分子印迹技术在药物分析中的研究与应用
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摘要
分子印迹的概念来源于人们对抗体—抗原及酶—底物的专一性的认识，著名的诺贝尔奖获得者Pauling最早提出以抗原为模板来合成抗体的设想。1949年Dickcy首次实验了染料在硅胶上的印迹，而直到20世纪70年代，人们才在有机聚合物上成功地实现了印迹。
分子印迹技术就是：第一、模板分子与功能单体按一定比例形成配合物。第二、加入交联剂形成聚合物从而配合物被固定。第三、采用一定的方法将模板分子提取出来，从而在聚合物中留下选择性的结合位点。这种高分子化合物为分子印迹聚合物(Molecularly Imprinted Polymer，简称MIP)。因为MIP具有预定性、识别性和实用性等特点，在对映体和异构体的分离、固相萃取、化学仿生传感器、模拟酶催化、临床药物分析和膜分离技术等领域展现了良好的应用前景。分子印迹聚合物在分子印迹过程中产生的识别位点常常依靠离子、氢键和金属配位等作用力选择性地结合模板分子。
本论文以分子印迹理论为指导，以不同的分子为模板合成了不同的分子印迹聚合物，系统研究了分子印迹聚合物的几个方面：
(1)MIP的评价和识别机理。以苯并三氮唑为模板分子，运用分子印迹技术合成分子印迹聚合物，通过Langmuir和Scatchard模型研究其识别机理，并讨论溶剂的极性和离子强度对识别特性的影响。同时还研究了金属配位分子印迹聚合物的识别机理。
(2)分子印迹聚合物的合成条件。不同的合成方法；不同种类和用量的功能单体、交联剂、引发剂和溶剂的加入对合成得到的分子印迹聚合物的性能有很大的影响。以络欣平为模板分子，运用大量溶剂合成微球形分子印迹聚合物，探讨合成的最佳条件，为合成性能良好的分子印迹聚合物提供实验方法。
(3)分子印迹聚合物的应用。因为MIP具有预定性、识别性和实用性等特点，所以得到越来越多的应用。以(S)-酮洛芬为模板分子利用分子印迹技术合成聚合物从而分离消旋体酮洛芬。以Zn(Ⅱ)和2，2’-联吡啶的配合物为模板合成分子印迹聚合物研究其对不同分子的选择性识别。
The opinion of molecularly imprinted polymer come from the recognition that antibody is single-minded to antigen, so is enzyme. Pauling, who ever had wined Noble prize, first planed synthesize antibody with antigen as template. Inl949, Dickey first experimented imprinting dye on silica gel. Until 1970s', people didn't succeed in imprinting molecular on organic polymer.
Molecularly imprinted polymer can be prepared as follows: First, template molecular and functional monomers are mixed in an appropriate solvent to form complex; Secondly, cross-linking monomer is added into the solution; and the complex is cross-linked; and thirdly, subsequent removal of the template molecular leaves behind cavities complementary to the template molecular that are expected to work as selective binding sites. Interactions between molecularly imprinted polymer and template molecular mostly are ionic bond, hydrogen bond or metal complex. With the characteristic of predetermination, recognition and practicability, MIPs are used in many fields, such as chiral separation of enantiomer and isomer, clinical drug analysis, enzyme-minic catalyzed reaction, solid phrase extraction, chemical bionics sensor and film separation.
With the direction of theory of molecularly imprinted polymer, this paper synthesize different polymer with template molecular and study several field of molecularly imprinted polymer in system.
(1) Evaluation and recognition mechanism of molecularly imprinted polymer. With Benztriazole as template molecular, we synthesized molecularly imprinted polymer in accord with the technology of molecular imprint; studied its recognition mechanism through the model of Langmuir and Scatchard and discussed the effects of solvent polarity and ionic intension on recognition mechanism. We also studied the recognition mechanism of metal complex molecularly imprinted polymer.
