Biodiesel is derived from renewable sources, such as vegetable oils, by means of a transesterification process in which triacylglycerols are transformed into smaller molecules of esters of fatty acids and glycerol. The transesterification reactions of 'Gefner' atemoya (Annona squamosa × A. cherimola) seed oil extracted by pressing (physical) and solvent (chemical) processes were studied, with analysis of the methyl esters produced. The reactions were monitored using gas chromatography coupled to mass spectrometry (GC-MS), as well as by hydrogen nuclear magnetic resonance spectroscopy ( 1 H-NMR). The methyl esters formed during the transesterification reaction with methanol were determined for each oil. The major methyl esters (16:0, 18:0, 18:1 and 18:2) formed during 50 min of reaction were similar to those reported in the literature for other biodiesels; the peak areas and retention times were also similar. No changes in signal intensity over time were observed for the oils obtained by the two extraction methods. It was also noted that the extraction method had no influence on the types of methyl esters formed during biodiesel production.