Development, Optimization, and Validation of HPLC Method for Quantification of Enzalutamide in Bulk and Pharmaceuticals

An isocratic high-performance liquid chromatography method for the quantitative determination of enzalutamide in pharmaceutical products has been developed, optimized, and validated. Optimization of the analytical method was performed using the response surface methodology. The flow rate (0.8 and 1.2 mL/min) and acetonitrile ratio of the mobile phase (45 and 55%) and detector wavelength (235 and 245 nm) were chosen as independent variables. The effects of three independent variables on the standard solution concentration were investigated. As a result of evaluation with the Box-Behnken Design model, chromatographic separation was performed using X-bridge phenyl (150 mm × 3.0 mm; 3.5 μm) column eluted with a mobile phase consisting of ultrapure water and acetonitrile at a ratio of 47.5:52.5 v/v, respectively, and a flow rate of 1.0 mL/min was optimized with a standard retention time of 3.05 min at a wavelength of 239 nm. The method was then validated using International Conference on Harmonization (ICH) guidelines with an analytical run time of 10 min. The technique has been validated for linearity, precision, accuracy and specificity, showing excellent linearity (R2 = 0.9998) in a concentration range of 5 – 30 μg/mL. The limits of detection and quantification were 0.70 and 2.00 μg/mL, respectively. The intraday and interday accuracy and precision data were recorded within acceptable limits. Experimental optimization design proved to be a useful tool for optimizing parameters of the proposed liquid chromatography method. The developed, optimized, and validated liquid chromatography method can be reliably used in quality control laboratories for determination of the content of enzalutamide in pharmaceutical products.