Central Composite Design (CCD) for the Optimisation of Ethosomal Gel Formulation of Punica granatum Extract: In Vitro and In Vivo Evaluations
Abstract
:1. Introduction
2. Results
2.1. Optimisation of Formulation
2.2. Response Analysis through Polynomial Equations
2.2.1. Effect of Variables on Particle Size
2.2.2. Effect of Variables on Entrapment Efficiency
2.3. Gel Punica granatum
2.3.1. Vesicle Size, Particle Size Distribution, and Zeta Potential (ZP)
2.3.2. Scanning Electron Microscopy (SEM)
2.3.3. FTIR
2.4. Gel Evaluations
2.5. In Vitro Diffusion Studies
2.6. Stability Studies
2.7. Formalin-Induced Rat Paw Oedema
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Plant Material and Chemicals
5.2. Instrument Details
5.3. Animals
5.4. Preparation of Peel Extract
5.5. Preparation of Ethosomes Suspensions
5.6. Preparation of Ethosomal Gel
5.7. Experimental Design
5.8. Punica granatum Gel Characterisation
5.8.1. Vesicle Size, Particle Size Distribution, and ZP
5.8.2. SEM
5.8.3. FTIR
5.9. Evaluations of Ethosomal Gel
5.9.1. Viscosity
5.9.2. Spreadability
5.9.3. pH Measurement
5.10. In Vitro Diffusion Studies
5.11. Stability Studies
5.12. Formalin-Induced Albino Rats Paw Oedema
5.13. Statistical Analysis of Results
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Coded Values Level | Independent Variables | |
---|---|---|
X1, Lecithin | X2, Ethanol | |
−1 | 1 | 15 |
0 | 1.5 | 20 |
+1 | 2 | 30 |
Formulation Code | X1, Lecithin | X2, Ethanol | Y1, Particle Size (nm) | Y2, Entrapment Efficiency (%) |
---|---|---|---|---|
F1 | 1 | 15 | 738 | 53.51 |
F2 | 1 | 20 | 762 | 57.57 |
F3 | 1 | 30 | 774 | 59.20 |
F4 | 1.5 | 15 | 794 | 57.74 |
F5 | 1.5 | 20 | 828 | 62.20 |
F6 | 1.5 | 30 | 832 | 64.25 |
F7 | 2 | 15 | 836 | 68.39 |
F8 | 2 | 20 | 854 | 72.20 |
F9 | 2 | 30 | 859 | 74.12 |
Formulation | pH | Viscosity (CPS) | Spreadability (g.cm/s) | Grittiness |
---|---|---|---|---|
PGE1 | 7.1 | 34,015 | 35.15 | No |
PGE2 | 7.0 | 35,125 | 33.58 | |
PGE3 | 6.9 | 38,478 | 31.35 | |
PGE4 | 6.9 | 30,975 | 36.28 | |
PGE5 | 6.8 | 35,750 | 34.36 | |
PGE6 | 6.8 | 36,435 | 33.65 | |
PGE7 | 7.1 | 34,019 | 35.95 | |
PGE8 | 7.1 | 33,735 | 32.18 | |
PGE9 | 7.1 | 32,158 | 31.55 |
Stability Study Period | Drug Content (%) | |
---|---|---|
Initial | 4 ± 2 °C | 98.5 |
30 ± 2 °C | 98.6 | |
After 3 weeks | 4 ± 2 °C | 97.4 |
30 ± 2 °C | 96.7 | |
After 6 weeks | 4 ± 2 °C | 97.2 |
30 ± 2 °C | 96.3 | |
After 9 weeks | 4 ± 2 °C | 97.6 |
30 ± 2 °C | 96.3 | |
After 12 weeks | 4 ± 2 °C | 97.5 |
30 ± 2 °C | 96.6 |
Groups | 0 h | 1 h | 2 h | 4 h | 8 h | 12 h |
---|---|---|---|---|---|---|
Control | 0.0 ± 0 | 0.0 ± 0 | 0.0 ± 0 | 0.0 ± 0 | 0.0 ± 0 | 0.0 ± 0 |
