Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry
In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples wer...
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MDPI AG
2021-08-01
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| Online Access: | https://www.mdpi.com/2304-8158/10/9/2025 |
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| author | Gabriele Rocchetti Francesca Ghilardelli Francesco Masoero Antonio Gallo |
| author_facet | Gabriele Rocchetti Francesca Ghilardelli Francesco Masoero Antonio Gallo |
| author_sort | Gabriele Rocchetti |
| collection | DOAJ |
| description | In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of <i>Aspergillus</i>- and <i>Penicillium</i>-mycotoxins; (2) low levels of fumonisins and other <i>Fusarium</i>-mycotoxins; (3) high levels of <i>Aspergillus</i>-mycotoxins; (4) high levels of non-regulated <i>Fusarium</i>-mycotoxins; (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some <i>Fusarium</i> mycotoxins, together with the tetrapeptide tentoxin (an <i>Alternaria</i> toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese. |
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| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T07:40:23Z |
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| spelling | doaj.art-2f1dc8b82d1f43549007f5d8cc7178182023-11-22T13:03:05ZengMDPI AGFoods2304-81582021-08-01109202510.3390/foods10092025Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass SpectrometryGabriele Rocchetti0Francesca Ghilardelli1Francesco Masoero2Antonio Gallo3Department of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyDepartment of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, ItalyIn this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of <i>Aspergillus</i>- and <i>Penicillium</i>-mycotoxins; (2) low levels of fumonisins and other <i>Fusarium</i>-mycotoxins; (3) high levels of <i>Aspergillus</i>-mycotoxins; (4) high levels of non-regulated <i>Fusarium</i>-mycotoxins; (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some <i>Fusarium</i> mycotoxins, together with the tetrapeptide tentoxin (an <i>Alternaria</i> toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese.https://www.mdpi.com/2304-8158/10/9/2025milk metabolomicsretrospective screeningUHPLC-Orbitrapmultivariate statisticsmycotoxins |
| spellingShingle | Gabriele Rocchetti Francesca Ghilardelli Francesco Masoero Antonio Gallo Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry Foods milk metabolomics retrospective screening UHPLC-Orbitrap multivariate statistics mycotoxins |
| title | Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry |
| title_full | Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry |
| title_fullStr | Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry |
| title_full_unstemmed | Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry |
| title_short | Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry |
| title_sort | screening of regulated and emerging mycotoxins in bulk milk samples by high resolution mass spectrometry |
| topic | milk metabolomics retrospective screening UHPLC-Orbitrap multivariate statistics mycotoxins |
| url | https://www.mdpi.com/2304-8158/10/9/2025 |
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