Background: Celiac disease, a complex, long-term autoimmune disorder and gluten intolerance, is estimated to affect from 1 to 5% of the world's population. Objective: As a consequence, to protect gluten-sensitive consumers, the development of reliable analytical methods allowing the detection of gluten in various food products is needed. Methods: Currently, ELISA is probably the most widespread used methodology. The method based on the R5 antibody has received type I status in Codex Alimentarius. However, the ELISA method suffers from some limitations, especially concerning quantification of nonwheat gluten. As a consequence, the development of another complementary methodology such as LC-tandem MS (MS/MS) is considered to be essential. Furthermore, this method could also be used for the simultaneous detection of gluten with other allergens, which will constitute a great additional benefit for producers of "free-from" food products and/or having a management policy integrated for several allergies and/or intolerances. Results: A multi-allergen and grain-specific ultra-HPLC coupled to MS/MS method allowing the identification and the discrimination of gluten from seven cereals, simultaneously with the detection and identification of 10 allergens in only one analysis, is thus described here. Conclusions: This method can be used for the analysis of a broad range of foodstuff matrices containing wheat and/or its derivatives, including cereals, flours, heat-treated and foodstuffs, but also more complex samples having undergone fermentation processes (such as beers).