This paper presents the Fourier transform infrared (FTIR) spectroscopic analysis of cholesterol, with the aim of highlighting the characteristic functional groups and confirming its chemical structure. The FTIR spectrum obtained shows significant absorption bands in the region of ~3400 cm-1, corresponding to the O–H stretching vibrations of the hydroxyl group. The intense bands in the region of ~2800–3000 cm-1 are associated with the C–H stretching vibrations of the aliphatic chains. Also, a weak band around ~1660 cm-1 is observed, attributed to the C=C double bond in the steroid nucleus. The bands in the region of ~1050–1150 cm-1 correspond to the C–O valence vibrations, confirming the presence of the secondary alcohol. The spectrum obtained is consistent with the known chemical structure of cholesterol and validates the purity of the analyzed sample. These results highlight the usefulness of the FTIR technique in the identification of lipid-based biological compounds and in their quality control.