High‐performance thin‐layer chromatography has the utmost separation efficiency. High‐performance thin‐layer chromatography methods can be used to separate, detect, and estimate the different phytochemicals and drug components along with additives. A simple, accurate, precise, and robust high‐performance thin‐layer chromatography method has been developed and validated for the simultaneous estimation of sulfacetamide sodium and hydrocortisone acetate in eye drops. The separation was carried out on pre‐coated silica gel aluminum plate 60 F254 as a stationary phase using ethyl acetate: toluene: methanol: triethylamine (6:2:2:0.1) as a mobile phase. The densitometric measurement of separated components was performed at 254 nm. The method was validated for accuracy, precision, reproducibility, robustness, the limit of detection, and the limit of quantification as per International Council for Harmonization (Q2[R1]) guidelines. The regression coefficients (r2) for sulfacetamide sodium and hydrocortisone acetate were found to be 0.9998 and 0.9994, respectively. The values for percentage recoveries of sulfacetamide sodium and hydrocortisone acetate were found to be 100.2%–100.9% and 101.1%–101.2%, respectively. The developed thin‐layer chromatography method can be used for the analysis of sulfacetamide sodium and hydrocortisone acetate in eye drops.