The carotenoids were separated using thin-layer chromatography (TLC) with activated 20×20 cm silica gel plates (Silica gel G. Merck, type 60, Darmstadt, Germany) following the modified procedure described by Sánchez-Camargo, Almeida Meireles, Lopes, and Cabral (2011). The extracted carotenoids were applied onto the plates and the separation was carried out using a mobile phase (acetone/hexane, (25:75%, v/v)). β-carotene and astaxanthin were used as standards. Rf of sample bands were calculated.