Validated Reverse-Phase HPLC Method for Quantification of Fluconazole and Quercetin in Pharmaceutical Formulation

Abstract

Objectives: Fluconazole (FCZ) and Quercetin (QCT) in pharmaceutical dosage form were simultaneously quantified using a sensitive, accurate, robust, and précised Reverse Phase-HPLC Method.Materials and Methods: RP-HPLC method was used to get the separation of FCZ and QCT using Imtakt® Unison-US-C18 column having a size of the particle, 5 µm (150 mm x 4.6 mm) operating at a 1.3 mL/min flow rate with an optimized mobile phase having Methanol-Water Trifluoroacetic acid (TFA) in the proportion of 50:50:1%, v/v/v, respectively. The quantification of FCZ and QCT was done at 258 nm wavelength. Results and Discussions:In the developed method FCZ was retained at a retention time of 1.896 min, while QCT was retained at a retention time of 5.637 min. The proposed method separates FCZ and QCT with a resolution of 13.261. The validation of the method was done with reference to the guideline of ICH Q2(R1). FCZ and QCT showed linearity at 4–20 μg/mL, and 1–5 μg/mL, respectively. The % RSD for the precision study was less than 2 and recovery was in the range of 98.69% to 102.14% for both drugs. In the robustness study, the proposed method has less than 2 % RSD.Conclusion: The Method can be used to quantify Fluconazole and Quercetin simultaneously in a newly developed thermosensitive in-situ mucoadhesive vaginal gel formulation.

Keywords: Simultaneous estimation; Fluconazole (FCZ); Quercetin (QCT); RP-HPLC; ICH Q2(R1) Guideline