摘要
目前环糊精化学已经发展成为超分子化学最重要的主体,与此后发展起来的冠醚、穴醚、杯芳等大环化合物相比,堪称超分子化学的先驱。环糊精有特殊的结构和稳定的化学性质,并且本身没有毒性可以食用,其在医药,食品,日用品,分析技术,化学化工和农业等领域中广为应用。环糊精外亲水内疏水,这种特性使其可同许多有机分子、无机离子、药物、染料及生物小分子等客体结合形成包合物。环糊精同客体分子形成包合物后可改变客体分子的物理性质、化学性质或光谱性质等,利用环糊精包合药物改变药物分子的性质可对环糊精和药物的包结作用进行研究,为环糊精在药物领域的发展奠定基础,同时还可借助环糊精包合药物改变药物分子的光谱学性质的原理,对药物进行直接检测。此外,染料具有吸收基团或荧光基团,经环糊精包合后,一般光谱学性质会发生改变,因此可设计染料-环糊精超分子体系,借助药物同染料的竞争包结作用来对药物进行间接检测。
本文借助分光光度法及荧光法,首先研究了β-环糊精同荧光素和中性红的包合作用,通过对β-环糊精对荧光素及中性红包合性能的对比选择了合适的染料作探针,并同β-环糊精组成超分子体系,然后借助此β-环糊精—染料超分子体系对合成样品中的胆酸进行了检测,同时研究了β-环糊精对槲皮素的包合性能。此外,还研究了牛血清白蛋白与槲皮素的相互作用。
本论文共分五章。各章介绍如下：
第一章,综述了环糊精在药物分析中的进展。简单介绍了环糊精的结构性质、物理性质、化学性质、环糊精在药物分析中的应用与研究及环糊精的研究趋势。
第二章,分别研究了β-环糊精对荧光素和中性红的包合作用,同时还研究了β-环糊精对荧光素的荧光猝灭机理,并求出了β-环糊精对荧光素和中性红的包结常数K、包结比n及各自的包合反应的热力学参数ΔG,ΔH及ΔS,并对β-环糊精对荧光素的包合性能及β-环糊精对中性红的包合性能进行了对比,从而选择出了合适的染料做为探针,同β-环糊精组成超分子体系,进而为下步的检测奠定了基础。
第三章,对合成样品中的胆酸进行了检测。β-环糊精包合中性红使中性红的荧光增强,当加入胆酸后,由于胆酸同中性红发生竞争反应,将中性红部分挤出β-环糊精的空腔,又使其荧光减弱。基于此原理,构建了中性红-β-环糊精超分子体系,并借助此超分子体系,建立了测定胆酸的新方法。
第四章,研究了β-环糊精对槲皮素的包合作用,计算出了包合常数K和包结比n,利用热力学方程及包合常数值求出了热力学参数ΔG,ΔH及ΔS,并基于热力学参数值确定了β-环糊精和槲皮素间的主要作用力为疏水作用力。
第五章,研究了牛血清白蛋白与槲皮素的相互作用。探讨了槲皮素对牛血清白蛋白荧光猝灭类型和机理,求算了牛血清白蛋白和槲皮素的结合常数和结合点数,并确定了其主要作用力,根据Forster非辐射能量转移理论,得到了槲皮素与牛血清白蛋白之间的能量转移效率和距离。中图分类号：0657.3
Currently Cyclodextrin Chemistry has been developed into the most important subject of Supramolecular chemistry and is the pioneer of Supramolecular Chemistry compared with the macrocyclic compound such as crown ether, cave ether and so on. Cyclodextrin is widely used in areas such as agriculture, food, analytical technique and chemical industry. Cyclodextrin with the property of hydrophobic inside at the same time hydrophilic the outer can react with many objects (drug molecules, biological molecules, organic molecules and inorganic ions) to form inclusion compounds and the physical property, chemical property or spectral property of inclusion compounds will be changed. Therefore the inclusion mechanism between drug molecules and cyclodextrin can be studied via this principle. Also this principle can be directly tested for drug molecules. Besides indirect detection of drugs can be realized by construction of dye-cyclodextrin supramolecular system.
In this paper, using molecule fluorescence spectroscopy and UV-Vis spectroscopy as research method, we have discussed the inclusion mechanism between dyes (fluorescein, neutral red), drug molecules (quercetin) and cyclodextrin. And also we have determined cholic acid with high sensibility and low detection limit. Simultaneously, we have discussed the reaction mechanism between quercetin and bovine serum albumin.
There are five chapters in this paper.
The first part reviews applications and progresses of cyclodextrin in the medication area.
In the second chapter, the inclusion interaction between fluorescein (FL), neutral red (NR) and P-cyclodextrin have been studied. The binding constant K and stoichiometric ratio n were obtained respectively. The thermodynamic parametersΔG,ΔH,ΔS of inclusion compounds were calculated respectively. Also the differences between the two inclusion reactions were summarized.
In the third part, a new, simple and highly sensitive P-cyclodextrin supramolecular system method for the determination of cholic acid has been developed. And we determined cholic acid with high sensibility and low detection limit based on the principle.
