Coeliac disease (CD) is a T-cell mediated autoimmune disorder triggered by ingestion of cereal gluten found in wheat (gliadins and glutenins), barley (hordeins), and rye (secalins). As the only treatment for CD is a lifelong gluten-free diet, the measurement of gluten in raw ingredients and processed food products is critical to protecting people with CD or gluten intolerance. The most commonly employed method is the enzyme-linked immunosorbent assay (ELISA), but more recently mass spectrometry has been employed wherein the extracted gluten proteins are digested to peptides that are then directly measured. To achieve the goal of accurate gluten quantitation, gluten must be efficiently extracted from the ingredient or food matrix and then digested to yield the peptides that are monitored by LC-MS. In this chapter, a rapid, simple, and reproducible protocol for extraction and digestion of gluten proteins is described.
Keywords: Chymotrypsin; Digestion; Flour; Gluten; Mass spectrometry; Trypsin.