Supplement Data

Supplement Data

Table S1 Experimental domain of central composite face centered design (Coded and actual values) for different selected solvents to extract polyphenols from whole mung bean (WMB)

Solvent / Independent variables / Levels
-1 / 0 / 1
Acetone / Solvent concentration (%v/v) / 30 / 60 / 90
Temperature (oC) / 40 / 50 / 60
Duration (min) / 40 / 80 / 120
Ethanol / Solvent concentration (%v/v) / 10 / 25 / 40
Temperature (oC) / 40 / 50 / 60
Duration (min) / 30 / 60 / 90
Methanol / Solvent concentration (%v/v) / 5 / 20 / 35
Temperature (oC) / 40 / 50 / 60
Duration (min) / 30 / 60 / 90
Water (100%) / Temperature (oC) / 40 / 50 / 60
Duration (min) / 120 / 180 / 240

Table S2 The experimental design of acetone in coded and actual value for the optimization of extraction variables using central composite face centered design

Standard ordera / Run orderb / Coded value / Actual value
Acetone
concentration (% v/v) / Temperature (oC) / Duration (min) / Acetone concentration (% v/v) / Temperature (oC) / Duration (min)
1 / 11 / -1 / -1 / -1 / 30 / 40 / 40
2 / 17 / 1 / -1 / -1 / 90 / 40 / 40
3 / 19 / -1 / 1 / -1 / 30 / 60 / 40
4 / 1 / 1 / 1 / -1 / 90 / 60 / 40
5 / 12 / -1 / -1 / 1 / 30 / 40 / 120
6 / 2 / 1 / -1 / 1 / 90 / 40 / 120
7 / 10 / -1 / 1 / 1 / 30 / 60 / 120
8 / 20 / 1 / 1 / 1 / 90 / 60 / 120
9 / 7 / -1 / 0 / 0 / 30 / 50 / 80
10 / 3 / 1 / 0 / 0 / 90 / 50 / 80
11 / 6 / 0 / -1 / 0 / 60 / 40 / 80
12 / 4 / 0 / 1 / 0 / 60 / 60 / 80
13 / 9 / 0 / 0 / -1 / 60 / 50 / 40
14 / 5 / 0 / 0 / 1 / 60 / 50 / 120
15 / 8 / 0 / 0 / 0 / 60 / 50 / 80
16 / 14 / 0 / 0 / 0 / 60 / 50 / 80
17 / 13 / 0 / 0 / 0 / 60 / 50 / 80
18 / 15 / 0 / 0 / 0 / 60 / 50 / 80
19 / 18 / 0 / 0 / 0 / 60 / 50 / 80
20 / 16 / 0 / 0 / 0 / 60 / 50 / 80

a No randomized; b Randomized.

Table S3 The experimental design of ethanol in coded and actual value for the optimization of extraction variables using central composite face centered design

Standard ordera / Run orderb / Coded value / Actual value
Ethanol concentration (% v/v) / Temperature(oC) / Duration (min) / Ethanol concentration (% v/v) / Temperature (oC) / Duration (min)
1 / 6 / -1 / -1 / -1 / 10 / 40 / 30
2 / 12 / 1 / -1 / -1 / 40 / 40 / 30
3 / 16 / -1 / 1 / -1 / 10 / 60 / 30
4 / 2 / 1 / 1 / -1 / 40 / 60 / 30
5 / 8 / -1 / -1 / 1 / 10 / 40 / 90
6 / 18 / 1 / -1 / 1 / 40 / 40 / 90
7 / 10 / -1 / 1 / 1 / 10 / 60 / 90
8 / 3 / 1 / 1 / 1 / 40 / 60 / 90
9 / 11 / -1 / 0 / 0 / 10 / 50 / 60
10 / 13 / 1 / 0 / 0 / 40 / 50 / 60
11 / 4 / 0 / -1 / 0 / 25 / 40 / 60
12 / 7 / 0 / 1 / 0 / 25 / 60 / 60
13 / 14 / 0 / 0 / -1 / 25 / 50 / 30
14 / 17 / 0 / 0 / 1 / 25 / 50 / 90
15 / 5 / 0 / 0 / 0 / 25 / 50 / 60
16 / 9 / 0 / 0 / 0 / 25 / 50 / 60
17 / 20 / 0 / 0 / 0 / 25 / 50 / 60
18 / 15 / 0 / 0 / 0 / 25 / 50 / 60
19 / 19 / 0 / 0 / 0 / 25 / 50 / 60
20 / 1 / 0 / 0 / 0 / 25 / 50 / 60

a No randomized; b Randomized.

