摘要
分子印迹技术(Molecular Imprinting Technology, MIT)是在仿生科学和模拟自然界中酶-底物及受体-抗体作用的基础之上发展起来的一项技术。其基本原理是通过模拟自然界存在的分子识别作用,在聚合物材料中引入印迹分子识别位点,制备在空间结构和结合位点上与目标分子(印迹分子)完全匹配的聚合物／膜,即分子印迹聚合物/膜(Molecularly Imprinted Polymers/Membranes, MIP/MIM),以实现对目标分子的特异性选择。随着MIT研究的不断深入和应用领域的不断扩展,MIT已经在手性拆分、固相萃取、传感器、催化和有机合成等领域得到了应用,但MIP/MIM的研究仍然存在许多问题和不足,其中之一就是制备MIP/MIM所能使用的功能单体种类还很有限。目前关于新型功能单体设计和合成的研究报道还较少,制备MIP/MIM一般都是采用十来种常规的功能单体,而可供选择的模板分子种类相对较多且结构多样,因此制备的MIP/MIM还不能很好满足分子识别的要求。
基于上述原因,本论文以非共价印迹法为基础,针对特定的目标分子(鬼臼毒素和靛玉红等)设计合成了一些可适用于分子印迹体系的新型功能单体,并把它们用来制备了一系列鬼臼毒素、靛玉红的MIP/MIM,通过MIP/MIM制备条件的优化及其性能测试,筛选出合适的功能单体。最后把优选出的MIP/MIM用于实际样品的富集分离,探讨了MIP/MIM的识别机理,为所合成的新功能单体在功能材料制备中的应用及所制备的MIP/MIM在复杂样品中的应用奠定了基础。其具体研究工作如下:
针对特定的目标分子(鬼臼毒素和靛玉红等),设计合成了14个应用于共价或非共价印迹体系的新型功能单体(分别是酰胺类功能单体、手性功能单体及具有β-酮酸酯结构和p-氨基酯单元的功能单体)并取得良好的收率,采用1HNMR及13CNMR谱等分别对其结构进行了鉴定。
以鬼臼毒素(PPT)为模板分子,自主设计合成的化合物(1-12)为新型功能单体,对鬼臼毒素分子印迹聚合物的合成条件及制备方法进行了优化。优化得到合成聚合物的最佳条件,即选用本体聚合法,以化合物2(L(-)-N-烯丙基吡咯烷基-2-羰基酰胺)为功能单体,氯仿为致孔溶剂,印迹分子、功能单体及交联剂比例为1：4:20。平衡结合实验表明,在氯仿中制备的分子印迹聚合物MIP14具有较大的吸附量和较高的印迹因子。接着实验结合超高效液相色谱(UPLC)分析方法,以MIP14为吸附剂成功用于混合样品溶液[含PPT结构类似物4’-去甲表鬼臼毒素(DMEP)]及桃儿七样品溶液中的PPT的固相萃取。在此基础上,采用表面接枝的方法(表面印迹法),考察了功能单体及交联剂用量、基膜种类、浸泡时间等因素对鬼臼毒素吸附性能的影响,制备了20个PPT分子印迹复合膜(MICMs)及相应的非印迹复合膜(NICMs)。平衡结合实验表明,以功能单体1-苯基-3-甲基-4-甲基丙烯酰基-5-吡啉酮(简称PMMP)制备的MICM对PPT具有较高的吸附容量和印迹因子。渗透实验表明,在其结构类似干扰物DMEP存在时,最佳膜MICM2对模板分子PPT表现出良好的渗透选择性。并在不同的pH条件下,研究了MICM2的渗透性能,实验研究表明MICM2具有良好的pH响应性(pH“色质”响应性)。通过扫描电镜(SEM)及氮气吸附等方法对MICM2进行了结构表征,并结合UPLC方法考察了MICM2对PPT和DMEP的甲醇混合标准样品及实际样品桃儿七甲醇提取液中鬼臼毒素的分离富集能力。
以靛玉红为模板分子,自主设计合成的化合物(1-12)为新型功能单体,通过MIP制备方法的筛选及制备条件的优化,确定了合成MIP最佳的实验条件是：采用牺牲硅胶法,以化合物4(L(-)-N-烯丙基-2-羟基酰胺)为功能单体,THF为致孔溶剂,印迹分子、功能单体及交联剂比例为1：4:30。作为对照,采用上述实验条件,以常用的甲基丙烯酸(MAA)、丙烯酰胺(AM)以及4-乙烯基吡啶(4-VP)为功能单体,合成了3个MIPs。平衡吸附实验研究表明,与实验室常用的功能单体MAA、AM及4-VP相比,以化合物4为功能单体制备的MIP20对靛玉红具有更大的印迹因子和吸附容量,表明化合物4对于模板分子靛玉红来说是合适的功能单体。以MIP20为固相萃取微柱(MISPE)的吸附剂成功实现了板蓝根样品溶液中靛玉红的分离富集。在此基础上,以靛玉红为模板分子,考察了功能单体及交联剂用量、基膜种类、浸泡时间等因素对靛玉红吸附性能的影响,制备了一系列靛玉红分子印迹复合膜(MICMs)。探讨了优化得到的分子印迹复合膜(MICM20)对靛玉红和靛蓝甲醇溶液混合标准样品及实际样品板蓝根甲醇提取液的渗透选择性,膜渗透实验结果表明,MICM20对印迹分子靛玉红具有一定的富集能力。
Molecular imprinting technique (MIT) is a technology developed on the basis of the bionic science, as well as the simulation of enzyme-substrate and antigen-antibody interactions in the nature. By introducing imprinting molecular recognition sites analogous to the natural effects of molecular recognition, a molecularly imprinted polymers/membranes (MIP/MIM) with spatial binding sites is artificially synthesized to achieve the specific selectivity to the target molecule (template molecule). With the development of the research and application concerning MIT, this technique has been widely used in many fields, such as the chiral separation, solid phase extraction, sensors, catalysis, organic synthesis, and so on. However, there are a number of unresolved problems in the MIP/MIM researches waiting for further efforts. One of them is that, while the template molecules are relatively diverse in terms of the types and structures, the number of available functional monomers is limited in the preparation of MIP/MIM. And there are few reports on the design and synthesis of new functional monomers up to now. Consequently, it is unable for the MIPs or MIMs to meet the requirements of recognition for template molecules with a wide range of molecular types and chemical structures.
