Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 400, Issue 5, Pages 1525-1531Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-011-4906-6
Keywords
MALDI-TOF; Milk; Staphylococcal enterotoxin A; Staphylococcus aureus; Tryptic digestion
Funding
- Conselleria de Sanitat-Generalitat Valenciana [072/2009]
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Staphylococcal enterotoxin A (SEA) is an exotoxin excreted mainly by Staphylococcus aureus and nowadays is the most prevalent compound in staphylococcal food poisoning worldwide. SEA is highly heat-resistant, and usual cooking times and temperatures are unlikely to completely inactivate it. A procedure for extraction of this toxin based on protein precipitation with a mixture of dichloromethane and acidified water was used before SDS-PAGE separation of soluble proteins. Finally, bands of interest were excised from the gel and in-gel enzymatic digestion was done. SEA from pasteurized milk was detected with matrix-assisted laser-desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Nineteen peptides (range 800-2400 Da) were identified as products of trypsin cleavage of the SEA standard with a score of 204 and 73% coverage of the protein sequence, whereas thirteen peptides were revealed for SEA extracted from milk with a score of 148 and 58% sequence coverage obtained. This procedure has been applied successfully for identification of SEA in milk.
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