Table S4 The experimental design of methanol in coded and actual value for the optimization of extraction variables using central composite face centered design

Standard ordera / Run orderb / Coded value / Actual value
Methanol concentration (% v/v) / Temperature(oC) / Duration
(min) / Methanol concentration (% v/v) / Temperature (oC) / Duration (min)
1 / 14 / -1 / -1 / -1 / 5 / 40 / 30
2 / 4 / 1 / -1 / -1 / 35 / 40 / 30
3 / 8 / -1 / 1 / -1 / 5 / 60 / 30
4 / 5 / 1 / 1 / -1 / 35 / 60 / 30
5 / 12 / -1 / -1 / 1 / 5 / 40 / 90
6 / 19 / 1 / -1 / 1 / 35 / 40 / 90
7 / 1 / -1 / 1 / 1 / 5 / 60 / 90
8 / 7 / 1 / 1 / 1 / 35 / 60 / 90
9 / 18 / -1 / 0 / 0 / 5 / 50 / 60
10 / 17 / 1 / 0 / 0 / 35 / 50 / 60
11 / 16 / 0 / -1 / 0 / 20 / 40 / 60
12 / 15 / 0 / 1 / 0 / 20 / 60 / 60
13 / 11 / 0 / 0 / -1 / 20 / 50 / 30
14 / 20 / 0 / 0 / 1 / 20 / 50 / 90
15 / 3 / 0 / 0 / 0 / 20 / 50 / 60
16 / 6 / 0 / 0 / 0 / 20 / 50 / 60
17 / 2 / 0 / 0 / 0 / 20 / 50 / 60
18 / 13 / 0 / 0 / 0 / 20 / 50 / 60
19 / 10 / 0 / 0 / 0 / 20 / 50 / 60
20 / 9 / 0 / 0 / 0 / 20 / 50 / 60

a No randomized; b Randomized.

Table S5 The experimental design of water (100%) in coded and actual value for the optimization of extraction variables using central composite face centered design

Standard ordera / Run orderb / Coded value / Actual value
Temperature
(oC) / Duration
(min) / Temperature
(oC) / Duration
(min)
1 / 2 / -1 / -1 / 40 / 120
2 / 12 / 1 / -1 / 60 / 120
3 / 10 / -1 / 1 / 40 / 240
4 / 3 / 1 / 1 / 60 / 240
5 / 11 / -1 / 0 / 40 / 180
6 / 5 / 1 / 0 / 60 / 180
7 / 1 / 0 / -1 / 50 / 120
8 / 7 / 0 / 1 / 50 / 240
9 / 9 / 0 / 0 / 50 / 180
10 / 13 / 0 / 0 / 50 / 180
11 / 4 / 0 / 0 / 50 / 180
12 / 6 / 0 / 0 / 50 / 180
13 / 8 / 0 / 0 / 50 / 180

a No randomized; b Randomized.

Table S6 Experimental and predicted values for the response total antioixdant activity (TAA) under different solvent extraction conditions

