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A precise measurement of drug levels both in a pharmaceutical industry‟ s perspective and health care setup is the need of the hour. Pharmaceutical companies spend extravagantly and also untiringly, day in and day out, to delineate a single successful drug moiety from thousands of lead compounds. In this pursuit, they rely on bioanalytical techniques which could help them in separating the most active elements from the crude mixtures. And also, later on in pre-clinical and clinical testing accurate measurement of drug levels in biological tissues using suitable analytical techniques are indispensable. Moreover, a treating physician depends on drug levels especially for those drugs with narrow therapeutic margin. Though the qualitative and quantitative analytical methods existing today are more sophisticated and complex, they actually originated and evolved from the roots of chromatography. Hence, chromatography still prevails as the most significant analytical method in molecular chemistry despite being primitive. This current indisputable status of chromatography is reflected by the fact that majority of the present techniques is based on the principle of chromatography. This review focusses on the various types of chromatography, their working principles and applications.