Numerous carotenoid-producing Gram positive bacteria have now been identified. Their carotenoid profiles are typically complex with C(30) diapocarotenoid skeletons being modified by glucosidation and esterification. In the present article the protocols necessary to identify and quantify these diapocarotenoids are described. Bacillus species capable of forming spores will be used as generic examples and the associated procedures will cover the cultivation of the bacteria, extraction, chromatographic separations, and online detection by Photo Diode Array (PDA) and Mass Spectrometry (MS).