Development and validation of a spectrofluorophotometric method for berberine hydrochloride quantification in diverse matrices

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Abstract
Berberine hydrochloride (BHC), a key isoquinoline alkaloid from the Berberis genus, is gaining prominence for its diverse pharmacological properties. The goal was to devise rapid and sensitive spectrofluorophotometric methods (Analytical and Bioanalytical) for precise BHC estimation in varied matrices, addressing crucial needs in pharmaceutical and biomedical research.Utilizing a luminescence spectrometer (Perkin Elmer, UK), BHC was detected with 1M Methanolic H2SO4, at excitation and emission wavelength of 350 nm, and 530 nm respectively. BHC extraction from spiked human plasma employed a protein precipitation method with acetonitrile. Validation studies, following ICH Q2 (R1) guidelines, demonstrated the efficacy of both methods. Linear responses were observed in the 200-800 ng/mL BHC concentration range, with precision (% RSD <5%) meeting acceptable limits. Recovery studies (90%-110%) affirmed reliability and reproducibility. The developed spectrofluorophotometric methods for BHC estimation offer simplicity, linearity, sensitivity, precision, and accuracy. Their robustness, confirmed through validation and comparison with existing methods, positions them as valuable tools for BHC quantification in pharmaceutical and biomedical research. These methods present a promising advancement in analytical approaches for researchers in the field.

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