{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T10:18:58Z","timestamp":1788430738906,"version":"build-2803163510"},"reference-count":42,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T00:00:00Z","timestamp":1788393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Among pathogenic microbes that can produce toxins, Bacillus cereus plays an important role in foodborne disorders. It is responsible for the production of cereulide, a cyclic dodecadepsipeptide toxin, very stable to heat and acidic pH, that can cause emetic syndromes. The study describes the secondary validation of the ISO 18465 method for food matrices in the concentration range 1\u2013100 \u03bcg\/kg and the in-house validation of a more sensitive liquid chromatography coupled with tandem mass spectrometry (LC-MS\/MS) method for infant formula at the lower concentration levels required by the EFSA since 2026. To improve the applicability of cereulide determination in infant formula, the solid matrix was reconstituted before extraction and the analysis was performed on the SCIEX QTRAP 7500 system. The method showed good linearity (0.010\u20131.000 \u03bcg\/L, R2 = 1000), a limit of quantification equal to the half-maximum level (LM = 0.054 \u03bcg\/L, LOQ = 0.025 \u03bcg\/L), good recovery (100\u2013109%) and acceptable within-laboratory reproducibility (RSD% &lt; 15%). This method provides a reliable analytical tool for cereulide monitoring in a broad variety of food commodities not considered before by ISO 18465 and makes it possible to reach the new lower limits established for infant formula. Furthermore, data on 332 samples involved in suspected food poisoning in Italy between 2019 and 2024 are presented in this retrospective study.<\/jats:p>","DOI":"10.3390\/foods15173129","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T09:20:43Z","timestamp":1788427243000},"page":"3129","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["LC-MS\/MS Determination of Cereulide in Food and Infant Formula: Method Validation and Six-Year Survey in Italy"],"prefix":"10.3390","volume":"15","author":[{"given":"Elena","family":"Torres","sequence":"first","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Tabata","family":"Bezzo Llufrio","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Stefania","family":"Massafra","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Gian Luca","family":"Ferro","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"conceptualization","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"Alessio","family":"Gasperin","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Alessia","family":"Ossino","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Daniela Manila","family":"Bianchi","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7556-6282","authenticated-orcid":false,"given":"Maria Cesarina","family":"Abete","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Marilena","family":"Gili","sequence":"additional","affiliation":[{"name":"Istituto Zooprofilattico Sperimentale Piemonte, Liguria e Valle d\u2019Aosta, Via Bologna 148, 10154 Turin, Italy"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,3]]},"reference":[{"key":"ref_1","unstructured":"Grumezescu, A.M., and Holban, A.M. (2018). Microbial Contamination, Prevention, and Early Detection in Food Industry. Handbook of Food Bioengineering: Microbial Contamination and Food Degradation, Academic Press."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority, and European Centre for Disease Prevention and Control (2018). The European Union summary report on trends and sources of zoonoses, zoonotic agents and foodborne outbreaks in 2017. EFSA J., 16, 5500.","DOI":"10.2903\/j.efsa.2018.5500"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority, and European Centre for Disease Prevention and Control (2023). The European Union One Health 2022 Zoonoses Report. EFSA J., 21, e8442.","DOI":"10.2903\/j.efsa.2023.8442"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority (EFSA), and European Centre for Disease Prevention and Control (ECDC) (2025). The European Union One Health 2024 Zoonoses Report. EFSA J., 23, e9759.","DOI":"10.2903\/j.efsa.2025.9759"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1111\/j.1574-6976.2008.00112.x","article-title":"From soil to gut: Bacillus cereus and its food poisoning toxins","volume":"32","author":"Fagerlund","year":"2008","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1128\/CMR.00073-09","article-title":"Bacillus cereus, a volatile human pathogen","volume":"23","author":"Bottone","year":"2010","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1111\/j.1574-6968.1997.tb12776.x","article-title":"Bacillus cereus and its food poisoning toxins","volume":"157","author":"Granum","year":"1997","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_8","first-page":"61S","article-title":"Bacillus cereus and its toxins","volume":"76","author":"Granum","year":"1994","journal-title":"J. Appl. Bacteriol. Symp. Suppl."