(2) Synthesizing conditions of molecularly imprinted polymer. Different synthetical
measures, various kinds and dosages of functional monomers, cross-linking monomer, initiator and solvent have different effects on the performance of molecularly imprinted polymer. With Mesylate as template molecular, we synthesized microsphere molecularly imprinted polymer with lots of solvent and discussed the optimal condition, which offered experimental measures for synthesizing best performance molecularly imprinted polymer.
(3) Appliances of molecularly imprinted polymer. With the characteristic of predetermination, recognition and practicability, MIPs are used in many fields. With (s)-ketoprofen as template molecular, we synthesized a kind of molecularly imprinted polymer, which can separate racemic ketoprofen. With Zn-2,2'-dipyriyl complex as template molecular, we also synthesized a kind of molecularly imprinted polymer and discussed its selective recognition to different molecules.
分子印迹技术就是：第一、模板分子与功能单体按一定比例形成配合物。第二、加入交联剂形成聚合物从而配合物被固定。第三、采用一定的方法将模板分子提取出来，从而在聚合物中留下选择性的结合位点。这种高分子化合物为分子印迹聚合物(Molecularly Imprinted Polymer，简称MIP)。因为MIP具有预定性、识别性和实用性等特点，在对映体和异构体的分离、固相萃取、化学仿生传感器、模拟酶催化、临床药物分析和膜分离技术等领域展现了良好的应用前景。分子印迹聚合物在分子印迹过程中产生的识别位点常常依靠离子、氢键和金属配位等作用力选择性地结合模板分子。
本论文以分子印迹理论为指导，以不同的分子为模板合成了不同的分子印迹聚合物，系统研究了分子印迹聚合物的几个方面：
(1)MIP的评价和识别机理。以苯并三氮唑为模板分子，运用分子印迹技术合成分子印迹聚合物，通过Langmuir和Scatchard模型研究其识别机理，并讨论溶剂的极性和离子强度对识别特性的影响。同时还研究了金属配位分子印迹聚合物的识别机理。
(2)分子印迹聚合物的合成条件。不同的合成方法；不同种类和用量的功能单体、交联剂、引发剂和溶剂的加入对合成得到的分子印迹聚合物的性能有很大的影响。以络欣平为模板分子，运用大量溶剂合成微球形分子印迹聚合物，探讨合成的最佳条件，为合成性能良好的分子印迹聚合物提供实验方法。
(3)分子印迹聚合物的应用。因为MIP具有预定性、识别性和实用性等特点，所以得到越来越多的应用。以(S)-酮洛芬为模板分子利用分子印迹技术合成聚合物从而分离消旋体酮洛芬。以Zn(Ⅱ)和2，2’-联吡啶的配合物为模板合成分子印迹聚合物研究其对不同分子的选择性识别。
The opinion of molecularly imprinted polymer come from the recognition that antibody is single-minded to antigen, so is enzyme. Pauling, who ever had wined Noble prize, first planed synthesize antibody with antigen as template. Inl949, Dickey first experimented imprinting dye on silica gel. Until 1970s', people didn't succeed in imprinting molecular on organic polymer.
Molecularly imprinted polymer can be prepared as follows: First, template molecular and functional monomers are mixed in an appropriate solvent to form complex; Secondly, cross-linking monomer is added into the solution; and the complex is cross-linked; and thirdly, subsequent removal of the template molecular leaves behind cavities complementary to the template molecular that are expected to work as selective binding sites. Interactions between molecularly imprinted polymer and template molecular mostly are ionic bond, hydrogen bond or metal complex. With the characteristic of predetermination, recognition and practicability, MIPs are used in many fields, such as chiral separation of enantiomer and isomer, clinical drug analysis, enzyme-minic catalyzed reaction, solid phrase extraction, chemical bionics sensor and film separation.
With the direction of theory of molecularly imprinted polymer, this paper synthesize different polymer with template molecular and study several field of molecularly imprinted polymer in system.
(1) Evaluation and recognition mechanism of molecularly imprinted polymer. With Benztriazole as template molecular, we synthesized molecularly imprinted polymer in accord with the technology of molecular imprint; studied its recognition mechanism through the model of Langmuir and Scatchard and discussed the effects of solvent polarity and ionic intension on recognition mechanism. We also studied the recognition mechanism of metal complex molecularly imprinted polymer.