The goal of this paper is to design and synthesize a number of novel monomers applicable to molecularly imprinted systems by the non-covalent imprinting method, and use them to prepare MIPs/MIMs specific to target molecules including podophyllotoxin and indirubin. After optimizing the conditions of preparation and testing the performance, the optimum MIP or MIM with suitable functional monomer was finally selected and used in the enrichment and separation of active components in complex samples. The recognition mechanism of MIP/MIM was further speculated. This study laid a solid foundation for the applications of these new functional monomers in preparation of functional materials, as well as the applications of this group of MIP/MIM for the separation of complex samples. The details of this research work are as follows:
Fourteen novel functional monomers belonging to three types, i.e. lactam functional monomer, chiral functional monomer, and functional monomer with β-keto ester structure or β-amino ester unit, were designed and synthesized for specific target molecules including podophyllotoxino, indirubin, etc. Their structures were identified by1HNMR and13CNMR spectra.
The self-designed compounds (1-12) were synthesized and used as functional monomers for the preparation of MIPs for podophyllotoxin (PPT). The preparation methods and conditions of MIPs for podophyllotoxin were optimized. The optimal conditions for preparation of polymers are as follow: the compound2used as the functional monomer, chloroform as the porogenic solvent, ratio of template molecule to functional monomer to crosslinker as1:4:20. The equilibrium binding experiments show that the molecularly imprinted polymer prepared in chloroform (MIP14) has a larger adsorption capacity and higher imprinting factor. As a result, MIP14was successfully used as an adsorbent in the solid-phase extraction for PPT in the complex sample solution. In addition, a series of molecularly imprinted composite membrane (MICMs) for PPT were also prepared by the surface grafting method (surface imprinting method). The effects of the amounts of functional monomer and crosslinker, the type of base membrane, soaking-time and other factors on the adsorption properties of PPT were investigated. The equilibrium binding experiments indicated that the MICM2prepared using the functional monomer1-phenyl-3-methyl-4-methyl propylene acyl-5-Pyrazoline ketone (PMMP) has a higher adsorption capacity and imprinting factor. Permeation experiments showed that, in the presence of its analogue DMEP, the optimum membrane MICM2exhibited a good permeating selectivity to the template molecule PPT. Especially, it was found that MICM2has good pH sensitivity (pH-responsibility) in the permeating tests. The structural characteristic of MICM2was investigated by scanning electron microscopy (SEM) and nitrogen adsorption method. Finally, the separation and enrichment capacity of MICM2in PPT-and DMEP-methanol mixture and actual samples of methanol extracts of Himalayan mayapple was also examined by ultra-high performance liquid chromatography (UPLC) method.
Compounds (1-12) were also used as the functional monomers for the preparation of MIPs for indirubin. The optimal experimental conditions of preparation of MIPs were determined as follow:Silica gel as sacrificial material, compound4as the functional monomer, THF as the porogenic solvent, ratio of template molecule to functional monomer to crosslinker as1:4:30. Moreover, methacrylic acid (MAA), acrylamide (AM) and4-vinylpyridine (4-VP) were used as functional monomer to prepareMIPs for indirubin, respectively. The equilibrium adsorption experimental studiesshowed that, compared with the general functional monomer MAA AM and4-VP, the MIP20prepared with the compound4as the functional monomer had a larger imprinting factor and adsorption capacity for indirubin. This implied that compound4is a suitable functional monomer for the template molecule indirubin. Using MIP20as sorbent in solid phase extraction micro-column (MISPE), indirubin was successfully separated and enriched from real sample solution. Amounts of functional monomer and crosslinker, the types of base membrane, soaking time duration and other factors were taken into account, a series of indirubin imprinted composite membrane (MICMs) and the control NICMs were also prepared. The selectivity of the optimum membrane (MICM20) in the standard mixture of indirubin and indigo, and in the sample solution of methanol extract of Radix Isatidis was investigated. The results of permeating experiments showed that MICM20is capable of enriching indirubin in complex solution.
引文
[1]姜忠义,喻应霞,吴洪,等,分子印迹聚合物膜的制备及应用[J],膜科学与技术,2006,26(1)：78-84.
[2]刘勤,周永新,刘荫棠,等,分子烙印传感器的研究进展[J],分析化学评述与进展1999,27(11)：1341-1347.
[3]雷建都,贺湘凌,谭天伟,等,分子印迹技术及应用研究进展[J],现代化工,2001,21(4)：17-20.
[4]Pauling, L. J., A theory of the structure and process of formation of antibodies [J], Journal of the American Chemical Society,1940,62(3):2643-2657.
[5]Wuff G., Sarhan A., Zabrocki K., Enzyme analogue built polymers and their use for the resolution of racemates [J], Tetrahedron Letters,1973,14 (44):4329-4332.
[6]Valatakis G., Andersson L., Mosbach K., et al, Drug assay using antibody mimics made by molecular imprinting [J], Nature,1993,361 (3):645-647.
[7]Luo X. B., Zhan Y. C., Huang Y. N., et al., Removal of water-soluble acid dyes from water environment using a novel magnetic molecularly imprinted polymer [J], Journal of Hazardous Materials,2011,187 (1-3):274-282.
[8]彭友元,分子印迹技术的原理与应用进展[J],泉州师范学院学报,2007,25(4) ：51-55.
[9]Dai C. M., Geissen S. U., Zhang Y. L., et al., Selective removal o f diclofenac from contaminated wate r using molecularlyimprinted polymer microspheres [J], Environmen-tal Pollution,2011,41 (2):1-7.
[10]谭淑珍,李革新,李再全,等,分子印迹技术的研究与应用[J],应用化工,2004,33(4)：4-6.
