In the present study, the interaction between cinchonine (CCN) and human serum albumin (HSA) was investigated using differential pulse polarography (DPP), cyclic voltammetry (CV) and spectroscopic techniques in Britton-Robinson (B-R) buffer pH 7.4. CCN displayed a main cathodic peak at -1.228 V (vs. Ag|AgCl|KClsat) on mercury working electrode. The addition of HSA into CCN sulfate solution resulted in the decrease of the main reduction peak current of CCN and no new peaks appeared. The decay in the peak current of CCN, after the addition of HSA, showed a decrease in free drug concentration and formation of a biocomplex. The peak current changes of CCN in the presence of HSA were followed by DPP to determine the binding parameters. The logarithm of binding constant and binding ratio between CCN and HSA were 6.128 and 1: 1, respectively. This interaction was also confirmed by UV-Vis and FTIR-ATR spectroscopic measurements.