Multiply protonated human hemoglobin ␣-chain species, ranging from [M ϩ 4H] 4ϩ to [M ϩ 20H] 20ϩ , have been subjected to ion trap collisional activation. Cleavages at 88 of the 140 peptide bonds were indicated, summed over all charge states, although most product ion signals were concentrated in a significantly smaller number of channels. Consistent with previous whole protein ion dissociation studies conducted under similar conditions, the structural information inherent to a given precursor ion was highly sensitive to charge state. A strongly dominant cleavage at D 75 /M 76 , also noted previously in beam-type collisional activation studies, was observed for the [M ϩ 8H] 8ϩ to [M ϩ 11H] 11ϩ precursor ions. At lower charge states, C-terminal aspartic acid cleavages were also prominent but the most abundant products did not arise from the D 75 /M 76 channel. The [M ϩ 12H] 12ϩ -[M ϩ 16H] 16ϩ precursor ions generally yielded the greatest variety of amide bond cleavages. With the exception of the [M ϩ 4H] 4ϩ ion, all charge states showed cleavage at the L 113 /P 114 bond. This cleavage proved to be the most prominent dissociation for charge states [M ϩ 14H] 14ϩ and higher. The diversity of dissociation channels observed within the charge state range studied potentially provides the opportunity to localize residues associated with variants via a top-down tandem mass spectrometry approach. (J Am Soc Mass Spectrom 2006, 17, 923-931)