Christ alpha 1 4 ld

The PTZ loaded Ethyl cellulose microspheres were prepared by oil in water emulsion solvent evaporation method reported by Kashif et al. [18 (link)]. The drug concentration was kept constant in all formulations (50 mg) while the polymer was taken in variant concentrations of 50, 100 and 150 mg, respectively, as suggested by design expert software. The internal phase was prepared by dissolving the drug and polymer in 5 mL DCM under gentle stirring. The aqueous phase was prepared by dissolving the varied concentrations of emulsifier (PVA) (0.5, 0.75, 1% w/v) in 50 mL distilled water and heated at 50°C on a hot plate magnetic stirrer. The internal phase was added in a dropwise manner in the aqueous phase and allowed to stir for 5 h. The organic solvent was allowed to evaporate from the mixture. Microspheres were filtered and washed several times (3 to 6) with 0.1 N HCl and finally with the water to remove the free drug. The microspheres were then dried in the freeze dryer (Christ alpha 1–4 LD, UK) and used for further investigation [27 (link)].

After the acid treatment, the remaining solid particles were filtered out using a cheese cloth, and the remaining liquid portion was transferred to round-bottom flasks and frozen by immersion in liquid nitrogen. The frozen samples were then loaded to a Christ Alpha 1–4 LD freeze-dryer (Martin Christ Gefriertrocknungsanlagen GmbH, Osterode, Germany). The freeze-dried gelatine samples were weighed and stored at room temperature in zip-lock pouches until further analysis.

Pomegranate rinds (PR) extraction from the varieties (L1, L2, L3, L4, I1, and I2) was performed by aqueous-based solid–liquid extraction in duplicate in a round-bottom flask with stirring (100 rpm) in ultra-pure water. For each rinds variety, a ratio of 1:10 (w/v) rind powder-to-heated ultrapure water (100 °C) was employed and extraction time was 12 h away from light at 20 °C. Afterwards, extracts obtained were first centrifuged at 5000 rpm for 15 min at 5 °C, the supernatant was vacuum filtered, frozen at −80 °C for 24 h, and finally lyophilized in a freeze dryer (Christ Alpha 1-4 LD plus (Martin Christ, Germany)) for 3 days. Extracts samples (L1, L2, L3, L4, I1, and I2) were then stored at 4 °C for the following studies.