In the fourth part, the inclusion interaction between quercetin and P-cyclodextrin has been studied using fluorescence spectroscopy. The binding constant K and stoichiometric ratio n were obtained. The thermodynamic parametersΔG,ΔH andΔS of inclusion compound were 3 calculated. The main action was confirmd by the value of hermodynamic parameters.
In the fifth part, the interaction between quercetin and bovine serum albumin (BSA) was studied by the fluorescence spectroscopy and the UV-visible absorption spectroscopy. The static fluorescence quenching process of quercetin to BSA was confirmed. The binding constant and the number of binding sites were obtained.
引文
[1]童林荟.环糊精化学[M].北京：科学出版社,2001：13-18.
[2]Breslow R, Steven D D. Biomimetic Reactions Catalyzed by Cyclodextrins and Their Derivatives [J]. Chem. Rev.,1998,98 (5):1997-2012.
[3]Osaki M, Takashima Y, Yamaguchi H, Harada A [J]. Macromolecules,2007,40 (9): 3154-3158.
[4]Bekiroglu S, Kenne L, Sandstrom C.1H NMR Studies of Maltose, Maltoheptaose, α-,β-, and y-Cyclodextrins, and Complexes in Aqueous Solutions with Hydroxy Protons as Structural Probes [J]. J. Org. Chem.,2003,68 (5):1671-1678.
[5]Hallberg F, Christoph F, Weise P V, Pettersson E T, Stilbs P, Fur I. Molecular Complexation and Binding Studied by Electrophoretic NMR [J].Spectroscopy.2008,130 (24):7550-7551.
[6]Freeman R, Finder T, Bahshi L, Willner I. P-Cyclodextrin-Modified CdSe/ZnS Quantum Dots for Sensing and Chiroselective Analysis [J]. Nano.Lett.,2009,9 (5):2073-2076.
[7]Nakashima H, Yoshida N. Fluorescent Detection for Cyclic and Acyclic Alcohol Guests by Naphthalene-Appended Amino-β-cyclodextrins [J]. Org. Lett.,2006,8 (22):4997-5000.
[8]Haina Yuan, Shanjing Yao, Lianqing Shen, Jianwei Mao. Preparation and Characterization of Inclusion Complexes of β-Cyclodextrin-BITC and β-Cyclodextrin-PEITC [J]. Ind. Eng. Chem. Res.,2009,48 (10):5070-5078.
[9]Park J W, Lee S Y, et al. Self-Inclusion Behavior and Circular Dichroism of Aliphatic Chain-Linked β-Cyclodextrin-Viologen Compounds and Their Reduced Forms Depending on the Side of Modification [J]. J. Org. Chem.,2005,70(23):9505-9513
[10]Deryabina M A, Hansen S H, Stergaard J, Jensen H. Effect of a-Cyclodextrin on Drug Distribution Studied by Electrochemistry at Interfaces between Immiscible Electrolyte Solutions [J]. J. Phys. Chem. B,2009,113 (20):7263-7269.
[11]Ionita G, Florent M, Goldfarb D, Chechik V. Studying Supramolecular Assemblies by ESEEM Spectroscopy:Inclusion Complexes of Cyclodextrins [J]. J. Phys. Chem. B,2009, 113 (17):5781-5787.
[12]Marzouqi A H, et al. Physicochemical properties of antifungal drug-cyclodextrin complexes prepared by supercritical carbon dioxide and by conventional techniques [J]. Journal of Pharmaceutical and Biomedical Analysis,2009,49 (2):227-233.
[13]Loftsson T, Hreinsdottir D, Masson M. Evaluation of cyclodextrin solubilization of drugs [J]. International Journal of Pharmaceutics,2005,302(1-2):18-28.
[14]Yu Liu,Yong Chen. Cooperative Binding and Multiple Recognition by Bridged Bis(β-cyclodextrin)s with Functional Linkers [J]. Accounts of Chemical Research.2006,39(10):681-691.
[15]Horvath G, Premkumar T,et al.Supramolecular Nanoencapsulation as a Tool:Solubilization of the Anticancer Drug trans-Dichloro(dipyridine)platinum(Ⅱ) by Complexation with β-Cyclodextrin[J]. Molecular Pharmaceutics.2008,5(2):358-363.
[16]. Juan M Benito, et al.Design, Synthesis, and Biological Evaluation of Glycodendrimer-Cyclodextrin Conjugates[J].J.Am. Chem.Soc.2004,126(23): 10355-10363.
[17]Bruno A. Rezende, Steyner F. Cortes, Frederico B. De Sousa, Ivana S. Lula, Martine Schmitt, Ruben D. Sinisterra, Virginia S. Lemos. Complexation with P-cyclodextrin confers oral activity on the flavonoid dioclein[J]. International Journal of Pharmaceutics, 2009,367 (1-2):133-139.
[18]Patricia S M,etal. Mode of action of β-cyclodextrin as an absorption enhancer of the water-soluble drug meglumine antimoniate [J]. International Journal of Pharmaceutics, 2006,325 (1-2):39-47.
[19]Torricelli C, Martini A, Muggetti L, et al. Stability studies on a steroidal drug/β-cyclodextrin coground mixture [J]. International Journal of Pharmaceutics,1991,71 (1-2):19-24.
