Influence of sample extraction solutions on the detection of wheat proteins by mass spectrometry.
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 34000 microg/g gluten were analyzed using the TRIS-HCI extraction buffer.
Eurofins Analytik GmbH, Grossmoorbogen 25, 21079 Hamburg, Germany. juliaheick@eurofins.de
Julia
J
Heick
Markus
M
Fischer
Bert
B
Popping
D005504:Food Analysis / Q000379:methods*; D059625:Liquid-Liquid Extraction / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry*; D015203:Reproducibility of Results; D014908:Triticum / Q000737:chemistry*
D010940:Plant Proteins