[11]Cirillo G., Curcio M., Parisi O. I., et al., Molecularly imprinted polymers for the selective extraction of glycyrrhizic acid from liquorice roots [J], Food Chemistry, 2011,125(3):1058-1063.
[12]Yang Y. Z., Liu X. G., Guo M. C., et al., Molecularly imprinted polymer on carbon microsphere surfaces for adsorbing dibenzothiophene [J], Colloids and Surfaces A,2011,377(1-3):379-385.
[13]黄健祥,胡玉斐,潘加亮,等,分子印迹样品前处理技术的研究进展[J],中国科学(B辑),2009,39(8)：733-746.
[14]曹丙庆,潘勇,赵建军,等,分子印迹聚合物的合成进展[J],现代科学仪器,2006,6：29-33.
[15]Hwang C. C., Lee W. C., Chromatographic resolution of the enantiomers of phenylpropanolamine by using molecularly imprinted polymers as the stationary phase [J], Journal of Chromatography B,2011,765 (1):45-53.
[16]Zhu X. F., Cao Q. E., Hou N. B., et al., The preparation and the recognition property of molecularly imprinted polymer of podophyllotoxin [J], Analytica Chimica Acta,2006,561 (1-2):171-177.
[17]Zhu X. F., Cao Q. E., Yang X. Q., et al., Preparation and Recognition Mechanism of gallic acid imprinted polymers [J], Helvetica Chimica Acta,2009,92:78-87.
[18]Wang G. S., Cao Q. E., Ding Z. T., et al, Preparation and Characteristics of Esculin-imprinted polymers [J], Helvetica Chimica Acta,2007,90 (6):1179-1189.
[19]Wang G. S., Cao Q. E., Xiong J., et al, Preparation and recognition properties of Vanillin-imprinted polymers [J], Helvetica Chimica Acta,2006,89(12):3032-3040.
[20]朱秀芳,曹秋娥,汪国松,等,以氢化阿魏酸为假模板制备的印迹聚合物对阿魏酸的识别[J],分析化学,2006,34(9)(特刊)：118-122.
[21]朱秀芳,汪国松,候能邦,等,咖啡因分子模板聚合物的合成及性能研究[J],云南大学学报(自然科学版),2004,26(6)：528-531.
[22]候能邦,刘院林,汪国松,等,香豆素分子模板聚合物的合成与性能研究[J],化学学报,2006,64(16)1705-1710.
[23]汪国松,朱秀芳,候能邦,等,邻香兰素分子模板聚合结合作用及选择性研究[J],云南大学学报(自然科学版),2005,27(4)：348-351.
[24]Wang G. S., Cao Q. E, Zhu, X. F., et al., Molecularly imprinted solid-phase extraction of Huperzine A from Huperzia Serrata [J], Journal of Applede Polymer Science,2009,113:3049-3058.
[25]朱秀芳,贺光艳,成会玲,等,橙皮苷印迹聚合物的识别性能及机理研究[J], 化学研究与应用，2009,21(11)：1516-1520.
[26]蒋旭红,涂伟萍,分子印迹聚合物微球制备方法研究进展[J],材料导报,2007,21(21)：52-57.
[27]Yang H. H., Zhang S. Q., Yang W., et al, Molecularly imprinted sol-gel nanotubes membrane for biochemical separations [J], Journal of America Chemistry Society,2004,126 (13):4054-4055.
[28]张学炜,孙慧等,分子印迹技术及其应用研究进展[J],天津农学院学报,2002,9(3)：23-27.
[29]Wuff G., Sarhan A., The use of polymers with enzyme-analogues structures for the resolution of racemates [J], Angewandte Chemie International Edition,1972,11 (4): 341-344.
[30]董文国,张敏莲,刘铮,等,分子印迹技术及其在生物化工领域的应用现状与展望[J],化工进展,2003,22(7)：683-688.
[31]Piletsky S. A., Piletskaya E. V., Yano K., ' et al., Biomimetic receptor system for sialic acid based on molecular imprinting [J], Analytical Letters,1996,29 (2): 157-170.
[32]张洪刚,陆书来,成国祥,等,悬浮聚合法制备磁性分子印迹聚合物微球[J],功能高分子学报,2007,19(3)：257-261.
[33]谢珍茗,何建峰,余林,等,冬凌草甲素分子印迹聚合物的制备及分子识别性能研究[J],化学研究与应用,2011,23(10)：1349-1352.
[34]Fan H. T., Li J., Li Z. C., et al., An ion-imprinted amino-functionalized silica gel sorbent prepared by hydrothermal assisted surface imprinting technique for selective removal of cadmium (Ⅱ) from aqueous solution [J], Applied Surface Science,2012, 258 (8):3815-382.
[35]陈超男,苏立强,赵珊,等,沉淀聚合法制备磺胺二甲嘧啶分子印迹聚合物[J],化工时刊，2011,25(12)：9-11.
[36]Hosoya K, Yoshizako K, Shirasu Y., et al., Uniform-size macroporous polymer-based stationary phase for high-performance liquid chromatography Structural contribution of cross-linked polymer network on specific molecular recognition [J], Journal of Chromatography A,1996,728 (1-2):139-147.
[37]李玲玲,分子印迹技术在固相萃取中的应用[J],广东化工，2011,38(11)71-78.
[38]王婧,陈元涛,王虹,等,柚皮苷分子印迹聚合物微球的制备及表征[J],安徽农业科学，2010,38(29)：16163-16165.
[39]Li Y., Li X., Chu J., et al., Synthesis of core-shell magnetic molecular imprinted polymer by the surface RAFT polymerization for the fast and selective removal of endocrine disrupting chemicals from aqueous solutions[J], Environmental Pollution, 2010,158 (6):2317-2323.
[40]R. Kryscio D., A. Peppas N., Surface imprinted thin polymer film systems with selective recognition for bovine serum albumin [J], Analytica Chimica Acta,2012, 718:109-115.
[41]Hu Y. L., Wang Y. Y., Chen X. G., et al, A novel molecularly imprinted soliid-phase microextraction fiber coupled with high performance liquid chromatography for analysis of Race estrogens in fishery samples [J], Talanta,2010, 80 (5):2099-2105.
