Advanced MALDI mass spectrometry imaging in pharmaceutical research and drug development

•Ideal mass spectrometer for MALDI MSI would satisfy the ‘4S-criteria for performance’ (speed, specificity, spatial resolution and sensitivity).•Quantification MALDI MSI is achieved by spotting calibration curves onto control tissue sections or by using mimetic tissue models.•Stable isotope standards are most commonly used for signal normalization in quantitative MSI.•Further Validation and multi-site studies are required for best practices in wet lab, ‘big data’ processing and reporting.•Besides drug disposition studies, investigation of target engagement, PK/PD and drug-induced toxicity is drawing more attention.