The extraction was performed with acetone (previously refrigerated
for 2 h) on 10–20 g of sample, using a pestle and mortar until
the residue became colourless, and after that the extract was partitioned
with petroleum ether. In the case of the C. maxima ‘Exposição’
samples, the extract was submitted to overnight saponification with
methanolic KOH (10%, w/v), while in the case of C. moschata ‘Menina
Brasileira’, where xanthophylls, which are oxy-carotenes, are in
lower concentrations, saponification was not performed in order
to minimise the loss which can occur in this step. The extracts were
washed with distilled water and concentrated at low pressure in a
rotoevaporator (Tecnal, TE-211, Piracicaba, Brazil), always at a temperature
below 35 C and using glass pearl for optimisation of the
recovery in the re-dissolving process. In order to avoid errors during
the carotenoid analysis, all the necessary precautions were taken as
recommended by Rodriguez-Amaya (1999).