<p>Nebivolol, a beta-blocker commonly prescribed for the treatment of hypertension and heart failure, functions by decelerating heart rate and promoting vasodilation, thereby enhancing blood flow and reducing blood pressure. Distinctively, nebivolol elevates nitric oxide levels, facilitating superior vasorelaxation compared to other beta-blockers. This dual mechanism renders it a potent option for hypertension management. Nebivolol is well-tolerated, particularly among patients with respiratory conditions, due to its minimal pulmonary impact. Administered once daily, it can be used as monotherapy or in combination with other antihypertensive agents to maintain optimal blood pressure control. This review comprehensively examines various analytical methodologies for the quantification of nebivolol and its combinations in pharmaceutical formulations and biological matrices. It encompasses both fundamental and advanced techniques for the estimation of nebivolol hydrochloride. Electrochemical techniques, high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC–MS/MS), UV–visible spectrophotometry, and spectrofluorometric method are some of the most common ways to do an assay. Among these, HPLC coupled with spectrophotometric detection emerges as the most extensively utilized method for nebivolol assay.</p> Graphical Abstract <p></p>