黄连叶片与须根中小檗碱的提取纯化研究
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摘要
以采自镇平县农科所五年生味连上的叶片与须根为试验材料,以紫外分光光度法测得的总生物碱含量作为考察指标,通过单因素试验和正交试验设计对影响乙醇回流提取法和纤维素酶解提取法提取效率的因素及水平进行了研究,同时考察了D101大孔树脂吸附法富集纯化黄连总生物碱的工艺条件。本试验旨在在试验室水平范围内找出从黄连叶片、须根中提取小檗碱的最佳条件及使用大孔树脂纯化黄连生物碱的工艺条件,为开发黄连叶片与须根、提高黄连综合利用水平提供参考。取得主要结论如下:
1.传统上利用很少但生物量比例很大的黄连叶片及须根中含有相当含量的生物碱,在最佳提取条件下黄连叶片和须根中总生物碱得率分别为5.48%和7.1%,其中小檗碱含量占黄连总生物碱的一半左右,因此其具有潜在的开发价值。
2.通过单因素试验和正交试验设计确定了黄连叶片的最佳回流提取条件为加入10X体积的70%乙醇回流提取3次,每次1.5h,生物碱得率为5.48%;而须根的最佳回流提取条件为加入12X体积的80%乙醇,回流提取3次,每次2.0h,生物碱得率为7.1%。
3.通过单因素试验和正交试验设计确定了黄连叶片的最佳纤维素酶酶解条件为加入样品质量2.0%的纤维素酶粉和10X体积的pH=2.6的缓冲液后在50℃水浴中反应150min;须根的最佳纤维素酶解条件为加入样品质量1.5%的纤维素酶粉和10X体积的pH=2.6的缓冲液后在55℃水浴中反应120min。从中得到的黄连生物碱得率分别为4.83%和6.01%。
4.无论用哪种方法提取,叶片与须根比较生物碱含量都较低,说明该种黄连的叶片中所含生物碱比须根中生物碱少。两种提取方法比较,无论材料是叶片还是须根都表现出乙醇回流法提取更彻底,生物碱得率更高,但是该法耗时长,且耗用了更多的试剂;而酶法省时、消耗少,生物碱得率相对较低,但这并不能完全说明酶解法不适应于黄连叶片及须根中生物碱的提取。
5.考察了非极性大孔树脂D101富集纯化黄连生物总碱的条件。试验结果表明,用D101大孔树脂纯化黄连生物碱的最佳条件为:0.0555g生物碱/g湿树脂的上样量,上样流速为1BV/h时具有最好的吸附效果,而用pH>12的30%乙醇以2BV/h的流速冲洗柱子时,能将更多的生物碱冲洗下来,在该条件下,黄连总生物碱回收率达到83.72%,结合重结晶法可以得到更高纯度的盐酸小檗碱。
Single factor experiments and orthogonal tests were used to secelect the optimum conditions of ethanol extraction and enzyme extraction from the leaves and fibrous roots of Coptis chinensis which had lived for five years.The test materials were from Zhenping agricultural science research institute and the extraction amount of alkaloid which detected by UV spectrophotometry was used as a marker.At the same time ,the process conditions of enrichment of coptis alkaloids by D101macroporous resin were studied.The purpose of the study was to find the best conditions of extraction and purification of coptis alkaloids from the leaves and fibrous roots of Coptis chinensis in laboratory and the data should be guidance for enhancing the level of comprehensive utilization of Coptis chinensis. Obtains the main conclusion to be as follows:
1.The leaves and fibrous roots of Coptis chinensis which were unutilized have large proportion of biomass.And the yield rate of alkaloids from the leaves and fibrous roots of Coptis chinensis were 5.48% and 7.1% under optimum extraction conditions.More than 50% of the alkaloid was berberine,so the leaves and fibrous roots of Coptis chinensis should be exploited.
2.The best extraction conditions of reflux extraction for leaves was: the solid-liquid ratio is 10,the ethanol concentration is 70%, the extraction time was 1.5 hours, and the extraction times were 3 times;and the fibrous roots’was: the solid-liquid ratio is 12,the ethanol concentration is 80%, the extraction time was 2 hours, and the extraction times were 3 times.The yield rates of coptis alkaloids were respectively 5.48% and 7.1%.
3.The best extraction conditions of enzyme extraction for leaves was:enzyme amount is 2.0%,the pH of buffer is 2.6 ,enzymolysis time was 150min in water bath which temperature was 50℃;and the fibrous roots’was: enzyme amount is 1.5%,the pH of buffer is 2.6 ,enzymolysis time was 120min in 55℃water bath . The yield rates of coptis alkaloids were respectively 4.83% and 6.01%.