[42]Yilmaz E., Ramstrom O., Moller P., et al., A facile method for preparing molecularly imprinted polymer spheres using spherical silica templates [J], Journal of Materials Chemistry,2002,12 (5):1577-1581.
[43]Gupta R., Kumar A., Molecular imprinting in sol-gel matrix [J], Journal of Biotechnology,2008,26:533-547.
[44]Sharma P. S., D'Souza F., Kutner W., Molecular imprinting for selective chemical sensing of hazardous compounds and drugs of abuse [J], Trends in Analytical Chemistry,2012,34:59-77.
[45]Zhang Y. Q., Jin Z. H., Wang Y.L., et al., Study on phosphorylated Zr-doped hybrid silicas/psf composite membranes for treatment of wastewater containing oil [J], Journal of Membrane Science,2010,361 (1-2):113-119.
[46]Ulbricht M., Membrane separations using molecularly imprinted polymers [J], Joumal of Chromatography B,2004,804 (1):113-125.
[47]Li H., Li Y. Z., Li Z. P., et al., Preparation and adsorption behavior of berberine hydrochloride imprinted polymers by using silica gel as sacrificed support material [J], Applied Surface Science,2012,258 (10):4314-4321.
[48]Jiang X. M., Tian W., Zhao C. D., et al., A novel sol-gel-material prepared by a surface imprinting technique for the selective solid-phase extraction of bisphenol A [J], Talanta,2007,72 (1):119-125.
[49]Spivak D. A., Simon R., Campbell J., Evidence for shape selectivity in non-convalently imprinted polymers [J], Analytica Chimica Acta,2004,504 (1):3-30.
[50]Yu C., Mosbach K., Influence of mobile phase composition and cross-linking density on the enantiomeric recognition properties of molecularly imprinted polymers [J], Joumal of Chromatography A,2000,888(1-2):63-72.
[51]陈立勇,骆玉涵,李红霞,等,用于纯化大黄素的分子印迹聚合物的制备与性能测试[J],河北理工大学学报(自然科学版),2011,33(4)：89-92.
[52]Hosoya K., Yoshizako K., Sasaki H., et al., Molecular recognition towards coplanar polychlorinated biphenyls based on the porogen imprinting effects of xylenes [J], Journal of Chromatography A,1998,828(1-2):91-94.
[53]Xu W. G., Su S. F., Jiang P., et al., Determination of sulfonamides in bovine milk with column—switching high performance liquid chromatography using surface imprinted silica with hydrophilic external layer as restricted access and selective extraction material[J], Joumal of Chromatography A,2010,1217(46):7198-7207.
[54]Tao J., Yan W., Qing D., et al., Preparation of mixed-templates molecularly imprinted polymers and investigation of the recognition ability for tetracycline antibiotics [J], Biosensors and Bioelectronics,2010,25:2219-2224.
[55]Deng F., Li Y. X., Luo X. B., et al., Preparation of conductive polypyrrole/TiO2 nanocomposite via surface molecular imprinting technique and its photocatalytic activity under simulated solar light irradiation [J], Colloids and Surfaces A: Physicochem. Eng. Aspects,2012,395:183-189.
[56]高晓非,邓英杰,曹金娜,等,六甲蜜胺脂质体的制备及影响因素考察[J],中国新药杂志,2008,17(15)：1326-1330.
[57]Li H. F., Xie C. G., Li S. Q. et al., Electropolymerized molecular imprinting on gold nanoparticle-carbon nanotube modified electrode for electrochemical detection of triazophos [J], Colloids and Surfaces B: Biointerfaces,2012,89:175-181.
[58]Fernandez-Llano L., Blanco-Lopez M. C., Lobo-Castanon J., et al., Determination of diclofenac in urine samples by molecularly-imprinted solid-phase extraction and adsorptive differential pulse voltammetry [J], Electroanalysis,2007,19 (15): 1555-1561.
[59]Kugimiya A., Takei H., Preparation of molecularly imprinted polymers with thiourea group for phosphate [J], Analytica Chimic aActa,2006,564:179-183.
[60]Rdvan, S., Arzu E., Hayrettin T., et al., Novel stereoselective molecularly imprinted polymers via ring-opening metathesis polymerisation [J], Analytica Chimica Acta.,2004,504:53-62.
[61]Yano K., Tanabe K., Takeuchi T., Matsui J., Ikebukuro K., Karube I, Molecularly imprinted polymers which mimic multiple hydrogen bonds between nucleotide bases [J],.Analytica Chimica Acta,1998,363:111-117.
[62]Hall J., Lanza-Sellergren A., Manesiotis F., Sellergren-Borje P., Non-covalent imprinting of phosphorous esters [J], Analytica Chimica Acta,2005,538:9-14.
[63]Chen C. B., Chen Y. J., Zhou J., Wu C.A.,9-vinyladenine-based molecularly imprinted polymeric membrane for the efficient recognition of plant hormone lH-indole-3-acetic acid [J], Analytica Chimica Acta,2006,569:58-65.
[64]周天佑,孟宪柱,唐倩,等,光和pH响应的功能单体的合成与表征[J],功能高分子学报,2009,22(3)：237-242.
[65]李福绵,叶卫平,冯新德,含胺基功能性单体的聚合研究-Ⅲ.甲基丙烯酸4-(N,N-二甲氨基)苄酯在过氧化月桂酰引发下的聚合动力学[J],高分子学报,1982,5:397-400.
[66]陈双基,李福绵,含胺基功能性单体的聚合研究-Ⅻ.N-丙烯酰-N′-苯基哌嗪类的合成、光聚合及其作为氧化还原引发体系组分的研究[J],高分子学报,1989,8(4)：503-508.
[67]蒋亚庆,宋宝东,宋丽,探讨2,2′-二羟甲基丁酸的合成方法[J],天津化 工,2007,21(5)：6-9.
[68]Joachim H. G., Hayrettin T, Patel A., et al., Novel stereoselective molecularly imprinted polymers via ring-opening metathesis polymerisation [J], Analytica Chimica Acta,2004,504 (1):53-62.
