Regulated Bioanalysis: Fundamentals and Practice

Abstract

The development of liquid chromatography-mass spectrometry technologies in the 1970s and 1980s had fundamentally changed the development of new drugs and the pharmacokinetic data quality generated during bioanalysis. A brief review of these historical developments of the interfacing of liquid chromatography with mass spectrometry transitions into the best use practices for those new to the field of bioanalysis. The chapter highlights the benefits of implementing ultra high-pressure liquid chromatography to provide improved sensitivity, selectivity, and analysis speed combined with triple quadrupole mass spectrometry. The use of appropriate solvents and solvent modifiers combined with the stationary phase appropriate for the analyte’s structure is discussed and provided in an easy to read and understand manner. The introduction of the eluate to the mass spectrometer ion source is described for the two most commonly used atmospheric pressure ionization techniques in use today. The process of electrospray ionization and atmospheric pressure chemical ionization are used to create charged molecules which are directed into the vacuum region of the mass spectrometer. A step-by-step guide is provided on the optimization of the mass spectrometer settings which provides selective and sensitive selected reaction monitoring signals that can be used to quantify compounds. Best practices are discussed for system maintenance, calibration, and tuning and appropriate documentation needed to support regulatory requirements.