分子印迹材料及量子点在分析化学中的应用
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摘要
随着生命科学、生物工程和环境科学等学科的迅速发展，以及各种工业生产过程中污染物排放日益增加，分析科学同时面临着新的机遇和挑战。分析对象日益复杂多样，对复杂基体中痕量和超痕量组分的分离和检测成为突出的问题。对于成分比较复杂的环境样品和生物样品，需要经过一定的分离及富集以后才能进入分析仪器进行准确的测定。而样品的富集分离通常需要借助选择性高、吸附容量大的各种吸附材料和合适的分离手段。因此，本论文针对成分复杂的环境样品，以建立痕量组分的富集分离及分析为目的，合成了几种功能化吸附分离材料，并对材料的选择性吸附性能开展了系统研究，主要进行了以下几方面的创新性研究工作。
1．以环境雌激素污染物双酚A为模板分子，以SiO_2颗粒为载体，以3-氨基丙基-三乙氧基硅烷为功能单体合成了表面分子印迹材料，并使用了FT-IR和静态吸附实验对材料进行了表征。将制备的印迹材料应用到固相萃取技术中，研究了分子印迹固相萃取柱与商品C8固相萃取柱富集分离双酚A分子的优劣性。并将分子印迹固相萃取与高效液相色谱仪器联用，分析了水样中的痕量双酚A。
2．以环境雌激素污染物己烯雌酚(DES)为模板分子，以SiO_2颗粒为载体，以3-氨基丙基-三乙氧基硅烷为功能单体合成了表面分子印迹材料，并使用了FT-IR和静态吸附试验对材料进行了表征。制备的己烯雌酚分子印迹材料对DES的最大静态吸附值达到62．58mg g~(-1)。当溶液中DES和干扰物的浓度为50 mg L~(-1)时，该材料对DES的选择因子达到61．7。吸附动力学实验显示，DES印迹材料对DES的吸附在10min内完成。将制备的分子印迹材料应用到固相萃取技术中，并与高效液相色谱仪器联用，测定了鱼肉样品中DES的含量。
3．以己烯雌酚(DES)为模板分子，以甲基丙烯酸为功能单体，以EGDMA为交联剂，分别以块聚合成方法和微乳聚合的方法合成了印迹块状材料(MIBs)和纳米印迹材料(MINs)。MINs材料的颗粒直接约在100nm左右。通过透射电镜和静态吸附试验显示，与传统块聚合方法制备的MIPs材料相比，乳液聚合不但实验方法简便，而且MINs材料具有更好的形貌和更高的吸附容量。
4．使用水相合成法、以巯基乙酸为稳定剂制备了三种不同发光的水溶性CdTe量子点溶液，分别使用透射电镜(TEM)、荧光光谱(FL)和紫外光谱(UV)技术对该量子点进行了表征。选取其中一种量子点与胰岛素分子的相互作用体系为研究对象，采用了荧光光谱技术对量子点和生物分子之间的相互作用进行了探讨。通过对胰岛素对CdTe的淬灭常数的计算，判断出胰岛素对CdTe的淬灭属于静态淬灭行为。另外，通过对不同盐浓度和酸度环境下淬灭效率的实验，得出以巯基乙酸为稳定剂的CdTe量子点和胰岛素之间的相互作用以静电作用为主。
With the rapid development of science and technology, analytical science isfacing the new opportunity and rigorous challenge. Sensitive, rapid, reproducible,simple and accurate analytical methods are required for the determination of tracecomponents in geological, biological and environmental samples. Although manyinstrument analysis methods possessing higher sensitivity and selectivity have beendeveloped recently, pre-concentration and analysis of trace elements is still necessarybefore the more accurate measurement by analytical instruments in reality. Thepre-concentration and analysis of trace elements will depend on different adsorptionmaterials with higher selectivity and larger adsorption capacity and appropriateseparation methods. In this research paper, several new adsorption materials aresynthesized in order to pre-concentration and separation of trace components. Theiradsorption and selectivity characteristics for trace components have been investigatedsystematically. The more detailed novelty of this research can be categorized asfollowing:
1. A novel and simple imprinted amino-functionalized silica gel material wassynthesized by combining a surface molecular imprinting technique with a sol-gelprocess on the supporter of activated silica gel for solid-phase extraction-highperformance liquid chromatography (SPE-HPLC) determination of bisphenol A(BPA). The BPA-imprinted silica sorbent and non-imprinted silica sorbent werecharacterized by FT-IR and the static adsorption experiments. The preparedBPA-imprinted silica sorbent showed high adsorption capacity, significant selectivityand good site accessibility for BPA. Furthermore, the difference of the retentioncharacteristics of BPA on the C8 SPE column and BPA-imprinted silica SPE(MIP-SPE) was compared. The MIP-SPE-HPLC method showed higher selectivity toBPA than the traditional SPE-HPLC method. At last, the BPA-imprinted polymerswere used as the sorbent in solid-phase extraction to determine BPA in water samples.
