Marosek, SEH, Antharam, V, and Dowlatshahi, K. Quantitative analysis of the acetic acid content in substances used by athletes for the possible prevention and alleviation of exercise-associated muscle cramps. J Strength Cond Res 34(6): 1539–1546, 2020—Athletes regularly consume commercially available food and sports shot products, carbohydrate beverages, and water to improve their physical exertion and to possibly prevent or relieve exercise-associated muscle cramps (EAMCs)—often experienced during practice, training, or competition. Acetic acid, a component of interest within these products, has been recognized for its potential role in cramp reduction. Acetic acid is postulated to mitigate cramping by decreasing alpha motor neuron activity through oropharyngeal stimulation and inhibitory neurotransmitter production, while aiding in the role acetylcholine plays in muscle contraction and relaxation. The purpose of this research is to analytically assess the most viable sources of acetic acid from substances that athletes ingest before or when experiencing these cramps. The range of samples investigated were based on their widespread use in the athletic world: dill and sweet pickle juices, yellow mustard, sweet relish, apple cider vinegar, Hot Shot, PJ Shot, PJ Sport, E-Lyte Sport, Powerade, Gatorade, Smartwater, and Propel (with electrolytes). As hypothesized, pH and enzymatic assay or spectroscopic analyses revealed that yellow mustard, sweet relish, all pickle juices, and the pickle juice products were composed of moderate amounts of acetic acid. Based on established studies resulting in EAMC relief, acetic acid consumption, and the appropriate serving size, the yellow mustard, PJ Shot, and all pickle juices would be the most practical and palatable sources of acetic acid for strength and conditioning professionals to recommend that athletes consume for the possible prevention or alleviation of muscle cramps.