Development and Validation of Analytical Method for Fluconazole and Fluconazole Related Compounds (A, B, and C) in Capsule Formulations by HPLC with UV Detection

A simple and stability-indicating liquid chromatographic method was developed and validated for the analysis of Fluconazole and its related compound (A, B, and C) in capsule formulations. Liquid chromatography with a UV detector at a wavelength of 260 nm using a reversed-phase C18 column was employed in this study. Isocratic elution was employed using a mixture of methanol and water (40:60, v/v). This new method was validated in accordance with USP requirements for new methods for assay determination, which include accuracy, precision, specificity, linearity and range. The current method demonstrates good linearity over the range of 0.05-0.15 mg/ml of Fluconazole. The accuracy of the method is 99.3%. The precision of this method reflected by relative standard deviation of replicates is 0.61%. Validation of the same method for Fluconazole related compounds analysis was also performed according to USP requirements for quantitative determination of impurities which include accuracy, precision, linearity and range, selectivity, and Limit of quantitation (LOQ). Low LOQ of the related compounds using this method enables the detection and quantitation of these impurities at low concentration.