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Cytokinins (CTKs) are a class of growth regulating hormones that play a central role in a wide range of biological processes in plants.Many challenges have been faced to develop a sensitive and accurate separation and analysis method to monitor trace CTKs in plants (approximately from pmol/g to fmol/g fresh sample).In this work, a novel molecularly imprinted solid-phase extraction column (MIP-SPEC) had been synthesized by in-situ polymerization method.Experimental results show that the prepared MIPs have selective enrichment capacity for trans-zeatin, trans-zeatin glucoside, kinetin and meta-topolin.The MIP-SPEC method was applied in the separation and enrichment the cytokinins from soybean sprouts and rape leaves.