In the SFE-EtOH procedure, EtOH was used as the co-solvent.
Seeds were prepared as described above and mixed with EtOH
(35mL/72 g of crushed seeds, approximately 1:2mL of EtOH:g
seeds) before the extraction. Next, the seeds were placed in
the extractor and the extraction was performed with a supercritical
pressure and temperature of 175 kgf/cm2 and 45 ◦C,
respectively. These conditions were those of the CCD central
point and produced the best per cent yield. Similar to the
supercritical extractions performed with no modifier polarities,
this extraction was also conducted for a standard period
of 16h to establish static equilibrium and the dynamic extraction
of period was 6h.
Purification of the extract from SFE-EtOH was performed
using the method described by Lhuillier et al. (2007) with modifications
(Oliveira et al., 2014)