博落回属植物中苄基异喹啉类生物碱代谢组学研究
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摘要
苄基异喹啉类生物碱由于其显著药理活性备受全世界研究者的重视,这类生物碱在生物合成上都包含在具有同一起源的代谢网络中。本论文以富含苄基异喹啉类生物碱的博落回为研究对象,整合高通量分析技术和复杂网络研究方法对异喹啉类生物碱代谢进行了较为全面系统的研究。本文研究内容如下:
1.基于化学分类学,植物代谢组学和苄基异喹啉类生物碱的次生代谢的基础上提出了苄基异喹啉类生物碱代谢组学概念。详细地研究了苄基异喹啉类生物碱代谢网络的结构,构建了描述苄基异喹啉类生物碱代谢网络的加权邻接矩阵。最后讨论其在苄基异喹啉类生物碱资源开发、化学分类学和道地药材等研究领域的意义。
2.建立了同时分析博落回中血根碱、白屈菜红碱、原阿片碱、别隐品碱四种生物碱的HPLC/UV分析方法,考察了博落回中在代谢上存在转化关系的四中生物碱在生长期中的含量动态变化规律。
3.针对目前生物碱分析中流动相体系与液质联用兼容性差的问题,综合考察了全氟羧酸和羧酸添加剂对生物碱在不同填料色谱柱上分离及对其质谱响应的影响。得到了高碳量色谱柱与甲酸水流动相的合理组合,建立了在分离和质谱响应上均达到了较好的效果的HPLC/DAD/ESI/MS分析方法。以此方法分析了博落回根部样品,并结合苄基异喹啉类生物碱代谢途径知识对博落回中苄基异喹啉类生物碱进行了初步鉴定。
4.建立了用于初步鉴定生物碱的数据库,数据库包含了存在于代谢途径中所有生物碱的分子量、质谱特征、紫外特征和还原特征。运用前一章建立的HPLC/DAD/ESI/MS方法对博落回和小果博落回不同部位进行了分析,并将获得的数据与数据库中生物碱进行匹配初步鉴定了其中苄基异喹啉类中生物碱,结合苄基异喹啉类生物碱代谢网络对两种植物的苄基异喹啉类生物碱代谢特点进行比较。
Due to benzylisoquinoline alkaloids pharmacological activity, the researchers all over the world have paid much attention to it. All of these alkaloids are biosynthesized in the metabolism network which have the same origin. This paper uses Macleaya cordata (Willd.) R. Br., in which many benzylisoquinoline alkaloids have been isolated, as research object, and a more comprehensive and systematic research for benzylisoquinoline alkaloids metabolism is carried out by integrating high-throughput analysis technology and complex metabolic network research methods. The details are summarized as follows:
1. The concept of benzylisoquinoline alkaloids metabonomics was proposed on the basis of plant chemotaxonomy, plant metabonomics and benzylisoquinoline alkaloids secondary metabolism. Structure of the benzylisoquinoline alkaloids metabolic network was studied in detail, and the weighted adjacent matrix was used to describe metabolism network, also its significance in the field of benzylisoquinoline alkaloids resource development, chemotaxonomy and authentic medicinal herbs was discussed.
2. A high-performace liquid chromatographic method coupled with UV detector was developed to simultaneously analyze protopine, allocry-ptopine, sanguinarine and chelerythrine in M cordata (Willd.) R. Br., which have transforming relations. The proposed method was applied to determine dynamic changes of the four alkaloids in different parts of M cordata (Willd.) R. Br. during growing period.
3. Given the poor compatibility between the mobile phase of the present system using for alkaloids analyzing, and HPLC/MS, we investigated the effect of perfluorinated carboxylic acid and carboxylic acid additive on the alkaloids separation in different column and mass spectrometry response in detail and a method of HPLC/DAD/ESI/MS was developed which the separation and mass spectrometry response can reach good results at the same time by the rational combination of high carbon content column and mobile phase with carboxylic acid, Combined with the knowledge of benzylisoquinoline alkaloids metabolic pathway, benzylisoquinoline alkaloids in M cordata ( Willd. ) R. Br. were identificated preliminaryly.
