Microalgae are autotrophs and CO2 fixers with great potential to produce biofuels in a sustainable way, however the high cost of biomass production is a challenge. Mixotrophic growth of microalgae has been presented as a great alternative to achieve economic sustainability. Thus, the present work reports the energetic characterization of S. platensis biomasses cultivated under autotrophic (A) and mixotrophic conditions using cheese whey waste at different concentrations, 2.5 (M2.5), 5.0 (M5) and 10.0% (M10), in order to analyze the potential production of valuable chemicals and bio-oil by TGA/DTG and Py-GC/MS. The biochemical compositions of the studied biomasses were different due to the influence of different culture mediums. As the whey concentration increased, there was an increase in the carbohydrate content and a decrease in the protein content, which influenced the elemental composition, calorific value, TGA and volatile compounds evaluated by Py-GC/MS at 450°C, 550°C and 650°C. Sample M10 had lower protein content and formed a smaller amount of nitrogenates compounds by pyrolysis at all temperatures evaluated. There was a reduction of 43.8% (450º), 45.6% (550ºC) and 23.8% (650ºC) in the formation of nitrogenates compounds in relation to sample A. Moreover, the temperature also showed a considerable effect in the formation of volatile compounds. The highest yields of nitrogenates compounds, phenols and aromatic and non-aromatic hydrocarbons were observed at 650ºC. The oxygenated, and N and O containing compounds decreased as the temperature increased. Hydrocarbons such as toluene, heptadecane and heneicosane were produced by S.platensis pyrolysis, which makes this biomass attractive for production of high quality bio-oil and valuable chemicals. Therefore, the results showed that it is possible to decrease the formation of nitrogen compounds via manipulation of growth conditions and temperature.