Leishmaniasis is one of the most important parasitic neglected disease. The electrochemical evaluation of the antileishmanial drug miltefosine-dsDNA interaction was investigated in incubated solutions and using dsDNA-electrochemical biosensors, following the changes in the oxidation peaks of guanosine and adenosine residues, and the occurrence of the free guanine residues, electrochemical signal. The electrochemical behaviour of miltefosine was also investigated, at a glassy carbon electrode, using cyclic, differential pulse and square wave voltammetry and no electrochemical redox processes were observed. The interaction mechanism of miltefosinedsDNA occurs in two ways: independent of the dsDNA sequence, and leading to the condensation/aggregation of DNA strands, producing a rigid miltefosine-dsDNA complex structure, and a preferential interaction between the guanine hydrogen atoms in the CÀG base pair and miltefosine, causing the release of guanine residues detected on the electrode surface. Miltefosine did not induce oxidative damage to DNA in the experimental conditions used.