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Confirmation of Peanut Protein Using Peptide Markers in Dark Chocolate Using Liquid Chromatography−Tandem Mass Spectrometry (LC-MS/MS)

Shefcheck K.J., Callahan J.H., Musser S.M.

Journal of Agricultural and Food Chemistry (2006), 54, 21, 7953--7959 DOI: 10.1021/jf060714e


Detection of peptides from the peanut allergen Ara h 1 by liquid chromatography-mass spectrometry (LC-MS) was used to identify and estimate total peanut protein levels in dark chocolate. A comparison of enzymatic digestion subsequent to and following extraction of Ara h 1 from the food matrix revealed better limits of detection (LOD) for the pre-extraction digestion (20 ppm) than for the postextraction digestion (50 ppm). Evaluation of LC-MS instruments and scan modes showed the LOD could be further reduced to 10 ppm via a triple-quadrupole and multiple-reaction monitoring. Improvements in extraction techniques combined with an increase in the amount of chocolate extracted (1 g) improved the LOD to 2 ppm of peanut protein. This method provides an unambiguous means of confirming the presence of the peanut protein in foods using peptide markers from a major allergen, Ara h 1, and can easily be modified to detect other food allergens. Keywords: Peanut protein; Ara h 1; dark chocolate; allergen