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Mass Spectrometric Detection of Bacterial Protein Toxins and Their Enzymatic Activity
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Toxins 2015, 7(9), 3637-3656; doi:10.3390/toxins7093637

Quantitative Analysis of Staphylococcal Enterotoxins A and B in Food Matrices Using Ultra High-Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS)

Science Department, National Food Agency, Box 622, Uppsala SE-751 26, Sweden
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Author to whom correspondence should be addressed.
Academic Editor: Xiaohua He
Received: 2 July 2015 / Revised: 2 September 2015 / Accepted: 6 September 2015 / Published: 11 September 2015
(This article belongs to the Collection Rapid Detection of Bacterial Toxins)
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Abstract

A method that uses mass spectrometry (MS) for identification and quantification of protein toxins, staphylococcal enterotoxins A and B (SEA and SEB), in milk and shrimp is described. The analysis was performed using a tryptic peptide, from each of the toxins, as the target analyte together with the corresponding 13C-labeled synthetic internal standard peptide. The performance of the method was evaluated by analyzing spiked samples in the quantification range 2.5–30 ng/g (R2 = 0.92–0.99). The limit of quantification (LOQ) in milk and the limit of detection (LOD) in shrimp was 2.5 ng/g, for both SEA and SEB toxins. The in-house reproducibility (RSD) was 8%–30% and 5%–41% at different concentrations for milk and shrimp, respectively. The method was compared to the ELISA method, used at the EU-RL (France), for milk samples spiked with SEA at low levels, in the quantification range of 2.5 to 5 ng/g. The comparison showed good coherence for the two methods: 2.9 (MS)/1.8 (ELISA) and 3.6 (MS)/3.8 (ELISA) ng/g. The major advantage of the developed method is that it allows direct confirmation of the molecular identity and quantitative analysis of SEA and SEB at low nanogram levels using a label and antibody free approach. Therefore, this method is an important step in the development of alternatives to the immune-assay tests currently used for staphylococcal enterotoxin analysis. View Full-Text
Keywords: staphylococcal enterotoxins; quantification; foods; UPLC-ESI-MS/MS staphylococcal enterotoxins; quantification; foods; UPLC-ESI-MS/MS
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Muratovic, A.Z.; Hagström, T.; Rosén, J.; Granelli, K.; Hellenäs, K.-E. Quantitative Analysis of Staphylococcal Enterotoxins A and B in Food Matrices Using Ultra High-Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). Toxins 2015, 7, 3637-3656.

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