| capillary zone electrophoresis (CE) |
Once the antioxidant butylated
hydroxytoluene hydrolyzed the acid, the isoflavones from a coffee
sample were extracted as well as purified using ether (BHT). An applied voltage of 25 kV, a buffer solution
of 20 mmol/L Na2HPO4, a hydrodynamic injection lasting 3 s at 30 mbar, and UV detection at 254 nm were experimental parameters
for the CE separation procedure. According to the findings, all three
substances may be examined within 10 min with a linearity of 0.5–50 g/mL. |
(17) |
| |
For DDZ, formononetin, and
genistein, 0.134, 0.0642, and 0.0825 g/mL were the limits of detection, respectively. |
|
| HPLC |
DDZ (15.2 min) and genistein, two isoflavonoids, were entirely
separated by the HPLC technique (17.3 min). First, genistein and DDZ concentrations in the analyzed milk prepared
from soy were calculated. Genistein (25.86 mg L–1 ± 0.66 SD) and DDZ (8.25 mg L–1 ± 1.13 SD) were
found in commercial soy milk samples when isoflavones were tested.
Genistein concentrations in soy milk were greater than DDZ concentrations.
On the basis of the major isoflavone concentration of soy milk, the study’s findings can be used to estimate how
much soy milk each individual can consume. |
(18) |
| radioimmunoassay |
Based on polyclonal antibodies
against DDZ-4′-O-(carboxymethyl)ether-BSA, radioimmunoassay was used. The assay’s intra- and interassay coefficients
of variation varied from 4.1 to 11.5% and from 5.6 to 21.7%, respectively,
and it had a sensitivity of 0.4 pg/tube based on the sample’s concentration
of DDZ and the technique (direct or extraction). Only 8% of the DDZ levels obtained by
direct radioimmunoassay were achieved after the removal of human sera
using diethyl ether. Mean and peak basal serum levels of free DDZ
were recorded through this experiment. It implies that the initial
DDZ unconjugated serial tests were made feasible by plasma as well
as the initial phytoestrogen immunoassay in bodily fluids from humans. |
(19) |
| extraction of supercritical
fluids |
An ideal environment
for
supercritical fluid extraction (SFE) was discovered. A solid–liquid extraction method using an aqueous methanol solution was tested
at various temperatures, pressures, and cosolvent concentrations (methanol, ethanol, and acetonitrile). The extraction conditions were 50–70 °C, 176–380 bar, and a modifier
of 0, 5, or 10 mol % cosolvents dissolved
in water. The greatest concentrations of DDZ and genistein were discovered
and extracted under the following conditions: static and dynamic extraction
for 15 min at 60 °C, 380 bar of
pressure, and 10% acetonitrile addition. It was found that, in comparison
to supercritical extraction, the amounts of DDZ and genistein produced
using solid–liquid extraction
were higher by 86 and 63%, respectively. |
(20) |
