Optimization of Ultrasound-Assisted Extraction of Flavonoids from Olive (Olea europaea) Leaves, and Evaluation of Their Antioxidant and Anticancer Activities
Abstract
:1. Introduction
2. Results and Discussion
2.1. Single Factor Experiment
2.1.1. Effect of Extraction Temperature
2.1.2. Effect of Ultrasound Power
2.1.3. Effect of Extraction Time
2.1.4. Effect of Liquid-Solid Ratio
2.2. Response Surface Optimization of Flavonoids
2.2.1. Analysis of Extraction Model
2.2.2. Analysis of Response Surface
2.2.3. Optimization of Extraction Conditions
2.3. Analysis of Flavonoid Compositions
2.4. Assay of Antioxidant Activity
2.4.1. Reducing Power
2.4.2. DPPH Radical Scavenging Activity
2.4.3. Superoxide Radical Scavenging Activity
2.5. Assay of Cell Viability
3. Material and Methods
3.1. Materials and Chemicals
3.2. Ultrasound-Assisted Extraction (UAE)
3.3. Response Surface Methodology (RSM) Design
3.4. Determination of Total Flavonoids
3.5. HPLC Analysis
3.6. Antioxidant Activity Assays
3.6.1. Reducing Power
3.6.2. DPPH Radical Scavenging Activity
3.6.3. Superoxide Radical-Scavenging Activity
3.7. Anticancer Activity Assays
3.7.1. Cell Culture
3.7.2. Cell Viability
3.8. Statistical Analysis
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the flavonoids extract are available from the authors. |
Run | Extraction Variables | Extraction Yield of Flavonoids (mg RE/g) | |||
---|---|---|---|---|---|
X1 Temperature (°C) | X2 Power (W) | X3 Time (min) | X4 Liquid-Solid Ratio (mL/g) | ||
1 | −1(45) | −1(240) | 0(50) | 0(40) | 66.12 |
2 | 0(50) | −1(240) | 0(50) | 1(50) | 69.84 |
3 | −1(45) | 0(270) | −1(40) | 0(40) | 66.72 |
4 | 0(50) | 1(300) | 0(50) | −1(30) | 66.23 |
5 | 1(55) | 0(270) | 1(60) | 0(40) | 66.87 |
6 | −1(45) | 0(270) | 0(50) | 1(50) | 68.26 |
7 | 1(55) | 0(270) | 0(50) | −1(30) | 66.16 |
8 | 0(50) | −1(240) | −1(40) | 0(40) | 68.77 |
9 | 0(50) | 0(270) | −1(40) | −1(30) | 66.15 |
10 | 0(50) | 0(270) | 1(60) | 1(50) | 67.52 |
11 | −1(45) | 0(270) | 1(60) | 0(40) | 62.98 |
12 | −1(45) | 1(300) | 0(50) | 0(40) | 64.89 |
13 | 0(50) | 0(270) | −1(40) | 1(50) | 67.84 |
14 | 0(50) | 0(270) | 0(50) | 0(40) | 74.91 |
15 | 0(50) | 1(300) | 1(60) | 0(40) | 66.30 |
16 | 0(50) | −1(240) | 1(60) | 0(40) | 67.31 |
17 | 1(55) | 1(300) | 0(50) | 0(40) | 66.24 |
18 | 0(50) | 0(270) | 0(50) | 0(40) | 74.90 |
19 | 0(50) | 0(270) | 1(60) | −1(30) | 65.16 |
20 | 1(55) | 0(270) | 0(50) | 1(50) | 66.30 |
21 | 1(55) | −1(240) | 0(50) | 0(40) | 66.37 |
22 | 0(50) | 1(300) | 0(50) | 1(50) | 67.40 |
23 | 0(50) | 0(270) | 0(50) | 0(40) | 74.57 |
24 | 1(55) | 0(270) | −1(60) | 0(40) | 64.67 |
25 | 0(50) | −1(240) | 0(50) | −1(30) | 66.00 |
26 | 0(50) | 0(270) | 0(50) | 0(40) | 74.91 |
27 | 0(50) | 1(300) | −1(40) | 0(40) | 66.60 |
28 | 0(50) | 0(270) | 0(50) | 0(40) | 74.67 |
29 | −1(45) | 0(270) | 0(50) | −1(30) | 62.82 |
Predicted | 50.10 | 267.32 | 49.62 | 41.64 | 74.93 |
Experimental | 50.00 | 270.00 | 50.00 | 41.00 | 74.95 |