4.Compared with leaves ,the fibrous roots of Coptis chinensis has more alkaloid no matter which extraction method was used.Ethanol reflux extraction has higher extraction efficiency but need more time and solvent.Enzymatic hydrolysis used less time but has lower extraction efficiency. Ethanol reflux extraction was better than enzymatic hydrolysis in extracting alkaloid from the leaves and fibrous roots of Coptis chinensis can not concluded by these experiments at all.
5.The enrichment conditions of coptis alkaloids by D101 macroporous resin were studied,the result showed that the best accumulation conditions were:the adsorption valus is 0.055526g/g(alkaloid/macroporous resin), sample flow rate is 1BV/h,the eluent is pH>12 30% ethanol with a 2BV/h elution speed.83.72% alkaloid was recovered under these conditions.Highly purified berberine should obtained by recrystallizing after enrichment by macroporous resin.
1.传统上利用很少但生物量比例很大的黄连叶片及须根中含有相当含量的生物碱,在最佳提取条件下黄连叶片和须根中总生物碱得率分别为5.48%和7.1%,其中小檗碱含量占黄连总生物碱的一半左右,因此其具有潜在的开发价值。
2.通过单因素试验和正交试验设计确定了黄连叶片的最佳回流提取条件为加入10X体积的70%乙醇回流提取3次,每次1.5h,生物碱得率为5.48%;而须根的最佳回流提取条件为加入12X体积的80%乙醇,回流提取3次,每次2.0h,生物碱得率为7.1%。
3.通过单因素试验和正交试验设计确定了黄连叶片的最佳纤维素酶酶解条件为加入样品质量2.0%的纤维素酶粉和10X体积的pH=2.6的缓冲液后在50℃水浴中反应150min;须根的最佳纤维素酶解条件为加入样品质量1.5%的纤维素酶粉和10X体积的pH=2.6的缓冲液后在55℃水浴中反应120min。从中得到的黄连生物碱得率分别为4.83%和6.01%。
4.无论用哪种方法提取,叶片与须根比较生物碱含量都较低,说明该种黄连的叶片中所含生物碱比须根中生物碱少。两种提取方法比较,无论材料是叶片还是须根都表现出乙醇回流法提取更彻底,生物碱得率更高,但是该法耗时长,且耗用了更多的试剂;而酶法省时、消耗少,生物碱得率相对较低,但这并不能完全说明酶解法不适应于黄连叶片及须根中生物碱的提取。
5.考察了非极性大孔树脂D101富集纯化黄连生物总碱的条件。试验结果表明,用D101大孔树脂纯化黄连生物碱的最佳条件为:0.0555g生物碱/g湿树脂的上样量,上样流速为1BV/h时具有最好的吸附效果,而用pH>12的30%乙醇以2BV/h的流速冲洗柱子时,能将更多的生物碱冲洗下来,在该条件下,黄连总生物碱回收率达到83.72%,结合重结晶法可以得到更高纯度的盐酸小檗碱。
Single factor experiments and orthogonal tests were used to secelect the optimum conditions of ethanol extraction and enzyme extraction from the leaves and fibrous roots of Coptis chinensis which had lived for five years.The test materials were from Zhenping agricultural science research institute and the extraction amount of alkaloid which detected by UV spectrophotometry was used as a marker.At the same time ,the process conditions of enrichment of coptis alkaloids by D101macroporous resin were studied.The purpose of the study was to find the best conditions of extraction and purification of coptis alkaloids from the leaves and fibrous roots of Coptis chinensis in laboratory and the data should be guidance for enhancing the level of comprehensive utilization of Coptis chinensis. Obtains the main conclusion to be as follows:
1.The leaves and fibrous roots of Coptis chinensis which were unutilized have large proportion of biomass.And the yield rate of alkaloids from the leaves and fibrous roots of Coptis chinensis were 5.48% and 7.1% under optimum extraction conditions.More than 50% of the alkaloid was berberine,so the leaves and fibrous roots of Coptis chinensis should be exploited.
2.The best extraction conditions of reflux extraction for leaves was: the solid-liquid ratio is 10,the ethanol concentration is 70%, the extraction time was 1.5 hours, and the extraction times were 3 times;and the fibrous roots’was: the solid-liquid ratio is 12,the ethanol concentration is 80%, the extraction time was 2 hours, and the extraction times were 3 times.The yield rates of coptis alkaloids were respectively 5.48% and 7.1%.
