Optimization of Extraction Condition of Bee Pollen Using Response Surface Methodology: Correlation between Anti-Melanogenesis, Antioxidant Activity, and Phenolic Content
Abstract
:1. Introduction
2. Results and Discussion
2.1. Anti-Melanogenesis and Antioxidant Activity of Bee Pollen Fractions
2.2. Optimization of Extraction Condition
2.2.1. Extraction Method Development
Run | Actual Variables | Observed Values | ||||
---|---|---|---|---|---|---|
EtOAc in MeOH (%) | Extraction Temperature (°C) | Extraction Time (h) | Tyrosinase Inhibition a (%) | Antioxidant Activity b (%) | Total Phenolic (μg GAE/mg Extract) | |
1 | 100 | 30 | 43 | 29.8 | 14.4 | 8.7 |
2 | 50 | 30 | 43 | 42.2 | 31.1 | 13.4 |
3 | 75 | 50 | 19 | 49.1 | 37.9 | 16.8 |
4 | 75 | 30 | 31 | 53.9 | 50.1 | 20.4 |
5 | 100 | 50 | 31 | 29.7 | 12.7 | 7.5 |
6 | 75 | 30 | 31 | 49.1 | 46.5 | 18.4 |
7 | 75 | 30 | 31 | 55.9 | 46.7 | 19.3 |
8 | 75 | 50 | 43 | 50.6 | 38.6 | 17.7 |
9 | 100 | 30 | 19 | 28.3 | 12.7 | 8.0 |
10 | 50 | 50 | 31 | 32.3 | 25.4 | 13.6 |
11 | 50 | 10 | 31 | 45.7 | 32.8 | 14.1 |
12 | 50 | 30 | 19 | 40.8 | 32.9 | 13.8 |
13 | 75 | 10 | 43 | 57.9 | 42.0 | 18.4 |
14 | 75 | 10 | 19 | 55.3 | 48.5 | 20.4 |
15 | 100 | 10 | 31 | 18.4 | 13.0 | 7.4 |
2.2.2. Fitting the Models
Coefficient | Standard Error | t Value | p Value | |
---|---|---|---|---|
(Tyrosinase Inhibition) | ||||
Intercept | 52.96 | 1.707 | 31.032 | <0.001 |
X1 | −6.868 | 1.045 | −6.571 | 0.001 |
X2 | 0.883 | 1.045 | 0.844 | 0.437 |
X3 | −1.943 | 1.045 | −1.859 | 0.122 |
−19.69 | 1.538 | −12.8 | <0.001 | |
2.015 | 1.538 | 1.31 | 0.247 | |
−1.755 | 1.538 | −1.141 | 0.306 | |
X1X2 | 0.04 | 1.478 | 0.027 | 0.979 |
X1X3 | 0.16 | 1.478 | 4.168 | 0.009 |
X2X3 | −0.315 | 1.478 | −0.213 | 0.84 |
(Antioxidant Activity) | ||||
Intercept | 47.473 | 1.15 | 41.534 | <0.001 |
X1 | −8.663 | 0.704 | −12.306 | <0.001 |
X2 | -0.75 | 0.704 | −1.065 | 0.335 |
X3 | −2.723 | 0.704 | −3.868 | 0.012 |
−22.884 | 1.036 | −22.086 | <0.001 | |
−2.089 | 1.036 | −2.016 | 0.1 | |
−3.889 | 1.036 | −3.754 | 0.013 | |
X1X2 | 0.9 | 0.996 | 0.904 | 0.407 |
X1X3 | 1.755 | 0.996 | 1.763 | 0.138 |
X2X3 | 1.8 | 0.996 | 1.808 | 0.13 |
Sum of Square | Degree of Freedom | Mean Square | F Value | p Value | |
---|---|---|---|---|---|
(Tyrosinase inhibition) | |||||
Regression | 2044.58 | 9 | 227.175 | 26 | 0.001 |
Linear | 413.72 | 3 | 137.906 | 15.78 | 0.006 |
Square | 1478.67 | 3 | 492.891 | 56.41 | <0.000 |
Interaction | 152.19 | 3 | 50.729 | 5.81 | 0.044 |
Residual error | 43.69 | 5 | 8.737 | ||
Lack-of-fit | 19.72 | 3 | 6.573 | 0.55 | 0.697 |
Pure error | 23.97 | 2 | 11.985 | ||
Total | 2088.26 | 14 | |||
R2 = 0.979, adjusted R2 = 0.941 | |||||
(Antioxidant activity) | |||||
Regression | 2643.18 | 9 | 293.687 | 74.09 | <0.001 |
Linear | 664.11 | 3 | 221.369 | 55.84 | <0.001 |
Square | 1950.56 | 3 | 650.185 | 164.02 | <0.001 |
Interaction | 28.52 | 3 | 9.507 | 2.4 | 0.184 |
Residual error | 19.82 | 5 | 3.964 | ||
Lack-of-fit | 11.54 | 3 | 3.848 | 0.93 | 0.556 |
Pure error | 8.28 | 2 | 4.138 | ||
Total | 2663 | 14 | |||
R2 = 0.993, adjusted R2 = 0.979 |
2.2.3. Effect of Extraction Variables on Tyrosinase Inhibition and Antioxidant Activity
2.3. Optimization of Extraction Parameters and Verification
Extraction Condition | Tyrosinase Inhibition a | Antioxidant Activity b | ||||
---|---|---|---|---|---|---|
EtOAc in MeOH (%) | Extraction Temperature (°C) | Extraction Time (h) | Predicted | Observed | Predicted | Observed |
69.6 | 10.0 | 24.2 | 55.0 | 57.9 | 48.6 | 49.3 |
2.4. Correlation between Activity and Phenolic Content
2.5. Discussion
3. Experimental Section
3.1. General Information
3.1.1. Bee Pollen
3.1.2. Preparation of Extract and Fractions
3.2. Response Surface Methodology
3.3. Evaluation of Anti-Melanogenesis and Antioxidant Activity
3.3.1. Preparation of Samples
3.3.2. Assessment of Tyrosinase Activity
3.3.3. Measurement of Antioxidant Activity
3.4. Measurement of Total Phenolic Content
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Kim, S.B.; Jo, Y.H.; Liu, Q.; Ahn, J.H.; Hong, I.P.; Han, S.M.; Hwang, B.Y.; Lee, M.K. Optimization of Extraction Condition of Bee Pollen Using Response Surface Methodology: Correlation between Anti-Melanogenesis, Antioxidant Activity, and Phenolic Content. Molecules 2015, 20, 19764-19774. https://doi.org/10.3390/molecules201119656
Kim SB, Jo YH, Liu Q, Ahn JH, Hong IP, Han SM, Hwang BY, Lee MK. Optimization of Extraction Condition of Bee Pollen Using Response Surface Methodology: Correlation between Anti-Melanogenesis, Antioxidant Activity, and Phenolic Content. Molecules. 2015; 20(11):19764-19774. https://doi.org/10.3390/molecules201119656
Chicago/Turabian StyleKim, Seon Beom, Yang Hee Jo, Qing Liu, Jong Hoon Ahn, In Pyo Hong, Sang Mi Han, Bang Yeon Hwang, and Mi Kyeong Lee. 2015. "Optimization of Extraction Condition of Bee Pollen Using Response Surface Methodology: Correlation between Anti-Melanogenesis, Antioxidant Activity, and Phenolic Content" Molecules 20, no. 11: 19764-19774. https://doi.org/10.3390/molecules201119656