Hexamethylphosphoramide (HMPA) is extensively used in various applications. However, it is categorized as carcinogenic and mutagenic substance that participates in adverse effects on public health. Herein, we developed a new Ru(bpy)32+ electrochemiluminescence (ECL) system for sensitive, cost-effective, rapid, and specific detection of HMPA. The developed ECL sensing platform enabled analysis of HMPA over a wide dynamic range of 0.1 μM to 1 mM with a detection limit of 0.08 μM (S/N = 3). The designed ECL sensor exhibited high percentage recoveries (98.0–102.5%) in real samples of lake water and waste water.