},{"key":"ref_9","first-page":"17","article-title":"A novel dodecadepsipeptide, cereulide, is an emetic toxin of Bacillus cereus","volume":"129","author":"Agata","year":"1995","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1099\/mic.0.27607-0","article-title":"Emetic toxin formation of Bacillus cereus is restricted to a single evolutionary lineage of closely related strains","volume":"151","author":"Svensson","year":"2005","journal-title":"Microbiology"},{"key":"ref_11","unstructured":"National Center for Biotechnology Information (NCBI) (2026, August 28). PubChem Compound Summary for Cereulide, CID 6441099; PubChem: Bethesda, MD, USA, 2026, Available online: https:\/\/pubchem.ncbi.nlm.nih.gov\/compound\/Cereulide."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1111\/j.1472-765X.2008.02350.x","article-title":"Heat resistance of Bacillus cereus emetic toxin, cereulide","volume":"46","author":"Rajkovic","year":"2008","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1292\/jvms.58.10_1027","article-title":"Mouse lethal activity of a HEp-2 vacuolation factor, cereulide, produced by Bacillus cereus isolated from vomiting-type food poisoning","volume":"58","author":"Shinagawa","year":"1996","journal-title":"J. Vet. Med. Sci."},{"key":"ref_14","first-page":"87","article-title":"Emesis of rhesus monkeys induced by intragastric administration with the HEp-2 vacuolation factor (cereulide) produced by Bacillus cereus","volume":"130","author":"Shinagawa","year":"1995","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1089\/fpd.2014.1788","article-title":"An emetic Bacillus cereus outbreak in a kindergarten: Detection and quantification of critical levels of cereulide toxin","volume":"12","author":"Delbrassinne","year":"2015","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"4277","DOI":"10.1128\/JCM.43.8.4277-4279.2005","article-title":"Fatal family outbreak of Bacillus cereus-associated food poisoning","volume":"43","author":"Dierick","year":"2005","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1142","DOI":"10.1056\/NEJM199704173361604","article-title":"Fulminant liver failure in association with the emetic toxin of Bacillus cereus","volume":"336","author":"Mahler","year":"1997","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Dietrich, R., Jessberger, N., Ehling-Schulz, M., M\u00e4rtlbauer, E., and Granum, P.E. (2021). The Food Poisoning Toxins of Bacillus cereus. Toxins, 13.","DOI":"10.3390\/toxins13020098"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Wang, Y., Liu, Y., Yang, S., Chen, Y., Liu, Y., Lu, D., Niu, H., Ren, F., Xu, A., and Dong, Q. (2024). Effect of Temperature, pH, and aw on Cereulide Synthesis and Regulator Genes Transcription with Respect to Bacillus cereus Growth and Cereulide Production. Toxins, 16.","DOI":"10.3390\/toxins16010032"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1046\/j.1472-765x.2000.00835.x","article-title":"Bacillus cereus produces most emetic toxin at lower temperatures","volume":"31","author":"Finlay","year":"2000","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0168-1605(01)00692-4","article-title":"Production of Bacillus cereus emetic toxin (cereulide) in various foods","volume":"73","author":"Agata","year":"2002","journal-title":"Int. J. Food Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"847","DOI":"10.4315\/0362-028X-69.4.847","article-title":"Influence of type of food on the kinetics and overall production of Bacillus cereus emetic toxin","volume":"69","author":"Rajkovic","year":"2006","journal-title":"J. Food Prot."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"EFSA, Eskes, C., Corti\u00f1as-Abrahantes, J., Bottex, B., Dorne, J.-L.C.M., Dujardin, B., Ferreira de Sousa, R., Horvath, Z., Kouloura, E., and Ramos Bordajandi, L. (2026). Rapid risk assessment on acute reference dose (ARfD) of cereulide in infants and information on acute consumption of infant formulae. EFSA J., 24, e9941.","DOI":"10.2903\/j.efsa.2026.9941"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1047","DOI":"10.4315\/0362-028X-66.6.1047","article-title":"Potential of Bacillus cereus for producing an emetic toxin, cereulide, in bakery products: Quantitative analysis by chemical and biological methods","volume":"66","author":"Andersson","year":"2003","journal-title":"J. Food Prot."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1056\/NEJMra035092","article-title":"Developmental Pharmacology\u2014Drug Disposition, Action, and Therapy in Infants and Children","volume":"349","author":"Kearns","year":"2003","journal-title":"N. Engl. J. