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The synthesis of a T-2 toxin imprinted polymer and its application in food analysis are reported for the first time. A molecularly imprinted polymer (MIP) for the selective recognition of T-2 toxin (T-2) was synthesized by bulk polymerization. Methacrylamide and ethyleneglycol dimethacrylate were applied as functional monomer and cross-linker, respectively. Molecularly imprinted solid-phase extraction (MISPE) procedures were optimized for further application in the analysis of T-2. Scatchard plot analysis revealed that two classes of imprinted binding sites were formed in the imprinted polymer. The dissociation constant (K(D)) of the higher affinity binding sites was 7.0 mu mol/l, while the K(D) of the lower affinity binding sites was 54.7 mu mol/l. The performance of the MW throughout the clean-up of spiked maize, barley and oat sample extracts was compared with the results obtained when using non-imprinted polymer, OASIS HLB (R) and immunoaffinity columns (IAC). Depending on the food matrix and the spiked concentration, recoveries after MISPE and non-imprinted solid-phase extraction varied respectively from 60% to 73% and from 21% to 57%. Recoveries obtained after clean-up using OASIS HLB (R) and IAC were in the range of 74-104% and 60-85%, respectively. Although highest recoveries were obtained with OASIS HLB (R) sorbents, the designed MIP and the IAC were superior regarding selectivity, cross-reactivity, matrix effect, limits of detection (LOD) and limits of quantification (LOQ). Depending on the matrix, LOD after MISPE ranged from 0.4 mu g/kg to 0.6 mu g/kg and LOQ from 1.4 mu g/kg to 1.9 mu g/kg. LOD and LOQ after OASIS HLB (R) clean-up varied from 0.9 mu g/kg to 3.5 mu g/kg and from 3.1 mu g/kg to 11.7 mu g/kg, respectively. The LOD and LOQ values obtained with IAC were in the range of 0.3-2.3 mu g/kg and 1.0-7.7 mu g/kg, respectively. Analysis of 39 naturally contaminated samples (maize, barley and oat) by liquid chromatography tandem mass spectrometry revealed that the MIP could be an excellent alternative for clean-up of contaminated food samples. (C) 2010 Elsevier B.V. All rights reserved.