A method for acquiring and analyzing polypeptide mass spectra comprising: creating first and second sets of fragmented ions using first and second fragmentation techniques; obtaining mass spectra of the first and second sets of fragmented ions; searching a database of fragments of a first fragment pair type and of a second fragment pair type to respectively provide a set of M ranked and a set of N ranked best matching polypeptide sequences; subtracting synthetic spectra corresponding to each of the M and N ranked sequences from the mass spectra to provide first and second sets of modified mass spectra; searching the database of fragments of the second fragment pair type and of the first fragment pair type to provide third and fourth sets of polypeptide sequences providing best matches to the first and second sets of modified spectra, respectively; and creating a cumulative ranking of the third and fourth sets.