Metabolomic analysis of serum has been successfully applied for the discovery of biomarkers notably in a wide variety of domains such as physiology, toxicology, or nutrition. This approach, minimally invasive, is considered as one important indicator of physiological or pathological states [1]. Indeed, serum contains hundreds or even thousands of metabolites, thus allowing giving an appropriate overview of the metabolism in the body. Regarding the complexity of this biofluid, a single analytical technique cannot cover the entire metabolome. Thus, multi-analytical platform approaches are required to extend the detection of metabolites, what makes it one of the main challenges in the field of metabolomics. Mass spectrometry (MS) coupled to liquid chromatography (LC) and gas chromatography (GC) are becoming the most used techniques for metabolomics analysis because of their sensitivity and selectivity. In this context, a multi-analytical platform based on LC-HRMS and GC-MS has been developed for serum metabolomics. Moreover, and considering the essential role of lipids in cell structure and organization, signaling events, protein regulations, and metabolic transformations and trafficking specific sample treatment has been applied to order to cover both polar metabolites and lipids. For analysis of small molecules such as amino acid and organic acids, two complementary LC-HRMS methods (reversed phase and HILIC) and GC-MS have been used [2]. GC-MS methodology was also applied for the analysis of fatty acids, while, LC-HRMS/MS was achieved for lipid speciation and lipid classes separation [3]. The developed methodology was successfully applied for the analysis of serum samples showing the complementarities and specificity of these techniques. [1] Psychogios, N., et al., PLoS ONE, 6(2) : e16957. [2] Gallart-Ayala, H., et al., Analytica Chimica Acta, 2013, 75-83. [3] Dunn, W.B., et al., Nature Protocol, 2011, 1060-1083.