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Tamoxifen (TAM) and Sulphoraphane (SFN) are well-known anti-estrogen drugs used for the treatment of breast cancer. Due to their synergistic therapeutic potential, their combination is preferred as it helps to minimize the drug-related toxicities and enhances therapeutic efficacy. A simple, robust and fast simultaneous reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed as well validated for the analysis of both the drugs based on their particular wavelength. The separation was performed on C₁₈ analytical column with dimensions of 4.6 × 250 mm, 5 μm using mobile phase methanol: water (pH 3.5) in the ratio 70:30 and flow rate of 0.8 min/mL. Box-Behnken experimental design was used to optimized these independent variables and analyze their effect on the response variables like retention time (RT), no. of theoretical plates and tailing factor of both analytes. Method …