Department of Chemistry, Carleton University, Ottawa, ON, Canada.
J Sep Sci. 2010 Sep;33(17-18):2796-803. doi: 10.1002/jssc.201000257.
Molecularly imprinted polymer (MIP) submicron particles were synthesized, using either ethylene glycol dimethacrylate or trimethylolpropane trimethacrylate as a cross-linker, specifically for recognition of 17β-estradiol (E2). HPLC with fluorescence detection (HPLC-FD) results showed that 90(±5)% of E2 bound onto these particles after 2 min of incubation, and 96(±3)% after long equilibrium. The binding capacity was 8(±3) μmol/g for MIP particles prepared using ethylene glycol dimethacrylate, and 33-43(±8) μmol/g for using trimethylolpropane trimethacrylate. CE separation of MIP and non-imprinted polymer particles was successful when 50 mM borate buffer (pH 8.5) containing 0.005% w/v EOTrol™ LN in reverse polarity (-30 kV) was used. The electrophoretic mobilities of MIP and non-imprinted polymer particles, together with dynamic light scattering measurement of particle sizes, allowed for an estimation of their surface charges. Automated injection of E2 and particles in mixture set a lower limit of 20(±1) s on incubation time for the study of fast binding kinetics. The presence of E2 and bisphenol A (BPA) together tested the selectivity of MIP particles, when the two compounds competed for available binding cavities or sites. Addition of E2 after BPA confirmed E2 occupation of the specific binding cavities, via displacement of BPA.
分子印迹聚合物（MIP）亚微米颗粒被合成，使用乙二醇二甲基丙烯酸酯或三羟甲基丙烷三甲基丙烯酸酯作为交联剂，专门用于识别 17β-雌二醇（E2）。高效液相色谱-荧光检测（HPLC-FD）结果表明，在孵育 2 分钟后，90（±5）%的 E2 结合到这些颗粒上，长时间平衡后达到 96（±3）%。使用乙二醇二甲基丙烯酸酯制备的 MIP 颗粒的结合能力为 8（±3）μmol/g，使用三羟甲基丙烷三甲基丙烯酸酯的结合能力为 33-43（±8）μmol/g。当使用反向极性（-30 kV）、50 mM 硼酸缓冲液（pH 8.5）中含有 0.005% w/v EOTrol™LN 时，MIP 和非印迹聚合物颗粒的 CE 分离是成功的。MIP 和非印迹聚合物颗粒的电泳迁移率，以及颗粒粒径的动态光散射测量，允许估计它们的表面电荷。E2 和颗粒混合物的自动进样将研究快速结合动力学的孵育时间下限设定为 20（±1）s。当两种化合物竞争可用的结合腔或位点时，E2 和双酚 A（BPA）的同时存在测试了 MIP 颗粒的选择性。在 BPA 之后添加 E2 确认了 E2 通过取代 BPA 占据特定的结合腔。
