Ji Xiang, Li Dan, Li Hua
College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China.
Biomed Chromatogr. 2015 Aug;29(8):1280-9. doi: 10.1002/bmc.3418. Epub 2015 Jan 23.
A novel molecularly imprinted polymer (MIP) monolith for highly selective extraction of cholecystokinin (CCK) neuropeptides was prepared in a micropipette tip. The MIPs were synthesized by epitope imprinting technique and the polymerization conditions were investigated and optimized. The synthesized MIPs were characterized by infrared spectroscopy, elemental analyzer and scanning electron microscope. A molecularly imprinted solid-phase microextraction (MI-μ-SPE) method was developed for the extraction of CCK neuropeptides in aqueous solutions. The parameters affecting MI-μ-SPE were optimized. The results indicated that this MIP monolith exhibited specific recognition capability and high enrichment efficiency for CCK neuropeptides. In addition, it showed excellent reusability. This MIP monolith was used for desalting and enrichment of CCK4, CCK5 and CCK8 from human cerebrospinal fluid prior to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, and the results show that this MIP monolith can be a useful tool for effective purification and highly selective enrichment of multiple homologous CCK neuropeptides in cerebrospinal fluid simultaneously. By employing MI-μ-SPE combined with HPLC-ESI-MS/MS analysis, endogenous CCK4 in human cerebrospinal fluid was quantified.
在微量移液器吸头中制备了一种用于高选择性萃取胆囊收缩素（CCK）神经肽的新型分子印迹聚合物（MIP）整体柱。通过表位印迹技术合成了MIP，并对聚合条件进行了研究和优化。采用红外光谱、元素分析仪和扫描电子显微镜对合成的MIP进行了表征。建立了一种分子印迹固相微萃取（MI-μ-SPE）方法用于萃取水溶液中的CCK神经肽。对影响MI-μ-SPE的参数进行了优化。结果表明，该MIP整体柱对CCK神经肽具有特异性识别能力和高富集效率。此外，它还具有出色的可重复使用性。该MIP整体柱用于在基质辅助激光解吸/电离飞行时间质谱分析之前对人脑脊液中的CCK4、CCK5和CCK8进行脱盐和富集，结果表明该MIP整体柱可作为同时有效纯化和高度选择性富集脑脊液中多种同源CCK神经肽的有用工具。通过采用MI-μ-SPE结合HPLC-ESI-MS/MS分析，对人脑脊液中的内源性CCK4进行了定量。
