Comparative LC‐MS–Based Untargeted Metabolomics of Acrylamide‐Induced Metabolic Alterations in SH‐SY5Y Human Neuroblastoma and Human Fibroblast Cell Lines

Acrylamide (ACR), a synthetic and food-derived contaminant, is widely recognized for its neurotoxic potential. However, the underlying metabolic alterations driving its toxicity remain poorly understood. In this study, we investigated ACR-induced metabolic alterations in SH-SY5Y human neuroblastoma cells and GM08680 human fibroblast cells using an untargeted metabolomics approach with liquid chromatography-Orbitrap high-resolution mass spectrometry (LC-Orbitrap HRMS). The altered profiles focused specifically on changes related to neurotoxicity. ACR cytotoxicity was assessed using MTT and live/dead cell assays in both control and ACR-treated cells. The half-maximal inhibitory concentration (IC₅₀) of ACR was found to be 1.32 mM for SH-SY5Y and 3.12 mM for GM08680. A concentration-dependent live/dead cell assay significantly revealed ACR-induced morphological changes and cell death. Untargeted metabolite profiling showed that ACR exposure significantly altered the metabolome of both cell lines. Of the 82 annotated metabolites identified, 45 displayed significant dysregulations in SH-SY5Y cells. Principal component analysis (PCA) demonstrated clear separation between control and treated groups, while heatmap clustering confirmed consistent metabolic shifts across both cell lines. Metabolite set pathway enrichment analysis showed that the altered metabolites mainly involved amino acid metabolism, glycolysis, nucleotide biosynthesis, and oxidative stress regulation. These alterations were more pronounced in SH-SY5Y cells, suggesting a higher neuronal susceptibility to ACR toxicity. In conclusion, this study highlights key metabolic disruptions associated with ACR exposure in both cell lines, providing insights into neuronal and nonneuronal differences. The disrupted pathways may serve as early biomarkers for acrylamide-induced neurotoxicity and neurodegeneration.