3.The best extraction conditions of enzyme extraction for leaves was:enzyme amount is 2.0%,the pH of buffer is 2.6 ,enzymolysis time was 150min in water bath which temperature was 50℃;and the fibrous roots’was: enzyme amount is 1.5%,the pH of buffer is 2.6 ,enzymolysis time was 120min in 55℃water bath . The yield rates of coptis alkaloids were respectively 4.83% and 6.01%.
4.Compared with leaves ,the fibrous roots of Coptis chinensis has more alkaloid no matter which extraction method was used.Ethanol reflux extraction has higher extraction efficiency but need more time and solvent.Enzymatic hydrolysis used less time but has lower extraction efficiency. Ethanol reflux extraction was better than enzymatic hydrolysis in extracting alkaloid from the leaves and fibrous roots of Coptis chinensis can not concluded by these experiments at all.
5.The enrichment conditions of coptis alkaloids by D101 macroporous resin were studied,the result showed that the best accumulation conditions were:the adsorption valus is 0.055526g/g(alkaloid/macroporous resin), sample flow rate is 1BV/h,the eluent is pH>12 30% ethanol with a 2BV/h elution speed.83.72% alkaloid was recovered under these conditions.Highly purified berberine should obtained by recrystallizing after enrichment by macroporous resin.
引文
艾志兵,何国厚,刘勇,王云甫,张晓东,李承晏,魏国耀. 2005.小檗碱对家兔颈动脉粥样硬化干预的实验研究.中药与神经杂质,22(2):138-140
陈爱萍,徐榕青,郑安,叶钦勇. 2005.高效毛细管区带电泳法在优选黄连提取工艺上的运用.福建医药杂质,27(1):97～99
陈馥馨,高晓山.1997.含黄连方剂及黄连配伍的文献统计.中成药, 19(8):40～41
陈小全,翟虎,邵辉莹. 2007.黄连素提取方法的改进.西南民族大学学报自然科学版,33(5):1121～1123
崔学军. 2006.黄连及其有效成分的药理研究进展.中国药师,9(5):469～470
付晓春,王敏伟. 2001.黄连解毒汤的抗血栓作用研究.沈阳药科大学学报,18 (6) : 425～427
傅立国.1992.中国植物红皮书-稀有濒危植物(第1册).北京:科学出版社:522
郭锦堂,李雄勇,杨俊红.2003.微波-索氏联合工艺提取盐酸小檗碱.化工进展,22(12):1338～1341
郭孝武.2002.超声提取与常规提取对部分中药碱类成分提出率的比较.世界科学技术—中药现代化,4(5):59～61
韩芳化,黄筑荣.2001.黄连素治疗室性早搏2000例.中国现代医学杂志,11(5):101～105
何贤辉,曾耀英,徐丽慧.2002.黄连素对T淋巴细胞活化和增殖的抑制作用.中国病理生理杂志,18(10):1183～1186
贺善安.1998.中国珍稀植物.上海:上海科学出版社:133
侯世祥,朱浩,孙毅毅,毛声俊. 2000.影响大孔吸附树脂吸附纯化黄连提取液因素的初步考察.中国中药杂志,25(11):666～668
胡冬华,袁绪富.2006.黄连素的提取及分子活性部位研究.长春中医学院学报,22(1):67
胡惠民,黄乾明. 1991.从黄连叶中提取黄连素.四川农业科技,(3):67
黄志强,周民杰. 2006.黄连中小檗碱的超声波提取工艺.化学工程师,(8):54～56