4. The database which contains molecular weight, mass spectrometry features, UV features, as well as reduction characteristics of alkaloids in biosynthesis pathway, was established for data matching. We analyzed the different parts of M cordata (Willd.) R. Br. and M microcarpa (Maxim.) Fedde using the method proposed the previous chapter, identificated preliminaryly benzylisoquinoline alkaloids by matching the data obtained and compared the differences of benzylisoquinoline alkaloids metabolism between two species by combining with benzylisoquinoline alkaloids metabolic network.
1.基于化学分类学,植物代谢组学和苄基异喹啉类生物碱的次生代谢的基础上提出了苄基异喹啉类生物碱代谢组学概念。详细地研究了苄基异喹啉类生物碱代谢网络的结构,构建了描述苄基异喹啉类生物碱代谢网络的加权邻接矩阵。最后讨论其在苄基异喹啉类生物碱资源开发、化学分类学和道地药材等研究领域的意义。
2.建立了同时分析博落回中血根碱、白屈菜红碱、原阿片碱、别隐品碱四种生物碱的HPLC/UV分析方法,考察了博落回中在代谢上存在转化关系的四中生物碱在生长期中的含量动态变化规律。
3.针对目前生物碱分析中流动相体系与液质联用兼容性差的问题,综合考察了全氟羧酸和羧酸添加剂对生物碱在不同填料色谱柱上分离及对其质谱响应的影响。得到了高碳量色谱柱与甲酸水流动相的合理组合,建立了在分离和质谱响应上均达到了较好的效果的HPLC/DAD/ESI/MS分析方法。以此方法分析了博落回根部样品,并结合苄基异喹啉类生物碱代谢途径知识对博落回中苄基异喹啉类生物碱进行了初步鉴定。
4.建立了用于初步鉴定生物碱的数据库,数据库包含了存在于代谢途径中所有生物碱的分子量、质谱特征、紫外特征和还原特征。运用前一章建立的HPLC/DAD/ESI/MS方法对博落回和小果博落回不同部位进行了分析,并将获得的数据与数据库中生物碱进行匹配初步鉴定了其中苄基异喹啉类中生物碱,结合苄基异喹啉类生物碱代谢网络对两种植物的苄基异喹啉类生物碱代谢特点进行比较。
Due to benzylisoquinoline alkaloids pharmacological activity, the researchers all over the world have paid much attention to it. All of these alkaloids are biosynthesized in the metabolism network which have the same origin. This paper uses Macleaya cordata (Willd.) R. Br., in which many benzylisoquinoline alkaloids have been isolated, as research object, and a more comprehensive and systematic research for benzylisoquinoline alkaloids metabolism is carried out by integrating high-throughput analysis technology and complex metabolic network research methods. The details are summarized as follows:
1. The concept of benzylisoquinoline alkaloids metabonomics was proposed on the basis of plant chemotaxonomy, plant metabonomics and benzylisoquinoline alkaloids secondary metabolism. Structure of the benzylisoquinoline alkaloids metabolic network was studied in detail, and the weighted adjacent matrix was used to describe metabolism network, also its significance in the field of benzylisoquinoline alkaloids resource development, chemotaxonomy and authentic medicinal herbs was discussed.
2. A high-performace liquid chromatographic method coupled with UV detector was developed to simultaneously analyze protopine, allocry-ptopine, sanguinarine and chelerythrine in M cordata (Willd.) R. Br., which have transforming relations. The proposed method was applied to determine dynamic changes of the four alkaloids in different parts of M cordata (Willd.) R. Br. during growing period.
3. Given the poor compatibility between the mobile phase of the present system using for alkaloids analyzing, and HPLC/MS, we investigated the effect of perfluorinated carboxylic acid and carboxylic acid additive on the alkaloids separation in different column and mass spectrometry response in detail and a method of HPLC/DAD/ESI/MS was developed which the separation and mass spectrometry response can reach good results at the same time by the rational combination of high carbon content column and mobile phase with carboxylic acid, Combined with the knowledge of benzylisoquinoline alkaloids metabolic pathway, benzylisoquinoline alkaloids in M cordata ( Willd. ) R. Br. were identificated preliminaryly.
4. The database which contains molecular weight, mass spectrometry features, UV features, as well as reduction characteristics of alkaloids in biosynthesis pathway, was established for data matching. We analyzed the different parts of M cordata (Willd.) R. Br. and M microcarpa (Maxim.) Fedde using the method proposed the previous chapter, identificated preliminaryly benzylisoquinoline alkaloids by matching the data obtained and compared the differences of benzylisoquinoline alkaloids metabolism between two species by combining with benzylisoquinoline alkaloids metabolic network.
