Introduction: Arising evidence suggests that resistance training has the potential to induce beneficial modulation of biomarker profile. To date, however, only immediate responses to resistance training have been investigated using high-throughput metabolomics whereas the effects of chronic resistance training on biomarker profile have not been studied in detail. Methods: A total of 86 recreationally active healthy men without previous systematic resistance training background were allocated into i) a resistance training (RT) group (n=68, age 33 ± 7 years, body mass index (BMI) 28 ± 3 kg/m 2) and ii) a non-RT group (n=18, age 31 ± 4 years, BMI 27 ± 3 kg/m 2). Blood samples were collected at baseline (PRE), after 4 weeks (POST-4wk), and after 16 weeks of resistance training intervention (POST-16wk), as well as baseline and after the non-RT period (20-24 weeks). Nuclear magnetic resonance (NMR)-metabolome platform was used to determine metabolomic responses to chronic resistance training. Results: Overall, the resistance training intervention resulted in favorable alterations (P < 0.05) in body composition with increased levels of lean mass (~2.8 %), decreased levels of android (~9.6 %), and total fat mass (~7.5 %). These changes in body composition were accompanied by anti-atherogenic alterations in serum metabolome profile (FDR < 0.05) as reductions in non-HDL cholesterol (e.g., free cholesterol, remnant cholesterol, IDL cholesterols, LDL cholesterols) and related apolipoprotein B, and increments in conjugated linoleic fatty acids levels were observed. Individuals with the poorest baseline status (i.e. body composition, metabolome profile) benefitted the most from the resistance training intervention.