Bioanalytical method development, validation and quantification of flutamide in spiked rat plasma by using high-performance liquid chromatography

Background A quick, low cost and precise method was developed and validated for flutamide in the rat plasma using a high-performance liquid chromatography. Acetanilide was used as internal standard in the study. Flutamide was extracted from rat plasma using methanol. The chromatographic analysis was carried out on Cromasil C18 column at 227 nm using methanol-water mixture as the mobile phase and flow rate of 0.8 ml/min. Results The developed method demonstrated good linearity over the 100-1000 ng/ml concentration range with a regression coefficient of 0.9947. The accuracy of the developed method was found to be 97-101%, while its precision was found to be less than 5%. Limits of detection and quantitation were determined to be 2.52 ng/ml and 7.66 ng/ml, respectively. Flutamide showed recovery of about 96-100% and the developed method was further used to investigate the pharmacokinetics of flutamide in the marketed formulation. Conclusions From the results, it can be concluded that the developed method has the potential to successfully determine flutamide in rat plasma and establish the pharmacokinetic parameters of other flutamide- containing formulations.