Development and Validation of an HPTLC Method for Simultaneous Estimation of Azithromycin, Fluconazole, and Tinidazole in Combined Dosage Form

A high-performance thin-layer chromatographic (HPTLC) method was developed and validated for the simultaneous estimation of azithromycin (AZM), fluconazole (FLZ), and tinidazole (TZ) in pharmaceutical formulations.Various mobile phase combinations were evaluated, and the optimized system-1,4-dioxane: methanol: toluene (2:3:5, v/v/v)-produced sharp, compact, and well-resolved spots with Rf values of 0.202, 0.382, and 0.522 for AZM, FLZ, and TZ, respectively.Densitometric detection was performed at 210 nm.The method exhibited excellent linearity (r 0.999) within the concentration ranges of 1,000-5,000 ng/band (AZM), 500-5,000 ng/band (FLZ), and 100-500 ng/band (TZ).Accuracy was confirmed by recovery rates between 98% and 102%, and precision was supported by %RSD values below 2% for intra-day, inter-day, and repeatability assessments.The method also demonstrated acceptable sensitivity with low limit of detection (LOD), and limit of quantification (LOQ).values,and system suitability parameters met all validation criteria.This validated HPTLC method is simple, accurate, precise, and robust, making it suitable for routine quality control of AZM, FLZ, and TZ in combined pharmaceutical dosage forms.