Min-Ho Song (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Ji-Ho Lee (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Min-Ho Song (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Ji-Woo Yu (Department of Crop Science, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea)

Jung-Hoon Lee (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Eun-Song Choi (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Ji-Ho Lee (School of Natural Resources and Environment Science College of Agriculture and Life Sciences, 1, Kangwondaehak-gil, Kangwon National University, Gangwon State 24341, Republic of Korea)

Pesticides such as α-cypermethrin are extensively used in modern agriculture; however, their broader impacts on non-target organisms and biochemical pathways remain insufficiently characterized. This study presents an integrated metabolomic assessment of α-cypermethrin exposure in two biological systems: Brussels sprouts (Brassica oleracea) and adult zebrafish (Danio rerio). In Brussels sprouts, a GC-MS/MS-based targeted metabolomics approach revealed significant perturbations in central carbon metabolism, amino acid turnover, and antioxidant defense pathways. Notably, accumulation of fumarate and malate in the TCA cycle, suppression of glycolate/glycerate in the glyoxylate pathway, and dynamic changes in stress-related amino acids were observed, suggesting adaptive responses to chemical stress. In parallel, adult zebrafish exposed to environmentally relevant concentrations of α-cypermethrin exhibited time- and dose-dependent metabolic disruptions, particularly in glycolysis, glycerolipid metabolism, and glutathione turnover. Early exposure led to the accumulation of glycolytic intermediates and 5-oxoproline, alongside glutamate and GABA depletion, indicating oxidative stress and redox imbalance. At 48 h, partial metabolic recovery suggested compensatory adaptation. Intriguingly, conserved alterations in core metabolic pathways were observed across both kingdoms, including TCA cycle involvement and amino acid metabolism, underscoring shared biochemical responses to xenobiotic stress.​