A reversed-phase high-performance liquid chromatographic (HPLC) procedure using ultra-violet (UV) detection for the analysis of edetate disodium (EDTA) via complexation with iron, in an ophthalmic cleanser, is reported. The method is selective, accurate, and reproducible. The peak area versus EDTA concentration is linear over the range of 50–150% of its label claim of 0.30 mg/mL, with a detection limit of 200 ng/mL. The mean absolute recovery of EDTA, using the described method, is 102.7 ± 0.4% (mean ± SD, n = 10).