High-speed HPLC/MS/MS analysis of biological fluids: a practical review

Over the course of the last three or four years, the need for increased throughput analytical methods in support of drug discovery operations has dramatically increased. Once a nicety, methods which can provide reliable quantitation of target analytes in biological fluids are now necessary to keep pace with biological screening. To date, the only successful approaches to direct high-speed quantitation of large numbers of diverse compounds are based on high-performance liquid chromatography/mass spectrometric (HPLC/MS/MS) techniques. Key challenges to this task include sample preparation, the automated development of methods and the rapid analysis of many samples. Accomplishing these tasks requires the use of generic methods, which can be applied across many structurally diverse compounds. In this review, the state-of-the-art in high-speed HPLC/MS/MS is reviewed with the aim of describing the most practical and effective approaches currently in use.