The carotenoid separation was carried out using high performance
liquid chromatography (HPLC) according to Azevedo-
Meleiro and Rodriguez-Amaya (2004), with the equipment Varian
PS-335 Photodiode Array Detector, PS-240 solvent delivery, PS-410
auto sampler and software Galaxie 1.9. The carotenoids were separated
in column C18 ODS-2 150 4.6 mm3 lm (Waters), using
acetonitrile (containing 0.05% triethylamine):methanol:ethyl acetate
95:5:0, during the initial 20 min of the running, and
60:20:20 between 20 and 45 min of the running. The flow used
was 0.5 mL/min and the injected volume was 10 lL. The identifications
were performed through the absorption spectra in UV–Vis
given by the photodiode array detector (PDA), according to the elution
order and comparison with standards.