Journal of Chromatography A, 1370 (2014) 105–114
Contents lists available at ScienceDirect
Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma
Using mass spectrometry to detect hydrolysed gluten in beer that is responsible for false negatives by ELISA Michelle L. Colgrave a,∗ , Hareshwar Goswami a , Malcolm Blundell b , Crispin A. Howitt b , Gregory J. Tanner b a b
CSIRO Agriculture Flagship, 306 Carmody Road, St Lucia, QLD 4067, Australia CSIRO Agriculture Flagship, GPO Box 1600, Canberra, ACT 2601, Australia
a r t i c l e
i n f o
Article history: Received 28 July 2014 Received in revised form 9 October 2014 Accepted 10 October 2014 Available online 18 October 2014 Keywords: Gluten ELISA Multiple reaction monitoring (MRM) Mass spectrometry (MS) Quantification
a b s t r a c t Gluten is the collective name for a class of proteins found in wheat, rye, barley and oats. Eating gluten triggers an inappropriate auto-immune reaction in ∼70 million people globally affected by coeliac disease, where the gut reacts to gluten proteins and this triggers an immune response, resulting in intestinal inflammation and damage. Gluten-free foods are now commonplace, however, it is difficult to accurately determine the gluten content of products claiming to be gluten-free using current methodologies as the antibodies are non-specific, show cross-reactivity and have different affinities for the different classes of gluten. The measurement of gluten in processed products is further confounded by modifications to the proteins that occur during processing and in some case hydrolysis of the proteins. In this study, LC–MS/MS was used to profile whole beer, and two beer fractions representing hydrolysed hordeins (
Contents lists available at ScienceDirect
Journal of Chromatography A journal homepage: www.elsevier.com/locate/chroma
Using mass spectrometry to detect hydrolysed gluten in beer that is responsible for false negatives by ELISA Michelle L. Colgrave a,∗ , Hareshwar Goswami a , Malcolm Blundell b , Crispin A. Howitt b , Gregory J. Tanner b a b
CSIRO Agriculture Flagship, 306 Carmody Road, St Lucia, QLD 4067, Australia CSIRO Agriculture Flagship, GPO Box 1600, Canberra, ACT 2601, Australia
a r t i c l e
i n f o
Article history: Received 28 July 2014 Received in revised form 9 October 2014 Accepted 10 October 2014 Available online 18 October 2014 Keywords: Gluten ELISA Multiple reaction monitoring (MRM) Mass spectrometry (MS) Quantification
a b s t r a c t Gluten is the collective name for a class of proteins found in wheat, rye, barley and oats. Eating gluten triggers an inappropriate auto-immune reaction in ∼70 million people globally affected by coeliac disease, where the gut reacts to gluten proteins and this triggers an immune response, resulting in intestinal inflammation and damage. Gluten-free foods are now commonplace, however, it is difficult to accurately determine the gluten content of products claiming to be gluten-free using current methodologies as the antibodies are non-specific, show cross-reactivity and have different affinities for the different classes of gluten. The measurement of gluten in processed products is further confounded by modifications to the proteins that occur during processing and in some case hydrolysis of the proteins. In this study, LC–MS/MS was used to profile whole beer, and two beer fractions representing hydrolysed hordeins (
