The aim of this article is to present a systematic review of magnetic resonance spectroscopy (MRS) studies of substance use disorders. As a noninvasive and nonionizing imaging technique, MRS is being widely used in substance abuse research to evaluate the effects substances of abuse have on brain chemistry. Nearly 40 peer-reviewed research articles that focused on the utility of MRS in alcohol, methamphetamine, 3,4-methylenedioxymethamphetamine, cocaine, opiates, opioids, marijuana, and nicotine use disorders were reviewed. Findings indicate inconsistencies with respect to alterations in brain chemistry within each substance of abuse, and the most consistent finding across substances was decreased N-acetylaspartate and choline levels with chronic alcohol, methamphetamine, and nicotine use. Variation in the brain regions studied, imaging technique, as well as small sample sizes might explain the discrepancies in findings within each substance. Future well-designed MRS studies offer promise in examining novel treatment approaches in substance use disorders.