Determination of methotrexate in human serum by high-performance liquid chromatography combined with pseudo template molecularly iniprinted polymer

A pseudo template molecularly Imprinted polymer (MIP) was prepared for methotrexate (MTX) and RP-HPLC method combined with the MIP was developed for the determination of m,l-x In human serum Because oft lie Poor solubility of MTX,I common MIP Preparation solvents, trimethopium (TMP), a molecule having the similar imprinting sites as MTX, is selected as the pseudo template The MIP was prepared using methacrylic acid (MAA) and ethylene glycol dimethacrylate as functional monomer and cross-linker. respectively. H-1 NMR study showed highly strong interaction between TMP and MAA with hydrogen bonds. Chromatographic behaviors indicated that the TMP-MIP possessed excellent affinity and selectivity for MTX. And the Imprinting factor for MTX was high LIP to 9 5 when 7 3 of acetonitrile methanol (v/v) was used as mobile phase Moreover,TMP-MIP was Used as the solid-phase extraction (SPE) material to enrich the larger compound MTX in human serum samples for HPLC analysis The SPE process was carefully optimized and good recoveries of MTX were obtained as 81.6-86.2% with RSD of 0.22-1.84% when the spiked concentration of MTX was 20-10.0 mu g mL(-1) an human serum samples The results indicated that the pseudo template MIP can be applied to preconcentration, purification and analysis of MTX In chnic samples. (C) 2009 Elsevier B.V. All rights reserved