[69]Kriz D., Ramstom O., Svensson A., Mosbach K., A biomimetic sensor based on a molecularly imprinted polymer as a recognition element combined with fiber-optic detection [J], Analytical Chemistry,1995,67:2142-2144.
[70]刑占芬,成洪达,苏立强,等,分子印迹手性功能单体的合成及红外光谱表征[J],化学工程与装备,2008,10(10)：41-44.
[71]Luo X. B., Zhan Y. C., Tu X. M., et al., Novel molecularly imprinted polymer using 1-(a-methylacrylate)-3-methylimidazolium bromide as functional monomer for simultaneous extraction and determination of water-soluble acid dyes in waste water and soft drink by solid phase extraction and high performance liquid chromatography [J], Journal of Chromatography A,2011,1218 (8):1115-1121.
[72]Luo X. B., Zhan Y. C., Huang Y. N., et al., Removal of water-soluble acid dyes from water environment using a novel magnetic molecularly imprinted polymer [J], Journal of Hazardous Materials,2011,187 (1-3):274-282.
[1]Matsui J., Doblhoff-Dier O., Takeuchi T,2-(trifluoromethyl)acrylic acid:a novel functional monomer in non-covalent molecular imprinting [J], Analytica Chimica Acta,1997,343 (1-2):1-4.
[2]Luo X. B., Zhan Y. C., Huan Y. N., et al., Removal of water-soluble acid dyes from water environment using a novel magnetic molecularly imprinted polymer [J], Journal of Hazardous Materials,2011,187 (1-3):274-282.
[3]苏立强,王远,韩爽,等,(2-甲基丙烯酰胺)乙氧基-2-甲基丙烯酸乙二醇单酯基质分子印迹手性分离介质的合成与表征[J],分析化尝2008,36(12):1732-1734.
[4]Zhu Q., Dai J., Gu X., et al., Synthesis and characteristics of imprinted 17-β-estradiol microparticle and nanopartilce with TFMAA as functional monomer [J], Journal of Applied Polymer Science,2007,104:1551-1558.
[5]高吉刚,陈长宝,王荣荣,等,新型长侧链双羧基功能单体N-(P-乙烯基苄基)-N,N-二[2-(3-羧基)乙基]胺的合成及其对牛血.清白蛋白印迹识别的应用[J],化学学报,2008,66(9)：1067-1073.
[6]Yano K., Nakagiri T., Takeuchi T., et al., Stereoselective recognition of dipeptide derivatives in molecularly imprinted polymers which incorporate anlvaline derivative as a novel functional monomer [J], Analytica Chimica Acta,1997,357(1-2):91-98.
[7]孙晶,苏立强,王俊涛,新型分子烙印手性功能单体的合成[J]，齐齐哈尔大学学报,2004,20(4)：1-4.
[8]Bumpus J. A., Lewis A., Stotts C., Characterization of high explosives and other energetic compounds by computational chemistry and molecular modeling [J], Journal of Chemical Education,2007,84 (2):329-332.
[9]Liu Y., Wang R., Tan T.W., et al., Study of the properties of molecularly imprinted polymers by computational and conformational analysis [J], Analytica Chimica Acta, 2007,581 (1):137-146.
[10]Li Z., Chen G.., Construction of new system for computational chemistry course [J], University Chemistry,2008,23 (1):11-16.
[11]白银娟,路军,甘海婴,等,无溶剂微波促进下1-苯基-3-甲基-5-吡唑啉酮与芳酰氯的反应研究[J],有机化学，2002,22(9):638-641.
[1]Jiang X. M., Zhao C. D., Jiang N., et al., Selective solid-phase extraction using molecular imprinted polymer for the analysis of diethylstilbestrol [J], Food Chemistry, 2008,108 (3):1061-1067.
[2]王晓慧,张艳梅,于浩强,等,分子印迹聚合物的制备及其应用[J],上海塑群,2011,3：1-4.
[3]Lin Z., Yang F., He X. W., et al., Preparation and evaluation of a macroporous molecularly imprinted hybrid silica monolithic column for recognition of proteins by high performance liquid chromatography [J], Journal of Chromatography A,2009, 1216 (49):8612-8622.
[4]徐志祥,方国臻,张燕,分子印迹免疫分析技术及其应用研究进展[J],现代化工,2008,28(4)：32-35.
[5]Puoci F., lemma R., Cirillo G., et al., New restricted access materials combined to molecularly imprinted polymers for selective recognition/release in water media [J], European Polymer Journal,2009,45(6):1634-1640.
[6]Benito-Pena E., Martins S., Orellana G., et al., Water-compatible molecularly imprinted polymer for the selective recognition of fluoroquinolone antibiotics in biological samples [J], Analytical and Bioanalytical Chemistry,2009,393:235-45.
[7]Cirillo G., lemma F., Puoci F., et al., Imprinted hydrophilic nanospheres as drug delivery systems for 5-fluorouracil sustained release [J]., Journal of Drug Targeting, 2009:17 (1):72-78.
[8]Sueyoshi Y., Fukushima C., Yoshikawa M., Molecularly imprinted nanofiber membranes from cellulose acetate aimed for chiral separation [J], Journal of Membrane Science,2010,357 (1-2):90-97.
[9]B. Altintas E., Turkmen D., Karakoc V., et al., Hemoglobin binding from human blood hemolysate with poly(glycidyl methacrylate) beads [J], Colloids and Surfaces B:Biointerfaces,2011,85 (2):235-240.
[10]Chai C. Y., Liu G. Y., Yao B., A portable optical sensor based on a one-off test strip for fast evaluation of bacterial contamination in raw tofu [J], Sensors and Actuators B,2011,152(1):1-7.
[11]陈燕,德吉,刘倩伶,等,桃儿七果实中鬼臼毒素的鉴别和含量测定[J],华西药学杂志,2009,24(1)：93-94.
[12]王宝玺,王日升,汪青,等,植物桃儿七中鬼臼毒素的提取及HPLC分析[J],中国医药学工业杂志,1999,11：484-486.