冀满丰. 2006.黄连中的盐酸小檗碱的提取、分离及分离物的含量测定研究.国际医药卫生导报,12(24):75～77
蒋德勋,蒋孝国.1994.南郑县黄连根茎干物质积累及小檗碱含量的变化规律.西北农业学报,3(3):90～93
可维,马春辉,季宇彬. 2008.不同产地川黄柏HPLC指纹图谱的研究.上海中医药大学学报,22(1):62～65
兰进,杨世林,郑玉权,邵家斌,李勇. 2001.黄连的研究进展.中草药,32(12):1139～1141
李平,果茵茵. 2008.凹凸棒对盐酸小檗碱吸附作用考察.中国药师,11(1):6～8
李英明,王立群,邓芬,孙群,孙素琴.2004.黄连药材不同部位、不同生长年限和不同海拔高度红外光谱法整体分析与评价.中国医学科学院学报,26(6):614～617
梁柏林,周民杰. 2006.酶法提取小檗碱工艺研究.应用化工,35(5):373～374,378
林菁. 1996.小檗碱对K562细胞生长的抑制作用.福建医学院学报,30(4)∶309～311
林志华,宋敏,李苗. 2004.反相高效液相色谱法测定黄连及制剂中小檗碱的含量.中国医院药学杂志,24(12):738～739
刘春海,刘西京,杨永华. 2006.黄连及其复方提取条件的优选.时珍国医国药,17(11):2249～2250
刘鸿雁,何津春,杨永建.2006.小檗碱降血脂作用的研究进展.卫生职业教育,24:144～146
刘新迎,周联,王培训.2006.小檗碱抗肿瘤机制研究进展.中医药学刊,24(8):1499～1501
刘映,李志忠. 2001.大孔吸附树脂预分离后测定黄连及左金丸含量.中国药师,4(2):113～114
刘永静,刘巧,于丽丽,陈丹. 2008.超临界流体萃取技术在中药分离分析中的应用进展.福建中医学院学报,18(2):60～62
刘志峰,李桂生,傅风华,刘珂. 2002.8种中药注射剂体外抗内毒素作用的观察.中草药,33(1):58～59
罗音久,曾中良,古淑英. 2002.黄连须中盐酸小檗碱的提取工艺研究.四川畜牧兽医学院院报,16(3):17～20
马桔云,赵晶岩,姜颖,于喜水.2000.纤维素酶在黄连提取工艺中的应用.中草药,31(2):103～104
毛理纳,杨玉华,胡新辉.2009.头孢他啶联合黄连的抗菌作用.中药材,32(6):865～866
濮社班. 1998.江苏省引种黄连的生长状况及生物碱积累.中国中药杂志,23(11):659～660
沈建斌.2006.黄连资源的利用与研究进展.现代医药卫生,22(7):1038～1039
史以菊,吴俊芳,刘天培. 1995.小檗碱对实验性脑缺血的改善作用.南京医科大学学报,15(2):256～259
唐睿,黄庆华,严志红. 2006.ICP-AES测定中药黄连中微量元素.光谱试验室,23(3):500～502
王传芬.2007.黄连中小檗碱的提取工艺.华西药学杂志,22(4):476～477
王立群,李英明,张丽萍,校合香,周群,孙素琴. 2006.生态技术栽培黄连的红外指纹图谱分析与表征.光谱学与光谱分析,26(6):1061～1066
王文雅,陈克敏,关永源.1999.盐酸小檗碱对毒蕈碱型受体的作用.药学学报,34 (4):260～263
韦藤幼,赵钟兴,童张法. 2006.解吸-内部沸腾两步法提取黄连小檗碱的工艺及机理.过程工程学报,6(3):380～383
魏金莹.2005.纤维素酶在中药材提取中的应用.[硕士学位论文].天津:天津大学
魏英勤,房海燕,袁久荣,容蓉. 2003.影响大孔树脂吸附黄连提取液的因素.中国药业,12(2):62～63
吴绍敏.2004.正交试验法优选湖北利川道地黄连栽培条件.中国民族民间医药杂志,(66):47～48
吴珍.1996.双波长薄层扫描法测定黄连丸盐酸小檗碱的含量.时珍国药研究,7(5): 282
武佳,谭桂莲,杨红.2007.黄连中盐酸小檗碱提取工艺探究.时珍国医国药,18(2):437～438
席国萍,何照范.2004.黄连中小檗碱的提取工艺.山地农业生物学报,23(6):502～506
席国萍.2005.黄连中小檗碱的提取、分离、纯化研究.[硕士学位论文].贵阳:贵州大学
谢培凤,周晖,高彦彬.2005.小檗碱治疗2型糖尿病40例疗效观察.中国临床保健杂志,8 (5) :402～403
徐锦堂.2004.黄连生态栽培技术研究与推广应用及前景展望.中国医学科学院学报,26(6):601～603
徐静华,于庆海,蔡爽.2003.黄连解毒汤对小鼠急性脑缺血、缺氧的影响.沈阳药科大学学报,2(3):132～134
徐文芬,黄勇其,何顺志,张天伦.1996.贵州省小檗属药用植物根中小檗碱含量的测定.中国中药杂志,21(3): 144
许沛虎,高媛,张雪琼,肖刚,刘莹,郭锋锋.2008.正交实验法优选黄连生物碱提取工艺.医药导报,27(4):446～448
许沛虎,高媛,张雪琼,许可,张良. 2009.大孔树脂纯化黄连总生物碱的研究.中成药,31(3):390～393
杨莉,刘亚娜.2001.酶法在中药提取制备中的应用.中药材,24(1):72～73
杨勇,叶小利,李学刚.2007.4种黄连生物碱的抑菌作用.时珍国医国药,18(12):3013～3014
