A simple procedure was developed to prepare a glassy carbon electrode modified with carbon nanotubes and Ruthenium (III) complexes. First, 25 μl of dimethyl sulfoxide–carbon nanotubes solutions (0.4 mg/ml) was cast on the surface of the glassy carbon electrode and dried in air to form a carbon nanotube film at the electrode surface. Then, the glassy carbon/carbon nanotube-modified electrode was immersed into a Ruthenium (III) complex solution (direct deposition) for a short period of time (10–20 s for multiwalled carbon nanotubes and 20–40 s for single-walled carbon nanotubes). The cyclic voltammograms of the modified electrode in aqueous solution shows a pair of well-defined, stable, and nearly reversible redox couple, Ru (III)/Ru(II), with surface-confined characteristics. The attractive mechanical and electrical characteristics of carbon nanostructures and unique properties and reactivity of Ru complexes are combined. The transfer coefficient (α), heterogeneous electron transfer rate constants (ks), and surface concentrations (Γ) for the glassy carbon/singlewalled carbon nanotubes/Ru(III) complex-, glassy carbon/ multiwalled carbon nanotubes/Ru(III) complex-, and glassy carbon/Ru(III) complex-modified electrodes were calculated using the cyclic voltammetry technique. The modified electrodes showed excellent catalytic activity, fast response time, and high sensitivity toward the reduction of nicotinamide adenine dinucleotide in phosphate buffer solutions at a pH range of 4–8. The catalytic cathodic current depends on the nicotinamide adenine dinucleotide concentration. In the presence of alcohol dehydrogenase, the modified electrode exhibited a response to addition of acetaldehyde. Therefore, the main product of nicotinamide adenine dinucleotide electroreduction at the Ru(III) complex/carbon nanotube-modified electrode was the enzymatically active NADH. The purposed sensor can be used for acetaldehyde determination.