Ultrasensitive Profiling of Arachidonic Acid Metabolites Based on 5-(Diisopropylamino)amylamine Derivatization–Ultraperformance Liquid Chromatography–Tandem Mass Spectrometry

Arachidonic acid (AA) and its metabolites play critical roles in inflammation and immune regulation, modulating the initiation, amplification, and resolution of inflammation. However, their comprehensive quantification remains a challenging endeavor owing to complex metabolic pathways and biological matrix effects. This study introduces a novel metabolomics method involving 5-(diisopropylamino)-amylamine (DIAAA) derivatization coupled with ultraperformance liquid chromatography-tandem mass spectrometry to address these issues. The method demonstrated high sensitivity and specificity, with limits of quantification meeting stringent criteria (relative standard deviation <20%; recovery rate, 85-115%, signal-to-noise ratio >10). It effectively quantified 14 key AA metabolites, including hydroxyeicosatetraenoic acids, prostaglandins, and leukotrienes, across a wide linear range (<i>R</i> ² > 0.98). The results of intra- and interassay precision tests exhibited low coefficients of variation (≤15%), underscoring the reproducibility of the method. DIAAA derivatization also mitigated matrix variability, improving the accuracy of metabolite detection in serum samples. The hallmark of allergic diseases is a disrupted AA metabolism, where elevated specific metabolites (AA, HETEs, LTB4, and PGD2) show strong diagnostic promise, and a unique metabolite signature in polysensitized patients indicates a link to inflammatory severity. This advanced analytical approach offers significant potential for elucidating the role of AA metabolism in allergic diseases and holds promise for applications in clinical diagnostics and therapeutic monitoring.