94
S. Ghosh et al.
2.3.1 DPPH Free Radical Scavenging Activity Assay
The mayonnaises were assessed using 1, 1-diphenyl 2-picrylhydrazyl (DPPH) radical
scavenging assay (Gorinstein et al. 2007). 0.1 mM solution of DPPH in methanol
was prepared. An aliquot of 0.2 ml of sample was added to 2.8 ml of this solution
and kept in the dark place for 30 min. The absorbance was measured at 517 nm. The
ability to scavenge the DPPH radical was calculated with the following equation.
Inhibition percentage (I%) = (A 0 − A 1 )/A 0 (A 0 = Absorbance of the control, A 1 =
Absorbance of the sample.)
2.3.2 Ferric Reducing Antioxidant Power (FRAP)
The FRAP assay of the test samples was determined (Benzie and Strain 1996).
The FRAP reagent consists of 10 mM TPTZ in 40 mM HCL, 250 mM sodium
acetate buffer (pH −3.6), and 20 mM FeCl 3 . The reagent was freshly prepared by
mixing TPTZ solution, FeCl 3 solution, and acetate buffer in a ratio of 1:1:10. An
extract solution (100 µl) was mixed with 900 µl of FRAP reagent. The mixture
was incubated at 37 °C temperature for 4 min and the absorbance was measured at
593 nm.
2.3.3 ABTS Free Radical Scavenging Activity
ABTS assay of mayonnaises were measured using the following method (Fatma
et al. 2016). A solution of ABTS (7 µM) was prepared in distilled water and mixed
with the solution of potassium per sulfate (2.45 µM). The mixture was kept in the
dark place for 16 h at room temperature. The resulting intense color matches the
ABTS radical cations. The solution subsequently diluted with distilled water and
absorbance was measured at 734 nm. 1 ml of ABTS diluted solution was mixed with
10 µl of sample at different concentration and the reaction mixture was kept for
6 min before measuring the absorbance. ABTS scavenging activity was calculated
by the following equation. Inhibition Percentage (I%) = (1 − A/A 0 ) × 100. (A =
Absorbance of the sample. A 0 = Absorbance of the ABTS solution.)
2.4 Fourier-Transformed Infrared Spectroscopy (FTIR)
Analysis
The functional groups, present in both sample and market mayonnaise, was analyzed
by FTIR (Siregar et al. 2018) equipped with a tri glycine sulfate (TGS) detector.
Absorbance intensity was measured at 4 cm
−1 resolution. 2–3 mg of sample was
S. Ghosh et al.
2.3.1 DPPH Free Radical Scavenging Activity Assay
The mayonnaises were assessed using 1, 1-diphenyl 2-picrylhydrazyl (DPPH) radical
scavenging assay (Gorinstein et al. 2007). 0.1 mM solution of DPPH in methanol
was prepared. An aliquot of 0.2 ml of sample was added to 2.8 ml of this solution
and kept in the dark place for 30 min. The absorbance was measured at 517 nm. The
ability to scavenge the DPPH radical was calculated with the following equation.
Inhibition percentage (I%) = (A 0 − A 1 )/A 0 (A 0 = Absorbance of the control, A 1 =
Absorbance of the sample.)
2.3.2 Ferric Reducing Antioxidant Power (FRAP)
The FRAP assay of the test samples was determined (Benzie and Strain 1996).
The FRAP reagent consists of 10 mM TPTZ in 40 mM HCL, 250 mM sodium
acetate buffer (pH −3.6), and 20 mM FeCl 3 . The reagent was freshly prepared by
mixing TPTZ solution, FeCl 3 solution, and acetate buffer in a ratio of 1:1:10. An
extract solution (100 µl) was mixed with 900 µl of FRAP reagent. The mixture
was incubated at 37 °C temperature for 4 min and the absorbance was measured at
593 nm.
2.3.3 ABTS Free Radical Scavenging Activity
ABTS assay of mayonnaises were measured using the following method (Fatma
et al. 2016). A solution of ABTS (7 µM) was prepared in distilled water and mixed
with the solution of potassium per sulfate (2.45 µM). The mixture was kept in the
dark place for 16 h at room temperature. The resulting intense color matches the
ABTS radical cations. The solution subsequently diluted with distilled water and
absorbance was measured at 734 nm. 1 ml of ABTS diluted solution was mixed with
10 µl of sample at different concentration and the reaction mixture was kept for
6 min before measuring the absorbance. ABTS scavenging activity was calculated
by the following equation. Inhibition Percentage (I%) = (1 − A/A 0 ) × 100. (A =
Absorbance of the sample. A 0 = Absorbance of the ABTS solution.)
2.4 Fourier-Transformed Infrared Spectroscopy (FTIR)
Analysis
The functional groups, present in both sample and market mayonnaise, was analyzed
by FTIR (Siregar et al. 2018) equipped with a tri glycine sulfate (TGS) detector.
Absorbance intensity was measured at 4 cm
−1 resolution. 2–3 mg of sample was