•A review on chromatographic methods for analysis of AA metabolites in brain tissue is presented.•A review on mass spectrometric methods for analysis of AA metabolites in brain tissue is presented.•A critical review of analytical and extraction techniques for brain tissue is demonstrated.•Recently published analytical methods are discussed and analytical challenges are highlighted.•Diagrams and tables are illustrated for comparison of methods. Eicosanoids are potent lipid mediators of inflammation and are known to play an important role in numerous pathophysiological processes. Furthermore, inflammation has been proven to be a mediator of diseases such as hypertension, atherosclerosis, Alzheimer's disease, cancer and rheumatoid arthritis. Hence, these lipid mediators have gained significant attention in recent years. This review focuses on chromatographic and mass spectrometric methods that have been used to analyze arachidonic acid and its metabolites in brain tissue. Recently published analytical methods such as LC–MS/MS and GC–MS/MS are discussed and compared in terms of limit of quantitation and sample preparation procedures, including solid phase extraction and derivatization. Analytical challenges are also highlighted.