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1338","DOI":"10.1128\/AEM.64.4.1338-1343.1998","article-title":"A novel sensitive bioassay for detection of Bacillus cereus emetic toxin and related depsipeptide ionophores","volume":"64","author":"Andersson","year":"1998","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_27","first-page":"31","article-title":"A novel dodecadepsipeptide, cereulide, isolated from Bacillus cereus causes vacuole formation in Hep-2 cells","volume":"121","author":"Agata","year":"1994","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2479","DOI":"10.1128\/AEM.68.5.2479-2483.2002","article-title":"Quantitative analysis of cereulide, the emetic toxin of Bacillus cereus, produced under various conditions","volume":"68","author":"Apetroaie","year":"2002","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2531","DOI":"10.1081\/JLC-200028373","article-title":"Quantification of Bacillus cereus emetic toxin (cereulide) in figs using LC\/MS","volume":"27","author":"Granum","year":"2004","journal-title":"J. Liq. Chromatogr. Relat. Technol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7466","DOI":"10.1128\/AEM.01659-10","article-title":"Quantification of the emetic toxin cereulide in food products by liquid chromatography-mass spectrometry using synthetic cereulide as a standard","volume":"76","author":"Reij","year":"2010","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3326","DOI":"10.3390\/toxins6123326","article-title":"Quantitative analysis of cereulide toxin from Bacillus cereus in rice and pasta using synthetic cereulide standard and 13C6-cereulide standard\u2014A short validation study","volume":"6","author":"Granelli","year":"2014","journal-title":"Toxins"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1021\/jf9033046","article-title":"Development of a stable isotope dilution analysis for the quantification of the Bacillus cereus toxin cereulide in foods","volume":"58","author":"Bauer","year":"2010","journal-title":"J. Agric. Food Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1007\/s00216-012-6485-6","article-title":"Mass spectrometric profiling of Bacillus cereus strains and quantitation of the emetic toxin cereulide by means of stable isotope dilution analysis and HEp-2 bioassay","volume":"405","author":"Stark","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_34","unstructured":"(2017). Microbiology of the Food Chain\u2014Quantitative Determination of Emetic Toxin (Cereulide) Using LC-MS\/MS (Standard No. ISO 18465:2017)."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Zhou, Z., Wang, Z., Mu, C., Qi, Y., Zhang, J., Cui, X., Fan, S., Xiao, J., and Zhao, R. (2026). Optimized SPE-UPLC-MS\/MS method for sensitive determination of cereulide in complex infant formula matrices. Toxins, 18.","DOI":"10.3390\/toxins18050222"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3019","DOI":"10.1021\/ac020361s","article-title":"Strategies for the Assessment of Matrix Effect in Quantitative Bioanalytical Methods Based on HPLC\u2013MS\/MS","volume":"75","author":"Matuszewski","year":"2003","journal-title":"Anal. Chem."},{"key":"ref_37","unstructured":"Youden, W.J., and Steiner, E.H. (1975). Statistical Manual of the Association of Official Analytical Chemists, Association of Official Analytical Chemists."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Schuck, P., Jeantet, R., and Dolivet, A. (2012). Determination of Rehydration Ability. Analytical Methods for Food and Dairy Powders, John Wiley & Sons, Ltd.","DOI":"10.1002\/9781118307397"},{"key":"#cr-split#-ref_39.1","unstructured":"European Commission (2021). Commission Implementing Regulation"},{"key":"#cr-split#-ref_39.2","unstructured":"(EU) 2021\/808 of 22 March 2021 on the Performance of Analytical Methods for Residues of Pharmacologically Active Substances Used in Food-Producing Animals and on the Interpretation of Results as Well as on the Methods to Be Used for Sampling and Repealing Decisions 2002\/657\/EC and 98\/179\/EC. Off. J. Eur. Union, L180, 84-109."},{"key":"ref_40","unstructured":"(2004). Accuracy (Trueness and Precision) of Measurement Methods and Results\u2014Part 6: Use in Practice of Accuracy Values (Standard No. ISO 5725-6:2004)."},{"key":"ref_41","unstructured":"European Commission (2023). Commission Implementing Regulation (EU) 2023\/2782 of 14 December 2023 laying down the methods of sampling and analysis for the control of the levels of mycotoxins in food and repealing Regulation (EC) No 401\/2006. Off. J. Eur. Union, L 2023\/2782, 1\u201344."}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/15\/17\/3129\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T09:52:11Z","timestamp":1788429131000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/15\/17\/3129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,3]]},"references-count":42,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["foods15173129"],"URL":"https:\/\/doi.org\/10.3390\/foods15173129","relation":{},"ISSN":["2304-8158"],"issn-type":[{"value":"2304-8158","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,3]]}}}