Safflower’s flowers were purchased from a local market
in Tehran and then grounded into powder using a
grinding machine and stored in plastic bags kept under
dry, cold and dark conditions for future analysis.
Cold solvent extraction of carotenoid compounds
(method of waterlogging)
In order to extract the color from the flower powders,
a mixture of three solvents including N-hexane, acetone,
and ethanol was used in the ratio of 2:1:1 and
with ratio of 1:10 sample to solvent according to the
method of Sadlr and co-workers (1990) and Shi (1999).
The extraction was done in a dark environment for 2 h
at room temperature on a magnetic stirrer. After the
desired time, samples were filtered by filter paper
Whatman No. 1, and then deionized distilled water was
added up to 20% of the volume of solvent, and then
samples were divided into organic and aqueous phases.
Soluble carotenoids and fat-soluble material were
extracted by air flow, but fat-soluble carotenoids solvent
was extracted by rotary evaporation under vacuum
conditions, cooled below 40° C. Afterwards the
extracts were placed inside a desiccator until constant
weight was observed. Finally the extract were poured
into dark-coloured containers and stored in a cool and
dry place.
Ultrasonic waves extraction of carotenoid compounds
After the time has elapsed, the mixture of sample and
solvent such as waterlogging prone method, ultrasonic
waves fixed in 24 kHz at room temperature between 5
and 15 minute time, and the intensity was 20, 40 and
60. After sonication, other steps such as filtration, addition
of deionized water, drying and packaging was
performed as waterlogging extracting carotenoids.