Source | Sum of Squares | df | Mean Square | F-value | p-Value |
---|---|---|---|---|---|
Model | 348.72 | 14 | 24.91 | 285.81 | 0.0001 *** |
X1 | 1.94 | 1 | 1.94 | 22.21 | 0.0003 *** |
X2 | 3.8 | 1 | 3.8 | 43.57 | 0.0001 *** |
X3 | 1.77 | 1 | 1.77 | 20.32 | 0.0005 *** |
X4 | 17.86 | 1 | 17.86 | 204.94 | 0.0001 *** |
X1X2 | 0.3 | 1 | 0.3 | 3.47 | 0.0836 ns |
X1X3 | 8.82 | 1 | 8.82 | 101.21 | 0.0001 *** |
X1X4 | 7.02 | 1 | 7.02 | 80.58 | 0.0001 *** |
X2X3 | 0.34 | 1 | 0.34 | 3.86 | 0.0696 ns |
X2X4 | 1.78 | 1 | 1.78 | 20.45 | 0.0005 *** |
X3X4 | 0.11 | 1 | 0.11 | 1.29 | 0.2755 ns |
X12 | 178.28 | 1 | 178.28 | 2045.69 | 0.0001 *** |
X22 | 81.01 | 1 | 81.01 | 929.49 | 0.0001 *** |
X32 | 113.41 | 1 | 113.41 | 1301.31 | 0.0001 *** |
X42 | 95.28 | 1 | 95.28 | 1093.29 | 0.0001 *** |
Lack of Fit | 1.12 | 10 | 0.11 | 4.31 | 0.0860 ns |
Residual | 1.22 | 14 | 0.087 | - | - |
Pure Error | 0.1 | 4 | 0.026 | - | - |
Cor Total | 349.94 | 28 | - | - | - |
R2 | 0.9965 | - | - | - | - |
R2adj | 0.9930 | - | - | - | - |
C.V.% | 0.44 | - | - | - | - |
CP | CE | CC | April | July | ||
---|---|---|---|---|---|---|
AL | YL | AL | YL | |||
Total flavonoids content | y=0.1793x − 0.0012 | 0.9993 | 74.81 ± 0.91a | 58.17 ± 2.12b | 48.29 ± 1.41c | 34.06 ± 4.02 d |
Rutin | y =1915.3x + 2.6063 | 0.9988 | 0.85 ± 0.02a | 0.59 ± 0.01c | 0.68 ± 0.01b | 0.43 ± 0.01d |
Luteolin-4’-O-glucoside | y =256.15x + 7.0455 | 0.9981 | 31.02 ± 0.48a | 19.16 ± 0.37c | 24.20 ± 0.36b | 23.15 ± 0.85b |
Apigenin-7-O-glucoside | y =2745.8x + 0.1299 | 1.0000 | 2.06 ± 0.04a | 1.00 ± 0.02d | 1.53 ± 0.01b | 1.11 ± 0.01c |
Luteolin | y =3157.00x − 5.0318 | 0.9997 | 0.16 ± 0.001b | 0.60 ± 0.006a | 0.07 ± 0.0010d | 0.13 ± 0.004c |
Quercetin | y = 4910.80x − 3.6071 | 0.9989 | nd | 0.0021 ± 0.00004 | nd | 0.0008 ± 0.00004 |
Apigenin | y = 378228x − 1.0121 | 0.9991 | nd | 0.038 ± 0.0005 | nd | nd |
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Wang, B.; Qu, J.; Luo, S.; Feng, S.; Li, T.; Yuan, M.; Huang, Y.; Liao, J.; Yang, R.; Ding, C. Optimization of Ultrasound-Assisted Extraction of Flavonoids from Olive (Olea europaea) Leaves, and Evaluation of Their Antioxidant and Anticancer Activities. Molecules 2018, 23, 2513. https://doi.org/10.3390/molecules23102513
Wang B, Qu J, Luo S, Feng S, Li T, Yuan M, Huang Y, Liao J, Yang R, Ding C. Optimization of Ultrasound-Assisted Extraction of Flavonoids from Olive (Olea europaea) Leaves, and Evaluation of Their Antioxidant and Anticancer Activities. Molecules. 2018; 23(10):2513. https://doi.org/10.3390/molecules23102513
Chicago/Turabian StyleWang, Bixia, Jipeng Qu, Siyuan Luo, Shiling Feng, Tian Li, Ming Yuan, Yan Huang, Jinqiu Liao, Ruiwu Yang, and Chunbang Ding. 2018. "Optimization of Ultrasound-Assisted Extraction of Flavonoids from Olive (Olea europaea) Leaves, and Evaluation of Their Antioxidant and Anticancer Activities" Molecules 23, no. 10: 2513. https://doi.org/10.3390/molecules23102513