The production of microalgae biomass shows wide valuable uses, in the aquaculture, biotechnology, and food science, among others. However, microalgae show fluctuations in their chemical profile generated mainly by the culture conditions. This study was designated to assess the effects of nitrogen starvation on growth, nutrient uptake, and gross chemical composition of Chlorella sp. and Nannochloropsis oculata. The control experiments were performed with Conway culture medium in 13-day batch cultures, 12 h photoperiod, and aeration. A second experimental condition was the addition of the nutrients except nitrogen, one week after the start of growth (experiments designed as N-). Cell yield were similar in the control and in the N- experiments for both species. Cell biovolumes did not vary over growth in the control, but both microalgae exhibited larger cell biovolumes in N- experiments, probably as a consequence of the higher accumulation of storage substances. Dissolved nitrogen was exhausted before the end of the experiments, but phosphorus was not totally consumed. Protein and total carotenoid did not vary from the exponential to the stationary growth phase of the control in both species. For Chlorella sp., concentrations of lipid did not vary in the control either, but there was a significant increase of carbohydrate over growth. In the N- experiment, concentrations of all substances varied throughout growth of Chlorella sp., except lipid. For N. oculata, all substances exhibited significant variations over growth, except protein and total carotenoid in the control. Protein and chlorophyll-a concentrations decreased over growth in N- experiments for both species. In contrast, concentrations of carbohydrate increased throughout growth in N- experiments, especially in Chlorella sp. Nitrogen starvation caused accumulation of carbohydrate, but increments of lipid were restricted to N. oculata. Both species showed a fast growth, but the small content of lipid in Chlorella sp. is unfavorable for its use as a food-species in a monospecific diet in mariculture, and as a feedstock for biodiesel production. N. oculata is a lipid-rich species, and its lipid content can be successfully incremented through nitrogen starvation. This species is promising in uses that demand high concentrations of lipid, such as the production of biodiesel.