Aim: The aim of the study was to determine the antifungal activity of six different concentrations of the ethanol extract of Bixa orellana (L) (Achiote) on the strains of Candida albicans (ATCC 90028). Materials and methods: The ethanol extract of Bixa orellana (L) (BEE) was prepared in six different concentrations (5%, 10%, 20%, 40%, 60%, and 100%), which were confronted with the positive control group (Nystatin 100,000 UI/mL). The total sample consisted of n = 174 wells inoculated with each concentration evaluated in the Petri dishes. The cultivation procedure was carried out at a temperature of 37°C, and then the 6 mm diameter wells on which the sowing of the Candida albicans in suspension was carried out. Subsequently, the BEE (Achiote) was inoculated in all the concentrations mentioned in the same way for Nystatin; finally, the inhibition halos were evaluated by the Kirby-Bauer method at the scheduled times of 24, 48, and 72 hours. Results: The highest antifungal activity was found at 24 hours and this increased directly proportional to the concentration of BEE (5%, 10%, 20%, 40%, 60%, and 100%): 11 ± 0, 11 ± 0, 13.1 ± 1.3, 17.2 ± 0.8, 19.1 ± 1.0, and 21.4 ± 1.3 mm, respectively. However, there was no statistically significant difference when compared with Nystatin, at p > 0.05 Conclusion: It was shown that 5%, 10%, 20%, 40%, 60%, and 100% BEE had an antifungal effect against Candida albicans. However, there were no statistically significant differences against the effect of Nystatin at 100,000 IU/mL, who has an inhibitory effect greater than BEE, with a significance level of p < 0.05. Clinical significance: This article describes the antifungal activity and the possible impact on dentistry that the use of Bixa orellana (L) may have because according to our results it had an antifungal effect against strains of Candida albicans. This opens a great line of research to potentially use this natural resource in dentistry.