DEVELOPMENT AND VALIDATION OF AN HPLC METHOD FOR THE QUANTIFICATION OF BENZOCAINE AND FUSIDIC ACID SIMULTANEOUSLY IN MICROEMULSION FORMULATION

Objective: The purpose of this study was to develop and validate an analytical technique for quantification in the microemulsion formulation containing fusidic acid and benzocaine developed for the treatment of mild and moderate wounds. Material and Method: Acetonitrile and acetic acid (0.1%) were used as a mobile phase in the HPLC method. The flow rate and injection volume were determined. The ICH Q2 guideline was followed in the validation of the developed method. Benzocaine (2%) and fusidic acid (2%) were added to the blank microemulsion prepared using ethyl oleate, propylene glycol, ethanol, and Cremophor EL, and the selectivity of the HPLC method for both these substances was investigated. Result and Discussion: The ratio of acetonitrile to acetic acid (0.1%) was determined to be 70:30 (v/v). The injection volume of 20 µl and the flow rate of 1 ml·min-1 were set. The developed method, which demonstrated a high correlation coefficient (0.9999) and a low variation coefficient (<2%), was simple, inexpensive, convenient, selective, and suitable for the analysis of both substances at the same wavelength (210 nm).