Rutin ion mobility spectra and MS spectra change with increasing transfer voltage parameter (TIMS Δ6) voltages. Increases of the TIMS Δ6 voltage cause changes to the ion mobility spectra of rutin. The 677 m/z adduct is no longer present when the TIMS Δ6 is increased to −175 V, and at −200 V, fragmentation of rutin occurs and a new aglycone ion at 300 m/z appears. This illustrates that increases in the TIMS Δ6 voltage can create collisions within the TIMS tunnel before TIMS analysis. This can be useful for the identification of adducts, reduction in space charge effects, and/or fragmentation of compounds for a three-step fragmentation method using TIMS Δ6, in-source CID, and collision cell CID.