The present study evaluated the applicability of 1H NMR and UV-Vis spectroscopies as analytical techniques for the characterization and determination of biodiesel conversion and for monitoring the oxidative stability of biodiesel samples with antioxidants. For this study, safflower and babassu biodiesels were obtained through transesterification, and physicochemical properties confirmed the success of both reactions. A bench-top accelerated oxidation system was used as an alternative to the Rancimat® method, with samples of 6.0 g heated at 110 ± 5 °C and collected every 2 h for 12 h. The agreement for biodiesel conversions was good, with divergences between 2% and 0.4% for safflower biodiesel and 1.9% for babassu biodiesel. As for UV-Vis spectroscopy, the technique showed the same trend as the Rancimat® method, showing efficiency in evaluating the oxidative stability of safflower biodiesel and in the performance of antioxidants BHT and DMP-30. The accuracy of NMR signals integration for mixtures of safflower oil and safflower biodiesel and the use of UV-Vis spectroscopy associated with a bench-top accelerated oxidation system to investigate the performance of phenolic and amine antioxidants in safflower and babassu biodiesel were explored for the first time, showing results close to the standard methods. Therefore, 1H NMR and UV-Vis spectroscopies could be applied as alternatives to the GC and Rancimat® methods to determine conversion and monitor the oxidative stability of biodiesel rapidly and practically.