<P><B>Abstract</B></P> <P>The present work evaluated the use of ultrasound as a pre-treatment to lipid and carotenoid extraction from the microalgae <I>Heterochlorella luteoviridis</I>. The pre-treatment was performed in the presence of ethanol (25%, v/v) with the ultrasound intensity varying from 0 to 100% (435kJkg<SUP>−1</SUP>). After the pre-treatment, a diffusive step was performed in order to evaluate different ethanol concentrations (50–75%, v/v). The results regarding carotenoid extraction showed that there is an optimal extraction region: 40–80% of ultrasound intensity and 60–75% of ethanol concentration. The lipid extraction was not influenced by the ultrasound pre-treatment and increased with the increase of ethanol concentration.</P> <P><B>Highlights</B></P> <P> <UL> <LI> An alternative carotenoid and lipid extraction method from microalgae was studied. </LI> <LI> Ultrasound was evaluated as a pre-treatment to the extraction. </LI> <LI> Ethanol at different concentrations was evaluated as extraction solvent. </LI> <LI> The methodology showed to be attractive for carotenoid extraction. </LI> </UL> </P>