[20]Garbow J R, Stephen A S, et al. Structure, Dynamics, and Stability of a-Cyclodextrin Inclusion Complexes of Aspartame and Neotame [J]. J. Agric. Food Chem.2001,49(4): 2053-2060.
[21]Castagnino E, Cangiotti M, Tongiani S, Ottaviani M F. A study of free radical release from (3-cyclodextrin-anticancer pro-drugs adducts in water[J]. Journal of Controlled Release, 2005,108 (2-3):215-225.
[22]Xinge Zhang, Zhongming Wu, Xiujun Gao, Shujun Shu, Huijie Zhang, Zhen Wang, Chaoxing Li. Chitosan bearing pendant cyclodextrin as a carrier for controlled protein release[J]. Carbohydrate Polymers,2009,77 (2):394-401.
[23]Cappello B, Rosa G D, Giannini L, et al. Cyclodextrin-containing poly(ethyleneoxide) tablets for the delivery of poorly soluble drugs:Potential as buccal delivery system [J].International Journal of Pharmaceutics,2006,319 (1-2):63-70.
[24]Jeulin H, Venard V, Carapito D, Finance C, Kedzierewicz F. Effective ribavirin concentration in mice brain using cyclodextrin as a drug carrier:Evaluation in a measles encephalitis model [J].Antiviral Research,2009,81 (3):261-266.
[25]Jinku Xu, Xinsong Li, Fuqian Sun. Cyclodextrin-containing hydrogels for contact lenses as a platform for drug incorporation and release [J].Acta Biomaterialia,2010,6(2):486-493.
[26]Gillet A, Grammenos A, et al. Development of a new topical system: Drug-in-cyclodextrin-in-deformable liposome [J].International Journal of Pharmaceutics, 2009,380 (1-2):174-180.
[27]Jianshu Li, Huining Xiao, Jiehua Li, YinPing Zhong. Drug carrier systems based on water-soluble cationic β-cyclodextrin polymers [J].International Journal of Pharmaceutics, 2004,278 (2):329-342.
[28]Domingues Z R, et al.Bioactive glass as a drug delivery system of tetracycline and tetracycline associated with β-cyclodextrin [J].Biomaterials,2004,25 (2):327-333.
[29]Oda Y, Yanagisawa H, et al. Design, synthesis and evaluation of d-galactose-β-cyclodextrin conjugates as drug-carrying molecules[J].Bioorganic & Medicinal Chemistry,2008,16 (19):8830-8840.
[30]Siemoneit U, Schmitt C, et al. Acrylic/cyclodextrin hydrogels with enhanced drug loading and sustained release capability [J]. International Journal of Pharmaceutics,2006,312 (1-2):66-74.
[31]Jie Chen, et al. Radiation synthesis of pH-sensitive hydrogels from P-cyclodextrin-grafted PEG and acrylic acid for drug delivery [J]. Materials Chemistry and Physics,2009,116(1): 148-152.
[32]Prabaharan M, R et al. Chitosan-graft-β-cyclodextrin scaffolds with controlled drug release capability for tissue engineering applications [J]. International Journal of Biological Macromolecules,2009,44 (4):320-325.
[33]Prabaharan M, Shaoqin Gong. Novel thiolated carboxymethyl chitosan-g-(3-cyclodextrin as mucoadhesive hydrophobic drug delivery carriers [J]. Carbohydrate Polymers,2008,73(1): 117-125.
[34]Bertolla C, Rolin S, Evrard B, et al. Synthesis and pharmacological evaluation of a new targeted drug carrier system:β-Cyclodextrin coupled to oxytocin [J]. Bioorganic & Medicinal Chemistry Letters,2008,18(6):1855-1858.
[35]Salmaso S, Semenzato A, et al. a-Cyclodextrin-Poly(ethylene Glycol)-Folic Acid Drug Delivery Bioconjugate[J]. Bioconjugate Chem.2004,15(5):997-1004.
[36]Layre A M, Amiel C, et al. Based on Cyclodextrin Polymer:A Model System for Drug Delivery Biomacromolecules [J].2009,10(12):3283-3289.
[37]Kralova J, Porphyrin-Cyclodextrin Conjugates as a Nanosystem for Versatile Drug Delivery and Multimodal Cancer Therapy [J]. J. Med. Chem.2010,53(1):128-138.
[38]Caliceti P, Salmaso S, et al. Synthesis and Physicochemical Characterization of Folate-Cyclodextrin Bioconjugate for Active Drug Delivery [J]. Bioconjugate Chem.2003, 14(5):899-908
[39]Xiaolei Zhu, Hongbo Wang, Qiong Chen, Wenchao Yang, Guangfu Yang Preparation and Characterization of Inclusion Complex of Iprodione and a-Cyclodextrin to Improve Fungicidal Activity [J]. J.Agric.Food.Chem.,2007,55 (9):3535-3539.
[40]Fermeglia M, Ferrone M, et al. Host-guest inclusion complexes between anticancer drugs and β-cyclodextrin:computational studies [J]. Carbohydrate Polymers,2003,53 (1):15-44.
[41]Xinming Liu, Andrew T. et al. β-cyclodextrin for local bone regeneration [J]. Biomaterials, 2008,29 (11):1686-1692.
