A novel silsesquioxane and cobalt nitroprusside compound was prepared from octa(aminopropyl)silsesquioxane, resulting in a nanocomposite (ACCoN), which was then characterized by different spectroscopic techniques. The cyclic voltammograms of the ACCoN‐modified graphite paste electrode indicated a redox pair with a formal potential (Eθ′)=0.38 V, assigned to the Co(II)Fe(II)(CN)5NO/Co(II)Fe(III)(CN)5NO redox couple. The ACCoN‐modified graphite paste electrode was sensitive to isoniazid concentrations, presenting a linear response at a concentration range from 6.0×10−7 to 1.0×10−5 mol L−1, with limit of detection and amperometric sensitivity of 5.53×10−7 mol L−1 and 0.17 A/mol L−1, respectively.