A sensitive competition enzyme‐immunoassay for quantification of gluten in foods was developed, using horseradish peroxidase‐labelled monoclonal antibodies. Selected antibodies specific for wheat omega‐gliadin components were used, and these antibodies bound proteins from the related cereals, rye and barley, which are also toxic to individuals with gluten‐intolerance (coeliac disease). Binding of these antibodies was not inhibited by heating of gluten during cooking or baking and the assay did not detect cereals not toxic in coeliac disease, such as maize or rice. Gluten could be quantified at higher levels in meat products or in cereal products such as flours or baked goods. Results were not affected by wheat variety. Quantitiative results could be obtained using simple extraction techniques and solvents (40% or 70% ethanol). Detection of gluten was quantitative in a wide range of foods, except for certain products containing gluten proteins that had been subjected to severe heat, enzymic or chemical treatment. In these products overestimates rather than underestimates were usually obtained.