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Attention-deficit/hyperactivity disorder (ADHD) affects up to 10% of children, often presenting with emotional dysregulation (irritability, anger). The kynurenine pathway, by which the amino acid tryptophan is converted into both neuroprotective and neurotoxic metabolites, was investigated for its role in ADHD pathophysiology. The MADDY study was an 8-week, multi-site, randomized controlled trial of a multinutrient supplement versus placebo in children 6-12 years old with symptoms of ADHD and emotional dysregulation. Participants from two U.S. sites (n = 84) provided baseline and week 8 urinary samples, analyzed via liquid chromatography-mass spectrometry for neurotransmitter levels. Linear mixed-effects models assessed change over time for seven analytes. Missing values were imputed, and results adjusted for multiple comparisons. Baseline neurotransmitter concentrations were not different between treatment groups. Changes over time in the seven analytes were not different between treatment groups. Nine participants had neurotransmitter levels below the lower limit of detection and were imputed; demographic characteristics were not different between these and other participants. Eight weeks of multinutrient supplementation did not change concentrations of urinary tryptophan, serotonin, or kynurenines in this sample of children 6-12 years old with ADHD and emotional dysregulation.