应懿,何志红,周世文,汤建林,黄永平,杨雪.2007.正交试验优化提取黄连中小檗碱型生物碱.中国药房,18(9):670～672
于淑萍,叶健.2007.微波提取小檗碱中提取温度的影响.皮革化工,24(4):37～39
于永福.1999.中国野生植物保护工作的里程碑.植物杂志,5(151):3
余园媛,王伯出,彭亮,王晶波,曾晨.2006.黄连的药理研究进展.重庆大学学报(自然科学版),29(2):107～111
翟显友,李隆云,钟国跃,陈仕江.2009.黄连栽培研究进展.重庆中草药研究,6(59):23～25
张红梅,程雪梅,王长虹,王峥涛.2008.黄连中生物碱提取工艺的正交试验研究.中国医药工业杂志,39(8):588～590
张金渝,王波,金航,潘俊,张智慧,赵振玲,杨维泽,李晚谊.2008.云南黄连不同部位有效成分及生物量的比较研究.中国中药杂志,33(3):311～313
张来新,杨琼,李小卫.2003.黄连中提取黄连素.贵州化工,28(2):30～32
张乐佳,夏新华.2001.黄连提取工艺的研究.中成药,23(6):398～400
张莉,张小平.2006.国产黄连属植物的研究现状.安徽师范大学学报(自然科学版),29(4):368～371
张希恩,杨磊.2003.小檗碱的临床新用途.人民军医,46(12) :730-732.
张晓丹,任宏敏,刘琳,渠永清.2008.小檗碱对DHF大鼠血流动力学和心肌细胞内钙离子浓度的影响.中国中药杂志,33(7):818～821
赵广琦,杜增平.2002.不同海拔高度与不同株龄和月份对黄连质量和产量的影响.中草药,33(12):1119～1121
赵国庆,吴素体,王颖,谢进,肖小河,贺承山.2001.不同品种和产地黄连的总生物碱含量测定.时珍国医国药,12(11):974
郑洪艳,徐为人.2004.小檗碱药理作用研究进展.中草药,35(6):708～711
周吉银,周世文.2007.小檗碱降糖调脂作用机制的研究进展.解放军药学学报,23(3):201～204
朱翠霞,张洪利,康大为.2009.不同炮制方法对黄连抗菌活性的影响.中药材,32(6):865～866
Brusq J M,Ancellin N , Grondin P , Guillard R,Martin S,Saintillan Y,Issandou M.2006.Inhibition of lipid synthesis through activation of AMP kinase : an additional mechanism for the hypolipidemic effects of berberine.J Lipid Res,47 (6) :1281～1288
Choi B H,Ahn I S,Kim Y H,et al. 2006.Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3 L1 adipocyte.Exp Mol Med,38 (6) :599～605
Hao Y,Qiu Q Y, Wu J , Wang J J.1999.Effect of berberine on lymphocyte endothelium adhesion and adhesion molecules expression.Chinese Journal Of Immunology,15 (11) : 523～525
Huang C,Zhang Y,Gong Z,Sheng X,Li Z,Zhang W,Qin Y.2006.Berberine inhibits 3T3-L1 adipocyte differentiation through the PPARgamma pathway .Biochem Biophys Res Commun,348 (2) :571～578
Jiang J Y,Geng D S,Tursonjan T,Liu F.1998.Anti-inflammatory effects and mechanism of berberine.Chinese Pharmacological Bulletin,14 (5) : 434～437
Ko W H, Yao X Q, Lau C W, et al.2000.Vasorelaxant and antiproliferative effects of berberine . Eur J Pharmacol,399 (2～3):187～196
Kobayashiy,Yoshinorik,Yoshineri,et al. 1995.Inhibitors of DNA topoisomeraseⅠandⅡisolated from the Coptis Rhizomes.Planta Med, 61(5):414～417
