Human Exhaled Breath Condensate (EBC) contains markers of several inflammatory diseases. Its analysis is of interest to a number of researchers. Nitrite ions (NO-), which are widely used in our daily lives, are nevertheless among these indicators. In this study, a simple, fast, portable, non-invasive and cheap electrochemical sensor is developed for the analysis of the nitrite profile in EBC. In this regard, sodium nitrite (NaNO) was first immobilized on self-assembled 2-aminothiophenol (2-ATP) on a screen-printed gold electrode (Au-SPE). Then, a polymer matrix composed of polyvinyl alcohol (PVA) crosslinked with glutaraldehyde (GA) was combined with gold nanoparticles (Au-NPs) to cover the modified Au-SPE and complete the fabrication of the Ion Imprinted Polymer (IIP) sensor. The electrochemical behaviour of the sensor was monitored using Cyclic Voltammetry (CV), Electrochemical Impedance Spectroscopy (EIS) and Differential Pulse Voltammetry (DPV) methods, while the morphology and chemical composition of its layers were observed by infrared Fourier transform (FTIR), Atomic Force Microscopy (AFM) and Scanning Electron Microscopy coupled with energy dispersion X-Ray spectroscopy (SEM-EDS) techniques. In addition, after a successful control test using a Non-Imprinted Ion Polymer (NIIP) sensor, the obtained results demonstrated satisfactory sensitivity and selectivity to nitrite compared to co-existing interfering substances in EBC, such as nitrate, acetate and ammonium nitrate. Under improved experimental conditions, the nitrite IIP sensor exhibits responses proportional to nitrite concentrations (R = 0.96) over a concentration range of 0.5-50 μg mL with a detection limit (LOD) of 4 μmol L (signal-to-noise ratio S/N = 3). The proposed approach was well applied for the nitrite determination in EBC samples with a relative standard deviation (RSD = 4%) and could open clinical applications in respiratory medicine.