2. A simple imprinted amino-functionalized silica gel material was synthesized bycombining a surface molecular imprinting technique with a sol-gel process forsolid-phase extraction-high performance liquid chromatography (SPE-HPLC)determination of diethylstilbestrol (DES). Activated silica gel was used as thesupporter and non-imprinted silica sorbent was synthesized without the addition ofDES using the same procedure as that of DES-imprinted silica sorbent. The maximumstatic adsorption capacity of the DES-imprinted and non-imprinted silica sorbent forDES was 62.58 mg g~(-1) and 19.89 mg g~(-1), respectively. The relatively selective factorvalue of this DES-imprinted silica sorbent was 61.7 at the level of 50 mg L~(-1). And theuptake kinetics was fairly rapid so that the adsorbent equilibrium was achieved within10 min. Furthermore, the DES-imprinted polymers were used as the sorbent insolid-phase extraction to determine DES in fish samples.
3. Molecularly imprinted bulk polymers (MIBs), prepared by bulk polymerizationand molecularly imprinted nanospheres (MINs), prepared by novel precipitationpolymerization method were synthesized in this work. The polymers were preparedwith diethylstilbestrol (DES) as the template molecule, methacrylic acid as thefunctional monomer and ethylene glycol dimethacrylate as the cross-linking monomerin the presence of acetonitrile solvent. The diameter of the MINs for DES wasapproximate 100 nm. These polymers were characterized by Fourier transforminfrared spectroscopy, transmission electron microscope and the static adsorptionexperiments. The prepared DES-imprinted nanospheres showed regularly shape andbetter adsorption capacity than DES-imprinted polymers.
4. Three CaTe quantum dots were synthesized in an aqueous phase usingthioglycolic acid as capped reagent. TEM, FL and UV were used to characterize theshape and optical properties. Insulin and CdTe QDs were used to study the interactionof protein with QDs as model samples. Fluorescence spectroscopy was used toinvestigate the stoichiometry and thermodynamic parameters of interaction. Theresults showed that the fluorescence of CdTe was quenched by insulin. The quenchingmechanism was discussed to be a static quenching procedure. Moreover, based on theevaluation of association constant of various the ionic strength and pH value, we speculated that the interaction of CdTe with insulin was mainly due to electrostaticattraction.
1．以环境雌激素污染物双酚A为模板分子，以SiO_2颗粒为载体，以3-氨基丙基-三乙氧基硅烷为功能单体合成了表面分子印迹材料，并使用了FT-IR和静态吸附实验对材料进行了表征。将制备的印迹材料应用到固相萃取技术中，研究了分子印迹固相萃取柱与商品C8固相萃取柱富集分离双酚A分子的优劣性。并将分子印迹固相萃取与高效液相色谱仪器联用，分析了水样中的痕量双酚A。
2．以环境雌激素污染物己烯雌酚(DES)为模板分子，以SiO_2颗粒为载体，以3-氨基丙基-三乙氧基硅烷为功能单体合成了表面分子印迹材料，并使用了FT-IR和静态吸附试验对材料进行了表征。制备的己烯雌酚分子印迹材料对DES的最大静态吸附值达到62．58mg g~(-1)。当溶液中DES和干扰物的浓度为50 mg L~(-1)时，该材料对DES的选择因子达到61．7。吸附动力学实验显示，DES印迹材料对DES的吸附在10min内完成。将制备的分子印迹材料应用到固相萃取技术中，并与高效液相色谱仪器联用，测定了鱼肉样品中DES的含量。
3．以己烯雌酚(DES)为模板分子，以甲基丙烯酸为功能单体，以EGDMA为交联剂，分别以块聚合成方法和微乳聚合的方法合成了印迹块状材料(MIBs)和纳米印迹材料(MINs)。MINs材料的颗粒直接约在100nm左右。通过透射电镜和静态吸附试验显示，与传统块聚合方法制备的MIPs材料相比，乳液聚合不但实验方法简便，而且MINs材料具有更好的形貌和更高的吸附容量。
4．使用水相合成法、以巯基乙酸为稳定剂制备了三种不同发光的水溶性CdTe量子点溶液，分别使用透射电镜(TEM)、荧光光谱(FL)和紫外光谱(UV)技术对该量子点进行了表征。选取其中一种量子点与胰岛素分子的相互作用体系为研究对象，采用了荧光光谱技术对量子点和生物分子之间的相互作用进行了探讨。通过对胰岛素对CdTe的淬灭常数的计算，判断出胰岛素对CdTe的淬灭属于静态淬灭行为。另外，通过对不同盐浓度和酸度环境下淬灭效率的实验，得出以巯基乙酸为稳定剂的CdTe量子点和胰岛素之间的相互作用以静电作用为主。
With the rapid development of science and technology, analytical science isfacing the new opportunity and rigorous challenge. Sensitive, rapid, reproducible,simple and accurate analytical methods are required for the determination of tracecomponents in geological, biological and environmental samples. Although manyinstrument analysis methods possessing higher sensitivity and selectivity have beendeveloped recently, pre-concentration and analysis of trace elements is still necessarybefore the more accurate measurement by analytical instruments in reality. Thepre-concentration and analysis of trace elements will depend on different adsorptionmaterials with higher selectivity and larger adsorption capacity and appropriateseparation methods. In this research paper, several new adsorption materials aresynthesized in order to pre-concentration and separation of trace components. Theiradsorption and selectivity characteristics for trace components have been investigatedsystematically. The more detailed novelty of this research can be categorized asfollowing:
1. A novel and simple imprinted amino-functionalized silica gel material wassynthesized by combining a surface molecular imprinting technique with a sol-gelprocess on the supporter of activated silica gel for solid-phase extraction-highperformance liquid chromatography (SPE-HPLC) determination of bisphenol A(BPA). The BPA-imprinted silica sorbent and non-imprinted silica sorbent werecharacterized by FT-IR and the static adsorption experiments. The preparedBPA-imprinted silica sorbent showed high adsorption capacity, significant selectivityand good site accessibility for BPA. Furthermore, the difference of the retentioncharacteristics of BPA on the C8 SPE column and BPA-imprinted silica SPE(MIP-SPE) was compared. The MIP-SPE-HPLC method showed higher selectivity toBPA than the traditional SPE-HPLC method. At last, the BPA-imprinted polymerswere used as the sorbent in solid-phase extraction to determine BPA in water samples.
2. A simple imprinted amino-functionalized silica gel material was synthesized bycombining a surface molecular imprinting technique with a sol-gel process forsolid-phase extraction-high performance liquid chromatography (SPE-HPLC)determination of diethylstilbestrol (DES). Activated silica gel was used as thesupporter and non-imprinted silica sorbent was synthesized without the addition ofDES using the same procedure as that of DES-imprinted silica sorbent. The maximumstatic adsorption capacity of the DES-imprinted and non-imprinted silica sorbent forDES was 62.58 mg g~(-1) and 19.89 mg g~(-1), respectively. The relatively selective factorvalue of this DES-imprinted silica sorbent was 61.7 at the level of 50 mg L~(-1). And theuptake kinetics was fairly rapid so that the adsorbent equilibrium was achieved within10 min. Furthermore, the DES-imprinted polymers were used as the sorbent insolid-phase extraction to determine DES in fish samples.
3. Molecularly imprinted bulk polymers (MIBs), prepared by bulk polymerizationand molecularly imprinted nanospheres (MINs), prepared by novel precipitationpolymerization method were synthesized in this work. The polymers were preparedwith diethylstilbestrol (DES) as the template molecule, methacrylic acid as thefunctional monomer and ethylene glycol dimethacrylate as the cross-linking monomerin the presence of acetonitrile solvent. The diameter of the MINs for DES wasapproximate 100 nm. These polymers were characterized by Fourier transforminfrared spectroscopy, transmission electron microscope and the static adsorptionexperiments. The prepared DES-imprinted nanospheres showed regularly shape andbetter adsorption capacity than DES-imprinted polymers.
4. Three CaTe quantum dots were synthesized in an aqueous phase usingthioglycolic acid as capped reagent. TEM, FL and UV were used to characterize theshape and optical properties. Insulin and CdTe QDs were used to study the interactionof protein with QDs as model samples. Fluorescence spectroscopy was used toinvestigate the stoichiometry and thermodynamic parameters of interaction. Theresults showed that the fluorescence of CdTe was quenched by insulin. The quenchingmechanism was discussed to be a static quenching procedure. Moreover, based on theevaluation of association constant of various the ionic strength and pH value, we speculated that the interaction of CdTe with insulin was mainly due to electrostaticattraction.
引文
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