引文
[1]庄璇.罂粟科植物的分类、进化与分布[J].云南植物研究.1993,15(2):137-148.
[2]吴征镒,庄璇,苏志云.中国植物志.北京:科学出版社,1999.77-79.
[3]余震生.草药号筒树治疗脓性指头炎[J].江西医药.1966,6(7):371.
[4]叶廷珖.博落回中毒一例报告[J].中华内科杂志.1958,(6):617.
[5]杨军,王静,刘信顺.博落回的药效研究[J].中药材.1999,22(2):88.
[6]徐国钧,强王,余伯明.抗肿瘤中草药彩色图鉴.福州:福建科学技术出版社,1997.759
[7]熊晓荣.三味博落回煎剂治疗痔疮合并感染[J].云南中医杂志.1988,23(4):18.
[8]王静,杨军,李小山.博落回粉的急性和长期毒性试验[J].安徽医学.1998,19(3):75.
[9]罗裕民.博落回为主外治皮肤病[J].福建中医药.1987,18(1):封四.
[10]李逢春.肌注博落回引起心源性脑缺血综合征3例报告[J].新医药资料.1972,(1):49.
[11]解放军366医院妇产科.号筒杆治疗滴虫性阴道炎的疗效观察[J].新医学.1971,(6-7):45.
[12]樊淑莲,峰焦,园张.博落回总生物碱对动物移植性肿瘤的作用研究[J].陕西肿瘤医学.2000,8(3):174.
[13]陈晓庄.有毒中草药大辞典.天津:天津科技翻译出版社,1992.542
[14]郭小青.中药博落回的研究进展[J].兽药研发.2006.
[15]郭小青.博落回的药理作用及其在动物保健中的应用[J].中国动物保健.2005.(5):34-35.
[16]朱永言.博落回(号筒秆)[J].湖南农业.1998,6:8.
[17]张世宣.博落回植物及其生物碱杀蛆作用的初步研究报告[J].中草药通报.1957,3(2):58.
[18]张胜菊.博落回抽提物对黄守瓜、菜青虫的田间药效评价[J].华中农业大学学报.2003,22(5):450-451.
[19]袁涛忠.博落回杀灭蛆蝇效果的实验观察[J].中国寄生虫学与寄生虫病杂志.1999,17(2):128.
[20]陈昭存.博落回防治梨木虱和二斑叶螨药效试验[J].果农之友.2003,8:30.
[21]王欣.博落回中生物碱成分的研究Ⅱ[D]:西北农林科技大学;2005.
[22]庞发根.博落回(Macleaya C.(willd)R.Br.)抗癌活性成分的研究[D]:沈阳药科大学:2005.
[23]毛鹏.博落回生物碱成分及杀螨活性研究[D]:西北农林科技大学;2003.
[24]林启寿.中草药成分化学.北京:科学出版社;1977:721.
[25]胡之壁.博落回果实中有效成分的研究[J].药学学报.1979,14(9):535-540.
[26]巩忠福.博落回杀螨活性成分的研究[D]:西北农林科技大学;2002.
[27]Tani C.,Takao S.Studies on the alkaloids of fumariaceous plants.Ⅸ.Alkaloids of Bocconia frutescens.[J].Yakugaku Zasshi.1967,87(6):699-701.
[28]Tani C.,Takao N.Studies on the alkaloids of fumariaceous plants.Ⅴ.Alkaloids of Bocconia cordata(Macleya cordata)[J].Jpn J Pharmacol.1962,82:755-759.
[29]Takao N.,Yasumoto Y.,Iwasa K.Studies on the alkaloids of papaveraceous plants.ⅩⅤ.On the alkaloids of Bocconia cordata.2[J].Yakugaku Zasshi.1973,93(2):242-244.
[30]Boit H G.Ergebnisse der Alkaloid-chemie Bis-1960.Berlin:Akademie-verlag,1961
[31]周天达.苯并菲啶类生物碱在中国罂粟科和芸香科一些植物一些属中分布初探[J].植物学通报.1986,4(1-2):38-41.
[32]周荣汉.药用植物化学分类学.上海:上海科技出版社,1988.542.
