Fabrication of Molecular Recognition Sensor based on Molecular Imprinting-Chemiluminescence Detection

分别以己烯雌酚（DES）和鲁米诺为探针分子，建立了DES和鲁米诺化学发光体系测定雌激素和邻苯二甲酸酯（PE）等内分泌干扰物（EDCs）。内分泌干扰物可能通过与雌激素受体结合而影响内分泌系统。采用分子印迹技术制备了分子印迹聚合物（molecularly imprinted polymer，MIP），并利用合成受体取代受体，测定了17 β-雌二醇（estradiol）、雌酮（estron）和雌三醇（estriol）等雌激素的含量。通过紫外线照射将DES转化为荧光化合物，然后使用另一个CL系统代替DES CL系统，使DES在10^9 × 10^6 M水平下的测定更加灵敏。因此，MIP可识别并测定低至10^7 M的17 β-雌二醇。分子印迹聚合物对雌激素的识别程度与其分子结构密切相关。由于这个原因，PE水解为邻苯二甲酸，这是由邻苯二甲酸从MIP中置换鲁米诺的能力决定的。因此，测定的总PE低至10^8 M水平，与PE的种类无关。该方法成功地应用于河水中邻苯二甲酸二乙酯（10^7M）的测定，并以4-乙烯基吡啶/Cu/L-Trp三维配合物为探针分子，建立了L-色氨酸的化学发光检测体系。L-Trp通过与相应MIP中的识别位点X（4-乙烯基吡啶/Cu/X）特异性结合而被识别，然后用过氧化氢发光氧化法测定。通过使用1-乙烯基咪唑代替4-乙烯基吡啶作为功能单体，改善了D-和L-Trp之间的识别。

The diethylstilbestrol(DES) and luminol CL systems were applied to the determination of the endocrine disrupting chemicals (EDCs) such as estrogens and phthalate esters(PEs) using DES and luminol, respectively, as probe molecules. The EDCs are suspected to affect the endocrine system by the binding to estrogen receptor. Instead of the receptor, we employed the synthetic receptor obtained by molecular imprinting, which was a technique to prepare the molecularly imprinted polymer(MIP).The estrogens such as 17 β-estradiol, estron and estriol were determined by the ability of the estrogens to displace DES from the MIP. The conversion of DES to a fluorescent compound by UV irradiation and then the employment of another CL system in place of the DES CL system made more sensitive determination of DES at 10^9〜10^6 M levels. Thus 17 β-estradiol as low as 10^7M was recognized and determined by the MIP. The method is expected to determine estrogens in river water.The extent of the recognition of PEs by the MIP was dependent strongly on the molecular structure of the PEs. For this reason, the PEs were hydrolyzed to phthalic acid, which was determined by the ability of the phthalic acid to displace luminol from MIP. Thus total PEs as low as 10^8M levels were determined with no dependence of the kinds of PEs. The method was successfully applied to the determination of diethylphthalate at 10^7M levels in river water.Moreover, the CL detection system for L-tryptophan was developed using a three-dimensional complex (4-vinylpyridine/Cu/L-Trp) as a probe molecule by the same method. L-Trp was recognized by the specific binding to the recognition site X(4-vinylpyridine/Cu/X) in the corresponding MIP and then determined by the luminous oxidation with hydrogen peroxide. The recognition between the D- and L-Trp was improved by the use of 1-vinylimidazol in place of 4-vinylpyridine as a functional monomer.