Kupeli E,Kosar M,Yesilada E,Husnu K,Baser C.2002.A comparative study on the anti-inflammatory ammatory ,antinociceptive and antipyretic effects of isoquino line alkaloids from the roots of Tu rkish berberis species.Life Sciences, 72 (6) : 645～657
Lee Y S,Kim W S,Kim K H,Yoon M J,Cho H J,Shen Y,Ye J M,Lee C H,Oh K W,Kim C T,Cordula H B,Alison G,Edward W,Kraegen,David E,Kim J B.2006.Berberine,a natural plant product,activates AMP-activated protein kinase with beneficial metabolic effects in diabetic and insulin-resistant states. Diabetes,55(8) :2256～2264
LüY N ,Qiu Q Y. 2000.Effects of berberine on the levels of DTH and several important cytokines in the mice induced by DNFB.Chinese Journal Of Immunology,16 (3) :139～141
Yin J , Hu R M , Tang J F, et al. 2001.Gluco se-lowering effect of berberine in vitro .A cta Univ Med Second Shang hai,21 (5) : 425～427
陈爱萍,徐榕青,郑安,叶钦勇. 2005.高效毛细管区带电泳法在优选黄连提取工艺上的运用.福建医药杂质,27(1):97～99
陈馥馨,高晓山.1997.含黄连方剂及黄连配伍的文献统计.中成药, 19(8):40～41
陈小全,翟虎,邵辉莹. 2007.黄连素提取方法的改进.西南民族大学学报自然科学版,33(5):1121～1123
崔学军. 2006.黄连及其有效成分的药理研究进展.中国药师,9(5):469～470
付晓春,王敏伟. 2001.黄连解毒汤的抗血栓作用研究.沈阳药科大学学报,18 (6) : 425～427
傅立国.1992.中国植物红皮书-稀有濒危植物(第1册).北京:科学出版社:522
郭锦堂,李雄勇,杨俊红.2003.微波-索氏联合工艺提取盐酸小檗碱.化工进展,22(12):1338～1341
郭孝武.2002.超声提取与常规提取对部分中药碱类成分提出率的比较.世界科学技术—中药现代化,4(5):59～61
韩芳化,黄筑荣.2001.黄连素治疗室性早搏2000例.中国现代医学杂志,11(5):101～105
何贤辉,曾耀英,徐丽慧.2002.黄连素对T淋巴细胞活化和增殖的抑制作用.中国病理生理杂志,18(10):1183～1186
贺善安.1998.中国珍稀植物.上海:上海科学出版社:133
侯世祥,朱浩,孙毅毅,毛声俊. 2000.影响大孔吸附树脂吸附纯化黄连提取液因素的初步考察.中国中药杂志,25(11):666～668
胡冬华,袁绪富.2006.黄连素的提取及分子活性部位研究.长春中医学院学报,22(1):67
胡惠民,黄乾明. 1991.从黄连叶中提取黄连素.四川农业科技,(3):67
黄志强,周民杰. 2006.黄连中小檗碱的超声波提取工艺.化学工程师,(8):54～56
冀满丰. 2006.黄连中的盐酸小檗碱的提取、分离及分离物的含量测定研究.国际医药卫生导报,12(24):75～77
蒋德勋,蒋孝国.1994.南郑县黄连根茎干物质积累及小檗碱含量的变化规律.西北农业学报,3(3):90～93
可维,马春辉,季宇彬. 2008.不同产地川黄柏HPLC指纹图谱的研究.上海中医药大学学报,22(1):62～65
兰进,杨世林,郑玉权,邵家斌,李勇. 2001.黄连的研究进展.中草药,32(12):1139～1141
李平,果茵茵. 2008.凹凸棒对盐酸小檗碱吸附作用考察.中国药师,11(1):6～8
李英明,王立群,邓芬,孙群,孙素琴.2004.黄连药材不同部位、不同生长年限和不同海拔高度红外光谱法整体分析与评价.中国医学科学院学报,26(6):614～617
梁柏林,周民杰. 2006.酶法提取小檗碱工艺研究.应用化工,35(5):373～374,378
林菁. 1996.小檗碱对K562细胞生长的抑制作用.福建医学院学报,30(4)∶309～311
林志华,宋敏,李苗. 2004.反相高效液相色谱法测定黄连及制剂中小檗碱的含量.中国医院药学杂志,24(12):738～739
刘春海,刘西京,杨永华. 2006.黄连及其复方提取条件的优选.时珍国医国药,17(11):2249～2250
刘鸿雁,何津春,杨永建.2006.小檗碱降血脂作用的研究进展.卫生职业教育,24:144～146
