Microwave-Assisted Extraction of Anticancer Flavonoid, 2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits
Abstract
:1. Introduction
2. Results and Discussion
2.1. Method Validation of HPLC Determination
2.2. Single Variables Experiment of DMC Microwave-Assisted Extraction
2.3. Fitting the Model
- Y is replaced by a DMC content (µg/g dry sample);
- X1 is replaced by a solid-to-liquid ratio (g/mL);
- X2 is replaced by a microwave power (W);
- X3 is replaced by a microwave time (min).
2.4. Analysis of Response Surface and Optimization of MAE Process by RSM
2.5. Verification of the DMC Predictive Model and Comparison of MAE with Conventional Methods
2.6. Cell Viability
3. Materials and Methods
3.1. Materials and Apparatuses
3.2. Isolation and Characterization of Standard DMC Compound
3.3. HPLC Conditions and Method Validation for Quantification of DMC
3.4. Microwave-Assisted Extraction (MAE) Approach of DMC-Rich Extract
3.5. Single Variable Investigation
3.6. Experimental Design
3.7. Verification of the DMC Predictive Model
3.8. Conventional Maceration and Heat Reflux Extraction (HRE)
3.9. Statistical Analysis
3.10. Cell Lines and MTT Assay
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Concentration of DMC (µg/mL) | %RSD | %Recovery (µg/mL ± SD) | |
---|---|---|---|
Intra-Day | Inter-Day | ||
0.25 | 5.75 | 7.39 | 85.3 ± 1.5 |
50 | 1.36 | 1.87 | 98.5 ± 2.5 |
100 | 0.67 | 1.22 | 97.3 ± 3.8 |
Standard Order | Extraction Variables | DMC Content (µg/g Dry Sample) | ||
---|---|---|---|---|
X1: Solid-to-Liquid Ratio (g/mL) | X2: Microwave Power (W) | X3: Microwave Time (min) | ||
2 | 1:35 | 350 | 25 | 1365 |
10 | 1:25 | 630 | 10 | 1205 |
5 | 1:15 | 490 | 10 | 1145 |
7 | 1:15 | 490 | 40 | 1264 |
13 | 1:25 | 490 | 25 | 1295 |
8 | 1:35 | 490 | 40 | 1355 |
4 | 1:35 | 630 | 25 | 1346 |
9 | 1:25 | 350 | 10 | 1209 |
11 | 1:25 | 350 | 40 | 1339 |
15 | 1:25 | 490 | 25 | 1283 |
1 | 1:15 | 350 | 25 | 1259 |
14 | 1:25 | 490 | 25 | 1285 |
6 | 1:35 | 490 | 10 | 1236 |
3 | 1:15 | 630 | 25 | 1230 |
12 | 1:25 | 630 | 40 | 1299 |
Source | Sum of Squares | Degree of Freedom | Mean Square | F-Value | p-Value | Remark |
---|---|---|---|---|---|---|
Model | 54,540.17 | 9 | 6060.02 | 98.01 | <0.0001 | significant |
X1-S-to-L Ratio | 20,402.00 | 1 | 20,402.00 | 329.95 | <0.0001 | significant |
X2-Microwave Power | 1058.00 | 1 | 1058.00 | 17.11 | 0.0090 | significant |
X3-Microwave Time | 26,680.50 | 1 | 26,680.50 | 431.49 | <0.0001 | significant |
X1X2 | 25.00 | 1 | 25.00 | 0.4043 | 0.5528 | |
X1X3 | 0.00 | 1 | 0.00 | 0.0000 | 1.0000 | |
X2X3 | 324.00 | 1 | 324.00 | 5.24 | 0.0707 | |
X1² | 0.4103 | 1 | 0.4103 | 0.0066 | 0.9382 | |
X2² | 592.41 | 1 | 592.41 | 9.58 | 0.0270 | significant |
X3² | 5146.26 | 1 | 5146.26 | 85.23 | 0.0003 | significant |
Residual | 309.17 | 5 | 61.83 | |||
Lack of Fit | 226.50 | 3 | 75.50 | 1.83 | 0.3729 | not significant |
Pure Error | 82.67 | 2 | 41.33 | |||
Cor Total | 54,849.33 | 14 | ||||
R2 | 0.9944 | |||||
R2 (adj) | 0.9842 | |||||
R2(pred) | 0.9305 | |||||
CV (%) | 0.6171 | |||||
Standard Deviation | 7.86 |
Extraction Method | Optimal MAE Conditions | Maximum Value (µg/g Dry Sample) | |||
---|---|---|---|---|---|
X1 | X2 | X3 | Predicted Value | Experimental Value | |
MAE | 1:35 g/mL | 350 W | 38 min | 1389 | 1409 ± 24 a |
HRE | 1:35 g/mL | - | 2 h | - | 1337 ± 37 a |
Maceration | 1:35 g/mL | - | 20 h | - | 1225 ± 81 b |
Independent Variable | Variable Symbol | Level | ||
---|---|---|---|---|
−1 | 0 | 1 | ||
Sample–Solvent ratio (g/mL) | X1 | 1:15 | 1:25 | 1:35 |
Microwave Power (W) | X2 | 350 | 490 | 630 |
Microwave Time (min) | X3 | 10 | 25 | 40 |
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Choommongkol, V.; Punturee, K.; Klumphu, P.; Rattanaburi, P.; Meepowpan, P.; Suttiarporn, P. Microwave-Assisted Extraction of Anticancer Flavonoid, 2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits. Molecules 2022, 27, 1397. https://doi.org/10.3390/molecules27041397
Choommongkol V, Punturee K, Klumphu P, Rattanaburi P, Meepowpan P, Suttiarporn P. Microwave-Assisted Extraction of Anticancer Flavonoid, 2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits. Molecules. 2022; 27(4):1397. https://doi.org/10.3390/molecules27041397
Chicago/Turabian StyleChoommongkol, Vachira, Khanittha Punturee, Piyatida Klumphu, Parintip Rattanaburi, Puttinan Meepowpan, and Panawan Suttiarporn. 2022. "Microwave-Assisted Extraction of Anticancer Flavonoid, 2′,4′-Dihydroxy-6′-methoxy-3′,5′-dimethyl Chalcone (DMC), Rich Extract from Syzygium nervosum Fruits" Molecules 27, no. 4: 1397. https://doi.org/10.3390/molecules27041397