(2) Synthesizing conditions of molecularly imprinted polymer. Different synthetical
measures, various kinds and dosages of functional monomers, cross-linking monomer, initiator and solvent have different effects on the performance of molecularly imprinted polymer. With Mesylate as template molecular, we synthesized microsphere molecularly imprinted polymer with lots of solvent and discussed the optimal condition, which offered experimental measures for synthesizing best performance molecularly imprinted polymer.
(3) Appliances of molecularly imprinted polymer. With the characteristic of predetermination, recognition and practicability, MIPs are used in many fields. With (s)-ketoprofen as template molecular, we synthesized a kind of molecularly imprinted polymer, which can separate racemic ketoprofen. With Zn-2,2'-dipyriyl complex as template molecular, we also synthesized a kind of molecularly imprinted polymer and discussed its selective recognition to different molecules.
引文
[1] Pauling L. J Am Chem Soc [J], 1944,62:264.
[2] Dickey F H. Proc Natl Acad Sci USA[J], 1949,36:277.
[3] Kriz D,Ramstrom O, Mosbach K. Anal Chem[J], 1997,69:165.
[4] 孟子晖，王清海，周良模等．分析化学[J]，1997，25(3)：169．
[5] Nicholls L A, Rastrom O,Mosbach K. J Chromatogr[J], I995,16:1812.
[6] Tan Z J, Remcho V T. Electrophoresis[J], 1998,19(12): 2055.
[7] Chirica G, Remcho V T. Electrophoresis[J], 1997,20(1)50.
[8] Whitecomb M J, Rodriguez M E,Villar p. J Am Chem Soci[J],1995,117:7107.
[9] Seinke J, Dunkin I R.. Adv. Polym Sci[J], 1995,123:95.
[10] Mastui J, Takeuchit. J. Anal Comm[J], 1997,16(7):199.
[11] Mastui J, Dier O D,Takeuchi T. Anal Chem Acta[J], 1997,162:1.
[12] 周良模，孟子晖，朱道乾等．分析化学[J]，1998，26(10)：1251．
[13] Vidyasankar S, Ru M, Arnold F H. J Chromatogr A[J],1997,775(1-2):51.
[14] Asanuma H, Kakazu M. Chem Comm[J], 1997,20(1):171.
[15] Kempe M. Anal Chem[J],1996,68:1948.
[16] Glad M,Reinholdsson J,Mosbach K. React Polym[J],1995,25(1):47.
[17] 孟子晖，王进防，周良模等．色谱[J]，1999，17(4)：323．
[18] Ansell R J,Mosbach K. J Chromatogr A[J],1997,787:55.
[19] Mayes A G,Mosbach K. Anal Chem[J],1996,68:3769.
[20] Schweitz L, Andersson L, Nilsson K. Anal Chem[J], 1997,69:1179.
[21] Matsui J, Kato J, Takeuchi T. Anal Chem[J],1993,65(17):2223.
[22] Nicholls I A. Chem Lett[J],1995,(11):1035.
[23] Sellergren B, Shea K J. J Chromatogr[J], 1995,690:29.
[24] Meng Z H, Wang J F, Zhou L M.Anal Sci[J], 1999,15(2): 141.
[25] Kempe M, Mosbach K. J Chromatogr A[J], 1994,664:276.
[26] Yoshikazo K, Hosoya K, Koshi Y Z. Anal Chem[J], 1998,70:386.
[27] Sellergren B, Dauwe C.J Chromatogr A[J], 1996,753:191.
[28] Fisher L, Muller R, Mosbach K. J Am Chem Soci[J], 1991,113:9358.
[29] Schweitz L, Andersson L I, Nilsson S. J Chromatogr A[J], 1997,792:441.
[30] Andersson L I, Shannessy D J, Mosbach K. J Chromatogr[J], 1990,513:167.
