Development and Validation of Three Different UV Spectrophotometric Methods for Determination of New Antifungal Combination Used in Treatment of External Fungal Infections and Their Comparison Using ANOVA

Abstract

Luliconazole and Terbinafine hydrochloride combination is used for external fungal infections and is newly introduced in market. Hence it is necessary to develop a appropriate method of analysis for rapid and accurate determination of these drugs. Three simple, sensitive, accurate, precise and economical UV-spectrophotometric methods (A,B,C) have been developed for simultaneous estimation of Luliconazole and Terbinafine hydrochloride in their pharmaceutical dosage form and their comparison by ANOVA. Method (A) is well based on first order derivative spectrophotometric method, in which zero crossing points for Luliconazole is 249.50 nm and Terbinafine hydrochloride is 258.50 nm. Linearity was found in range of 3-18 µg/ ml for both the drugs using methanol as a common solvent. Method (B) is constructed on the principle of dual wavelength method using absorbance difference at 281 and 306 nm for Luliconazole & 262 and 296 nm for Terbinafine hydrochloride. Method (C) is based on norm of Q-absorbance ratio method where iso-absorptive point was obtained at 250 nm and the λ max selected was of Terbinafine hydrochloride 282 nm. The developed methods were validated as per ICH guideline. Accuracy and precision of methods were determined and validated. All the developed methods showed good reproducibility and recovery with % RSD <2. These three methods developed were compared using one way ANOVA and the fcal value was found to be less than ftab value indicating that there is no significant difference in the assay results of the three methods. All three methods were found to be rapid, specific, precise and accurate and found no interferences from the cream excipients so it can be used for routine investigation of both drugs in quality control laboratories.