2.3. Sample extraction
100 g of black and orange fresh carrots were peeled and finely
chopped, to avoid the traces of carotenes in the black carrots. These
samples were subjected to extraction with 100 mL EtOH:H2O (1:1,
v/v), containing 0.01% HCl (37% v/v) and left with stirring at room
temperature (25 8C) for a period of 2 h. Then, the extracts were
filtered through a 0.45 mm polytetrafluoroethylene (PTFE) membrane,
and the ethanol was evaporated under vacuum. The
resulting extracts were purified by liquid–liquid extraction with
chloroform (4 50 mL), pentane (4 50 mL) and cyclohexane (4
50 mL). The resulting aqueous fractions were concentrated in a
vacuum rotary evaporator until 2 mL at a temperature of 30 8C and
stored at 4 8C in the absence of light. The extraction was carried out
in triplicate for each sample, and the further analyses were
performed within 3 days.
2.3. Sample extraction100 g of black and orange fresh carrots were peeled and finelychopped, to avoid the traces of carotenes in the black carrots. Thesesamples were subjected to extraction with 100 mL EtOH:H2O (1:1,v/v), containing 0.01% HCl (37% v/v) and left with stirring at roomtemperature (25 8C) for a period of 2 h. Then, the extracts werefiltered through a 0.45 mm polytetrafluoroethylene (PTFE) membrane,and the ethanol was evaporated under vacuum. Theresulting extracts were purified by liquid–liquid extraction withchloroform (4 50 mL), pentane (4 50 mL) and cyclohexane (450 mL). The resulting aqueous fractions were concentrated in avacuum rotary evaporator until 2 mL at a temperature of 30 8C andstored at 4 8C in the absence of light. The extraction was carried outin triplicate for each sample, and the further analyses wereperformed within 3 days.
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