The variability in the stable C, N, and H isotopic composition caused by different beef processing operations (boiling, frying, and roasting) was studied. The aim was to evaluate the stability of stable isotopic fingerprint information during the beef cooking process. The δ13C, δ15N, and δ2H values for raw, boiled, fried and roasted beef were measured, and the differences in the stable isotopic composition between raw and processed beef products were assessed. The results indicated that the δ13C and δ15N values in raw beef were not significantly different compared with processed beef, but theδ2H values were significantly higher in processed beef than in raw beef. In general, boiling, frying, and roasting had no significant effect on δ13C and δ15N values, but theδ2H value of processed beef increased.