The final aim of metabolomics is the comprehensive and holistic study of the metabolome in biological samples. Therefore, the use of instruments that enable the analysis of metabolites belonging to various chemical classes in a wide range of concentrations is essential, without compromising on robustness, resolution, sensitivity, specificity, and metabolite annotation. These characteristics are crucial for the analysis of very complex samples, such as wine, whose metabolome is the result of the sum of metabolites derived from grapes, yeast(s), bacteria(s), and chemical or physical modification during winemaking. In recent years, a big advantage, in this direction, was the hardware developments on hyphenated instruments that enable the integration of liquid chromatography (LC), ion mobility spectrometry (IMS), and mass spectrometry (MS). This chapter describes an LC-IMS-MS protocol for the analysis of wine and grape samples as well as the use of IMS data in metabolite annotation.