Quality Control Analysis of Neomycin, Gramicidin and Triamcinolone in Ointment Formulation using HPLC Methodology: Methodology Development, Validation and Application

Abstract

The combination of gramicidin (GMN), a bactericidal agent; neomycin (NMN), an aminoglycoside; and triamcinolone (TAA), a glucocorticoid, is used for treating eye infections that are accompanied by inflammation. The contents of GMN, NMN, and TAA in eye ointments must be monitored for quality. In this research, a quick, selective, and robust stability-indicating HPLC method has been developed for the concurrent assay of GMN, NMN, and TAA in bulk and ointment formulations. Analysis was performed using ODS, a250mm × 4.6mm column and mobile phase having 0.1% aqueous formic acid-acetonitrile with a ratio of 20:80 v/v; PDA analysis was performed at 225nm. The elution times for GMN, NMN, and TAA were 2.597min, 4.440min, and 3.251 min, respectively. The GMN, NMN, and TAA linear ranges were 0.625 µg/ml to 3.75 µg/ml, 6.25 µg/ml to 37.50µg/ml and 2.5µg/ml to 15.0µg/ml, respectively. The method is precise with 0.68-0.96% (GMN), 0.52-1.02% (NMN), and 0.26-0.62% (TAA) RSD values. The method is accurate with 98.75-101.25% (GMN), 98.61-100.33% (NMN), and 98.67-100.23% (TAA) recovery values. In degradation investigation, the degradant’s peak elution times are different from the elution times of GMN, NMN, and TAA. Thus, the proved specificity and stability indicating the power of the method. Finally, this developed analytical approach was efficaciously applied to a commercial ointment formulation containing afixed dose of GMN, NMN, and TAA, demonstrating its usefulness for quality control and degradation investigations on GMN, NMN, and TAA.