Serotonin (5-hydroxytryptamine (5-HT)) is one of the important neurotransmitters which is released from the endocrine system. An abnormal level of this biomarker leads to several neurological diseases. The accurate assessment of serotonin is the utmost option to start treatment in the early stages of the disease. The current work is focused on the development of a disposable, screen-printed electrochemical sensor for the depression biomarker, serotonin in the physiological pH medium (pH 7.4) with the aid of a hexagonal, Ni(OH)2-nanoplate (NH-HNP)-embedded chitosan (Chit) and modified, screen-printed carbon electrode (SPCE). Initially, hexagonal nanoplates of Ni(OH)2 were synthesized by an eco-friendly and simple hydrothermal method. The prepared materials were well characterized by advanced analytical techniques to examine the physicochemical properties of the synthesized Ni(OH)2 hexagonal nanoplates. From the cyclic voltametric (CV) analysis, it was found that the oxidative current response of 5-HT at a NH-HNP-modified SPCE has about fivefold higher current values than over bare SPCE. The scan rate studies of NH-HNP-Chit/SPCE electrodes revealed that the oxidation mechanism of 5-HT is controlled by the diffusion behavior of the analyte. Differential pulse voltammetric tests of the NH-HNP-Chit/SPCE electrode exhibited a linear response in the dynamic concentration range of 0.1 to 30 µM, with a detection limit of about 60 nM. The sensor response is very reproducible from electrode to electrode, and the deactivation or surface-fouling of the sensor was not observed within the several experimental measurements. The sensor exhibited excellent storage stability over a period of twenty days. Finally, the fabricated, disposable SPCE sensor has shown respectable activity for the detection of depression biomarker 5-HT from synthetic urine and saliva samples. © 2023 by the authors.