Chloroquine is the first-line therapy against the asexual stages of Plasmodium vivax . There is a high variation of chloroquine plasma levels after therapeutic doses, which can lead to inadequate exposure to the drug. The gender influence was low regarding the disposition of the drug, which is relevant as there are significant physiological variations between male and female patients. The objective of the study was to investigate whether gender modifies the pharmacokinetics parameters of chloroquine in patients with malaria vivax. A prospective study was performed in male and female adult patients using chloroquine (total dose of 25 mg/kg for three days) combined with primaquine. Serial blood samples were collected at admission and up to 672 h post-administration of the drugs. Chloroquine was measured in plasma samples by high-performance liquid chromatography with fluorescence detection. A non-compartmental analysis was used for modeling the data. A total of 26 male and 25 female patients were enrolled in the study. The pharmacokinetic parameters of chloroquine were similar between male and female patients: a half-life of 9.5 days and 10.2 days, maximum concentration (Cmax) of 1295 ng/ml and 1220 ng/ml, area-under-the-curve (AUC 0–28) of 241 µg/mL h and 237 µg/mL h, observed clearance (CL/f) of 5.8 and 5.5 L/h and the volume of distribution (V/f) of 1869 L and 1936 L. The study results suggest that a similar dose regimen of chloroquine combined with primaquine provides a comparable pattern of exposure in male and female patients.