Creatinine thin films were synthesised by matrix assisted pulsed laser deposition (PLD) techniques for enzyme-based biosensor applications. An UV KrF* (lambda=248 nm, tau approximately 10 ns) excimer laser source was used for the irradiation of the targets at incident fluence values in the 0.3-0.5 J/cm2 range. For the matrix assisted PLD the targets consisted on a frozen composite obtained by dissolving the biomaterials in distilled water. The surface morphology, chemical composition and structure of the obtained biomaterial thin films were investigated by scanning electron microscopy, Fourier transform infrared spectroscopy, and electron dispersive X-ray spectroscopy as a function of the target preparation procedure and incident laser fluence.