Profiling at GCs level is an important issue since it is a key factor to determine administration intent for either therapeutic or doping purpose. In this study, the sensing platform which is based on a combination of surface plasmon resonance system with molecularly imprinted nanoparticles was developed for monitoring of prednisolone level as a selected GCs analyte in plasma and urine. Fabricated sensing platform was utilized with ellipsometer and contact angle measurements followed by characterization of nanoparticles (NPs) by the means of structurally (i.e. FTIR and elemental analysis) and morphologically (i.e. TEM and Zeta-sizer analysis). The lowest limit of detection by PIN2 encoded molecularly imprinted nanoparticle was achieved as at 5.5 ppb and 8.9 ppb at pH 6.6 and 7.4, respectively. Competition kinetic assays were performed by using 17-alpha-ethynyl estradiol (EEST) and estradiol (EST). The reusability studies have been applied to the sensor with the best results and 92.8% recovery is achieved. Validation studies of the system were performed according to the standard procedure by high performance liquid chromatography (HPLC). The method was successfully applied to monitor PRD levels in body fluids (urine and plasma) as a natural sample. (c) 2017 Elsevier B.V. All rights reserved.