The carotenoid analysis in foods includes several steps: sampling, subsampling, and size reduction to obtain a homogenized size-laboratory sample, followed by exhaustive extraction usually with acetone and liquid-liquid partition to ether/diethyl ether. At this point, the carotenoid extract is optionally saponified with 10% KOH in methanol (MeOH) when carotenoid esters are not the target, followed by alkali washout, complete solvent evaporation, and storage under low temperature. Before chromatographic separation, the extract is dissolved in methyl t-butyl ether (MTBE)/MeOH and analyzed by high-performance liquid chromatography (LC) on a C30 column with gradient elution of MeOH/MTBE for saponified samples and water/MeOH/MTBE for non-saponified ones. Detection is achieved by a diode array detector followed by a mass spectrometer with atmospheric pressure chemical ionization at positive mode. Identification is based on the combined information provided by elution order on the C30 column, UV-visible spectrum features (maximum wavelengths, spectral fine structure, and cis-peak intensity), and mass spectrum (protonated molecule and tandem mass fragments from both polyene chain and end-groups). Quantification is carried out with external calibration curves. Finally, selectivity, recovery, repeatability, and limits of quantification and detection are determined to evaluate the method suitability and result reliability.