The use of specific microalgae as sources of polyunsaturated fatty acids (PUFA), for incorporation in either aquaculture or human diets, has been receiving increasing interest. The fatty acid profile of intracellular fats in a given species can, within certain limits, be modulated via manipulation of the culture growth conditions, namely nutrient availability. In attempts to shed further light on the effects of chemical parameters upon microalgal metabolism, correlations between the components of a medium and the resulting biomass yield, as well as the eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) contents are presented, using Pavlova lutheri as model system. Although several studies of individual nutrients have been published previously, the effects of variation of all medium components upon fatty acid production are systematically presented here, apparently for the first time.The optimum formulation to optimise EPA and DHA contents should consider the buffering agent, since some of them are toxic to the cells; P. lutheri was able to use several nitrogen sources, yielding equivalent amounts of EPA and DHA; synthesis of EPA was induced in culture media under low salinity, whereas a trend could be noticed between decreasing yields of EPA and increasing concentrations of N, at 0.15 g L −1 of P. Regarding the micronutrient composition, a culture medium lacking sulphur was unable to sustain cellular growth, whereas cultures deprived of either calcium, iron or manganese revealed a statistically significant decrease in cellular yields relative to those possessing such minerals; conversely, those cultures without boron produced higher cell numbers than those containing this element. Media deprived of boron, molybdenum or copper led to increases of the relative amounts of both EPA and DHA, whereas calcium deficiency decreased the levels of those PUFA.