Measurements of the radiocarbon abundance in 5 samples of bis(2–ethylhexyl) phthalate (DEHP) isolated from Stilton cheese were made by accelerator mass spectrometry (AMS) to determine the fraction of carbon originating from contemporary biogenic sources. DEHP is classified as a “priority hazardous substance” by the European Union, a probable human carcinogen by the United States Environmental Protection Agency, and is suspected to be a human endocrine disrupter. Measurement of its 14C abundance in a specific food indicates whether its presence is due to contamination from industrially synthesized DEHP or a naturally inherent component. A method was developed to determine the contemporary carbon fraction of DEHP in a fatty food matrix at concentrations of 0.14 mg/kg. Five 90–μg quantities of DEHP were extracted from 12 kg of Stilton cheese and isolated by silica gel, size exclusion, and high-performance liquid chromatography (HPLC). Masses of samples were determined by gas chromatography mass spectrometry (GC-MS) analyses prior to combustion and manometry afterwards. The purity of DEHP carbon mass in each isolate was determined by multivariate deconvolution of GCMS fragmentation spectra obtained from measurements of standards and isolates, and ranged from 88.0 ± 1.8% to 92.3 ± 1.1% (n = 5, 1σ). Concurrently processed isolation method blanks contained from 0.15 ± 0.04 to 1.52 ± 0.06 μg (n = 3, 1σ) DEHP per sample and significant quantities of pre- and post-chromatographic extraneous carbon contamination. The mean 14C-corrected contemporary carbon fraction of DEHP in the isolates was 0.235 ± 0.073 (1σ; and ± 0.091 at the 95% confidence level), revealing that the majority of DEHP in Stilton cheese results from anthropogenic sources, but with a significant naturally occurring component.