[33]王宗训.从异喹啉生物碱试论双子叶一些科与罂粟科的一些属的亲缘关系[J].植物学报.1965,13(3):283-296.
[34]苏中武.被子植物的几个科的化学分类学(五)罂粟科的植物化学分类学[J].国外医学药学分册.1981,(02):65-70.
[35]黄松.罂粟科白屈菜族的化学成分及其植物化学分类依据的研究进展[J].湖南中医学导报.2002,8(10):582-584.
[36]陈孝泉.植物化学分类学.北京:高等教育出版社,1990.542.
[37]Liscombe D.K.,MacLeod B.P.,Loukanina N..Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms[J].Phytochemistry.2005,66(20):2501-2520.
[38]Park S.U.,Yu M.,Facchini P.J.Antisense RNA-mediated suppression of benzophenanthridine alkaloid biosynthesis in transgenic cell cultures of California poppy[J].Plant Physiol.2002,128(2):696-706.
[39]Kutchan T.M.,Dittrich H.Characterization and mechanism of the berberine bridge enzyme,a covalently flavinylated oxidase of benzophenanthridine alkaloid biosynthesis in plants[J].J Biol Chem.1995,270(41):24475-24481.
[40]Otani M.,Shitan N.,Sakai K..Characterization of vacuolar transport of the endogenous alkaloid berberine in Coptis japonica[J].Plant Physiol.2005,138(4):1939-1946.
[41]Sato F.,Hashimoto T.,Hachiya A..Metabolic engineering of plant alkaloid biosynthesis[J].Proc Natl Acad Sci U S A.2001,98(1):367-372.
[42]Morishige T.,Tsujita T.,Yamada Y..Molecular characterization of the S-adenosyl-L-methionine:3'-hydroxy-Nomethylcoclaurine 4'-O-methyltransferase involved in isoquinoline alkaloid biosynthesis in Coptis japonica[J].J Biol Chem.2000,275(30):23398-23405.
[43]Facchini J.P.,Luca V.D.Phloem-Specific Expression of Tyrosine/Dopa Decarboxylase Genes and the Biosynthesis of Isoquinoline Alkaloids in Opium Poppy[J].Plant Cell.1995,7(11):1811-1821.
[44]Alcantara J.,Bird D.A.,Franceschi V.R..Sanguinarine biosynthesis is associated with the endoplasmic reticulum in cultured opium poppy cells after elicitor treatment[J].Plant Physiol.2005,138(1):173-183.
[45]曾建国,张胜,候团章.不同生长期博落回果实中血根碱与白屈菜红碱的含量[J].中药材.1999,22(5):229
[46]Abizov E.A.,Tolkachev O.N.,Kopylova I.E..Distribution of the Sum of Sanguinarine and Chelerythrine in the Above-Ground Part of Macleaya Microcarpa[J].Pharmaceutical Chemistry Journal.2003,37(8):413-414.
[47]安彩贤.RP-HPLC法同时测定小果博落回中血根碱和白屈菜红碱的含量研究[J].中成药.2001,23(11):824-825.
[48]Nicholson J.K.,Lindon J.C.,Holmes E.'Metabonomics':understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J].Xenobiotica.1999,29(11):1181-1189.
[49]刘祥东.代谢物组学应用的领域之二-植物代谢物组学[J].中成药.2006,28(10):1515-1517.
[50]陈晓亚.植物次生代谢研究[J].世界科技研究与发展.2006,28(5):1-4.
[51]Fiehn O.Metabolic networks of Cucurbita maxima phloem[J].Phytochemistry.2003,62(6):875-886.
[52]Janet T,King R.D.,Thomas A.Application of metabolomics to plant genotype discrimination using statistics and machine learning[J].Bioinformatics.2002,18S241-248.
[53]Le G.Go,DuPont M.S.,Mellon F.A..Characterization and content of flavonoid glycosides in genetically modified tomato(Lycopersicon esculentum)fruits[J].J Agric Food Chem.2003,51(9):2438-2446.
[54]Choi H.K.,Choi Y.H.,Verberne M..Metabolic fingerprinting of wild type and transgenic tobacco plants by 1H NMR and multivariate analysis technique[J].Phytochemistry.2004,65(7):857-864.
