Rapid Evaporative Ionisation Mass Spectrometry (REIMS) was used to determine the impact of in-bag ageing regimes (stepwise-ageing at different air velocities and straight-dry-ageing) and trimming on the metabolic profile of dry-aged lean beef. Orthogonal projection to latent structures-discriminant analysis (OPLS-DA) models based on 1705 tentatively identified m/z features were found for ageing methods (Q2 = 0.85), ageing time (0 vs. 21 days, Q2 = 0.95) and sampling locations (surface meat vs. trimmings, Q2 = 0.94). No significant (P > 0.05) difference in metabolites due to air velocities. Small metabolites such as dipeptides and amino acids were more abundant, especially on the surface of untrimmed lean beef, following 21 days of straight-dry-ageing. Stepwise-ageing produced different metabolic profiles from straight-dry-ageing, suggesting that the two methods may differ in dry-aged meat quality and flavour. This work demonstrates REIMS's potential for real time differentiation of meat on processing parameters.