In this study, the method for detection of chloramphenicol was investigated by electrochemical sensor; the sensor was constructed by biomass derived porous carbon. At first, porous carbon doped with hetroatoms (nitrogen, sulfur, phosphorus) was synthesized based on the use of pyrolysis and high temperature carbonization methods. Elaeagnusangustifolia L. gum was used as the carbon source in the facile template-free process. The biomass derived porous carbon was then used as the active electrode material for antibiotic sensing. The chemically modified electrodes properties were studied with the cyclic voltammetry and differential pulse voltammetry methods. The effects of the scan rate, accumulation time and pH, were carefully considered. Comparison with other working electrodes at the optimized conditions indicated that the N, S, P triple doped porous carbon modified glassy carbon electrode appeared a well-defined reduction peak towards chloramphenicol. The linear concentration response of chloramphenicol ranged from 1 to 40 mu M (R = 0.9903) and 50 to 500 mu M (R = 0.9923), and a low detection limit of 0.01 mu M (S/N = 3). Furthermore, the constructed novel electrochemical sensor was used for detection of chloramphenicol in real samples and achieved satisfactory recovers.