http://grasasyaceites.revistas.csic.es/index.

php/grasasyaceites/article/view/1477/1517

The DPPH assay of radical scavenging activity was performed according to the procedure reported by Morales and

Jiménez–Pérez (2001). For the methanolic extract assay, an aliquot of 1 mL methanolic extract of Sacha Inchi oil was added

to 4 ml of DPPH (40 mg·L -1) in ethyl acetate. The mixture was vigorously stirred for a few seconds and kept in the dark for 1

h. Absorbance was measured at 515 nm against ethyl acetate. For Sacha Inchi oil samples, 1 g of oil was diluted with 10 mL

of n–hexane and an aliquote of 1mL of the dilution was added to 4 mL of DPPH solution which was previously diluted with

ethyl acetate (Minioti and Georgiou, 2010). In both measurements, methanolic solutions of gallic acid were used for

calibration ranging from 5 to 60 mM

y=– 0.0939x+0.086

being x=absorbance and y=μmol GAE·mL-1 sample.

The results were expressed as mmol equivalent of gallic acid per kg of sample.

Minioti KS, Georgiou CA. 2010. Comparison of different tests used in mapping the Greek virgin
olive oil production for the determination of its total antioxidant capacity. Grasas Aceites 61, 45–
51

A 1.3 104 M working solution of the DPPH radical in ethyl acetate which shows an absorbance of
approximately 1.2 at 515 nm was prepared daily. Olive oil aliquots of 20, 80, 120 and 180 mg or
170 µL of olive oil hydrophilic extract were added to 4.0 mL DPPH working solution. Mixtures were
vigorously stirred for a few seconds and kept in the dark for 1 h. Absorbencies were measured at
515 nm against ethyl acetate. Olive oil antioxidants scavenge the DPPH cation radical, resulting in
decolorization of its purple solution. Total antioxidant capacity, expressed as mmol L1 of gallic acid
equivalents (GAE) per kilogram of oil, is calculated using the appropriate amount that shows 50%
absorbance inhibition as determined by plotting absorbencies against the amount of olive oil .

0.50 g of olive oil sample was diluted 1:1 (v:v) in n-hexane. Diluted samples were extracted by two
0.50 mL portions of methanol:water 80:20 (v:v) solvent. After separation from the lipidic fraction
by 5 min of centrifugation at 5000 rpm, the two hydrophilic extracts were combined.

https://www.sciencedirect.com/science/article/pii/S2221169117310134

The radical scavenging activity of sacha inchi oil and its emulsion was
assayed by using DPPH method [14]. This method is based on a
single electron transfer mechanism and measures the ability of the
antioxidants of oil to reduce a stable DPPH radical [15]. The
DPPH solution was prepared at 0.1 mM. The standard solution was
prepared by dissolving 1 mg of vitamin C in 1 mL absolute alcohol. The
standard solution 300 μL was mixed with DPPH 1 000 μL. Control
solution was prepared by adding 300 μL of absolute alcohol then mixed
with DPPH 1 000 μL in test tube. The oil sample was prepared at 67 ppm
and 167 ppm in absolute alcohol. The sacha inchi oil emulsion (5% w/w
of seed oil in formulation) 1 g was extracted by dissolving in 3 mL
absolute alcohol and the mixture was then centrifuged at 5 000 rpm for
5 min. The clear solution of samples (300 μL) was mixed with DPPH
1 000 μL in test tube. Each mixture was kept in the dark for 30 min and
the absorbance was measured at 517 nm against a blank using UV–VIS
spectrophotometer. The ability of sacha inchi oil and its emulsion to
scavenge DPPH radical was calculated as % inhibition by the following
equation:

where A control is s the absorbance of the control reaction (containing all

reagents except the test compound), and A sample is absorbance of test
sample.