Analytical method development and validation of antifungal drugs in updated ointment formulation using UV spectroscopy and RP-HPLC

Abstract

A precise, simple, and validated Reverse-phase high-performance liquid chromatography (RP-HPLC) method alongside spectrophotometric analysis has been established for the simultaneous quantification of clobetasol propionate, miconazole nitrate, and salicylic acid in an updated ointment formulation. The impact of organic modifiers on the retention of the target compounds was assessed. The chromatographic analysis was conducted using a simple low-pressure gradient method with UV detection at 282 nm on a C18 column (5 µm, 4.6 × 250 mm HSS). The mobile phase consisted of a mixture of methanol, acetonitrile, and tetra ethylamine acetate buffer at pH 4, adjusted with acetic acid in a 40:40:20 v/v ratio, at a flow rate of 1.2 mL min^-1. The method demonstrated linearity in the 5-15 μg mL^-1 concentration range for clobetasol propionate, 30-90 μg mL^-1 for miconazole nitrate, and 15-45 μg mL^-1 for salicylic acid. The limits of detection and quantification were determined to be 1.49 μg mL^-1 and 4.53 μg mL^-1 for clobetasol propionate, 8.72 μg mL^-1 and 26.43 μg mL^-1 for miconazole nitrate, and 3.37 μg mL^-1 and 10.22 μg mL^-1 for salicylic acid, respectively. The recoveries for drugs in formulation range from 95 % to 99 %.