[13]张杰,周春山,向大雄,等,高效液相色谱法测定小八角莲中鬼臼毒素的含量[J],湖南中医杂志,2007,23(4)：101-102.
[14]肖昌钱,杨淑贞,单文,等,HPLC比较不同产地生药八角莲中鬼臼类成分含量[J],中国现代应用药学杂志,2009,26(8)：659-661.
[15]王晓玲,杨伯伦,刘谦光,等,鬼臼毒素的提取及其溶剂加合物的表征[J],中草药，2003,3(4)：330-331.
[16]温普红,桃儿七中鬼臼毒素的分离与纯化[J],宝鸡文理学院学报(自然科学版,2002,22(4)：277-282.
[17]Liu B. M., Yan H. Y., Qiao F.X., et al., Determination of clenbuterol in porcine tissues using solid-phase extraction combined with ultrasound-assisted dispersive liquid-liquid microextraction and HPLC-UV detection [J], Journal of Chromatography B,2011,879 (1):90-94.
[18]Ran D., Wang Y. Z., Jia X. P., et al., Bovine serum albumin recognition via thermosensitive molecular imprinted macroporous hydrogels prepared at two different temperatures [J], Analytica Chimica Acta,2012,723:45-53.
[19]Pedrouzo M., Borrull F., Marce R. M., et al., Analytical methods for personal-care products in environmental waters [J], Trends in Analytical Chemistry, 2011,30 (5):749-760.
[20]Bozzetti F., Yu W., Baratti D., et al., Locoregional treatment of peritoneal carcinomatosis from gastric cancer [J], Journal of Surgical Oncology,2008,98: 273-276.
[21]Barni S., Cabiddu M., Ghilardi M., et al., A novel perspective for an orphan problem:Old and new drugs for the medical management of malignant ascites [J], Critical Reviews in Oncology/Hematology,2011,79 (2):144-153.
[22]Esquivel J., Elias D., Baratti D., et al., Consensus statement on the loco regional treatment of colorectal cancer with peritoneal dissemination [J], Journal of Surgical Oncology,2008,98:263-267.
[23]Wu H., Zhao Y. Y., Nie M. C., et al., Molecularly imprinted organic-inorganic hybrid membranes for selective separation of phenylalanine isomers and its analogue [J], Separation and Purification Technology,2009,68 (1):97-104.
[24]王颖,李楠,分子印迹技术及其应用[J],化工进展,2010,29(12)：2315-2323.
[25]Qi P. P., Wang J. C., Wang L.D., et al., Molecularly imprinted polymers synthesized via semi-covalent imprinting with sacrificial spacer for imprinting phenols [J], Polymer,2010,51:5417-5423.
[26]戴玉杰,卢战胜,焦良,等,二烯丙基胺为功能单体的分子印迹聚合物合成及分子识别性能[J],天津科技大学学报,2009,24(3)：1-5.
[27]Tekin K., Uzun L., Denizli A., et al., Preparation and characterization of composite cryogels containing imidazole group and use in heavy metal removal [J], Reactive and Functional Polymers,2011,71 (10):985-993.
[28]He J. F., Zhu Q. H., Deng Q. Y., Investigation of imprinting parameters and their recognition nature for quinine-molecularly imprinted polymer [J], Spectrochimica Acta Part A:Molecular and Biomolecular Spectroscopy,2007,67 (5):1297-1305.
[29]Baniceru M., Manda C. V., Popescu S. M., Chromatographic analysis of local anesthetics in biological samples [J], Journal of Pharmaceutical and Biomedical Analysis,2011,54 (1):1-12.
[30]Hoizey G., Lamiable D., Gozalo C., et al., Millart H., Mauprivez, C., Determination of articaine in human plasma by liq-uid chromatography-mass spectrometry and its application in a preliminary pharmacokinetic study [J], Journal of Pharmaceutical and Biomedical Analysis,2009,49 (4):1082-1087.
[31]Ulbricht M., Membrane separations using molecularly imprinted polymers [J], Journal of Chomatography B,2004,804 (1):113-125.
[32]邱增英,钟世安,烟酸分子印迹复合膜的制备及其分离性能研究[J],化学学报,2010,68(3)：246-250.
[33]董佳,高晓磊,李亮晨,等,对氯苯酚分子印迹复合膜的制备及其表征[J],上海师范学学报自然科学版),2011,40(6)：615-618.
[34]彭晓霞,迟栋,龚来觐,分子印迹复合膜在中草药及天然药物提取分离中的应用概述[J],中国药事，2009,23(10)：1011-1014.
[35]Juang R. S., Huang W. C., Hsu Y. H., Treatment of phenol in synthetic saline wastewater by solvent extraction and two-phase membrane biodegradation[J], Journal of Hazardous Material,2009,164 (1):46-52.
[36]Yoshimi Y., Arai R., Nakayama S., Influence of the solvent on nature of gate effect in molecularly imprinted membrane [J], Analytica Chimica Acta,2010,682 (1-2):110-116.
[37]陈晖,分子印迹复合膜在中药活性成分分离纯化中的应用[J],西北药学杂志,2009,24(4)：336-338.
[38]] Huang X. D., Zou N. F., Chen X. M., et al., Molecularly imprinted monolithic stationary oboes for liquid chromatographic · separation of enantiomers and diastereomers [J], Journal of Chromatography A,2003,984 (2):273-282.
[39]卢春阳,马向霞,何锡文,等,邻香草醛分子印迹聚合物膜的制备及其透过选择性质的研究[J],化学学报,2005,63(6)：479-483.
[40]张春静,钟世安,醋酸纤维-奎宁分子印迹复合膜的制备及分离性能研究[J],膜科学与技术,2008,28(4)：59-62.
[41]Jiang Z. Y., Ying Y. X., Wu H., Preparation of CS/GPTEMS hybrid molecularly imprinted membrane for efficient chiral resolution of phenylalanine isomers, Journal of Membrane Science,2006,280 (1-2):876-882.
[42]Liang X. Q., Pu X. M., Zhou H. W., et al., Keto-enol tautomerization of cyanuric acid in the gas phase and in water and methanol [J], Journal of Molecular Structure, 2007,816 (1-3):125-136.