[55]Walker T.S.,Bais H.P.,Halligan K.M..Metabolic profiling of root exudates of Arabidopsis thaliana[J].J Agric Food Chem.2003,51(9):2548-2554.
[56]Fiehn O.,Kopka J.,Dormann P..Metabolite profiling for plant functional genomics[J].Nat Biotechnol.2000,18(11):1157-1161.
[57]Choi Y.H.,Kim H.K.,Hazekamp A.Metabolomic Differentiation of Cannabis sativa Cultivars Using 1H NMR Spectroscopy and Principal Component Analysis[J].J Nat Prod.2004,67(6):953-957.
[58]Wang Y.,Tang H.,Nicholson J.K..Metabolomic strategy for the classification and quality control of phytomedicine:a case study of chamomile flower(Matricaria recutita L.)[J].Planta Med.2004,70(3):250-255.
[59]沈一丁.代谢组学促进中医药现代化[J].精细化工.2005,22(6):444-446.
[60]Narasimhan K,Basheer C,Bajic V.B..Enhancement of Plant-Microbe Interactions Using a Rhizosphere Metabolomics-Driven Approach and Its Application in the Removal of Polychlorinated Biphenyls[J].Plant Physiol.2003,132:146-153
[61]Verdonk J.C.,Verhoeven H.A..Regulation of floral scent production in petunia revealed by targeted metabolomics[J].Phytochemistry.2003,62(6):997-1008.
[62]Tolstikov V.V.,Fiehn O.Analysis of highly polar compounds of plant origin:combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry[J].Anal Biochem.2002,301(2):298-307.
[63]Popl M,Fahnrich J,Tatar V.Chromatographic analysis of alkaloids.In. NewYork:Marcel Dekker;1990
[64]Haberlein P,Tschiersch K.Effects of surface structure of reversed phases on.the separation of alkaloids by HPLC[J].Pharmazie.1994,49(4):769-775.
[65]McCalley V.D.Reversed-phases HPLC of basic samples-an update 《LC-GC Asia Pacific》[J].MCCALLEY D V.1999,2(03):37-44
[66]Theodoridis G.,Papadoyannis I.,Hermans-Lokkerbol A..A study of the behaviour of some new column materials in the chromatographic analysis of catharanthus alkaloids[J].Chromatographia.1997,45:52-58.
[67]Gennaro M C.Reversed-phase ion-pair and ion-interaction chromatography.NewYork:Marcel Dekker,1995.37.
[68]朱龙,封满良,万秀琴.吗啡和海洛因等生物碱的高效液相色谱化学光测定[J].分析化学.1996,24(11):1295-1297.
[69]Wu C.C,Young D.S,Liu L.K.Liquid chromatographic-electrospray mass spectrometric analysis of Coptidis Rhizoma[J].Journal of Chromatography A.1996,755(1):19-26.
[70]Ma H.W,Zeng A.P..Reconstruction of metabolic networks from genome data and analysis of their global structure for various organisms[J].Bioinformatics 2003,19(2):270-277
[71]Wagner A.,Fell D.A.The small world inside large metabolic net- works[J].Proc Biol Sci.2001,268(1478):1803-1810.
[72]孙之荣.后基因组信息学.北京:清华大学出版社,2002.45.
[73]王冰.基于复杂网络理论的代谢网络结构研究进展[J].中国生物工程杂志.2005,25(6):10-14.
[74]Ziegler J.,Voigtlander S.,Schmidt J..Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis[J].Plant J.2006,48(2):177-192.
[75]Zenk M.H.The formation of benzophenanthridine alkaloids[J].Pure and Applied Chemistry.1994,66(10):2023-2028.
[76]Weiss D.,Baumert A.,Vogel M..Sanguinarine reductase,a key enzyme of benzophenanthridine detoxification[J].Plant Cell Environ.2006,29(2):291-302.
[77]王树禾.图论.北京:科学出版社,2004.167.
[78]孙丽艳,魏雄辉,韦宇.博落回中总生物碱提取方法的研究[J].药物分析杂志.2003,(23):15-18.
[79]罗旭彪.生物碱的液相色谱分析方法学及其液相色谱保留机理的研究[D].长沙:湖南大学;2005.
