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In recent years there have been major developments in the field of mass spectrometry (MS) that permit the analysis and characterization of peptides and proteins at the femtomolar level (
1
–
8
). This chapter deals with the use of MS for the elucidation of peptide sequences of gliadin- and/or glutenin-derived fragments. It is meant to give an overview of what modern mass spectrometric methods can mean for your research in the biomolecular field rather than an in-depth step-by-step practical protocol. Important aspects are covered to give the reader an idea of what is needed to perform, and what can be expected from, this mass spectrometric application.