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Abstract This study aimed to evaluate muscle oxidative function during exercise in amyotrophic lateral sclerosis patients (pALS) with non-invasive methods in order to assess if determinants of reduced exercise tolerance might match ALS clinical heterogeneity. 17 pALS, who were followed for 4 months, were compared with 13 healthy controls (CTRL). Exercise tolerance was assessed by an incremental exercise test on cycle ergometer measuring peak O2 uptake ( $$\dot{{\rm{V}}}$$ V ̇ O2peak), vastus lateralis oxidative function by near infrared spectroscopy (NIRS) and breathing pattern ( $$\dot{{\rm{V}}}$$ V ̇ E peak). pALS displayed: (1) 44% lower $$\dot{{\rm{V}}}$$ V ̇ O2peak vs. CTRL (p