[80]Suchomelova J.,Bochorakova H.,Paulova H..HPLC quantification of seven quaternary benzo[c]phenanthridine alkaloids in six species of the family Papaveraceae[J].J Pharm Biomed Anal.2007,44(1):283-287.
[2]吴征镒,庄璇,苏志云.中国植物志.北京:科学出版社,1999.77-79.
[3]余震生.草药号筒树治疗脓性指头炎[J].江西医药.1966,6(7):371.
[4]叶廷珖.博落回中毒一例报告[J].中华内科杂志.1958,(6):617.
[5]杨军,王静,刘信顺.博落回的药效研究[J].中药材.1999,22(2):88.
[6]徐国钧,强王,余伯明.抗肿瘤中草药彩色图鉴.福州:福建科学技术出版社,1997.759
[7]熊晓荣.三味博落回煎剂治疗痔疮合并感染[J].云南中医杂志.1988,23(4):18.
[8]王静,杨军,李小山.博落回粉的急性和长期毒性试验[J].安徽医学.1998,19(3):75.
[9]罗裕民.博落回为主外治皮肤病[J].福建中医药.1987,18(1):封四.
[10]李逢春.肌注博落回引起心源性脑缺血综合征3例报告[J].新医药资料.1972,(1):49.
[11]解放军366医院妇产科.号筒杆治疗滴虫性阴道炎的疗效观察[J].新医学.1971,(6-7):45.
[12]樊淑莲,峰焦,园张.博落回总生物碱对动物移植性肿瘤的作用研究[J].陕西肿瘤医学.2000,8(3):174.
[13]陈晓庄.有毒中草药大辞典.天津:天津科技翻译出版社,1992.542
[14]郭小青.中药博落回的研究进展[J].兽药研发.2006.
[15]郭小青.博落回的药理作用及其在动物保健中的应用[J].中国动物保健.2005.(5):34-35.
[16]朱永言.博落回(号筒秆)[J].湖南农业.1998,6:8.
[17]张世宣.博落回植物及其生物碱杀蛆作用的初步研究报告[J].中草药通报.1957,3(2):58.
[18]张胜菊.博落回抽提物对黄守瓜、菜青虫的田间药效评价[J].华中农业大学学报.2003,22(5):450-451.
[19]袁涛忠.博落回杀灭蛆蝇效果的实验观察[J].中国寄生虫学与寄生虫病杂志.1999,17(2):128.
[20]陈昭存.博落回防治梨木虱和二斑叶螨药效试验[J].果农之友.2003,8:30.
[21]王欣.博落回中生物碱成分的研究Ⅱ[D]:西北农林科技大学;2005.
[22]庞发根.博落回(Macleaya C.(willd)R.Br.)抗癌活性成分的研究[D]:沈阳药科大学:2005.
[23]毛鹏.博落回生物碱成分及杀螨活性研究[D]:西北农林科技大学;2003.
[24]林启寿.中草药成分化学.北京:科学出版社;1977:721.
[25]胡之壁.博落回果实中有效成分的研究[J].药学学报.1979,14(9):535-540.
[26]巩忠福.博落回杀螨活性成分的研究[D]:西北农林科技大学;2002.
[27]Tani C.,Takao S.Studies on the alkaloids of fumariaceous plants.Ⅸ.Alkaloids of Bocconia frutescens.[J].Yakugaku Zasshi.1967,87(6):699-701.
[28]Tani C.,Takao N.Studies on the alkaloids of fumariaceous plants.Ⅴ.Alkaloids of Bocconia cordata(Macleya cordata)[J].Jpn J Pharmacol.1962,82:755-759.
[29]Takao N.,Yasumoto Y.,Iwasa K.Studies on the alkaloids of papaveraceous plants.ⅩⅤ.On the alkaloids of Bocconia cordata.2[J].Yakugaku Zasshi.1973,93(2):242-244.
[30]Boit H G.Ergebnisse der Alkaloid-chemie Bis-1960.Berlin:Akademie-verlag,1961
[31]周天达.苯并菲啶类生物碱在中国罂粟科和芸香科一些植物一些属中分布初探[J].植物学通报.1986,4(1-2):38-41.
[32]周荣汉.药用植物化学分类学.上海:上海科技出版社,1988.542.
[33]王宗训.从异喹啉生物碱试论双子叶一些科与罂粟科的一些属的亲缘关系[J].植物学报.1965,13(3):283-296.
