ABSTRACT:
Hypothesis: The precise measurement of Donepezil HCl in intricate pharmaceutical and biological matrices is crucial for assessing its pharmacokinetic properties, therapeutic effectiveness, and formulation quality. Current analytical techniques frequently exhibit restricted sensitivity, insufficient specificity in biological matrices, and inadequate reproducibility. We proposed the establishment & verification of a high-performance liquid chromatography system that uses reverse phase separation method to ensure reliable, accurate, and sensitive quantification of Donepezil HCl in diverse matrices, including emulsomal formulations, in vitro release media, plasma, and brain tissue. Experiments: Acetonitrile and 0.01 M ammonium formate buffer with a pH of 6.4 was used as mobile phase to develop a RP-HPLC method, C18 column of a Waters Nova-Pak (3.9 × 150mm) and packing 4 µm was utilized in a 62:38 v/v ratio. The methodology was confirmed in accordance with the requirements described in ICH Q2(R1), encompassing evaluations of specificity, linearity (0.05–5µg/mL), accuracy, reliability, precision, limit of detection (LOD), & limit of quantification (LOQ) are all important aspects to consider. Injection volume of 20µL and 1.0mL/min flow rate of resulted in detection at 268nm. The method's suitability was confirmed across several concentrations and matrices, including placebo, plasma, and brain homogenate. Results: The retention period of Donepezil HCl was around 6.0 minutes, exhibiting an excellent peak shape and negligible matrix interference. Within the range of 0.05–5µg/mL (R² = 0.9984) linearity was detected. The approach exhibited significant specificity without interference at the retention time of Donepezil HCl. Recovery rates varied between 97.8% and 98.52%, while %RSD values for precision experiments were consistently below 0.05%. The detection limit was established to be 0.037 µg/mL and quantitation limit to be 0.113 µg/mL. Robustness testing validated the dependability of method and slight fluctuations in operational parameters. This validated RP -HPLC method is reproducible, specific, and sensitive for routinely quantifying donepezil HCl in biological and pharmaceutical applications.
Cite this article:
Rupanshi Sahu, Rakhee Kapadia. Analytical Method Development and Validation of Donepezil HCl by RP-HPLC. Research Journal of Pharmacy and Technology. 2026;19(7):3007-2. doi: 10.52711/0974-360X.2026.00428
Cite(Electronic):
Rupanshi Sahu, Rakhee Kapadia. Analytical Method Development and Validation of Donepezil HCl by RP-HPLC. Research Journal of Pharmacy and Technology. 2026;19(7):3007-2. doi: 10.52711/0974-360X.2026.00428 Available on: https://rjptonline.org/AbstractView.aspx?PID=2026-19-7-15
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