The development of the bio analytical techniques brought a progressive discipline that the longer term holds many exciting opportunities to further improvement. The main impact of biolysis within the pharmaceutical industry is to get a quantitative measure of the drug and its metabolites. The purpose is to perform the pharmacokinetics, toxic kinetics, and bioequivalence and exposure response like pharmacokinetic/pharmacodynamics studies. Various bio analytical techniques are performed in bio analytical studies such as hyphenated techniques, chromatographic techniques, and ligand binding assays. This review extensively highlights the role of bio analytical techniques and hyphenated instruments in assessing the bio analysis of the drugs.