Abstract

Mass spectrometry (MS) has been the basis of phytochemical study, yet the area is dramatically transitioning to high- throughput workflows that can disentangle the extreme complexity of biological systems. The present review deals with the shift that occurs Mass Spectrometry Technologies and their integration, which is leading to a system-level phytochemical analysis. We discuss cutting-edge concepts with emphasis on ambient ionization methods (for example, LAESI-MS, DESI-MSI, PS-MS) that by direct in-situ access of metabolites from the plant facilitate fast spatially resolved analysis with critical spatial resolution and minimum sample preparation. The transition to advanced hyphenation is a main idea. The coupling of chromatographic methods (LC, GC) with Ion Mobility Spectrometry (IMS) and high-resolution MS (HRMS) creates a separation space that can solve the problem of isobaric along with isomeric resolution, which is the main problem in natural product extracts. Besides that, we are acknowledging the emergence of methods of interpretation of data, such as Parallel Covariance Two-Dimensional MS (pC-2DMS) and the joining of MS with NMR spectroscopy and AI-based bioinformatics tools (like molecular networking) that are necessary to make more rapid the process of unknown compound annotation and establish a connection with the chemical structures and the biosynthetic pathways. The problems of matrix effects and accessibility are still there, yet the future of phyto- chemical MS is marked by miniaturization, portability, and advanced data processing, all of these facilitating a rapid pace of dis- covery of new bioactive molecules and leading to the dynamic understanding of plant metabolism.