.黄芩中黄酮类生物活性的研究进展[J].中国药学杂志.1998,(12):705-707.
[66]赵雪梅,朱大元,叶兴乾等.柑桔属中类黄酮的研究进展[J].天然产物研究与开发,2002,14(1):89-91.
[67]Yoshikazu Sakagami,Hiroko Murata,Tsutomu Nakanishi,etal.lnhibitory effect of plant Extracts on production of verotoxni by enterohemorrhagic Escherichia coliO157:H7[J].J.Health Sci,2001,47(4):473-477.
[68]Peterson Jetal.Flavoncids:dierary occurrence and biochemical activity[J].Nutr.Res.1998,18(12):1995-2018
[69]焦晶晶,张英.植物类黄酮作为护肤因子在化妆品领域的研究进展[J].精细化工,2004,24(1):98-102.
[70]武雪芬,景小琪等.金银花叶药用成分的提取及抑菌试验[J].中药材,2001,13(3):43.
[71]肖崇厚主编.中药化学[M].上海:上海科学技术出版社,2002,276-277.
[72]中国科学院上海药物研究所植物化学研究室.黄酮体化合物鉴定手册[M].北京:科学技术出版社,1981,14.
[73]于世林编,高效液相色谱方法及应用[M],化学工业出版社,2002.1,2-10.
[74]陈战国,吕九如,中草药有效成分定量分析方法研究新进展[J],陕西师范大学学报,自然科学版.2002,30(4).-116-121.
[75]汪正范,色谱定性与定量[M],化学工业出版社,2000:11-122.
[76]陈国珍,分光光度法[M],科学出版社,1961.6,1-100.
[77]吕丽爽,黄健花,周庆,等.芦蒿中黄酮类化合物提取工艺研究[J].食品工业科技,2002,23(9):48.
[78]王冬梅.2002.大孔吸附树脂在药用植物有效成分分离中的应用.西北林学院报.17(1):60.
[79]陈会良,顾有方,王月雷.中草药化学成分与抗氧化活性的研究进展[J].中国中医药科技,2006,13(1):63-64.
[80]李秀信,王荣花,李萍.紫苏黄酮对食用油脂抗氧化作用的研究[J].中国粮油学报,2007(2):63-64.
[81]陈丛瑾,黄克瀛,李德良,等.香椿叶总黄酮的超声波辅助提取及其清除DPPH自由基能力的研究[J].食品与机械,2007(1):76-77.
[82]王丽玲,焦必林,曾凡坤.保健功能因子-生物类黄酮[J].粮食与油脂,2004(9):18-19.
[83]王婷,张金超,陈瑶,等.6种淫羊藿黄酮抗氧化和抗肿瘤活性的比较[J].中国中药杂志,2007(8):715-717.
[84]许钢.红薯中黄酮提取及抗氧化研究[J].食品与生物技术学报,2007,Vol.26,No.4:22-27