[42]Robert O. Williams III, et al. Haracterization of an inclusion complex of cholesterol and hydroxypropyl-β-cyclodextrin[J]. European Journal of Pharmaceutics and Biopharmaceutics,1998,46(3):355-360.
[43]Naomi O, et al. Determination of stability constant of β-cyclodextrin complexes using the membrane permeation technique and the permeation behavior of drug-competing agent-β-cyclodextrin ternary systems [J]. European Journal of Pharmaceutical Sciences, 1999,8(2):133-139.
[44]Nozawa Y, et al. Effects of shear rate in roll mixing on mechanical complex formation of ibuprofen with P-cyclodextrin [J]. Pharmaceutica Acta Helvetiae,1994,69(3):135-139.
[45]Letellier S, et al. Interaction of floctafenine with methyl-β-cyclodextrin in aqueous solution [J]. Pharmaceutica Acta Helvetiae,1994,68(4):229-233.
[46]Figueiras A, Juan J, et al.Solid-state characterization and dissolution profiles of the inclusion complexes of omeprazole with native and chemically modified β-cyclodextrin [J]. European Journal of Pharmaceutics and Biopharmaceutics,2007,67 (2):531-539.
[47]Niya A, et al. The role of water in the thermodynamics of drug binding to cyclodextrin [J]. The Journal of Chemical Thermodynamics,2007,39(7):1038-1048.
[48]Spamer E, Muller D G., et al. Characterization of the complexes of furosemide with 2-hydroxypropyl-β-cyclodextrin and sulfobutyl ether-7-p-cyclodextrin. [J] European Journal of Pharmaceutical Sciences,2002,16 (4-5):247-253.
[49]Comini S, Olivier P, et al. Interaction of β-cyclodextrin with bile acids and their competition with vitamins A and D3 as determined by'H-NMR spectrometry [J]. Clinica Chimica Acta, 1994,228 (2):181-194.
[1]Breslow R, Steven D D. Biomimetic Reactions Catalyzed by Cyclodextrins and Their Derivatives [J]. Chem. Rev.,1998,98 (5):1997-2012.
[2]Osaki M, Takashima Y, Yamaguchi H, Harada A. Polymerization of Lactones Initiated by Cyclodextrins:Effects of Cyclodextrins on the Initiation and Propagation Reactions[J]. Macromolecules,2007,40 (9):3154-3158.
[3]Bekiroglu S, Kenne L, Sandstrom C. 1H NMR Studies of Maltose, Maltoheptaose, α-, β-, and y-Cyclodextrins, and Complexes in Aqueous Solutions with Hydroxy Protons as Structural Probes [J]. J. Org. Chem.,2003,68 (5):1671-1678.
[4]Hallberg F, Christoph F, Weise P V, Pettersson E T, Stilbs P, Fur I. Molecular Complexation and Binding Studied by Electrophoretic NMR Spectroscopy [J]. J.Am.Chem.Soc.,2008,130 (24):7550-7551.
[5]Freeman R, Finder T, Bahshi L, Willner I. β-Cyclodextrin-Modified CdSe/ZnS Quantum Dots for Sensing and Chiroselective Analysis [J]. Nano.Lett.,2009,9 (5):2073-2076.
[6]Nakashima H, Yoshida N. Fluorescent Detection for Cyclic and Acyclic Alcohol Guests by Naphthalene-Appended Amino-β-cyclodextrins [J]. Org. Lett.,2006,8 (22):4997-5000.
[7]Haina Yuan, Shanjing Yao, Lianqing Shen, Jianwei Mao. Preparation and Characterization of Inclusion Complexes of β-Cyclodextrin-BITC and β-Cyclodextrin-PEITC [J]. hid. Eng. Chem. Res.,2009,48 (10):5070-5078.
[8]Park J W, Lee S Y, Song H J, Park K K.. Self-Inclusion Behavior and Circular Dichroism of Aliphatic Chain-Linked β-Cyclodextrin-Viologen Compounds and Their Reduced Forms Depending on the Side of Modification [J]. J. Org. Chem.,2005,70(23):9505-9513
[9]Deryabina M A, Hansen S H, Stergaard J, Jensen H. Effect of a-Cyclodextrin on Drug Distribution Studied by Electrochemistry at Interfaces between Immiscible Electrolyte Solutions [J]. J. Phys. Chem. B,2009,113 (20):7263-7269.
[10]Ionita G, Florent M, Goldfarb D, Chechik V. Studying Supramolecular Assemblies by ESEEM Spectroscopy:Inclusion Complexes of Cyclodextrins [J]. J. Phys. Chem. B,2009, 113 (17):5781-5787.
[11]Yu Liu, Bin Li, Li Li, Hengyi Zhang. Synthesis of Organoselenium-Modified β-Cyclodextrins Possessing a 1,2-Benzisoselenazol-3(2H)-one Moiety and Their Enzyme-Mimic Study [J]. Helv. Chim. Actam,2002,85 (1):9-18.
[12]Naughton M M, Engman L, Birmingham A, Powis G, Cotgreave I A. Cyclodextrin-Derived Diorganyl Tellurides as Glutathione Peroxidase Mimics and Inhibitors of Thioredoxin Reductase and Cancer Cell Growth [J]. J. Med. Chem.2004, 47(1):233-239.
