Herein, palladium-cobalt supported on ketjenblack (PCKB) electrocatalyst is synthesized and tested i) for the reaction of glucose electrooxidation (GOR) in presence of dopamine and uric acid and ii) for the simultaneous detection of the above three substances. The electrocatalyst, prepared using the sulphite complex method, is physicochemically characterized by X-Ray diffraction (XRD), transmission electron microscopy (TEM) and X-Ray photoelectron spectroscopy (XPS) and electrochemically characterized by cyclic voltammetry, potentiometry and chronoamperometry. It is found that the electrocatalytic activity towards GOR: i) is favoured in case of ketjenblack support, ii) is enhanced more when Co is added into Pd, iii) is enhanced with temperature increment from 26.0 to 42.0 °C, iv) is enhanced when glucose concentration increases up to 1.5 mM, while at higher concentration values is suppressed. The electrocatalyst ability of simultaneous glucose, dopamine and uric acid electrooxidation is attributed to the bi-functional role of the PCKB due to cobalt presence. •Glucose oxidation reaction (GOR) is more favoured using as support ketjenblack (KB) than vulcan-XC72.•Pd10Co1/KB favours GOR, even in presence of dopamine (DA) and uric acid (UA).•Dopamine (DA) and uric acid (UA) are also electrooxidated on Pd10Co1/KB.•Pd10Co1/KB is able to co-detect dopamine (DA), uric acid (UA) and glucose (Glu).