title: Multiplex quantification of protein toxins in human biofluids and food matrices using immunoextraction and high-resolution targeted mass spectrometry.
abstract: The development of rapid methods for unambiguous identification and precise quantification of protein toxins in various matrices is essential for public health surveillance. Nowadays, analytical strategies classically rely on sensitive immunological assays, but mass spectrometry constitutes an attractive complementary approach thanks to direct measurement and protein characterization ability. We developed here an innovative multiplex immuno-LC-MS/MS method for the simultaneous and specific quantification of the three potential biological warfare agents, ricin, staphylococcal enterotoxin B, and epsilon toxin, in complex human biofluids and food matrices. At least 7 peptides were targeted for each toxin (43 peptides in total) with a quadrupole-Orbitrap high-resolution instrument for exquisite detection specificity. Quantification was performed using stable isotope-labeled toxin standards spiked early in the sample. Lower limits of quantification were determined at or close to 1 ng·mL(-1). The whole process was successfully applied to the quantitative analysis of toxins in complex samples such as milk, human urine, and plasma. Finally, we report new data on toxin stability with no evidence of toxin degradation in milk in a 48 h time frame, allowing relevant quantitative toxin analysis for samples collected in this time range.
authors.affiliation: †CEA, DSV, iBiTec-S, Laboratoire d'études du métabolisme des médicaments, 91191 Gif-sur-Yvette, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Dupré
authors.affiliation: §Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France.|∥CEA, iRTSV-BGE, F-38000 Grenoble, France.|⊥INSERM, BGE, F-38000 Grenoble, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Gilquin
authors.affiliation: †CEA, DSV, iBiTec-S, Laboratoire d'études du métabolisme des médicaments, 91191 Gif-sur-Yvette, France.
authors.forename: François
authors.initials: F
authors.lastname: Fenaille
authors.affiliation: ‡CEA, DSV, iBiTec-S, Laboratoire d'études et de recherches en immunoanalyse, 91191 Gif-sur-Yvette, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Feraudet-Tarisse
authors.affiliation: ‡CEA, DSV, iBiTec-S, Laboratoire d'études et de recherches en immunoanalyse, 91191 Gif-sur-Yvette, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Dano
authors.affiliation: §Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France.|∥CEA, iRTSV-BGE, F-38000 Grenoble, France.|⊥INSERM, BGE, F-38000 Grenoble, France.
authors.forename: Myriam
authors.initials: M
authors.lastname: Ferro
authors.affiliation: ‡CEA, DSV, iBiTec-S, Laboratoire d'études et de recherches en immunoanalyse, 91191 Gif-sur-Yvette, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Simon
authors.affiliation: †CEA, DSV, iBiTec-S, Laboratoire d'études du métabolisme des médicaments, 91191 Gif-sur-Yvette, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Junot
authors.affiliation: §Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France.|∥CEA, iRTSV-BGE, F-38000 Grenoble, France.|⊥INSERM, BGE, F-38000 Grenoble, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Brun
authors.affiliation: †CEA, DSV, iBiTec-S, Laboratoire d'études du métabolisme des médicaments, 91191 Gif-sur-Yvette, France.
authors.forename: François
authors.initials: F
authors.lastname: Becher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004768:Enterotoxins / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D007553:Isotope Labeling; D008892:Milk / Q000737:chemistry; D009587:Nitrogen Isotopes / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000592:standards; D012276:Ricin / Q000032:analysis* / Q000097:blood / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000592:standards
chemical_list: D002247:Carbon Isotopes; D004768:Enterotoxins; D009587:Nitrogen Isotopes; D010455:Peptides; C031818:enterotoxin B, staphylococcal; D012276:Ricin