Confirmation of gluten-free status of wheatgrass (Triticum aestivum)

Main Article Content

S.L. Adrianos
B. Mattioni
M. Tilley


wheatgrass, gluten analysis, ELISA, toxic gliadin peptide


Young wheat (Triticum aestivum) leaves contain concentrated nutrients that may be beneficial as a nutritional supplement to those with malabsorption disorders such as celiac disease (CD) if both gluten-free, and free of gluten contamination. The tender 8-10 day leaves are juiced and the juice is consumed, added to smoothies, or consumed as a purchased wheatgrass dietary supplement in tablet, capsule and liquid forms. CD and other gluten related disorders cause both malabsorption of nutrients and an abnormal immune reaction to gluten. The only effective therapy is a lifelong gluten-free diet. Codex Alimentarius sets the gluten threshold level at 20 mg/kg for gluten-free foods. This threshold has been adopted by many regulatory agencies such as European Commission and the US Food and Drug Administration. Consumption of wheatgrass extract has been reported to alleviate symptoms of ulcerative colitis and anaemia which are often symptoms of CD. This investigation utilised the R5 and G12 sandwich enzyme-linked immuno sorbent assays (ELISAs) to examine the potential gluten content of commercial and home-grown wheatgrass. Samples were spiked with commercial gliadin to assess recovery rates. Acceptable recovery rates ranging from 91 to 133% from gluten spiked samples were found. In all preparations the gluten content of wheatgrass leaf tissue was lower than the limit of detection and limit of quantification for both ELISAs as well as the thresholds defined by Codex Alimentarius standards. According to results presented here appropriately prepared wheatgrass contains no gluten and may have a role as a dietary supplement in a gluten-free diet.

