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The on-line MISPE-HPLC method for selective determination of seven coumarins in various samples was developed.Developed and optimized method showed good validation results.Method was succesfully applied to analysis of Cassia cinnamon, chamomile tea and Tokaj specialty wines.Total time of whole analytical run, including extraction step was less than 13.50 min.In this work, an on-line SPE-HPLC method with spectrophotometric detection was developed for the determination of coumarins in complex samples. For the on-line cleanup of samples, a molecularly imprinted polymer was packed into the column cartridge and coupled directly with HPLC (MISPE-HPLC) using a column switching system. The separation of coumarins was performed on a C18 core-shell column (100 × 4.6 mm, 5 μm) with a mobile phase consisting of 0.3% acetic acid/acetonitrile with gradient elution at a flow-rate of 1 mL min−1. The total time of the whole analytical run, including the extraction step, was 13.25 min. The on-line MISPE-HPLC method was optimized and validated. The results showed good linearity (0.10–100 μg mL−1) with correlation coefficients higher than 0.99. The LOD values were from 0.03 to 0.15 μg mL−1. The proposed method was successfully applied for analysis of real samples (Cassia cinnamon, chamomile tea, and Tokaj specialty wines) and obtained recoveries varied from 78.7% to 112.2% with an RSD less than 9%.