In this study, a method for the sensitive, rapid, efficient and simultaneous determination of the riboflavin (VB2) and pyridoxine (VB6) by voltammetry technique was developed. For this purpose, a novel electrochemical sensor (44-DABP/GC) based on GC electrode modified with 4,4'-diamino benzophenone was fabricated by the cyclic voltammetry. Best analytical performance was obtained in 0.1 M, pH 4.5 acetate buffer solution, with a scanning rate of 100mVs(-1), a puls time of 10 ms, a scan increment of 10 mV and a pulse amplitude of 125 mV. 44-DABP/GC electrode exhibited good analytical performance for the determination of VB2 and VB6. Three linear ranges in wide concentration ranges for VB2 from 13 nM to 660 mu M, and two linear ranges for VB6 from 15.7 mu M to 2210 mu M were obtained with the detection limit of 8.52 nM and 4.70 mu M, respectively. Besides, repeatability, reproducibility and interference studies were performed. The proposed sensor was applied successfully in food and pharmacological samples.