Formalin | 1.0 ± 0.01 | 1.6 ± 0.02 | 1.8 ± 0.03 | 1.9 ± 0.03 | 1.4 ± 0.02 | 1.2 ± 0.02 |
PG Extract | 0.9 ± 0.01 | 1.4 ± 0.01 | 1.3 ± 0.01 * | 1.2 ± 0.01 * | 1.0 ± 0.01 * | 0.9 ± 0.01 * |
PG EthoGel | 0.7 ± 0.01 | 0.9 ± 0.01 ** | 0.7 ± 0.01 ** | 0.6 ± 0.01 *** | 0.5 ± 0.01 *** | 0.5 ± 0.01 ** |
Standard | 0.5 ± 0.01 | 0.7 ± 0.01 ** | 0.6 ± 0.01 *** | 0.5 ± 0.01 *** | 0.7 ± 0.01 ** | 0.6 ± 0.01 *** |
Groups | 0 h | 1 h | 3 h | 5 h | 12 h |
---|---|---|---|---|---|
Control | 0.0% | 0.0% | 0.0% | 0.0% | 0.0% |
PG EXTRACT | 0.0% | 24.6% | 39.50% | 46.13% | 10.10% |
PG ET GEL | 0.0% | 67.40% | 74.10% | 86.34% | 82.64% |
Standard | 0.0% | 71.20% | 84.30% | 70.12% | 56.24% |
Quantities in w/w % (100 gm) | ||||||||
---|---|---|---|---|---|---|---|---|
Code | EAP Extract | Carbapol 934 (% w/v) gms | Lecithin (w/v) (%) | Ethanol (v/v) mL | Cholesterol (%) | PEG 400 (v/v) mL | TEM (w/v) (%) | Water QS mL |
EPG1 | 10 | 1.5 | 1 | 15 | 0.2 | 1 | 0.5 | 100 |
EPG2 | 10 | 1.5 | 1.5 | 15 | 0.2 | 1 | 0.5 | 100 |
EPG3 | 10 | 1.5 | 2 | 15 | 0.2 | 1 | 0.5 | 100 |
EPG4 | 10 | 1.5 | 1 | 20 | 0.2 | 1 | 0.5 | 100 |
EPG5 | 10 | 1.5 | 1.5 | 20 | 0.2 | 1 | 0.5 | 100 |
EPG6 | 10 | 1.5 | 2 | 20 | 0.2 | 1 | 0.5 | 100 |
EPG7 | 10 | 1.5 | 1 | 30 | 0.2 | 1 | 0.5 | 100 |
EPG8 | 10 | 1.5 | 1.5 | 30 | 0.2 | 1 | 0.5 | 100 |
EPG9 | 10 | 1.5 | 2 | 30 | 0.2 | 1 | 0.5 | 100 |
Formulation Code | Lecithin (% w/v) | Ethanol (mL) | Cholesterol (%) |
---|---|---|---|
F1 | 1 | 15 | 0.2 |
F2 | 1 | 20 | 0.2 |
F3 | 1 | 30 | 0.2 |
F4 | 1.5 | 15 | 0.2 |
F5 | 1.5 | 20 | 0.2 |
F6 | 1.5 | 30 | 0.2 |
F7 | 2 | 15 | 0.2 |
F8 | 2 | 20 | 0.2 |
F9 | 2 | 30 | 0.2 |
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Alam, P.; Shakeel, F.; Foudah, A.I.; Alshehri, S.; Salfi, R.; Alqarni, M.H.; Aljarba, T.M. Central Composite Design (CCD) for the Optimisation of Ethosomal Gel Formulation of Punica granatum Extract: In Vitro and In Vivo Evaluations. Gels 2022, 8, 511. https://doi.org/10.3390/gels8080511
Alam P, Shakeel F, Foudah AI, Alshehri S, Salfi R, Alqarni MH, Aljarba TM. Central Composite Design (CCD) for the Optimisation of Ethosomal Gel Formulation of Punica granatum Extract: In Vitro and In Vivo Evaluations. Gels. 2022; 8(8):511. https://doi.org/10.3390/gels8080511
Chicago/Turabian StyleAlam, Prawez, Faiyaz Shakeel, Ahmed I. Foudah, Sultan Alshehri, Roshan Salfi, Mohammed H. Alqarni, and Tariq M. Aljarba. 2022. "Central Composite Design (CCD) for the Optimisation of Ethosomal Gel Formulation of Punica granatum Extract: In Vitro and In Vivo Evaluations" Gels 8, no. 8: 511. https://doi.org/10.3390/gels8080511