刘新迎,周联,王培训.2006.小檗碱抗肿瘤机制研究进展.中医药学刊,24(8):1499～1501
刘映,李志忠. 2001.大孔吸附树脂预分离后测定黄连及左金丸含量.中国药师,4(2):113～114
刘永静,刘巧,于丽丽,陈丹. 2008.超临界流体萃取技术在中药分离分析中的应用进展.福建中医学院学报,18(2):60～62
刘志峰,李桂生,傅风华,刘珂. 2002.8种中药注射剂体外抗内毒素作用的观察.中草药,33(1):58～59
罗音久,曾中良,古淑英. 2002.黄连须中盐酸小檗碱的提取工艺研究.四川畜牧兽医学院院报,16(3):17～20
马桔云,赵晶岩,姜颖,于喜水.2000.纤维素酶在黄连提取工艺中的应用.中草药,31(2):103～104
毛理纳,杨玉华,胡新辉.2009.头孢他啶联合黄连的抗菌作用.中药材,32(6):865～866
濮社班. 1998.江苏省引种黄连的生长状况及生物碱积累.中国中药杂志,23(11):659～660
沈建斌.2006.黄连资源的利用与研究进展.现代医药卫生,22(7):1038～1039
史以菊,吴俊芳,刘天培. 1995.小檗碱对实验性脑缺血的改善作用.南京医科大学学报,15(2):256～259
唐睿,黄庆华,严志红. 2006.ICP-AES测定中药黄连中微量元素.光谱试验室,23(3):500～502
王传芬.2007.黄连中小檗碱的提取工艺.华西药学杂志,22(4):476～477
王立群,李英明,张丽萍,校合香,周群,孙素琴. 2006.生态技术栽培黄连的红外指纹图谱分析与表征.光谱学与光谱分析,26(6):1061～1066
王文雅,陈克敏,关永源.1999.盐酸小檗碱对毒蕈碱型受体的作用.药学学报,34 (4):260～263
韦藤幼,赵钟兴,童张法. 2006.解吸-内部沸腾两步法提取黄连小檗碱的工艺及机理.过程工程学报,6(3):380～383
魏金莹.2005.纤维素酶在中药材提取中的应用.[硕士学位论文].天津:天津大学
魏英勤,房海燕,袁久荣,容蓉. 2003.影响大孔树脂吸附黄连提取液的因素.中国药业,12(2):62～63
吴绍敏.2004.正交试验法优选湖北利川道地黄连栽培条件.中国民族民间医药杂志,(66):47～48
吴珍.1996.双波长薄层扫描法测定黄连丸盐酸小檗碱的含量.时珍国药研究,7(5): 282
武佳,谭桂莲,杨红.2007.黄连中盐酸小檗碱提取工艺探究.时珍国医国药,18(2):437～438
席国萍,何照范.2004.黄连中小檗碱的提取工艺.山地农业生物学报,23(6):502～506
席国萍.2005.黄连中小檗碱的提取、分离、纯化研究.[硕士学位论文].贵阳:贵州大学
谢培凤,周晖,高彦彬.2005.小檗碱治疗2型糖尿病40例疗效观察.中国临床保健杂志,8 (5) :402～403
徐锦堂.2004.黄连生态栽培技术研究与推广应用及前景展望.中国医学科学院学报,26(6):601～603
徐静华,于庆海,蔡爽.2003.黄连解毒汤对小鼠急性脑缺血、缺氧的影响.沈阳药科大学学报,2(3):132～134
徐文芬,黄勇其,何顺志,张天伦.1996.贵州省小檗属药用植物根中小檗碱含量的测定.中国中药杂志,21(3): 144
许沛虎,高媛,张雪琼,肖刚,刘莹,郭锋锋.2008.正交实验法优选黄连生物碱提取工艺.医药导报,27(4):446～448
许沛虎,高媛,张雪琼,许可,张良. 2009.大孔树脂纯化黄连总生物碱的研究.中成药,31(3):390～393
杨莉,刘亚娜.2001.酶法在中药提取制备中的应用.中药材,24(1):72～73
杨勇,叶小利,李学刚.2007.4种黄连生物碱的抑菌作用.时珍国医国药,18(12):3013～3014
应懿,何志红,周世文,汤建林,黄永平,杨雪.2007.正交试验优化提取黄连中小檗碱型生物碱.中国药房,18(9):670～672
于淑萍,叶健.2007.微波提取小檗碱中提取温度的影响.皮革化工,24(4):37～39
于永福.1999.中国野生植物保护工作的里程碑.植物杂志,5(151):3
余园媛,王伯出,彭亮,王晶波,曾晨.2006.黄连的药理研究进展.重庆大学学报(自然科学版),29(2):107～111
翟显友,李隆云,钟国跃,陈仕江.2009.黄连栽培研究进展.重庆中草药研究,6(59):23～25
张红梅,程雪梅,王长虹,王峥涛.2008.黄连中生物碱提取工艺的正交试验研究.中国医药工业杂志,39(8):588～590
张金渝,王波,金航,潘俊,张智慧,赵振玲,杨维泽,李晚谊.2008.云南黄连不同部位有效成分及生物量的比较研究.中国中药杂志,33(3):311～313
张来新,杨琼,李小卫.2003.黄连中提取黄连素.贵州化工,28(2):30～32
张乐佳,夏新华.2001.黄连提取工艺的研究.中成药,23(6):398～400
张莉,张小平.2006.国产黄连属植物的研究现状.安徽师范大学学报(自然科学版),29(4):368～371
张希恩,杨磊.2003.小檗碱的临床新用途.人民军医,46(12) :730-732.