Std.a order / TAAb (μmol of AAE/g of sample)
Acetone / Ethanol / Methanol / Water
Exp.c / Pred.d / Exp.c / Pred.d / Exp.c / Pred.d / Exp.c / Pred.d
1 / 31.92 / 31.47 / 34.68 / 35.00 / 23.87 / 24.16 / 25.88 / 25.93
2 / 9.73 / 8.28 / 37.42 / 36.96 / 32.49 / 32.43 / 33.23 / 32.71
3 / 34.14 / 33.50 / 40.01 / 39.58 / 36.05 / 35.40 / 33.56 / 33.64
4 / 11.38 / 12.14 / 41.19 / 41.24 / 38.19 / 37.57 / 37.16 / 36.67
5 / 34.62 / 33.59 / 39.51 / 39.22 / 26.61 / 27.01 / 34.52 / 34.38
6 / 11.21 / 11.57 / 40.84 / 41.01 / 34.50 / 34.95 / 38.28 / 39.29
7 / 47.22 / 48.40 / 39.52 / 39.73 / 37.02 / 36.87 / 27.74 / 28.20
8 / 28.03 / 28.21 / 41.79 / 41.23 / 39.19 / 38.70 / 33.61 / 34.02
9 / 36.56 / 37.49 / 41.27 / 41.45 / 35.01 / 35.11 / 35.26 / 35.71
10 / 15.66 / 15.80 / 42.37 / 43.17 / 39.44 / 40.16 / 36.21 / 35.71
11 / 32.26 / 34.82 / 43.61 / 43.87 / 35.81 / 34.73 / 35.88 / 35.71
12 / 45.64 / 44.15 / 45.55 / 46.27 / 40.32 / 42.23 / 35.10 / 35.71
13 / 39.86 / 41.64 / 43.48 / 43.99 / 40.89 / 41.93 / 36.97 / 35.71
14 / 51.43 / 50.73 / 45.63 / 46.09 / 44.13 / 43.92 / NA / NA
15 / 43.66 / 43.21 / 47.21 / 46.59 / 43.61 / 42.83 / NA / NA
16 / 44.92 / 43.21 / 45.35 / 46.59 / 42.96 / 42.83 / NA / NA
17 / 43.62 / 43.21 / 46.89 / 46.59 / 43.59 / 42.83 / NA / NA
18 / 43.46 / 43.21 / 47.56 / 46.59 / 42.71 / 42.83 / NA / NA
19 / 42.04 / 43.21 / 47.89 / 46.59 / 41.68 / 42.83 / NA / NA
20 / 43.72 / 43.21 / 46.59 / 46.59 / 44.09 / 42.83 / NA / NA

a Standard run - non randomized; b Total antioxidant activity in μmol AAE/g of sample; c Experimental value; d Predicted value; AAE, ascorbic acid equivalent

Table S7 Analysis of variance of the second-order polynomial model of different selected solvents for the total antioxidant activity (TAA) of WMB

Source of variation / SS / df / MS / F value / p value
Acetone
Model / 3178.01 / 9 / 353.11 / 146.01 / < 0.0001
Lack of fit / 19.97 / 5 / 3.99 / 4.74 / 0.0563
Pure error / 4.21 / 5 / 0.84
Total / 3202.19 / 19
Ethanol
Model / 248.87 / 9 / 27.65 / 37.61 / < 0.0001
Lack of fit / 3.35 / 5 / 0.67 / 0.84 / 0.5740
Pure error / 4.00 / 5 / 0.80
Total / 256.22 / 19
Methanol
Model / 593.17 / 9 / 65.91 / 54.39 / < 0.0001
Lack of fit / 8.46 / 5 / 1.69 / 2.31 / 0.1897
Pure error / 3.66 / 5 / 0.73
Total / 605.28 / 19
Water
Model / 150.19 / 5 / 30.04 / 48.09 / < 0.0001
Lack of fit / 2.09 / 3 / 0.70 / 1.22 / 0.4110
Pure error / 2.28 / 4 / 0.57
Total / 154.57 / 12

SS, Sum of square; MS, Mean square; df, Degree of freedom

Table S8 Estimated regression coefficients for the independent extraction variables of the second order polynomial equation for total antioxidant activity (TAA) of WMB

Parameter / Acetone / Ethanol / Methanol / Water
Constant / 43.21 / 46.59 / 42.83 / 35.71
X1 / -10.84*** / 0.86* / 2.53*** / NA
X2 / 4.67*** / 1.20** / 3.75*** / 2.45***
X3 / 4.55*** / 1.05** / 1.00* / 2.91***
X1X2 / 0.46 / -0.076 / - 1.53** / NA
X1X3 / 0.29 / - 0.040 / - 0.085 / NA
X2X3 / 3.19*** / -1.02* / - 0.35 / - 0.94*
X12 / -16.56*** / -4.28*** / - 5.19*** / NA
X22 / -3.72** / -1.52* / - 4.35*** / 1.13*
X32 / 2.97* / -1.55* / 0.097 / - 4.60***
p / 0.0001 / 0.0001 / 0.0001 / 0.0001
R2 / 99.24 / 97.13 / 98 / 97.17
Adjusted R2 / 98.57 / 94.55 / 96.20 / 95.15