[13]Huaiyou Wang, Juan Han, Xiaguang Feng, Yanling Pang. Study of inclusion complex formation between tropaeolin OO and β-cyclodextrin by spectrophotometry and Infrared spectroscopy [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2006,65 (1):100-105.
[14]Huai You Wang, Juan Han, Xia Guang Feng Spectroscopic study of orange G-β-cyclodextrin complex and its analytical application [J].Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy,2007,66 (3):578-585.
[15]Hui Xie, Huaiyou Wang, Liying Ma, Yan Xiao, Juan Han. Spectrophotometric study of the inclusion complex between β-cyclodextrin and dibenzoyl peroxide and its analytical application [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2005,62(1-3):197-202.
[16]Yu Liu, Yong Chen, Shuangxi Liu, Xudong Guan, Wada T, Inoue Y. Organic.Letters [J]., 2001,3 (11):1657-1660.
[17]Guangchao Zhao, Junjie Zhu, Hongyuan Chen. Spectroscopic studies of the interactive model of methylene blue with DNA by means of P-cyclodextrin [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy.1999,55 (5):1109-1117.
[18]Guangchao Zhao, Junjie Zhu, Hongyuan Chen. Voltammetric studies of the interaction of methylene blue with DNA by means of β-cyclodextrin [J]. Analytica Chimica. Acta., 1999,394 (2-3):337-344.
[19]Davies J E D, etal. Physical Methods in Supramolecular Chemistry (8th). Oxford: Pergamon Press,1996,426.
[20]Catena G. C, Bright F V. Thermodynamic study on the effects of.beta.-cyclodextrin inclusion with anilinonaphthalenesulfonates [J]. Anal. Chim.,1989,61(8):905-909.
[21]童林荟.环糊精化学[M].北京：科学出版社,2001：164-176.
[22]许金钩,王尊本.荧光分析法(第三版)[M].北京：科学出版社,2006：64-70.
[23]Lakowicz J R, Principles of fluorescence spectroscopy (2ed) [M]. Newyork:Plenum Press,1999:243-249.
[1]Yinxue Song, Jun Gong. Effect of bile salts and bile acids on human gastric mucosal epithelial cells [J]. Journal of Nanjing Medical University,2008,22 (4):217-223.
[2]Halpern M D, Holubec H, et al. Bile Acids Induce Ileal Damage During Experimental Necrotizing Enterocolitis [J].Gastroenterology,2006,130 (2):359-372.
[3]Batta A K, Salen G.Gas chromatography of bile acids [J]. Journal of Chromatography B: Biomedical Sciences and Applications,1999,723 (1):1-16.
[4]Sylvia K, Gerhard J.Determination of underivatised sterols and bile acid trimethyl silyl ether methyl esters by gas chromatography-mass spectrometry-single ion monitoring in faeces [J]. Journal of chromatography. B, Analytical technologies in the biomedical and life faeces [J]. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences,2004,813 (1-2):199-207.
[5]Sakakura H, Suzuki M, Kimura N, Takeda H, et al. Simultaneous determination of bile acids in rat bile and serum by high-performance liquid chromatography [J]. Journal of Chromatography B:Biomedical Sciences and Applications,1993,621 (2):123-131.
[6]Tagliacozzi D, Mozzi A F, Casetta B et al. Quantitative Analysis of Bile Acids in Human Plasma by Liquid Chromatography-Electrospray Tandem Mass Spectrometry:A Simple and Rapid One-Step Method [J]. Clin Chem Lab Med.2003,41 (12):1633-1641.
[7]Burkard I, Eckardstein A V, et al. Differentiated quantification of human bile acids in serum by high-performance liquid chromatography-tandem mass spectrometry [J]. Journal of Chromatography B,2005,826 (1-2):147-159.
[8]Bentayeb K, Batlle R, Sanchez C, et al. Determination of bile acids in human serum by on-line restricted access material-ultra high-performance liquid chrorriatography-mass spectrometry [J].Journal of Chromatography B,2008,869 (1-2):1-8.
[9]Ando M, Kaneko T, Watanabe R, Kikuchi S, Goto T, et al. High sensitive analysis of rat serum bile acids by liquid chromatography/electrospray ionization tandem mass spectrometry [J]. Journal of Pharmaceutical and Biomedical Analysis,2006,40 (5):1179-1186.
[10]Perwaiz S, Tuchweber B, Mignault D, Gilat D, Yousef I M. Determination of bile acids in biological fluids by liquid chromatography-electrospray tandem mass spectrometry [J]. Journal of Lipid Research,2001,42 (1):114-119.
[11]Porter JL, Fordtran J S et al.Accurate enzymatic measurement of fecal bile acids in patients with malabsorption [J]. Journal of Laboratory and Clinical Medicine,2003,141 (6): 411-418.
[12]Guohua Zhang, Airi Cong, Guobing Xu, Chuanbao Li, Ruifeng Yang, Tiean Xia. An enzymatic cycling method for the determination of serum total bile acids with recombinant 3a-hydroxysteroid dehydrogenase [J]. Biochemical and Biophysical Research Communications,2005,326 (1):87-92.