张晓丹,任宏敏,刘琳,渠永清.2008.小檗碱对DHF大鼠血流动力学和心肌细胞内钙离子浓度的影响.中国中药杂志,33(7):818～821
赵广琦,杜增平.2002.不同海拔高度与不同株龄和月份对黄连质量和产量的影响.中草药,33(12):1119～1121
赵国庆,吴素体,王颖,谢进,肖小河,贺承山.2001.不同品种和产地黄连的总生物碱含量测定.时珍国医国药,12(11):974
郑洪艳,徐为人.2004.小檗碱药理作用研究进展.中草药,35(6):708～711
周吉银,周世文.2007.小檗碱降糖调脂作用机制的研究进展.解放军药学学报,23(3):201～204
朱翠霞,张洪利,康大为.2009.不同炮制方法对黄连抗菌活性的影响.中药材,32(6):865～866
Brusq J M,Ancellin N , Grondin P , Guillard R,Martin S,Saintillan Y,Issandou M.2006.Inhibition of lipid synthesis through activation of AMP kinase : an additional mechanism for the hypolipidemic effects of berberine.J Lipid Res,47 (6) :1281～1288
Choi B H,Ahn I S,Kim Y H,et al. 2006.Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3 L1 adipocyte.Exp Mol Med,38 (6) :599～605
Hao Y,Qiu Q Y, Wu J , Wang J J.1999.Effect of berberine on lymphocyte endothelium adhesion and adhesion molecules expression.Chinese Journal Of Immunology,15 (11) : 523～525
Huang C,Zhang Y,Gong Z,Sheng X,Li Z,Zhang W,Qin Y.2006.Berberine inhibits 3T3-L1 adipocyte differentiation through the PPARgamma pathway .Biochem Biophys Res Commun,348 (2) :571～578
Jiang J Y,Geng D S,Tursonjan T,Liu F.1998.Anti-inflammatory effects and mechanism of berberine.Chinese Pharmacological Bulletin,14 (5) : 434～437
Ko W H, Yao X Q, Lau C W, et al.2000.Vasorelaxant and antiproliferative effects of berberine . Eur J Pharmacol,399 (2～3):187～196
Kobayashiy,Yoshinorik,Yoshineri,et al. 1995.Inhibitors of DNA topoisomeraseⅠandⅡisolated from the Coptis Rhizomes.Planta Med, 61(5):414～417
Kupeli E,Kosar M,Yesilada E,Husnu K,Baser C.2002.A comparative study on the anti-inflammatory ammatory ,antinociceptive and antipyretic effects of isoquino line alkaloids from the roots of Tu rkish berberis species.Life Sciences, 72 (6) : 645～657
Lee Y S,Kim W S,Kim K H,Yoon M J,Cho H J,Shen Y,Ye J M,Lee C H,Oh K W,Kim C T,Cordula H B,Alison G,Edward W,Kraegen,David E,Kim J B.2006.Berberine,a natural plant product,activates AMP-activated protein kinase with beneficial metabolic effects in diabetic and insulin-resistant states. Diabetes,55(8) :2256～2264
LüY N ,Qiu Q Y. 2000.Effects of berberine on the levels of DTH and several important cytokines in the mice induced by DNFB.Chinese Journal Of Immunology,16 (3) :139～141
Yin J , Hu R M , Tang J F, et al. 2001.Gluco se-lowering effect of berberine in vitro .A cta Univ Med Second Shang hai,21 (5) : 425～427