Development and Validation of New UV Spectrophotometric Methods for Simultaneous Estimation of Alcaftadine in Combination with Ketorolac Tromethamine in a Pharmaceutical Dosage Form and Statistical Analysis using Analysis of Variance

Abstract

Alcaftadine and Ketorolac tromethamine combination is newly introduced in the market for the treatment of conjunctivitis. Hence, it is essential to develop a rapid, accurate, sensitive and precise method (in accordance with ICH guidelines) to estimate drugs. Three simple and cost-effective UV-spectrophotometric methods (First-order derivative (A), Dual Wavelength (B) and Q- absorbance ratio method (C) have been developed for simultaneous estimation of Alcaftadine and Ketorolac tromethamine in their pharmaceutical dosage form and statistical comparison using analysis of variance (ANOVA). Method (A) is based on the first-order derivative spectrophotometric method, in which zero-crossing points (ZCP) for Alcaftadine is 281.50 nm and Ketorolac tromethamine is 268 nm. Linearity was found in the 4–14 µg/mL range for Alcaftadine and 6.4–22.4 µg/mL for Ketorolac tromethamine using methanol as a common solvent. Method (B) is based on the principle of the dual-wavelength method using absorbance difference at 336 and 299.50 nm for Alcaftadine; 268 and 293.50 nm for Ketorolac tromethamine. Method (C) is constructed on Q-absorbance ratio method where the iso-absorptive point was obtained at 296 nm and the max selected was of Alcaftadine 282 nm. The accuracy, precision, Limit of Detection (LOD), and Limit of Quantification (LOQ) 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 the statistical method 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 excipients, so it can be used for routine investigation of both drugs in quality control laboratories.