A new and simple strategy based on nanostructured CaCO3-poly(ethyleneimine) (PEI) microparticles (MPs) for phenol sensing using PDMS/glass fluidic microchip is developed. This fluidic microsystem including integrated screen-printed electrodes modified with CaCO3-PEI MPs and tyrosinase (Tyr) through cross-linking with glutaraldehyde, represents a low-cost platform for phenol detection. The designed fluidic microsystem improves the sensitivity of the biosensor allowing the detection of very low concentrations of phenol (up to 10 nM). This device shows high repeatability and low detection limit, is easy to be fabricated, inexpensive, disposable, and amenable to mass production. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.