As a common flavonoid glycoside, rutin owns excellent physiologic activities and has been widely used in clinical chemistry and human health. Therefore the establishment of sensitive and reliable methods for rutin determination is highly important. Black phosphorene (BP) nanoshcet modified glassy carbon electrode (GCE) was fabricated by using poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) as the film and the stabilizer. The incorporation of BP with PEDOT:PSS resulted in a stable nanocomposite (BP-PEDOT:PSS) and its performance were characterized by SEM, EDS and Raman. BP-PEDOT:PSS/GCE showed excellent conductivity and stability, which was applied for the sensitive voltammetric determination of rutin, a commonly used electroactive drug. On cyclic voltammogram a pair of well-defined redox peak of rutin could be obviously observed with enhanced voltammetric response, which could be due to the presence of BP nanosheet that accelerated the electrode reaction. Electrochemical parameters were calculated with the electron transfer coefficient (alpha) as 0.55 and the apparent heterogeneous electron transfer rate constant (k(s)) as 4.22 s(-1). Under the selected conditions differential pulse voltammetric oxidation peak current of rutin showed linear dependence on its concentration within two sections of 0.02-15.0 mu mol/L and 15.0-80.0 mu mol/L with a low detection limit of 0.007 mu mol/L (3S(0)/S). Rutin tablet samples were successfully examined by the as-proposed method with satisfactory results, showing the practical applications.