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Abstract

DEVELOPMENT OF ANALYTICAL METHOD AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF CLOTRIMAZOLE, MOMETASONE FUROATE AND FUSIDIC ACID IN PHARMACEUTICAL CREAM FORMULATION BY RP-HPLC METHOD

Harikishor Barange* and Suhail Asghar

ABSTRACT

A reversed-phase liquid chromatographic method has been developed and validated for estimation of Clotrimazole, Mometasone Furoate and Fusidic Acid in a pharmaceutical cream formulation. The chromatographic separation was achieved on Analytical Technology HPLC instrument (Software: HPLC Work Station) equipped with Deuterium lamp as detector, HPLC pump and manual Injecting facility programmed at 20μL capacity per injection was used. The stationary phase was Zodiac Column C8 (250 mm x 4.6 mm, 5μm) at ambient temperature. The mobile phase containing Acetonitrile: Methanol Water: Glacial Acetic Acid (60:10:30:0.1) adjust pH 3.5 by 10% orthophosphoric acid. Detection was carried out at 240nm using UV Detector. The flow rate was 1.5mL/min and retention time was about 1.7±0.1mins, 5.4±0.1mins and 8.2±0.1mins for Clotrimazole, Mometasone Furoate and Fusidic Acid. The linearity was obtained in the concentration range of 48-112μg/mL, 4.8-11.2μg/mL and 96-224μg/mL for Clotrimazole, Mometasone Furoate and Fusidic Acid respectively. Mean percentage recoveries were 99.15% for Clotrimazole, 99.17% for Mometasone Furoate and 99.87% for Fusidic Acid. The LOD of Clotrimazole, Mometasone Furoate and Fusidic Acid was found to be 1.0μg/mL, 0.30μg/mL and 0.2μg/mL where as the LOQ was 10μg/mL, 1.2μg/mL and 1μg/mL respectively. The assay values of both the analytes was found to be well within the limits that is 99.82%, 99.17% and 100.23% for Clotrimazole, Mometasone Furoate and Fusidic Acid respectively. Percentage relative standard deviation of percent assay values for replicate sample preparation was 0.33% for Clotrimazole, 0.51% for Mometasone Furoate and 0.43% for Fusidic Acid. The method was robust with respect to change in flow rate and composition of mobile phase.

Keywords: Cream formulation Clotrimazole, Mometasone Furoate, Fusidic acid, HPLC, analytical method development, validation.


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