[34]苏中武.被子植物的几个科的化学分类学(五)罂粟科的植物化学分类学[J].国外医学药学分册.1981,(02):65-70.
[35]黄松.罂粟科白屈菜族的化学成分及其植物化学分类依据的研究进展[J].湖南中医学导报.2002,8(10):582-584.
[36]陈孝泉.植物化学分类学.北京:高等教育出版社,1990.542.
[37]Liscombe D.K.,MacLeod B.P.,Loukanina N..Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms[J].Phytochemistry.2005,66(20):2501-2520.
[38]Park S.U.,Yu M.,Facchini P.J.Antisense RNA-mediated suppression of benzophenanthridine alkaloid biosynthesis in transgenic cell cultures of California poppy[J].Plant Physiol.2002,128(2):696-706.
[39]Kutchan T.M.,Dittrich H.Characterization and mechanism of the berberine bridge enzyme,a covalently flavinylated oxidase of benzophenanthridine alkaloid biosynthesis in plants[J].J Biol Chem.1995,270(41):24475-24481.
[40]Otani M.,Shitan N.,Sakai K..Characterization of vacuolar transport of the endogenous alkaloid berberine in Coptis japonica[J].Plant Physiol.2005,138(4):1939-1946.
[41]Sato F.,Hashimoto T.,Hachiya A..Metabolic engineering of plant alkaloid biosynthesis[J].Proc Natl Acad Sci U S A.2001,98(1):367-372.
[42]Morishige T.,Tsujita T.,Yamada Y..Molecular characterization of the S-adenosyl-L-methionine:3'-hydroxy-Nomethylcoclaurine 4'-O-methyltransferase involved in isoquinoline alkaloid biosynthesis in Coptis japonica[J].J Biol Chem.2000,275(30):23398-23405.
[43]Facchini J.P.,Luca V.D.Phloem-Specific Expression of Tyrosine/Dopa Decarboxylase Genes and the Biosynthesis of Isoquinoline Alkaloids in Opium Poppy[J].Plant Cell.1995,7(11):1811-1821.
[44]Alcantara J.,Bird D.A.,Franceschi V.R..Sanguinarine biosynthesis is associated with the endoplasmic reticulum in cultured opium poppy cells after elicitor treatment[J].Plant Physiol.2005,138(1):173-183.
[45]曾建国,张胜,候团章.不同生长期博落回果实中血根碱与白屈菜红碱的含量[J].中药材.1999,22(5):229
[46]Abizov E.A.,Tolkachev O.N.,Kopylova I.E..Distribution of the Sum of Sanguinarine and Chelerythrine in the Above-Ground Part of Macleaya Microcarpa[J].Pharmaceutical Chemistry Journal.2003,37(8):413-414.
[47]安彩贤.RP-HPLC法同时测定小果博落回中血根碱和白屈菜红碱的含量研究[J].中成药.2001,23(11):824-825.
[48]Nicholson J.K.,Lindon J.C.,Holmes E.'Metabonomics':understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data[J].Xenobiotica.1999,29(11):1181-1189.
[49]刘祥东.代谢物组学应用的领域之二-植物代谢物组学[J].中成药.2006,28(10):1515-1517.
[50]陈晓亚.植物次生代谢研究[J].世界科技研究与发展.2006,28(5):1-4.
[51]Fiehn O.Metabolic networks of Cucurbita maxima phloem[J].Phytochemistry.2003,62(6):875-886.
[52]Janet T,King R.D.,Thomas A.Application of metabolomics to plant genotype discrimination using statistics and machine learning[J].Bioinformatics.2002,18S241-248.
[53]Le G.Go,DuPont M.S.,Mellon F.A..Characterization and content of flavonoid glycosides in genetically modified tomato(Lycopersicon esculentum)fruits[J].J Agric Food Chem.2003,51(9):2438-2446.
[54]Choi H.K.,Choi Y.H.,Verberne M..Metabolic fingerprinting of wild type and transgenic tobacco plants by 1H NMR and multivariate analysis technique[J].Phytochemistry.2004,65(7):857-864.
[55]Walker T.S.,Bais H.P.,Halligan K.M..Metabolic profiling of root exudates of Arabidopsis thaliana[J].J Agric Food Chem.2003,51(9):2548-2554.
