Confirmation of peanut protein using peptide markers in dark chocolate using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
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.
Shefcheck
Kevin J
KJ
U.S. Food and Drug Administration, 5100 Paint Branch Parkway, College Park, MD 20740, USA. kshefche@cfsan.fda.edu
Callahan
John H
JH
Musser
Steven M
SM
D000485:Allergens / Q000032:analysis*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D002099:Cacao / Q000737:chemistry*; D002853:Chromatography, Liquid; D005506:Food Contamination / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D013058:Mass Spectrometry; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D010447:Peptide Hydrolases / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity
D000485:Allergens; D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D006023:Glycoproteins; D008565:Membrane Proteins; D010446:Peptide Fragments; D010940:Plant Proteins; D010447:Peptide Hydrolases