Optimisation of Polyphenols Extraction from Wild Bilberry Leaves—Antimicrobial Properties and Stability Studies
Abstract
:1. Introduction
2. Results and Discussion
2.1. Design of Experiments
2.2. Spectrophotometric Characterisation of the Extracts
2.3. Experiments Modelling and Analysis of Variance (ANOVA)
2.4. Numerical Optimisation
2.5. Extracts Composition Determined by HPLC–PDA
2.6. Evaluation of the Extract Chemical Stability
2.7. Biological Evaluation of the E7×3 Extract
2.7.1. Antibacterial Activity on Planktonic Growth
2.7.2. The Antibacterial Activity on Biofilm Formation
3. Materials and Methods
3.1. Materials
3.2. Methods
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Factor | Factor Code | Coded Level | Level |
---|---|---|---|
Temperature | A | −1 | 30 °C |
+1 | 60 °C | ||
Solvent | B | −1 | EtOH 100% |
+1 | EtOH 50% | ||
Extraction method | C | −1 | Conv. |
+1 | US |
Run | A | B | C |
---|---|---|---|
1 | −1 | −1 | −1 |
2 | −1 | −1 | +1 |
3 | −1 | +1 | −1 |
4 | −1 | +1 | +1 |
5 | +1 | −1 | −1 |
6 | +1 | −1 | +1 |
7 | +1 | +1 | −1 |
8 | +1 | +1 | +1 |
(R2 = 0.9948, p = 0.0130) | |
Conventional extraction | Extraction yield (%) = 44.05 + 0.035833·A − 0.442·B + 0.002367·AB |
Ultrasound extraction | Extraction yield (%) = 47.15 − 0.055833·A − 0.442·B + 0.002367·AB |
(R2 = 0.9924, p = 0.0188) | |
Conventional extraction | TPC (mgCAE/g) = 271.15750 + 0.322167·A − 0.0469·B − 0.017443·AB |
Ultrasound extraction | TPC (mgCAE/g) = 226.10250 + 1.05217·A − 0.0469·B − 0.017443·AB |
(R2·=·0.9912, p = 0.0218) | |
Conventional extraction | TFC (mgRHE/g) = 78.815 − 0.58075·A + 0.32·B − 0.00923 AB |
Ultrasound extraction | TFC (mgRHE/g) = 53.58 + 0.92525·A + 0.32 B − 0.00923 AB |
(R2 = 0.9915, p = 0.0212) | |
Conventional extraction | TCh (mgCht/g) = −4.9575 + 0.0315·A + 0.1117·B − 0.00084·AB |
Ultrasound extraction | TCh (mgCht/g) = −6.0625 + 0.05383·A + 0.1117·B − 0.00084·AB |
(R2 = 0.9827, p = 0.0426) | |
Conventional extraction | RSADPPH (mg/TE/g) = 474.5125 + 1.82183·A − 0.9277·B − 0.038793·AB |
Ultrasound extraction | RSADPPH (mg/TE/g) = 389.8575 + 3.22917·A − 0.9277·B − 0.038793·AB |
(R2 = 0.9898, p = 0.0252) | |
Conventional extraction | RSAABTS (mg/TE/g) = 936.08 + 0.52833·A − 2.5778·B − 0.034867·AB |
Ultrasound extraction | RSAABTS (mg/TE/g) = 825.96 + 1.50317·A − 2.5778·B − 0.034867·AB |
Optimal Extraction Conditions | Temperature (°C) | Solvent | Extraction Method | Desirability |
---|---|---|---|---|
Extraction yield (%) | 60 | EtOH–H2O | Conv. | 1.000 |
TPC | 30 | EtOH–H2O | Conv. | 0.964 |
TFC | 60 | EtOH–H2O | Conv. | 0.962 |
TCh | 30 | EtOH | Conv | 0.998 |
RSAABTS | 60 | EtOH–H2O | Conv. | 1.000 |
RSADPPH | 30 | EtOH–H2O | Conv. | 1.000 |
All responses | 60 | EtOH–H2O | Conv. | 0.784 |
Catechin Hydrate | Chlorogenic Acid | Caffeic Acid | (-) Epicatechin | Rutin Hydrate | |
---|---|---|---|---|---|
RT (min) | 12.751 ± 0.009 | 13.469 ± 0.011 | 15.521 ± 0.006 | 17.686 ± 0.011 | 27.435 ± 0.012 |
Run 1 | 4.462 ± 0.044 | 75.207 ± 0.222 | nd | 14.851 ± 0.094 | 16.372 ± 0.016 |
Run 2 | 4.156 ± 0.025 | 61.849 ± 0.233 | nd | 14.110 ± 0.098 | 14.739 ± 0.083 |
Run 3 | 4.918 ± 0.004 | 65.866 ± 0.457 | nd | 14.824 ± 0.095 | 15.454 ± 0.104 |
Run 4 | 5.517 ± 0.022 | 74.039 ± 0.017 | nd | 15.498 ± 0.008 | 14.814 ± 0.007 |
Run 5 | 4.456 ± 0.010 | 69.304 ± 0.036 | 0.744 ± 0.000 | 14.002 ± 0.082 | 38.232 ± 0.023 |
Run 6 | 3.683 ± 0.064 | 60.901 ± 0.068 | 0.575 ± 0.004 | 10.988 ± 0.036 | 32.271 ± 0.102 |
Run 7 | 5.799 ± 0.057 | 87.980 ± 0.012 | 1.882 ± 0.009 | 12.482 ± 0.045 | 46.650 ± 0.096 |
Run 8 | 4.002 ± 0.089 | 62.521 ± 0.007 | 1.429 ± 0.003 | 10.362 ± 0.019 | 33.901 ± 0.010 |
Extract | Time at 40 °C at 75% RHu (Days) | Volatile Components | Organics Combustion | Dried Residue |
---|---|---|---|---|
(% wt vs. Dry Mass) | ||||
E7×3 | 0 | 44.4 | 51.1 | 2.1 |
7 | 34.4 | 49.2 | 1.6 | |
14 | 33.5 | 46.8 | 2.0 | |
28 | 34.9 | 45.0 | 3.4 |
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Brezoiu, A.-M.; Deaconu, M.; Mitran, R.-A.; Prelipcean, A.-M.; Matei, C.; Berger, D. Optimisation of Polyphenols Extraction from Wild Bilberry Leaves—Antimicrobial Properties and Stability Studies. Molecules 2023, 28, 5795. https://doi.org/10.3390/molecules28155795
Brezoiu A-M, Deaconu M, Mitran R-A, Prelipcean A-M, Matei C, Berger D. Optimisation of Polyphenols Extraction from Wild Bilberry Leaves—Antimicrobial Properties and Stability Studies. Molecules. 2023; 28(15):5795. https://doi.org/10.3390/molecules28155795
Chicago/Turabian StyleBrezoiu, Ana-Maria, Mihaela Deaconu, Raul-Augustin Mitran, Ana-Maria Prelipcean, Cristian Matei, and Daniela Berger. 2023. "Optimisation of Polyphenols Extraction from Wild Bilberry Leaves—Antimicrobial Properties and Stability Studies" Molecules 28, no. 15: 5795. https://doi.org/10.3390/molecules28155795