Where, X1 , solvent concentration; X2 , temperature; X3 , duration; NA, not applicable

*p≤ 0.05; **p≤0.005; *** p≤ 0.0005;

Table S9 F values from ANOVA analysis of data (solvent vs. fraction, solvent vs. treatment and fraction vs. treatment) reported in table 3

DF / TPC / TFC / TAA / FRP / DPPH
Solvent / 3 / 57.22** / 7.45** / 25.46** / 66.85** / 4.81**
Fraction / 2 / 286.7** / 109.06** / 60.05** / 507.9** / 259.06**
Interaction / 6 / 31.03** / 3.65** / 8.58** / 68.67** / ND
Solvent / 3 / 4.75** / ND / 9.20** / 2.97* / ND
Treatment / 1 / 3.46* / 8.68** / 14.11** / ND / 4.58*
Interaction / 3 / ND / ND / ND / ND / ND
Fraction / 2 / 51.06** / 140.0** / 26.86** / 53.88** / 426.8**
Treatment / 1 / 7.41* / 45.78** / 17.76** / 4.69* / 63.75**
Interaction / 2 / ND / 11.15** / ND / ND / 3.21*

DF, degree of freedom; TPC, Total phenolic content, TFC, total flavonoids content; TAA, Total antioxidant activity; FRP, Ferric reducing power; DPPH, 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity; * p≤ 0.05; ** p≤ 0.005

Table S10 F values from ANOVA analysis of data (solvent vs. fraction, solvent vs. treatment and fraction vs. treatment) reported in table 4.

DF / Gallic acid / PCA / Chlorogenic acid / Vanillic acid / Caffeic acid / p Coumaric / Ferulic acid / Sinapic acid / Quercetin / Catechin / Luteolin / Resveratrol / Trans-stilbene
Solvent / 3 / 5.80** / 12.26** / 4.65** / 6.28** / 6.75** / ND / 2.97* / 3.50* / 10.67** / 5.42** / ND / 19.08** / 3.69*
Fraction / 2 / 14.92* / ND / 47.86** / 24.12** / 4.62* / 111.4** / 96.08** / 104** / 19.60** / 8.57** / 24.55** / 198.5** / 5.49**
Interaction / 6 / 4.33** / ND / 3.22* / 4.47** / 2.87* / ND / ND / ND / 4.40** / 2.36* / ND / 11.74** / 2.72*
Solvent / 3 / 4.35* / 13.24** / ND / 3.77* / 6.78** / ND / ND / ND / 7.84** / 6.31** / ND / 2.46* / 4.15**
Treatment / 1 / 21.13** / 3.35* / 3.99* / 4.09* / 6.31* / 4.10* / 7.40* / 5.81* / 19.38** / 22.45** / 14.11** / ND / 21.34**
Interaction / 3 / ND / ND / 3.17* / 4.32** / 5.53** / ND / ND / ND / 2.97* / 6.62** / ND / ND / 4.25**
Fraction / 2 / 13.48** / ND / 39.01** / 19.03** / 3.92* / 213.6** / 165.8** / 153.3** / 14.57** / 9.55** / 44.99** / 75.45** / 6.59**
Treatment / 1 / 25.43** / ND / 7.34* / 5.38* / 5.32* / 31.69** / 44.14** / 32.27** / 19.61** / 21.46** / 36.79** / 3.20* / 22.82**
Interaction / 2 / ND / 4.19* / ND / 5.98** / 5.65** / 16.08** / 9.73** / 7.98** / ND / 6.60** / 11.86* / ND / 8.10**

DF, degree of freedom; PCA, Protocatechuic acid; *p≤0.05; ** p≤ 0.005; ND, Not detected

Fig. S1 Boxplot of TPC, TFC, TAA, FRP, DPPH vs Solvent