[13]Yongsheng Li, Weiping Liu, Xiufeng Gao, Dandan Chen, Weiguang Li Immobilized enzymatic fluorescence capillary biosensor for determination of sulfated bile acid in urine[J]. Biosensors and Bioelectronics,2008,24(4):538-544.
[14]Brandon B, Lu Li, et al..Determination of total bile acid levels using a thick-film screen-printed Ir/C sensor for the detection of liver disease [J].Analyst.2009,134 (5): 973-979.
[15]Henegouwen V B, et al. Quantitative analysis of bile acids in serum and bile, using gas-liquid chromatography [J]. Clinica Chimica Acta,1974,54 (3):249-261.
[16]Que A H, Konse T, Baker A G, Novotny M V. Analysis of Bile Acids and Their Conjugates by Capillary Electrochromatography/Electrospray Ion Trap Mass Spectrometry [J]. Anal. Chem.2000,72 (13):2703-2710.
[17]Yang, Z. Breslow, R. Very Strong Binding of Lithocholic Acid to P-Cyclodextrin [J]. Tetrahedron Letters,1997,38 (35):6171-6172.
[18]Yu Liu, Jun Shi, and Dong-Sheng Guo. Novel permethylated beta-cyclodextrin derivatives appended with chromophores as efficient fluorescent sensors for the molecular recognition of bile salts [J].The Journal of organic chemistry,2007,72 (22):8227-8234.
[19]Xiashi Zhu, Jing Sun, Jun Wu. Study on the inclusion interactions of β-cyclodextrin and its derivative with dyes by spectrofluorimetry and its analytical application [J].Talanta, 2007,72 (1):237-242.
[20]Velic D, Knapp M, Kohler G. Supramolecular inclusion complexes between a coumarin dye and β-cyclodextrin, and attachment kinetics of thiolated β-cyclodextrin to gold surface [J]. Journal of Molecular Structure,2001,598 (1):49-56.
[21]Huarong Zhang, Gang Chen, Ling Wang, Lan Ding, Yuan Tian, Weiqun Jin, Hanqi Zhang. Study on the inclusion complexes of cyclodextrin and sulphonated azo dyes by electrospray ionization mass spectrometry [J].International Journal of Mass Spectrometry, 2006,252 (1):1-10.
[22]Moritz E D, Sahyun M R V. Spectroscopic studies of β-cyclodextrin-complexed cyanine dyes [J]. Journal of Photochemistry and Photobiology A:Chemistry,2005,169 (3): 211-220.
[23]Shannigrahi M, BagchiS. Time resolved fluorescence study of ketocyanine dye-β cyclodextrin interactions in aqueous and non-aqueous media [J].Chemical Physics Letters, 2005,403 (1-3):55-61.
[24]Huaiyou Wang, Juanhan, Xiaguang Feng. Spectroscopic study of orange G-β-cyclodextrin complex and its analytical application [J].Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2007,66 (3):578-585.
[25]Huaiyou Wang, Juanhan, Xiaguang Feng, Yanling Pang. Study of inclusion complex formation between tropaeolin 00 and β-cyclodextrin by spectrophotometry and Infrared spectroscopy [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy, 2006,65 (1):100-105.
[26]Guomei Zhang, Shaomin Shuang, Chuan Dong, Jinghao Pan. Study on the interaction of methylene blue with cyclodextrin derivatives by absorption and fluorescence spectroscopy [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2003,59 (13): 2935-2941.
[27]Guomei Zhang, Shaomin Shuang, Zhenming Dong, Chuan Dong, Jinghao Pan Investigation on the inclusion behavior of neutral red with β-cyclodextrin, hydroxypropyl-β-cyclodextrin and sulfobutylether-β-cyclodextrin [J].Analytica Chimica Acta,2002,474 (1-2):189-195.
[28]Guomei Zhang, Shaomin Shuang, Chuan Dong, Diansheng Liu, et al. Investigation on DNA assembly to neutral red-cyclodextrin complex by molecular spectroscopy [J]. Journal of Photochemistry and Photobiology B:Biology,2004,74 (2-3):127-134.
[1]Rezende B A, Cortes SF, Sousa F B D, Lula I S, Schmitt M, Sinisterra R D, Lemos V S. Complexation with β-cyclodextrin confers oral activity on the flavonoid dioclein [J].International Journal of Pharmaceutics,2009,367 (1-2):133-139.
[2]Martins P S, Ochoa R, Pimenta A M C, Ferreira L A M, Melo AL, et al. Mode of action of P-cyclodextrin as an absorption enhancer of the water-soluble drug meglumine antimoniate [J]. International Journal of Pharmaceutics,2006,325 (1-2):39-47.
[3]Al-Marzouqi A H, Elwy H M, Shehadi I, Adem A. Physicochemical properties of antifungal drug-cyclodextrin complexes prepared by supercritical carbon dioxide and by conventional techniques [J]. Journal of Pharmaceutical and Biomedical Analysis,2009,49 (2):227-233.
[4]Torricelli C, Martini A, Muggetti L, Ponti R D. Stability studies on a steroidal drug/p-cyclodextrin coground mixture [J].International Journal of Pharmaceutics,1991,71 (1-2):19-24.
