Development and validation of analytical (RP-HPLC) method for estimation of Fluconazole & Ivermectin in pharmaceutical formulation

ABSTRACTA rapid, accurate, and reliable Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of fluconazole andivermectin in bulk drug and tablet dosage forms. Method optimization was performed throughdifferent chromatographic trials using various mobile phase compositions and gradient elutionprograms to achieve acceptable retention, peak symmetry, and resolution. The optimizedchromatographic conditions employed an Agilent Poroshell C18 column with 10 mMammonium formate as solvent A and a mixture of ammonium formate–methanol–acetonitrile assolvent B under gradient elution at a flow rate of 0.3 mL/min with UV detection at 210 nm.Under optimized conditions, fluconazole and ivermectin were eluted at retention times of 8.607min and 12.075 min, respectively, with satisfactory system suitability parameters includingtheoretical plates, tailing factor, and resolution. The developed method exhibited excellentlinearity over concentration ranges of 1.65–25 μg/mL for fluconazole and 3.75–50 μg/mL forivermectin with correlation coefficients (R2) of 0.9993 and 0.9995, respectively. The limits ofdetection and quantification confirmed adequate sensitivity of the method.The validated method demonstrated acceptable accuracy, precision, robustness, and repeatabilityaccording to ICH guidelines. Recovery studies showed percentage recoveries within acceptablelimits and %RSD values below 2%. Assay results for marketed formulation indicated percentagedrug content within pharmaceutical acceptance criteria. The developed RP-HPLC method wasfound to be simple, precise, robust, and suitable for routine quality control analysis offluconazole and ivermectin in combined pharmaceutical dosage forms.