Formulation and in vitro evaluation of hesperidin phytosomal gel for anti-inflammatory potential

2.4 Preparation of phytosomal gel

The optimized phytosomal dispersion was incorporated into a topical gel base containing Carbopol 934. The prepared gel was evaluated for pH, viscosity and spreadability and subsequently investigated for morphology, anti-inflammatory activity, in-vitro drug release, release kinetics, permeation and stability.

3. EVALUATION METHODS

Particle size and zeta potential were determined for the optimized phytosomes. Entrapment efficiency and drug content were evaluated to assess incorporation of hesperidin into the phytosomal system. FTIR was used to assess compatibility and DSC/XRD were used as physicochemical characterization tools.

For anti-inflammatory evaluation, the protein denaturation method was used. A reaction mixture containing egg albumin and phosphate-buffered saline was treated with test samples at different concentrations. Following incubation and heating, absorbance was measured at 660 nm and percentage inhibition of protein denaturation was calculated.

In-vitro drug release was studied using a dialysis technique in phosphate-buffered saline at controlled temperature with continuous stirring. Samples were withdrawn at predetermined time points and analyzed spectrophotometrically. Release data were fitted to zero-order, first-order, Higuchi, Korsmeyer–Peppas and Hixson–Crowell models as described in the dissertation.

4. RESULTS AND DISCUSSION

4.1 Characterization and optimization

The optimized F9 phytosome exhibited a particle size of 133.8 nm and a zeta potential of −22.6 mV. The dissertation reports zeta-potential values from −22.6 to −10.6 mV across the formulations, with F9 showing the most negative value. Entrapment efficiency ranged from 77.15% to 88.35%, and F9 showed the highest value, indicating efficient incorporation of hesperidin in the selected formulation composition.

The F9 phytosomes were described as predominantly spherical under projection microscopy, with moderate dispersion and slight aggregation. TEM was also used for morphology assessment. The physicochemical characterization supported formation of the intended lipid-based delivery system.

4.2 Anti-inflammatory activity

At 20 µg/mL, pure hesperidin showed higher inhibition than the gel. At higher concentrations, the phytosomal gel showed greater inhibition, reaching 55.19% at 100 µg/mL compared with 43.50% for hesperidin. The reported IC?? for the phytosomal gel was 94.81 µg/mL, whereas pure hesperidin was reported as not effective within the tested concentration range for IC?? determination.

4.3 In-vitro drug release and kinetics

The phytosomal gel demonstrated progressive drug release, with cumulative release reported up to 92.31% over 8 h. The release profile indicated sustained delivery from the formulation. The dissertation reports the best fit with zero-order and Korsmeyer–Peppas models, with R² = 0.999, and interprets the release as controlled and non-Fickian diffusion.

4.4 Stability

Stability evaluation was conducted over 30 days. The dissertation reports that the formulation remained stable over this period based on the evaluated physicochemical parameters. Longer-term and accelerated stability studies were identified as future requirements.

CONCLUSION

The study developed and evaluated a hesperidin-loaded phytosomal gel for topical delivery. The optimized F9 phytosomes showed a particle size of 133.8 nm, zeta potential of −22.6 mV, entrapment efficiency of 88.35% and drug content of 97.23%. The gel showed an acceptable pH of 6.90 and suitable topical characteristics. The protein denaturation assay demonstrated measurable anti-inflammatory activity, with an IC?? of 94.81 µg/mL for the phytosomal gel. Drug release reached 92.31% over 8 h and the formulation showed the reported zero-order and Korsmeyer–Peppas release behavior. Overall, the results indicate that phytosomal incorporation followed by gel formulation can provide a useful approach for topical delivery of hesperidin.

FUTURE SCOPE

Further work should include in-vivo pharmacokinetic and pharmacodynamic studies, long-term and accelerated stability studies, scale-up studies and clinical evaluation. Advanced morphology and surface-characterization techniques may also be used. Comparative studies with conventional hesperidin formulations could further characterize the delivery performance of the phytosomal gel.

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