Preparation and characterization of molecularly imprinted solid-phase extraction column coupled with high-performance liquid chromatography for selective determination of melamine.

We synthesized a selective molecularly imprinted solid-phase extraction (MIP-SPE) column and established an extraction and enrichment method using this MIP-SPE column. By coupling with HPLC, we developed a new method to detect trace amounts of melamine in eggs. The MIP-SPE column was synthesized by thermal-initiated polymerization using melamine as the template, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker and azodiisobutyronitrile as the initiator. HPLC was used to evaluate the identification and enrichment capability of the MIP-SPE column and for the measurement of melamine in the sample. The melamine concentration exhibited an excellent linear relationship in the range of 0.1-25.0 µg ml ( = 0.9983). The identification capability of the MIP-SPE column was apparently superior to that of the non-imprinted polymer solid-phase extraction column; an average enrichment factor of 46.8-fold (RSD = 3.5%) was obtained for 0.4 µg ml melamine by the MIP-SPE column. When the MIP-SPE HPLC method was applied to the detection of melamine in eggs, an average recovery rate of 93.5-102.0% (RSD = 3.6-4.9%) and a limit of detection of 0.05 µg kg were obtained. This method is simple, fast and cost-effective; thus, it can greatly simplify the pre-treatment of complex samples and can be used in the detection of residual melamine in eggs and other products.