以局部结构类似物为类模板的分子印迹整体柱在线检测三甲氧苄啶

On-line Determination of Trimethoprim in Human Serum by Molecularly Imprinted Monolithic Column with Melamine Employed as Analogue Template

以三聚氰胺(MAM)为类模板分子,甲基丙烯酸(MAA)为功能单体,二甲基丙烯酸乙二醇酯(ED-MA)为交联剂,聚乙二醇(PEG400)为致孔剂,原位聚合法制备了对三甲氧苄啶(TMP)有较强选择性吸附作用的分子印迹(MIP)整体柱。以该MIP整体柱为高效液相色谱柱,考察了其在不同色谱流动相组成条件下对TMP的识别性能。结果显示,MIP整体柱在甲醇、乙腈、水等条件下能够吸附TMP而不出峰。可以利用MIP整体柱在甲醇-水(80∶20,V/V)中在线选择性吸附(或富集)TMP,然后将流动相转换为甲醇进一步除去疏水性杂质,最后用强洗脱剂洗脱TMP出峰。MIP整体柱线检测人血清中TMP的工作曲线为A=42.8c-3.03(r=0.9994);线性范围为8.3～93.8mg/L;检出限为0.14mg/L。

A molecularly imprinted monolithic column being capable of selectively adsorbing trimethoprim（TMP） was synthesized by in situ polymerization method.During the polymerization process,melamine was employed as analogue template,and methacrylic acid as functional monomer,ethylene glycol dimethacrylate as cross linker,polyethylene glycol 400（PEG400） as porogenic agent,respectively.The change of recognition ability for MIP column towards TMP was validated under different kinds of mobile phase compositions in a high performance liquid chromatographic（HPLC） system.Experimental results showed that MIP column could completely retain TMP under mobile phases of methanol,acetonitrile or water.TMP was selectively adsorbed in MIP column under methanol-water（80∶20,V/V）,then the hydrophobic impurities were further erased by changing the mobile phase to methanol,and finaly,the TMP peak was obtained in the elution step of with ammonia water-methanol（2.5∶97.5,V/V）.Online determination of TMP in human serum was performed in HPLC system with the imprinted column employed as chromatography column.The linear regression equation obtained was determined as A=42.8-3.03（r=0.9994） with the linear range of 8.32-93.8 mg/L and a limit of detection of 0.145 mg/L.