High-resolution LC-MS/MS-based Metabolite Profiling of Medicinal Herbs and Spices: Innovations in Untargeted and Targeted Phytochemical Characterization

High-resolution liquid chromatography–tandem mass spectrometry (LC-MS/MS) has reshaped phytochemical characterization by enabling the sensitive and structurally resolved profiling of metabolites in medicinal herbs and spices. This chapter outlines methodological innovations spanning sample preparation, class-aware extraction strategies, and chromatographic modes optimized for complex botanical matrices. Advances in ionization chemistry, adaptive acquisition logic, and ion-mobility integration have increased spectral richness while improving discrimination among isomeric or low-abundance metabolites. Post-processing frameworks that combine spectral libraries, fragmentation, retention-time prediction, and CCS filtering now support transparent, multi-evidence annotation consistent with modern confidence-level standards. Case studies illustrate how LC-MS/MS enhances botanical authentication, quality assessment, and mechanistic interpretation relevant to pharmacological and nutraceutical applications. By uniting high-resolution analytics with open spectral resources and machine-assisted annotation, LC-MS/MS provides a scalable and reproducible foundation for research, regulatory standardization, and functional-food innovation.