[5]Castagnino E, Cangiotti M, Tongiani S, Ottaviani M F. A study of free radical release from β-cyclodextrin-anticancer pro-drugs adducts in water [J]. Journal of Controlled Release, 2005,108 (2-3):215-225.
[6]Xinge Zhang, Zhongming Wu, Xiujun Gao, Shujun Shu, Huijie Zhang, Zhen Wang, Chaoxing Li. Chitosan bearing pendant cyclodextrin as a carrier for controlled protein release[J]. Carbohydrate Polymers,2009,77 (2):394-401.
[7]Cappello B, Rosa G D, Giannini L, et al.Cyclodextrin-containing poly(ethyleneoxide) tablets for the delivery of poorly soluble drugs:Potential as buccal delivery system [J].International Journal of Pharmaceutics,2006,319 (1-2):63-70.
[8]Jeulin H, Venard V, Carapito D, Finance C, Kedzierewicz F. Effective ribavirin concentration in mice brain using cyclodextrin as a drug carrier:Evaluation in a measles encephalitis model [J].Antiviral Research,2009,81 (3):261-266.
[9]Jinku Xu, Xinsong Li, Fuqian Sun. Cyclodextrin-containing hydrogels for contact lenses as a platform for drug incorporation and release [J].Acta Biomaterialia,2010,6(2):486-493.
[10]Gillet A, Grammenos A, Compere P, Evrard B, Piel G. Development of a new topical system:Drug-in-cyclodextrin-in-deformable liposome [J].International Journal of Pharmaceutics,2009,380 (1-2):174-180.
[11]Jianshu Li, Huining Xiao, Jiehua Li, Yinping Zhong. Drug carrier systems based on water-soluble cationic P-cyclodextrin polymers [J].International Journal of Pharmaceutics, 2004,278 (2):329-342.
[12]Domingues Z R, Cortes M E, Gomes T A, Diniz H F, Freitas C S, et al.Bioactive glass as a drug delivery system of tetracycline and tetracycline associated with β-cyclodextrin [J].Biomaterials,2004,25 (2):327-333.
[13]Oda Y, Yanagisawa H, Maruyama M, Hattori K, Yamanoi T. Design, synthesis and evaluation of d-galactose-P-cyclodextrin conjugates as drug-carrying molecules [J].Bioorganic & Medicinal Chemistry,2008,16(19):8830-8840.
[14]Siemoneit U, Schmitt C, Lorenzo C A, Luzardo A, et al. Acrylic/cyclodextrin hydrogels with enhanced drug loading and sustained release capability [J]. International Journal of Pharmaceutics,2006,312 (1-2):66-74.
[15]Li Sun, Stenken J A.The effect of P-cyclodextrin on liquid chromatography/electrospray-mass spectrometry analysis of hydrophobic drug molecules [J].Journal of Chromatography A,2007,1161 (1-2):261-268.
[16]Papiez M A, Cierniak A, Krzysciak W, Bzowska M, et al.The changes of antioxidant defense system caused by quercetin administration do not lead to DNA damage and apoptosis in the spleen and bone marrow cells of rats [J]. Food and Chemical Toxicology, 2008,46 (9):3053-3058.
[17]Robaszkiewicz A, Balcerczyk A, Bartosz G. Antioxidative and prooxidative effects of quercetin on A549 cells [J]. Cell Biology International,2007,31 (10):1245-1250.
[18]Woude H V D, Boersma M G, Alink G M, et al.Consequences of quercetin methylation for its covalent glutathione and DNA adduct formation [J]. Chemico-Biological Interactions,2006,160 (3):193-203.
[19]Huang S L, Hsu C L, Yen G C. Growth inhibitory effect of quercetin on SW 872 human liposarcoma cells [J]. Life Sciences,2006,79 (2):203-209.
[20]Woude H V D, Struijs K, Smeets A, et al. Biphasic modulation of cell proliferation by quercetin at concentrations physiologically relevant in humans [J]. Cancer Letters,2003, 200(1):41-47.
[21]Braganhol E, Alessandra S K, et al.. Ecto-5'-nucleotidase/CD73 inhibition by quercetin in the human U138MG glioma cell line Biochimicaet [J]. Biophysica Acta (BBA)-General Subjects,2007,1770 (9):1352-1359.
[22]Brookes P S, Digerness S B, Parks D A, etal.Mitochondrial function in response to cardiac ischemia-reperfusion after oral treatment with quercetin [J]. Free Radical Biology and Medicine,2002,32 (11):1220-1228.
[23]Moon J H, Tsushida T, Nakahara K, Terao J. Identification of quercetin 3-O-β-D-glucuronide as an antioxidative metabolite in rat plasma after oral administration of quercetin [J]. Free Radical Biology and Medicine,2001,30 (11):1274-1285.
[24]Ping Yao, Nussler A, Liegang Liu, Liping Hao, Fangfang Song, Schirmeier A, Nussler N. Quercetin protects human hepatocytes from ethanol-derived oxidative stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways [J]. Journal of Hepatology, 2007,47 (2):253-261.
