Detection, confirmation, and quantification of staphylococcal enterotoxin B in food matrixes using liquid chromatography--mass spectrometry.
Although immunoassay-based methods are sensitive and widely used for measuring protein toxins in food matrixes, there is a need for methods that can directly confirm the molecular identity of the toxin in situations where immunoassay tests yield a positive result. A method has been developed that uses mass spectrometry to identify a protein toxin, staphylococcal enterotoxin B (SEB), in a model food matrix, apple juice. The approach employs ultrafiltration to remove low molecular weight components from the sample, after which the remaining high molecular weight fraction, containing the protein, is digested with trypsin. The tryptic fragments are separated from residual biopolymers and analyzed by liquid chromatography-electrospray mass spectrometry. The background is still sufficiently complex that tandem mass spectrometry (MS/MS) is used to confirm the identity of target peptides. Limits of detection are 80 ng of SEB for MS and 100 ng for full scan MS/MS, using a tryptic fragment as the analytical target. Lower detection limits can be obtained using selected ion monitoring and multiple reaction monitoring. The presence of SEB can be confirmed at concentrations as low as 5 parts-per-billion by increasing the size of the sample to 10 mL. The method is applicable to the detection of SEB in other water-soluble food matrixes.
Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, 5100 Paint Branch Parkway, College Park, Maryland 20740, USA. john.callahan@cfsan.fda.gov
John H
JH
Callahan
Kevin J
KJ
Shefcheck
Tracie L
TL
Williams
Steven M
SM
Musser
D002853:Chromatography, Liquid; D004768:Enterotoxins / Q000032:analysis*; D005504:Food Analysis; D005638:Fruit / Q000737:chemistry*; D013058:Mass Spectrometry; D012680:Sensitivity and Specificity; D013997:Time Factors; D014675:Vegetables / Q000737:chemistry*
D004768:Enterotoxins; C031818:enterotoxin B, staphylococcal