STRATEGIC OPTIMIZATION OF HPTLC METHOD FOR BERBERINE QUANTIFICATION IN Β-CYCLODEXTRIN INCLUSION COMPLEX USING DESIGN OF EXPERT.

Wound healing involves complex stages like hemostasis and inflammation. Berberine, a natural compound, exhibits antimicrobial and anti-inflammatory properties, aiding healing. Nanotechnology enhances the delivery of berberine, highlighting the need for a reliable analytical method for its quantification. In this study, we developed and validated a High-Performance Thin-Layer Chromatography (HPTLC) method for the simultaneous quantification of berberine in β-cyclodextrin inclusion complex nanoparticles. The method development employed a Box-Behnken Design (BBD) to optimize chromatographic conditions, focusing on the volume of methanol, saturation time, and migration distance. The optimized HPTLC conditions utilized a mobile phase of ethyl acetate, methanol, and water (7:2:1 v/v), achieving a precise separation of berberine with an Rf value of 0.52 ± 0.01. Method validation, following ICH guidelines, confirmed linearity (200-1000 ng/band, R² = 0.999), precision (intra-day and inter-day %RSD < 2%), accuracy (recoveries between 97.1% and 98.5%), robustness, and solution stability (%RSD < 2% after 48 hours). This validated method effectively quantified berberine in nanoformulations, highlighting its potential as a robust analytical tool for evaluating berberine-based wound healing therapies. [ABSTRACT FROM AUTHOR]