[43]Yao X. S., in:L. J. Wu (Ed.), Natural products chemistry [M], People's Medical Publishing House, Beijing, China, 2000,256-257.
[44]Alfredson T., Sheehan T., Lenert T., et al., Determination of compound identity by group type and class using diode-array detection in high-performance liquid chromatography [J], Journal of Chromagraphy A,1987,385:213-223.
[45]Deng F., Li Y. X., Luo X. B., et al., Preparation of conductive polypyrrole/TiO2 nanocomposite via surface molecular imprinting technique and its photocatalytic activity under simulated sola light irradiation [J], Colloids and Surfaces A: Physicochem. Eng. Aspects,2012,395:183-189.
[46]Songa G., Kellam M. E., Lianga D., et al., Influence of processing conditions on the microstmcture and permeability of BCC V-Ni membranes [J], Journal of Membrane Science,2010,363 (1-2):309-315.
[47]Liu J. M., Lin L. P., Wang X. X., et al., A highly sensitive coupling technique for the determination of trace quercetin based on solid substrate room temperature phosphorimetry and poly (vinylalcohol) complex imprinting [J], Analytica Chimica Acta,2012,723:76-82.
[48]Kusunoki T., Kobayashi T., Molecular imprinting micropolymer beads having cooperative effect of both surfactant and inosine template [J], Journal of Applied Polymer Science,2010,117:565-571.
[49]Qi P. P., Wang J. C., Wang L.D., et al., Molecularly imprinted polymers synthesized via semi-covalent imprinting with sacrificial spacer for imprinting phenols [J], Polymer,2010,51 (23):5417-5423.
[50]Hillberg A. L., Brain K. R., Allender C. J., Design and evaluation of thin and flexible theophylline imprinted polymer membrane materials [J], Journal of Molecular Recognition,2009,22:223-231.
[51]韦情情,李怀祥,武怀娣,等,多孔硅表面组装胆固醇分子印迹膜的光电化学[J],纳米科技,2011,8(6)：5-9.
[52]张杰,周春山,向大雄,等,高效液相色谱法测定小八角莲中鬼臼毒素的含量[J],湖南中医杂志,2007,23((4)：101-110.
[53]Sloop J. C., Bumgardner C. L., Washington G., et al., Keto-enol and enol-enol tautomerism in trifluoromethyl-β-diketones [J], Journal Fluorine Chemistry,2006, 127 (6):780-786.
[1]谢芳,靛玉红提取分离及定量测定的研究[J],云南中医中药杂志，2008,29(4)：46-47.
[2]杜萍,刘少娟,刘永俊,复方板蓝根颗粒中靛玉红的含量测定[J],齐鲁药事，2010,29(6)：338-339.
[3]朱丽君,穆建生,HPLC法测定牛黄消炎片中靛玉红的含量[J],中国民康医学,2010,22(12)：1522-1523.
[4]谢国祥,张立国,邱明丰,等,大青叶中靛玉红的提取分离工艺研究[J],中成药，2006,28(6)：791-793.
[5]马莉,孙琴,李友,等,HPLC法测定板蓝根药材及制剂中靛蓝和靛玉红含量[J],药物分析杂志，2010,30(9)：1642-1645.
[6]颜艺周,邹品文,HPLC法测定板蓝根颗粒中靛玉红的含量[J],中国药房,2008,19(3)：211-213.
[7]白宗利,任玉珍,陈彦琳,等,HPLC测定市售青黛中靛蓝和靛玉红的含量[J],中国现代中药，2010,12(8):27-32.
[8]Hoshino Y., Koide H., Urakami T., et al., Recognition, neutralization, and clearance of target peptides in the blood-stream of living mice by molecularly imprinted polymer nanoparticles:a plastic antibody [J], Journal of American Chemistry Society,2010,132 (19):6644-6645.
[9]Claude B., Morin P., Lafosse M., et al., Selective solid-phase extraction of a triterpene acid from a plant extract by molecularly imprinted polymer [J], Talanta, 2008,75 (2):344-350.
[10]Janiak D. S., Ayyub O. B., Kofinas P., Effects of charge density on the recognition properties of molecularly imprinted polyampholyte hydrogels [J], Polymer Science, 2010,51 (3):665-670.
[11]Zhao P. N., Yu J. H., Liu S. Q., et al., One novel chemiluminescence sensor for determination of fenpropathrin based on molecularly imprinted porous hollow microsphere [J], Sensors and Actuators B,2012,162(1):166-172.
[12]Pan J., Xue X. H., Wang J. H., et al., Recognition property and preparation of Staphylococcus aureus protein A-imprinted polyacrylamide polymers by inverse-phase suspension and bulk polymerization [J], Polymer,2009,50(11):2365-2372.
[13]Li Y., Li X., Dong C. K., et al., Selective recognition and removal of chlorophenols from aqueous solution using molecularly imprinted polymer prepared by reversible addition-fragmentation chain transfer polymerization [J], Biosensors and Bioelectronics,2009,25(2):306-312.
[14]Hirayama K., Sakai Y., Kameoka K., Synthesis of polymer particles with specific lysozyme recognition sites by a molecular.imprinting technique [J], Journal of Applied Polymer Science,2001,81 (14):3378-3387.
[15]Fu G. Q., Yu H., Zhu J., Imprinting effect of protein-imprinted polymers composed of chitosan and polyacrylamide:a re-examination [J], Biomaterials,2008, 29 (13):2138-2142.
[16]Pakade V., Lindahl S., Chimuka L.. et al., Molecularly imprinted polymers targeting quercetin in high-temperature aqueous solutions [J], Journal of Chromatography A,2012,1230:15-23.
[17]朱林敏,朱文佳,周锋伟,等,紫杉醇分子印迹聚合物的制备及其吸附性能[J],大连工业大学学报，2011,30(4):246-249.
[18]Gao J. F., Tian H. J., Wang Y., et al., The design of protein-imprinted polymers as antibody substitutes forinvestigating protein eprotein interactions [J], Biomaterials, 2012,33 (11):3344-3352.
