Aspartame, methyl-L-α-aspartyl-L-phenylalaninate, is used worldwide as a sweetener in foods and drinks and is considered to be safe at an acceptable daily intake (ADI) of 40 mg per kg of body weight. This compound is completely hydrolyzed in the gastrointestinal tract to aspartic acid, phenylalanine and methanol, each being toxic at high levels. The objective of the present study was to quantify the volatile methanol component in the exhaled breath of ten healthy volunteers following the ingestion of a single ADI dose of aspartame. Direct on-line measurements of methanol concentration were made in the mouth and nose breath exhalations using selected ion flow tube mass spectrometry, SIFT-MS, several times before aspartame ingestion in order to establish individual pre-dose (baseline) levels and then during two hours post-ingestion to track their initial increase and subsequent decrease. The results show that breath methanol concentrations increased in all volunteers by 1082 ± 205 parts-per-billion by volume (ppbv) from their pre-ingestion values, which ranged from 193 to 436 ppbv to peak values ranging from 981-1622 ppbv, from which they slowly decreased. These observations agree quantitatively with a predicted increase of 1030 ppbv estimated using a one-compartment model of uniform dilution of the methanol generated from a known amount of aspartame throughout the total body water (including blood). In summary, an ADI dose of aspartame leads to a 3-6 fold increase of blood methanol concentration above the individual baseline values.