[31] Andersson L I, Mosbach K. J Chromatogr[J], 1990,516:313.
[32] Yu C, Mosbach K. J Org Chem[J], 1997,62(12): 4457.
[33] Bromann O, Feitag R, Whitecom M J. J Chromatogr A[J],1997,781:43.
[34] Svenson J, Andersson H S, Piletsky S A. J Mol Recognit[J], 1998,11:83.
[35] Sabourin L, Ansell R J, Mosbach K. Anal Commun[J],1998,35:285.
[36] Nisson G I, Sakaguchi K, Mosbach K. J Chromatogr A[J],1995,707:199.
[37] Sellergren B, Shea K J. J Chromatogr[J],1993,654:17.
[38] Zhou J,He X, Liy. Anal Commun[J],1999,36:243.
[39] Joshi V P, Kulkarni M G,Mashelkar R A. J Chromatogr[J],1999,849(2):319.
[40] Ramstrom O, Andersson L I, Mosbach K. J Org Chem[J],1993,58(26):7562.
[41] Nilsson K, Sakaguchi K, Mosbach K. J Chromatogr A[J],1995,707:199.
[42] jainamma M K,Shea K J,J. Am. Chem. Soc., 1996,118,8154～8155.
[43] Asanuma H,Kajiya K,Hishiya T. Chem. Lett.,1999,32(7):665～666.
[44] Matsui J,Tachibana Y, Takeuchi T. Anal.Commun., 1998,35:225～227.
[45] Spivak D,Gilmore M A,Shea K J.J.Am. Chem. Soc.,1997:4388～4393.
[46] Kriz D,Ramstrom O,Mobach K,Anal,Chem., 1995,67,2141～2144.
[47] Dicker F L,Besenbook H,Tortschanoff G,Adv. Mater., 1998,10,149.
[48] Andersson L I,Anal. Chem., 1996,68,111～117.
[49] 郭洪声、何锡文、邓昌辉．高等学校分析学报．2000，21(3)，391～396．
[50] Levi R,Piletsky S A,Cheong S H,Yano K,Anal. Chem.,1997,69(11):2017.
[51] matsai J,Okada M,Tsuruoka M,Takeuchi T, Anal. Comm.,1997,34,85～87.
[52] Baggiani C,Trotta F, Giraudi G,Giovannoli C,Anal. Commun.,1999,36(7):263～266.
[53] Olsen J,Martin P, Wilson I D,Jones G R.Analyst, 1999,124:467～471.
[54] 怀其勇，杨俊佼，雷荣等．分析测试学报，2001，20(6)：84．
[2] Dickey F H. Proc Natl Acad Sci USA[J], 1949,36:277.
[3] Kriz D,Ramstrom O, Mosbach K. Anal Chem[J], 1997,69:165.
[4] 孟子晖，王清海，周良模等．分析化学[J]，1997，25(3)：169．
[5] Nicholls L A, Rastrom O,Mosbach K. J Chromatogr[J], I995,16:1812.
[6] Tan Z J, Remcho V T. Electrophoresis[J], 1998,19(12): 2055.
[7] Chirica G, Remcho V T. Electrophoresis[J], 1997,20(1)50.
[8] Whitecomb M J, Rodriguez M E,Villar p. J Am Chem Soci[J],1995,117:7107.
[9] Seinke J, Dunkin I R.. Adv. Polym Sci[J], 1995,123:95.
[10] Mastui J, Takeuchit. J. Anal Comm[J], 1997,16(7):199.
[11] Mastui J, Dier O D,Takeuchi T. Anal Chem Acta[J], 1997,162:1.
[12] 周良模，孟子晖，朱道乾等．分析化学[J]，1998，26(10)：1251．
[13] Vidyasankar S, Ru M, Arnold F H. J Chromatogr A[J],1997,775(1-2):51.
[14] Asanuma H, Kakazu M. Chem Comm[J], 1997,20(1):171.
[15] Kempe M. Anal Chem[J],1996,68:1948.
