Influence of sample extraction solutions on the detection of wheat proteins by mass spectrometry
Abstract:Routinely used methods for the detection of gluten mostly use denaturing agents and high salt concentrations to increase the extractability of the gluten fraction. These work well in combination with ELISA methods, but may have a negative effect on MS methods due to their influence on the ionization of the analyte leading to a significant reduction of signal intensities. A newly developed HPLC/MS/MS method was used to assess this influence. Four different extraction buffers were compared: 70% ethanol, TRIS-HCl, TRIS-HCl with dithiothreitol, and a commercially available cocktail solution. Unprocessed and processed wheat samples were analyzed. When analyzing unprocessed samples, a negative effect on ionization could be observed. Considering extraction capabilities and signal intensities, TRIS-HCl seemed to be the most suitable buffer in combination with the MS method. To assess whether the method was capable of detecting hidden wheat protein in different kinds of food, different food samples containing 0 to 34 000 μg/g gluten were analyzed using the TRIS-HCl extraction buffer.
Document Type: Research Article
Publication date: March 1, 2012
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- The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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