When danger is perceived, the human body responds to overcome obstacles and survive a stressful situation; however, sustained levels of stress are associated with health disorders and diminished life quality. Hence, stress biomarkers are monitored to control stress quantitatively. Herein, a porous sensor (4l-COP/p) composed of poly(3,4-ethylenedioxythiophene) (PEDOT) and poly(3,4-ethylenedioxythiophene-co-N-methylpyrrole) (COP), which is prepared in a four-layered fashion to detect dopamine (DA) and serotonin (5-HT), is presented. Specifically, the detection is conducted in phosphate-buffered saline (PBS), as well as artificial urine and sweat, by applying cyclic voltammetry. The limit of detection values obtained are as low as 5.7 × 10−6 and 1.4 × 10−6 m for DA and 5-HT, respectively, when assessed individually in artificial urine. When mixed in PBS, 4l-COP/p detects both biomarkers with a resolution of 0.18 V and a sensitivity of 40 and 30 μA mm−1 for DA and 5-HT, respectively. Additionally, by theoretical calculations, the interaction pattern that each stress biomarker establishes with the PEDOT outer layer is elucidated. Whereas DA interacts with the 𝝅������-system of PEDOT, 5-HT forms specific hydrogen bonds with the conducting polymer chains. The resolution value obtained depends upon such interactions. Overall, 4l-COP/p electrodes display potential as stress sensing devices for healthcare technologies.