Background: Vitamin A (retinol)-deficiency and falciparum malaria are two major public health problems in developing countries. Falciparum malaria is associated with significant destruction of erythrocytes and can lead to severe anaemia. Methodology: The present study was designed to estimate the serum retinol concentration in 150 acute falciparum-malaria patients (aged two to five years) by High Performance Liquid Chromatography (HPLC). Serum retinol concentrations of 20 healthy (age and sex matched, population based) volunteers were included as controls. Results: The mean serum retinol concentration of healthy controls was 34.31 ± 1.274 µg/dl and that of diseased cohort was 12.562 ± 0.276 µg/dl. The mean parasitemia was 1239.2 ± 33.609 per µL. Conclusions: The diseased cohort demonstrated significant reduction in concentrations of retinol in comparison to healthy controls (p < 0.001) and there was an inverse relationship (coefficient of correlation r = -0.899) between parasitemia and serum retinol concentration.