A portable and non-invasive sensor presents an innovative way to measure inflammation biomarkers in exhaled breath condensate (EBC). This research is focused on developing a miniaturized bipolar electrochemical sensor that can be connected to a smartphone app. This device will be able to detect adding known amounts of nitrite (spikes) to a salt solution and small amounts of nitrite ions in collected real samples in EBC. The sensor was fabricated and tested for its rapid electron transfer capability and ability to detect nitrite ions even at very low concentrations and low real sample levels. In the proposed setup, when the required potential was applied by using a direct power supply, the nitrite ions were oxidized electrocatalytically at amine-functionalized graphene oxide (AGO) decorated with gold nanoparticles on a carbon paper anodic pole. On the other hand, the reduction reaction of Prussian blue occurred at the cathodic pole of the bipolar electrode simultaneously. This strategy led to a change in color from blue to white as a result of the reduction process and the color change is proportional to the concentration of nitrite ions in the analytical solution. The combination of smartphones with the colorimetric method has resulted in a platform for the detection of test strips that is more visual and convenient. The amperometry and voltammetric methods of nitrite detection showed a linear range of up to 1230 μM. The bipolar electrochemical sensor was able to detect the clinically relevant range of nitrite from 0.5 to 85 μM in a buffer with an ultralow detection limit (LOD) of 250 nM (S/N = 3), fast response and excellent selectivity. It was benchmarked by utilizing pre-characterized real EBC samples to differentiate patients with respiratory diseases from healthy volunteers. By tracking the results of nitrite measurements over time, it has become possible to detect trends and changes in an individual's nitrite ion concentration and to potentially identify lung inflammation earlier.