Mass spectrometry (MS) has the inherent advantages of being the most specific and sensitive of generally applicable analytical methods. In a traditional mass spectrometry experiment a sample is dissolved and/or extracted using an MS friendly solvent and introduced into a vacuum. This talk will emphasize a new class of ionization methods (ambient ionization methods) that have surfaced over the last decade, and are now being applied broadly. In ambient ionization methods the analysis of the sample is carried out outside of the mass spectrometer in its native environment with minimal sample handling and often in a single step. The scope of ambient ionization methods and the specific application of desorption electrospray ionization (DESI) and its broad utility across the pharma pipeline will be highlighted. Paper spray ionization (PS) and its advantages and limitations in drug discovery and pharmacokinetic applications will also be discussed. The second half of the talk will focus on imaging applications by mass spectrometry using both ambient and traditional MALDI based approaches. A review of the multiple platforms (DESI, MALDI, liquid extraction surface analysis LESA, Flow-probe, Laser Capture Micro-dissection LCM) that are redefining the D (distribution) in ADME (absorption, distribution, metabolism and excretion) will be presented.