The purpose of this study was to assess the vitamin A status of lactating women in southern Ethiopia. A total of 108 women participated in this cross-sectional investigation. Anthropometric measurements, blood samples, breast milk, dietary habits, socioeconomic data, and dark adaptation were analyzed. Pearson correlation coefficients were calculated for all appropriate variables. Significance was set at p < 0.05. The mean serum retinol level in participants as determined by HPLC and fluorometry was 1.49 � 0.04 and 1.68 � 0.04 ?mol/L, respectively. Results from analysis of serum retinol concentrations by HPLC and fluorometry were highly correlated (r = 0.63, p < 0.0001). Forty one percent of the participants had low levels of breast milk vitamin A. Pupillary threshold scores and serum retinol concentrations tended to be related, as did pupillary threshold scores and breast milk vitamin A.