Hypertension is a multifaceted cardiovascular disease, a significant risk factor for stroke, heart attack, heart failure, and renal damage. An essential phase in the drug development process is the exploration of effective bioanalytical approaches to investigate drug metabolism and pharmacokinetics precisely. The use of LC-MS has increased significantly over the last 10 years; numerous researchers have made contributions to the field and enhanced the technical capabilities of workflows based on LC-MS. This review provides a critical analysis of the method development and validation of bioanalytical methods using Liquid Chromatography-Mass Spectrometry (LC-MS) of a few antihypertensive drugs, focusing on extraction techniques and validation parameters. Furthermore, a fleeting look at the GLP, regulatory guidelines, machine learning and artificial intelligence in bioanalysis. Despite these advancements, the document identifies gaps in current regulatory guidelines and advocates areas for further research, predominantly concerning matrix effects and the impact of co-medications. The integration of AI tools in LC-MS has shown the potential to revolutionize bioanalytical methods, yet there is still an imperative for global harmonization. We assume that this review will offer a foundation for the research of new strategies and assist in the identification of the optimum relevant methodology parameters for known and emerging antihypertensive drugs.