Impact of a CoQ10, Alpha-Lipoic Acid, and Chaga Extract Formulation on Mitochondrial Dysfunction and Oxidative Stress: NMR Metabolomic Insights into Cellular Energy and Neurotransmission

Objectives: The aim of this study was to evaluate the impact of a novel antioxidant formulation (RP-25) con-taining CoQ10, alpha-lipoic acid, and Chaga extract on mitochondrial dysfunction and oxidative stress. The formulation’s potential to modulate cellular energy metabolism and neurotransmission was explored using a combination of NMR-based metabolomics and UHPLC-HRMS analytical techniques. Methods: SH-SY5Y neu-roblastoma cells were treated with RP-25, and cell viability was assessed via CCK-8 assay. Metabolomic profiles of the treated and untreated cells were analyzed by 1D-NMR, providing insights into the endo- and esometab-olomes. Additionally, a UHPLC-HRMS method was developed for quality control and analysis of the RP-25 formulation. Multivariate statistical approaches, including PLS-DA and volcano plot analysis, were used to identify key metabolic changes. Results: RP-25 treatment did not affect cell viability but significantly modulat-ed metabolic pathways, including amino acid biosynthesis, oxidative phosphorylation, and glycolysis. Increased levels of ATP, glutamate, tyrosine, and proline were observed in treated cells, indicating enhanced cellular en-ergy production and neurotransmission activity. Pathway enrichment analysis highlighted the formulation's influence on mitochondrial energy metabolism and oxidative stress response. Conclusions: RP-25 demonstrated potential as a therapeutic strategy for mitigating mitochondrial dysfunction and oxidative stress, with implica-tions for conditions associated with aging and neurodegenerative diseases.