DEVELOPMENT AND VALIDATION OF AN REVERSE PHASE-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS DETERMINATION OF OLANZAPINE AND FLUOXETINE HYDROCHLORIDE IN TABLET DOSAGE FORM
DOI:
https://doi.org/10.22159/ajpcr.2026v19i3.57694Keywords:
Reverse phase-high performance liquid chromatography, Olanzapine, Fluoxetine hydrochloride, Validation, ICH guidelines, tablet dosage form, Simultaneous estimationAbstract
Objectives: To develop and validate a novel reverse phase-high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of olanzapine and fluoxetine hydrochloride in a fixed-dose combination tablet formulation.
Methods: Chromatographic separation was carried out on a Waters Symmetry C18 column (250 mm × 4.6 mm, 5 μm) using a mobile phase of phosphate buffer (pH 3.0) and acetonitrile in the ratio of 55:45 (v/v), delivered at a flow rate of 1.0 mL/min. Detection was performed at 235 nm using a ultraviolet detector. The method was validated in accordance with ICH Q2(R1) guidelines.
Results: The method exhibited linearity over the concentration range of 2–12 μg/mL for olanzapine and 4–24 μg/mL for fluoxetine hydrochloride, with correlation coefficients (r2) of 0.999 for both drugs. Precision studies showed percentage relative standard deviation (%RSD) values below 1% for intra-day and inter-day measurements. Accuracy was confirmed by recovery studies, yielding average recoveries of 99.44% for olanzapine and 99.59% for fluoxetine hydrochloride. The method demonstrated robustness at lower flow rates, while sensitivity was observed at higher flow variations. Ruggedness studies showed %RSD values within acceptable limits. Assay of marketed tablets resulted in average assay values of 100.22% for olanzapine and 99.66% for fluoxetine hydrochloride.
Conclusion: The developed RP-HPLC method is simple, precise, accurate, and suitable for routine quality control analysis of olanzapine and fluoxetine hydrochloride in combined dosage forms.
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