Eplerenone (EPL) and spironolactone (SPR) belong to the class of potassium-sparing diuretics. Both drugs are prescribed for the treatment of hypertension and heart failure. Spironolactone is structurally like progesterone and binds to progesterone, androgen and mineralocorticoid receptors. Eplerenone is a selective mineralocorticoid receptor antagonist, so it lacks the anti-androgenic side effects of spironolactone. Using a thorough computer assisted literature survey; this review article touches upon the reported analytical methods for the quantification of both drugs in raw materials, pharmaceutical dosage forms and biological fluids. Bioanalytical methods are widely used for quantitative estimation of drugs and their metabolites in biological matrices. Various analytical methods such as spectrophotometry, spectrofluorimetry, HPLC, TLC, and UPLC have been used in laboratories for the quantitative analysis of these drugs in biological samples and pharmaceutical preparations. Therefore, the aim of this review is to give a summary of the current analytical methods used for the assay of both drugs. The review represents a guide for the QC labs and the bioequivalence centers to analyze the studied drugs.