缩胆囊素分子印迹整体柱的制备及固相微萃取-高效液相色谱分析研究

Preparation of Cholecystokinin Molecularly Imprinted Polymer Monolith and Its Application in Solid Phase Microextraction and High Performance Liquid Chromatographic Analysis

采用抗原决定簇法,开展了对缩胆囊素(CCK)神经肽具有特异性识别和富集作用的新的分子印迹聚合物(MIP)整体柱合成与制备的研究。对MIP整体柱合成条件进行了系统的选择和优化,以缩胆囊素五肽(CCK5)为模板分子,在小移液器吸头中原位聚合制备了可以特异性识别和富集CCK神经肽的MIP整体柱。用扫描电镜和红外光谱对该分子印迹聚合物进行了表征。将此MIP整体柱作为固相微萃取小柱,研究并优化了固相微萃取条件,结合高效液相色谱-紫外检测法,建立了脑脊液中CCK5和CCK8的分析检测方法;本方法成功应用于加标脑脊液中CCK神经肽的分离富集,CCK5和CCK8的检测限分别为4.38和15.95 ng/m L,平均加标回收率分别为90.2%和87.6%,相对标准偏差分别为3.2%和3.9%。此CCK-MIP整体柱具有特异性强、经久耐用的特点。

The synthesis and preparation of molecularly imprinted polymer （MIP） monolith for selective recognition of cholecystokinin were carried out by epitope imprinting technique. The MIP was synthesized by using CCK5 as template in a micropipette tip, and the polymerization conditions were investigated and optimized. The synthesized MIPs were characterized by scanning electron microscope （SEM） and infrared spectroscopy. By using this MIP monolith as solid phase microextraetion （SPME） column and coupling to high performance liquid chromatography-ultraviolet （HPLC/UV） detection, an analytical method was developed for the determination of CCK, and the parameters affecting SPME were optimized. The proposed method was successfully applied to determination of CCK5 and CCK8 in spiked cerebrospinal fluid （CSF） simultaneously. The detection limit of CCK5 and CCK8 were 4. 38 ng/mL and 15. 95 ng/mL, respectively; the mean recoveries were 90.2% and 87.6% with the relative standard deviations of 3.2% and 3.9%. This CCK-MIP monolith showed high specificity, excellent reusability.