HarvestPlus Method (19). Dried ground maize (3 g) was hydrated
at room temperature in water (10 mL) for 30 min followed by addition
of acetone (20 mL) for 15 min. Carotenoids were extracted by
grinding the mixture in a mortar and pestle with 50 mL acetone. The
residue was vacuum filtered in a Buchner funnel equipped with filter
paper (Whatman #2 filter paper, Middlesex, U.K.). The residue was
returned to the mortar, and the procedure was repeated until the residue
was nearly colorless, usually once more. One-third of the filtrate was
transferred to a separatory funnel containing 20 mL of petroleum ether,
to which 300 mL of distilled water was added. After the aqueous and
organic layers separated, the aqueous layer was discarded. The
procedure was repeated for the remaining filtrate. The organic phase
was washed 3 times with 200 mL of distilled water and passed through
anhydrous sodium sulfate (15 g) into a round-bottom flask. The
sample was concentrated with a rotary evaporator and dried under argon.
Samples, extracted and prepared using the original procedure, were
also analyzed on a Resolve C18 column (5 ím, 3.9 300 mm, Waters
Corporation, Milford, MA) according to published procedures (20, 21).
To determine the effect of extraction solvent, the HarvestPlus method
was also performed using hexanes and hexanes/ether mixture (25:75
v/v) instead of acetone.
HarvestPlus Method (19). Dried ground maize (3 g) was hydrated
at room temperature in water (10 mL) for 30 min followed by addition
of acetone (20 mL) for 15 min. Carotenoids were extracted by
grinding the mixture in a mortar and pestle with 50 mL acetone. The
residue was vacuum filtered in a Buchner funnel equipped with filter
paper (Whatman #2 filter paper, Middlesex, U.K.). The residue was
returned to the mortar, and the procedure was repeated until the residue
was nearly colorless, usually once more. One-third of the filtrate was
transferred to a separatory funnel containing 20 mL of petroleum ether,
to which 300 mL of distilled water was added. After the aqueous and
organic layers separated, the aqueous layer was discarded. The
procedure was repeated for the remaining filtrate. The organic phase
was washed 3 times with 200 mL of distilled water and passed through
anhydrous sodium sulfate (15 g) into a round-bottom flask. The
sample was concentrated with a rotary evaporator and dried under argon.
Samples, extracted and prepared using the original procedure, were
also analyzed on a Resolve C18 column (5 ím, 3.9 300 mm, Waters
Corporation, Milford, MA) according to published procedures (20, 21).
To determine the effect of extraction solvent, the HarvestPlus method
was also performed using hexanes and hexanes/ether mixture (25:75
v/v) instead of acetone.
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