This work presents a simple and fast method for colchicine (CO) determination in pharmaceutical formulations and urine by multiple pulse amperometry (MPA) with flow injection analysis (FIA) system using a boron-doped diamond electrode. In optimized conditions, it was possible to quantify CO in urine samples without interference of uric acid and ascorbic acid. The working linear range for CO quantification was achieved from 1.0 × 10-7 to 0.5 × 10-3mol L-1 with a low limit of detection of 2.14 × 10-8mol L-1. Furthermore, the proposed method showed high repeatability for 10 consecutive injections of 1.0 × 10-4mol L-1 CO (relative standard deviation = 1.28%) and good analytical frequency (30 determinations per hour). The addition and recovery studies in all pharmaceutical samples were approximately 100% and the results for CO determination were compared by high performance liquid chromatography (HPLC) with UV detection.