[16] Glad M,Reinholdsson J,Mosbach K. React Polym[J],1995,25(1):47.
[17] 孟子晖，王进防，周良模等．色谱[J]，1999，17(4)：323．
[18] Ansell R J,Mosbach K. J Chromatogr A[J],1997,787:55.
[19] Mayes A G,Mosbach K. Anal Chem[J],1996,68:3769.
[20] Schweitz L, Andersson L, Nilsson K. Anal Chem[J], 1997,69:1179.
[21] Matsui J, Kato J, Takeuchi T. Anal Chem[J],1993,65(17):2223.
[22] Nicholls I A. Chem Lett[J],1995,(11):1035.
[23] Sellergren B, Shea K J. J Chromatogr[J], 1995,690:29.
[24] Meng Z H, Wang J F, Zhou L M.Anal Sci[J], 1999,15(2): 141.
[25] Kempe M, Mosbach K. J Chromatogr A[J], 1994,664:276.
[26] Yoshikazo K, Hosoya K, Koshi Y Z. Anal Chem[J], 1998,70:386.
[27] Sellergren B, Dauwe C.J Chromatogr A[J], 1996,753:191.
[28] Fisher L, Muller R, Mosbach K. J Am Chem Soci[J], 1991,113:9358.
[29] Schweitz L, Andersson L I, Nilsson S. J Chromatogr A[J], 1997,792:441.
[30] Andersson L I, Shannessy D J, Mosbach K. J Chromatogr[J], 1990,513:167.
[31] Andersson L I, Mosbach K. J Chromatogr[J], 1990,516:313.
[32] Yu C, Mosbach K. J Org Chem[J], 1997,62(12): 4457.
[33] Bromann O, Feitag R, Whitecom M J. J Chromatogr A[J],1997,781:43.
[34] Svenson J, Andersson H S, Piletsky S A. J Mol Recognit[J], 1998,11:83.
[35] Sabourin L, Ansell R J, Mosbach K. Anal Commun[J],1998,35:285.
[36] Nisson G I, Sakaguchi K, Mosbach K. J Chromatogr A[J],1995,707:199.
[37] Sellergren B, Shea K J. J Chromatogr[J],1993,654:17.
[38] Zhou J,He X, Liy. Anal Commun[J],1999,36:243.
[39] Joshi V P, Kulkarni M G,Mashelkar R A. J Chromatogr[J],1999,849(2):319.
[40] Ramstrom O, Andersson L I, Mosbach K. J Org Chem[J],1993,58(26):7562.
[41] Nilsson K, Sakaguchi K, Mosbach K. J Chromatogr A[J],1995,707:199.
[42] jainamma M K,Shea K J,J. Am. Chem. Soc., 1996,118,8154～8155.
[43] Asanuma H,Kajiya K,Hishiya T. Chem. Lett.,1999,32(7):665～666.
[44] Matsui J,Tachibana Y, Takeuchi T. Anal.Commun., 1998,35:225～227.
[45] Spivak D,Gilmore M A,Shea K J.J.Am. Chem. Soc.,1997:4388～4393.
[46] Kriz D,Ramstrom O,Mobach K,Anal,Chem., 1995,67,2141～2144.
[47] Dicker F L,Besenbook H,Tortschanoff G,Adv. Mater., 1998,10,149.
[48] Andersson L I,Anal. Chem., 1996,68,111～117.
[49] 郭洪声、何锡文、邓昌辉．高等学校分析学报．2000，21(3)，391～396．
[50] Levi R,Piletsky S A,Cheong S H,Yano K,Anal. Chem.,1997,69(11):2017.
[51] matsai J,Okada M,Tsuruoka M,Takeuchi T, Anal. Comm.,1997,34,85～87.
[52] Baggiani C,Trotta F, Giraudi G,Giovannoli C,Anal. Commun.,1999,36(7):263～266.
[53] Olsen J,Martin P, Wilson I D,Jones G R.Analyst, 1999,124:467～471.
[54] 怀其勇，杨俊佼，雷荣等．分析测试学报，2001，20(6)：84．