[25]Catena G. C, Bright F V. Thermodynamic study on the effects of beta.-cyclodextrin inclusion with anilinonaphthalenesulfonates[J]. Anal Chim,1989,61(8):905-909.
[26]童林荟.环糊精化学[M].北京：科学出版社,2001：164-176.
[1]Harwood M, Nikiel B D, Borzelleca J F, Flamm G W, Williams G M, Lines T C. A critical review of the data related to the safety of quercetin and lack of evidence of in vivo toxicity, including lack of genotoxic/carcinogenic properties [J].Food and Chemical Toxicology, 2007,45(11):2179-2205.
[2]Papiez M A, Krzysciak A C W, et al. The changes of antioxidant defense system caused by quercetin administration do not lead to DNA damage and apoptosis in the spleen and bone marrow cells of rats [J]. Food and Chemical Toxicology,2008,46(9):3053-3058
[3]Robaszkiewicz A, Balcerczyk A, Bartosz G. Antioxidative and prooxidative effects of quercetin on A549 cells [J]. Cell Biology International,2007,31(10):1245-1250
[4]Woude H V D, Boersma M G, Alink G M, et al. Consequences of quercetin methylation for its covalent glutathione and DNA adduct formation [J]. Chemico-Biological Interactions, April 2006,160(3):193-203.
[5]Huang S L, Hsu C L, Yen G C. Growth inhibitory effect of quercetin on SW 872 human liposarcoma cells [J]. Life Sciences,2006,79 (2):203-209.
[6]Woude H V D, et al. Biphasic modulation of cell proliferation by quercetin at concentrations physiologically relevant in humans [J]. Cancer Letters,2003,200(1):41-47
[7]Braganhol E,Tamajusuku A S K, Bernardi A, Wink M R,Battastini A M O. Ecto-5'-nucleotidase/CD73 inhibition by quercetin in the human U138MG glioma cell line [J]. Biochimica et Biophysica Acta (BBA)-General Subjects,2007,1770 (9):1352-1359
[8]Brookes P S, Digerness S B, Parks D A, et al. Mitochondrial function in response to cardiac ischemia-reperfusion after oral treatment with quercetin [J]. Free Radical Biology and Medicine,2002,32 (11):1220-1228
[9]Moon J K, Tsushida T, Nakahara K, Terao J. Identification of quercetin 3-O-β-D-glucuronide as an antioxidative metabolite in rat plasma after oral administration of quercetin [J]. Free Radical Biology and Medicine,2001,30(11):1274-1285
[10]Ping Yao, Nussler A, Liegang Liu, Liping Hao, Fangfang Song, Schirmeier A, Nussler N. Quercetin protects human hepatocytes from ethanol-derived oxidative stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways [J]. Journal of Hepatology, Volume 47, Issue 2, August 2007,47(2):253-261.
[11]Sengupta B, Sengupta P K. The interaction of quercetin with human serum albumin:a fluorescence spectroscopic study [J]. Biochemical and Biophysical Research Communications,2002,299(3):400-403
[12]Rasoulzadeh F, Jabary H N, Naseri A, Rashidi M R. Fluorescence quenching study of quercetin interaction with bovine milk xanthine oxidase [J].Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy,2009,72(1):190-193
[13]Walle T, Vincent T S, Walle U K. Evidence of covalent binding of the dietary flavonoid quercetin to DNA and protein in human intestinal and hepatic cells [J]. Biochemical Pharmacology,2003,65(10):1603-1610
[14]Zsila F, Bikadi Z, Simonyi M. Probing the binding of the flavonoid, quercetin to human serum albumin by circular dichroism, electronic absorption spectroscopy and molecular modelling methods [J]. Biochemical Pharmacology,2003,65 (31):447-456
[15]Kim M K, Park K S, Yeo W S, Choo H, Chong Y. In vitro solubility, stability and permeability of novel quercetin-amino acid conjugates [J]. Bioorganic & Medicinal Chemistry,2009,17(3):1164-1171
[16]许金钩,王尊本[M].荧光分析法(第三版),北京：科学出版社2006：64-69.
[17]Joseph R L. Principles of fluorescence spectroscopy(2nd), [M]. New York:Plenum Press, 1999,243.
[18]Rahman M H, Maruyama T, Okada T, Yamasaki K, Otagiri M. Study of interaction of carprofen and its enantiomers with human serum albumin-Ⅰ:Mechanism of binding studied by dialysis and spectroscopic methods [J]. Biochemical Pharmacology,1993,46 (10):1721-1731.
[19]Shuyun Bi, Daqian Song, Yuan Tian, Xin Zhou, Zhongying Liu, Hanqi Zhang. Molecular spectroscopic study on the interaction of tetracyclines with serum albumins [J]. Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy.2005,61(4): 629-636.
[20]Subramanian S,. Ross PD. Calorimetric investigation of NAD binding to some dehydrogenases [J]. Biochemical and Biophysical Research Communication, 1977,78(1):461-466.
[21]Ross P D, Subramanian S. Thermodynamics of protein association reactions:forces contributing to stability [J]. Biochemistry and Biophysical Research Communication, 1981,20(11):3096-3102.
[22]Joseph R L. Principle of Fluorescence Spectroscopy [M]. New York:Plenum Press,1999: 305-309.