Amyotrophic lateral sclerosis (ALS) is a devastating disease of motor neurons with unknown etiology. Evidence suggests that the brain undergoes degenerative changes in ALS, particularly within areas of the descending motor pathway. Identifying robust and non-invasive biomarkers that are sensitive to neurodegeneration in ALS is essential for improving clinical trial design and assessment of treatment effectiveness. This study evaluated 1H-MRS-measured metabolite levels as biomarkers of disease severity. Ultra-high field (7 tesla) 1H-MRS revealed metabolic abnormalities in the motor cortex and brainstem of humans with ALS that are dependent on disease stage.