Analysis of the component basis for anti-fatigue active ingredients in fermented black mulberry juice based on the...

This study aimed to investigate the anti-fatigue activity of fermented black mulberry juice and elucidate its component basis. A fatigue model was established using C2C12 mouse myoblasts induced by using a combined treatment of H₂O₂ induction and electrical stimulation. The ethyl acetate, butan-1-ol, and water extracts enhanced cell viability and mitochondrial membrane potential while reducing lactate dehydrogenase and reactive oxygen species levels. RT-qPCR indicated these extracts promote mitochondrial biogenesis, with ethyl acetate extract showing the most significant effect. Compositional analysis via liquid chromatography-mass spectrometry revealed that the ethyl acetate extract primarily contained phenolic acids such as caffeic acid, while the butan-1-ol fraction was rich in flavonoids including rutin and quercetin. Although the water extract shared a similar compositional profile, its antioxidant activity was lower due to insufficient levels of characteristic active substances. The butan-1-ol extract exhibited the highest chemical richness, with 2972 compounds tentatively identified.