[56]Fiehn O.,Kopka J.,Dormann P..Metabolite profiling for plant functional genomics[J].Nat Biotechnol.2000,18(11):1157-1161.
[57]Choi Y.H.,Kim H.K.,Hazekamp A.Metabolomic Differentiation of Cannabis sativa Cultivars Using 1H NMR Spectroscopy and Principal Component Analysis[J].J Nat Prod.2004,67(6):953-957.
[58]Wang Y.,Tang H.,Nicholson J.K..Metabolomic strategy for the classification and quality control of phytomedicine:a case study of chamomile flower(Matricaria recutita L.)[J].Planta Med.2004,70(3):250-255.
[59]沈一丁.代谢组学促进中医药现代化[J].精细化工.2005,22(6):444-446.
[60]Narasimhan K,Basheer C,Bajic V.B..Enhancement of Plant-Microbe Interactions Using a Rhizosphere Metabolomics-Driven Approach and Its Application in the Removal of Polychlorinated Biphenyls[J].Plant Physiol.2003,132:146-153
[61]Verdonk J.C.,Verhoeven H.A..Regulation of floral scent production in petunia revealed by targeted metabolomics[J].Phytochemistry.2003,62(6):997-1008.
[62]Tolstikov V.V.,Fiehn O.Analysis of highly polar compounds of plant origin:combination of hydrophilic interaction chromatography and electrospray ion trap mass spectrometry[J].Anal Biochem.2002,301(2):298-307.
[63]Popl M,Fahnrich J,Tatar V.Chromatographic analysis of alkaloids.In. NewYork:Marcel Dekker;1990
[64]Haberlein P,Tschiersch K.Effects of surface structure of reversed phases on.the separation of alkaloids by HPLC[J].Pharmazie.1994,49(4):769-775.
[65]McCalley V.D.Reversed-phases HPLC of basic samples-an update 《LC-GC Asia Pacific》[J].MCCALLEY D V.1999,2(03):37-44
[66]Theodoridis G.,Papadoyannis I.,Hermans-Lokkerbol A..A study of the behaviour of some new column materials in the chromatographic analysis of catharanthus alkaloids[J].Chromatographia.1997,45:52-58.
[67]Gennaro M C.Reversed-phase ion-pair and ion-interaction chromatography.NewYork:Marcel Dekker,1995.37.
[68]朱龙,封满良,万秀琴.吗啡和海洛因等生物碱的高效液相色谱化学光测定[J].分析化学.1996,24(11):1295-1297.
[69]Wu C.C,Young D.S,Liu L.K.Liquid chromatographic-electrospray mass spectrometric analysis of Coptidis Rhizoma[J].Journal of Chromatography A.1996,755(1):19-26.
[70]Ma H.W,Zeng A.P..Reconstruction of metabolic networks from genome data and analysis of their global structure for various organisms[J].Bioinformatics 2003,19(2):270-277
[71]Wagner A.,Fell D.A.The small world inside large metabolic net- works[J].Proc Biol Sci.2001,268(1478):1803-1810.
[72]孙之荣.后基因组信息学.北京:清华大学出版社,2002.45.
[73]王冰.基于复杂网络理论的代谢网络结构研究进展[J].中国生物工程杂志.2005,25(6):10-14.
[74]Ziegler J.,Voigtlander S.,Schmidt J..Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis[J].Plant J.2006,48(2):177-192.
[75]Zenk M.H.The formation of benzophenanthridine alkaloids[J].Pure and Applied Chemistry.1994,66(10):2023-2028.
[76]Weiss D.,Baumert A.,Vogel M..Sanguinarine reductase,a key enzyme of benzophenanthridine detoxification[J].Plant Cell Environ.2006,29(2):291-302.
[77]王树禾.图论.北京:科学出版社,2004.167.
[78]孙丽艳,魏雄辉,韦宇.博落回中总生物碱提取方法的研究[J].药物分析杂志.2003,(23):15-18.
[79]罗旭彪.生物碱的液相色谱分析方法学及其液相色谱保留机理的研究[D].长沙:湖南大学;2005.
[80]Suchomelova J.,Bochorakova H.,Paulova H..HPLC quantification of seven quaternary benzo[c]phenanthridine alkaloids in six species of the family Papaveraceae[J].J Pharm Biomed Anal.2007,44(1):283-287.