[19]Tov O. Y., Luvitch S., Bianco-Peled H., Molecularly imprinted hydrogel displaying reduced non-specific binding and improved protein recognition [J], Journal of Separation Science,2010,33(11):1673-1681.
[20]Wang Z. H., Li H., Chen J., et al., Acetylsalicylic acid electrochemical sensor based on PATP-AuNPs modified molecularly imprinted polymer film [J], Talanta, 2011,85 (3):1672-1679.
[21]Silvestri D., Barbani N., Cristallini C., et al., Molecularly imprinted polymer membranes for an improved recognition of biomolecules in aqueous medium [J], Journal of membrane science,2006,282 (1-2):284-295.
[22]Wang J. Y., Xu Z.L., Wu P., et al., Chemical binding constant and transport property of S-Naproxen molecularly imprinted composite membrane [J], Journal of Membrane Science,2009,331 (1-2):84-90.
[23]Tasselli F., Donato L., Drioli E., Evaluation of molecularly imprinted membranes based on different acrylic copolymers [J], Journal of Membrane Science,2008,320 (1-2):167-172.
[24]Takeda K., Uemura K., Kobayashi T., Hybrid molecular imprinted membranes having selectivity and separation behavior to targeted indole derivatives [J], Analytica Chimica Acta,2007,591 (1):40-48.
[25]张茂升,黄佳蓉,郑向华,等,孔雀石绿分子印迹膜的制备和渗透性[J],应用化花，2010,27(1)：107-111.
[26]Faizal C.K., Hoshina Y., Kobayashi T., Scaffold membranes for selective adsorption of a-tocopherol by phase inversion covalently imprinting technique [J], Journal of Membrane Science,2008,322 (2):503-511.
[27]Sergeyeva T.A., Matuschewski H., Piletsky S.A. et al., Molecularly imprinted polymer membranes for substance-selective solid-phase extraction from water by surface photo-grafting polymerization [J], Journal of Chromatography A,2001,907 (1-2):89-99.
[28]刘美华,邱彬,陈国南,等,17p-雌二醇乙酸纤维分子印迹复合膜的制备及性能研究[J],福建分析测试，2011,20(6)：1-5.
[29]管萍,胡小玲,郝明燕,红霉素分子印迹膜的制备及性能研究[J],功能材料,2010,379-381.
[30]范培民,王兵,槲皮素金属配位印迹聚合物膜的制备及性能研究[J],化学学报,2010,68(24)：2543-2550.
[31]熊远钦,杨晓静,刘含茂,等,相转移法制备熊果酸分子印迹膜[J],湖南大学学报(自然科学版),2010,37(2)：60-63.
[32]Trotta F., Baggiani C., Luda M. P., et al, A molecular imprinted membrane for molecular discrimination of tetracycline hydro-Chloride [J], Journal of Membrane Science,2005,254 (1):13-19.
[33]Wang C. Y., Chen Y. C., Sheu D. C., et al, Molecularly imprinted polymers for the recognition of sodium dodecyl sulfate denatured creatine kinase [J], Journal of the Taiwan Institute of Chemical Engineers,2012,43:188-194.
[34]Sibrian-Vazquez M., Spivak D. A., Enhance enantioselectivity of molecularly imprinted polymers formulated with novel cross-linking monomers [J] Macromolecules,2003,36:5105-5113.
[35]Norrlow O., Glad M., Mosbach K., Acrylic polymer preparation containing recognition sites obtained by imprinted with substrates [J], Journal of Chromatograp-hy A,1984,299 (1):29-41.
[36]Viswanathan S., Rani C., Ribeiro S., et al, Molecular imprinted nanoelectrodes for ultra sensitive detection of ovarian cancer marker [J], Biosensors and Bioelectronics,2012,33 (1):179-183.
[37]Hu Y. L., Wang Y. Y., Hu Y. F., et al., Liquid-liquid-solid microextraction based on membrane-protected molecularly imprinted polymer fiber for trace analysis of triazines in complex aqueous samples [J], Journal of Chromatography A,2009, 1216(47):8304-8311.
[38]Chimuka L., Pinxteren M. V., Billing J., el al., Selective extraction of triazine herbicides based on a combination of membrane assisted solvent extraction and molecularly imprinted solid phase extraction [J], Journal of Chromatography A,2011, 1218(5):647-653.
[39]Mhaka B., Cukrowska E., Bui B. T. S., et al., Selective extraction of triazine herbicides from food samples based on a combination of a liquid membrane and molecularly imprinted polymer [J], Journal of Chromatography A,2009,1216 (40): 6796-6801.
[40]Wang J. Y., Xu Z. L., Wu P., et al., Binding constant and transport property of S-Naproxen molecularly imprinted composite membrane [J], Journal of Membrane Science,2009,331 (1):84-90.
[41]Balamurugan K., Gokulakrishnan K., Prakasam T., Preparation and evaluation of molecularly imprinted polymer liquid chromatography column for the separation of Cathine enantiomers [J], Saudi Pharmaceutical Journal,2012,20:53-61.
[42]谢友良,何百寅,李远彬,等,青黛药材中的靛蓝和靛玉红含量的同时测定[J],中药新药与临床药理，2011,22(4)：452-455.
[43]陈冬金,孙世坤,张红云,复方板蓝根提取工艺与靛玉红含量髓[J],福建畜牧兽医，2011,33(5)：8-10.
[44]吴彦,超临界C02萃取板蓝根靛玉红的工艺优化研究[J],光谱实验室，2010,27(2)：732-736.
[45]黄以钟,潘大仁,王占成,等,不同生长时期对马蓝药效成分的影响[J],中国农学通报,2009,25(16)：75-78.
[46]罗丹冬,丘振文,HPLC法对比不同产地南板蓝根药材靛蓝靛玉红含量的研究[J],现代生物医学进展,2010,10(14)：2720-2722.
[47]Asliyuce S., Uzun L., Rad A. Y., et al., Denizli A., Molecular imprinting based composite cryogel membranes for purification of anti-hepatitis B surface antibody by fast protein liquid chromatography [J], Journal of Chromatography B,2012,889-890: 95-102.