Abstract 84 | PDF Downloads 87


Abbott, M., Hayward, S., Ross, W., Godefroy, S.B., Ulberth, F., Van Hengel, A.J., Roberts, J., Akiyama, H., Popping, B., Yeung, J.M., Wehling, P., Taylor, S.L., Poms, R.E. and Delahaut, P., 2010. Validation procedures for quantitative food allergen ELISA methods: community guidance and best practices. Journal of AOAC International 93: 442­-450.
Altenbach, S.B., Kothari, K.M. and Dao Lieu, D., 2002. Environmental conditions during wheat grain development alter temporal regulation of major gluten protein genes. Cereal Chemistry 79: 279­-285.
Bar-­Sela, G., Tsalic, M., Fried, G. and Goldberg, H., 2007. Wheat grass juice may improve hematological toxicity related to chemotherapy in breast cancer patients: a pilot study. Nutrition and Cancer 58: 43­-48.
Bar­-Sela, G., Cohen, M., Ben­Arye, E. and Epelbaum, R., 2015. The medical use of wheatgrass: review of the gap between basic and clinical applications. Mini Reviews in Medicinal Chemistry 15: 1002-1010.
Baudo, M.M., Lyons, R., Powers, M.J.S., Pastori, G.M., Edwards, K.J., Holdsworth, M.J. and Shewry, P.R., 2006. Transgenesis has less impact on the transcriptome of wheat grain than conventional breeding. Plant Biotechnology Journal 4: 369-380.
Bauer, B.A., 2014. Healthy lifestyles: Q&A: what is wheatgrass, and should I add it to my smoothies for better health? Mayo Clinic, 22 January 2014. Available at:
Ben­-Arye, E., Goldin, E., Wengrower, D., Stamper, A., Kohn, R. and Berry, E., 2002. Wheatgrass juice in the treatment of active distal ulcerative colitis: a randomized double­blind placebo­controlled trial. Scandinavian Journal of Gastroenterology 37: 444­-449.
Benincasa, P., Galieni, A., Manetta, A.C., Pace, R., Guiducci, M., Pisante, M. and Stagnari, F., 2015. Phenolic compounds in grains, sprouts, and wheatgrass of hulled and non­hulled wheat species. Journal of the Science of Food and Agriculture 95: 1795-1803.
Bugyi, Z., Török, K., Hajas, L., Adonyi, Z., Popping, B. and Tömösközi, S., 2013. Comparative study of commercially available gluten ELISA kits using an incurred reference material. Quality Assurance and Safety of Crops & Foods 5: 79­-87.
Catassi, C., Bai, J.C., Bonaz, B., Bouma, G., Calabrò, A., Carroccio, A., Castillejo, G., Ciacci, C., Cristofori, F., Dolinsek, J., Francavilla, R., Elli, L., Green, P., Holtmeier, W., Koehler, P., Koletzko, S., Meinhold, C., Sanders, D., Schumann, M., Schuppan, D., Ullrich, R., Vécsei, A., Volta, U., Zevallos, V., Sapone, A. and Fasano, A., 2013. Non-­celiac gluten sensitivity: the new frontier of gluten related disorders. Nutrients 5: 3839­-3853.
Chu, P.T. and Wen, H.W., 2013. Sensitive detection and quantification of gliadin contamination in gluten­free food with immunomagnetic beads based liposomal fluorescence immunoassay. Analytica Chimica Acta 787: 246­-253.
Codex Alimentarius Commission, 1979 (revised 2008). Codex standard for foods for special dietary use for persons intolerant to gluten. Codex Stan 118, 3 pp.
Codex Alimentarius Commission, 2014. Codex Standard 234­1999. Recommended methods of analysis and sampling. Gluten-­free foods: enzyme-­linked immunoassay R5 Mendez (ELISA) method. FAO/WHO, Rome, Italy.
Commission of the European Communities, 2009. Commission Regulation (EC) no. 41/2009 of 20 January 2009 concerning the composition and labelling of foodstuffs suitable for people intolerant to gluten. Official Journal of the European Union 52: 3-5.
Cosnes, J., Cellier, C., Viola, S., Colombel, J.­-F., Michaud, L., Sarles, J., Hugot, J.-­P., Ginies, J.-­L., Dabadie, A., Mouterde, O., Allez, M. and Nion-­Larmurier, I., 2008. Incidence of autoimmune diseases in celiac disease: protective effect of the gluten-­free diet. Clinical Gastroenterology and Hepatology 6: 753­-758.
Department of Health and Human Services – Food and Drug Administration, 2013. Food labeling; gluten-­free labeling of foods; final rule. Federal Register 78: 47154-47179.
Dickey, W., 2007. A case of sequential development of celiac disease and ulcerative colitis. Nature Clinical Practice Gastroenterology and Hepatology 4: 463­-467.
El­Salhy, M., Hatlebakk, J.G., Gilja, O.H. and Hausken, T., 2015. The relation between celiac disease, nonceliac gluten sensitivity and irritable bowel syndrome. Nutrition Journal 14: 92.
Fasano, A., 2011. Zonulin and it's regulation of intestinal barrier function: the biological door to inflammation, autoimmunity, and cancer. Physiological Reviews 91: 151-175.
Food and Drug Administration (FDA), 2013. Gluten-­free labeling of foods; final rule. Federal Register, Vol. 78, No. 150. Rules and regulations. FDA, College Park, MD, USA, pp. 47154-47179.
Halbmayr-­Jech, E., Hammer, E., Fielder, R., Coutts, J., Rogers, A. and Cornish, M., 2012. Characterization of G12 sandwich ELISA, a next­-generation immunoassay for gluten toxicity. Journal of AOAC International 95: 372­-376.
Halbmayr-­Jech, E., Rogers, A., Don, C. and Prinster, M., 2015. Gluten in rice flour and baked rice products by G12 sandwich ELISA: First Action 2014.03. Journal of AOAC International 98: 103­-111.
Koehler, P., Hartmann, G., Wieser, H. and Rychlik, M., 2007. Changes of folates, dietary fiber, and proteins in wheat as affected by germination. Journal of Agricultural and Food Chemistry 55: 4678­-4683.
Kulkarni, S.D., Acharya, R., Nair, A.G.C., Rajurkar, N.S. and Reddy, A.V.R., 2006. Determination of elemental concentration profiles in tender wheatgrass (Triticum aestivum L.) using instrumental neutron activation analysis. Food Chemistry 95: 699­-707.
Kulkarni, S.D., Acharya, R., Rajurkar, N.S. and Reddy, A.V.R., 2007. Evaluation of bioaccessibility of some essential elements from wheatgrass (Triticum aestivum L.) by in vitro digestion method. Food Chemistry 103: 681­-688.
Lewis, N.R., Hubbard, R.B., Sanders, D.S., Logan, R.F., Holmes, G.K. and West, J., 2011. Treating occult coeliac disease with a gluten-­free diet is associated with a significant improvement in quality of life. Gut 60: A1­-A2.
Michalcova, E., Potocka, E., Chmelova, D. and Ondrejovic, M., 2012. Study of wheat protein degradation during germination. Journal of Microbiology, Biotechnology and Food Sciences 1: 1439­-1447.
Murray, J.A., 1999. The widening spectrum of celiac disease. American Journal of Clinical Nutrition 69: 354­-365.
Ng, S.C., Lam, Y.T., Tsoi, K.K.F., Chan, F.K.L., Sung, J.J.Y. and Wu, J.C.Y., 2013. Systematic review: the efficacy of herbal therapy in inflammatory bowel disease. Alimentary Pharmacology and Therapeutics 38: 854­-863.
Osman, A.A., Uhlig, H.H., Valdes, I., Amin, M., Mendez, E. and Mothes, T., 2001. A monoclonal antibody that recognizes a potential coeliac­toxic repetitive pentapeptide epitope in gliadins. European Journal of Gastroenterology and Hepatology 13: 1189­-1193.
Riddle, M.S., Murray, J.A. and Porter, C.K., 2012. The incidence and risk of celiac disease in a healthy US adult population. American Journal of Gastroenterology 107: 1248­-1255.
Sapone, A., Bai, J.C., Ciacci, C., Dolinsek, J., Green, P.H., Hadjivassiliou, M., Kaukinen, K., Rostami, K., Sanders, D.S., Schumann, M., Ullrich, R., Villalta, D., Volta, U., Catassi, C. and Fasano, A., 2012. Spectrum of gluten­related disorders: consensus on new nomenclature and classification. BMC Medicine 10: 13.
Silano, M., Penas Pozo, E., Uberti, F., Manferdelli, S., Del Pinto, T., Felli, C., Budelli, A., Vincentini, O. and Restani, P., 2014. Diversity of oat varieties in eliciting the early inflammatory events in celiac disease. European Journal of Nutrition 53: 1177­-1186.
Valdes, I., García, E., Llorente, M. and Mendez, E., 2003. Innovative approach to low­level gluten determination in foods using a novel sandwich ELISA protocol. European Journal of Gastroenterology 15: 465­-474.