Electrochemical behaviors of bovine hemoglobin (Hb) at an acetylene black paste electrode based on the enhancement effect of sodium dodecyl sulfate (SDS) were investigated. In the optimal conditions, a very weak reduction peak was observed at an acetylene black paste electrode for hemoglobin in the absence of SDS. However, the reduction peak current increased remarkably after the addition of 4.0 × 10?4 mol L?1 SDS, suggesting that SDS exhibits obvious enhancement effect to the determination of hemoglobin. All the experimental parameters, such as pH value, concentration of SDS, accumulation time and accumulation potential were optimized for hemoglobin analysis. The proposed method possesses high sensitivity (detection limit is 3.0 × 10?9 mol L?1), wide linearity (6.0 × 10?9 to 6.0 × 10?7 mol L?1), rapid response and low cost. Finally, the method was successfully employed to determine hemoglobin in a spiked sample.