The analytical technique of High Performance Liquid Chromatography (HPLC) is used extensively thought the pharmaceutical industry. It allows simultaneously both qualitative and quantitative information of a drug. It is used to provide information on the composition of drug related samples. The information obtained may be qualitative, indicating what compounds are present in the sample. The information obtained may be qualitative, providing the actual amount of compounds in the sample. HPLC is used in all the different stages in the creation of new drug discoveries and also used routinely in the drug manufacturing process. The HPLC techniques is highly accessible to the author's knowledge this review represents the most exhaust description of instrumentation, method development, recent advancement, area of application. This article reviews the most recent advances in sample preparation, separation and the mass spectrometric aspects of high-throughput for method development.