Nakamura Yukari, Masumoto Shizuka, Kubo Arisa, Matsunaga Hisami, Haginaka Jun
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68, Koshien Kyuban-cho, Nishinomiya 663-8179, Japan.
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University, 11-68, Koshien Kyuban-cho, Nishinomiya 663-8179, Japan; Institute for Biosciences, Mukogawa Women's University, 11-68, Koshien Kyuban-cho, Nishinomiya 663-8179, Japan.
J Chromatogr A. 2017 Sep 22;1516:71-78. doi: 10.1016/j.chroma.2017.08.016. Epub 2017 Aug 8.
Monodisperse molecularly imprinted polymers (MIPs) for warfarin (WF) and coumachlor (CC), MIP and MIP, respectively, were prepared using 4-vinylpyridine (4-VPY) as a functional monomer and ethylene glycole dimethacrylate (EDMA) as a crosslinker by multi-step swelling and polymerization. Six kinds of MIP, MIP - MIP, were prepared varying the concentrations of WF and 4-VPY, while maintaining the EDMA concentration constant, and their retention and molecular recognition properties were evaluated using a mixture of sodium phosphate buffer and acetonitrile as a mobile phase in LC. In addition to shape recognition, hydrogen bonding, ionic and hydrophobic interactions could affect the retention and molecular recognition of WF on MIP, and ionic interactions seem to govern the retention and molecular recognition of WF above mobile phase pH 6 associated with higher molar ratio of 4-VPY to EDMA. Furthermore, MIP was prepared under the same conditions with MIP, which gave the highest imprinting factor for WF. WF could be recognized more strongly on MIP than MIP, and the imprinting factors of WF on MIP and MIP, respectively, are 2.68 and 5.03 using 20mM sodium phosphate buffer - acetonitrile (30/70, v/v)(final pH 6.1) as the mobile phase. This result indicates that the use of CC as a template molecule instead of WF could be useful for getting a higher imprinting factor for WF and for avoiding the leakage problem in the assay of WF in LC.
分别以4-乙烯基吡啶（4-VPY）为功能单体、乙二醇二甲基丙烯酸酯（EDMA）为交联剂，通过多步溶胀和聚合反应制备了用于华法林（WF）和氯灭鼠灵（CC）的单分散分子印迹聚合物（MIP），即分别为针对WF的MIP和针对CC的MIP。在保持EDMA浓度恒定的情况下，通过改变WF和4-VPY的浓度制备了六种MIP，即MIP - MIP，并在液相色谱中使用磷酸钠缓冲液和乙腈的混合物作为流动相评估了它们的保留和分子识别性能。除了形状识别外，氢键、离子和疏水相互作用都可能影响WF在MIP上的保留和分子识别，并且在流动相pH值高于6且4-VPY与EDMA摩尔比更高时，离子相互作用似乎主导着WF的保留和分子识别。此外，在与针对WF给出最高印迹因子的MIP相同的条件下制备了针对CC的MIP。WF在针对CC的MIP上的识别比在针对WF的MIP上更强，以20mM磷酸钠缓冲液 - 乙腈（30/70，v/v）（最终pH 6.1）作为流动相时，WF在针对CC的MIP和针对WF的MIP上的印迹因子分别为2.68和5.03。该结果表明，使用CC作为模板分子而非WF可有助于获得更高的WF印迹因子，并避免液相色谱法测定WF时的渗漏问题。
