# Determination of mycotoxins and plant toxins in raw cow milk from 5 EU countries using a validated LC-MS/MS Multi-Method

Ahmed H. El-Khatib, Arnold Bahlmann, Michael Weiss, Carolin Hoyer, Marine Gele, L. Rey-Cadilhac, Rennie Eppenstein, Marcon Berton, Donagh Berry, Christoph Hutzler, et al.

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To cite this version:

Ahmed H. El-Khatib, Arnold Bahlmann, Michael Weiss, Carolin Hoyer, Marine Gele, et al. Determination of mycotoxins and plant toxins in raw cow milk from 5 EU countries using a validated LC-MS/MS Multi-Method. 46. Mycotoxin Workshop, May 2025, Martina Franca, Italy. hal-05009212

HAL Id: hal-05009212

https://hal.inrae.fr/hal-05009212v1

Submitted on 27 Mar 2025

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# Determination of mycotoxins and plant toxins in raw cow milk from 5 EU countries using a validated LC-MS/MS Multi-Method

Ahmed H. El-Khatib¹*, Arnold Bahlmann¹, Michael Weiss¹, Carolin Hoyer¹, Marine Gele², Lucille Rey-Cadilhac³, Rennie Eppenstein⁴, Marco Berton⁵, Donagh Berry⁶, Christoph Hutzler¹, Stefan Weigel¹

¹German Federal Institute for Risk Assessment (BfR), Berlin, Germany, ²ACTA IDELE (Paris, France), ³INRAE (Paris, France), ⁴FIBL (Frick, Switzerland), ⁵UNIPD (Padova, Italy), ⁶TEAGASC (Carlow, Ireland)

*Corresponding author: ahmed.el-khatib@bfr.bund.de

Milk is one of the most highly consumed food products in all age groups and forms the basis of dairy products. The quality and safety of milk are significantly influenced by the animal's diet, which is composed of a complex mix of individual feed components (e.g. forages and concentrates). Each raw feed component can carry different contaminants, leading to multi-toxin exposure in the overall animal diet. This multitude of toxins could finally find its way into the milk. The presence of contaminants such as mycotoxins and plant toxins can pose health risks to consumers.

Currently, only aflatoxin M1 is regulated by the European Union in raw milk, heat-treated milk and milk for the manufacture of milk-based products. However, there have been reports on the presence of beauvericin and enniatin B [1, 2], T-2 and HT-2 toxins [2], pyrrolizidine and tropane alkaloids [3] and quinolizidine alkaloids [4] in raw milk samples.

Within the framework of the EU project INTAQT, an LC-MS/MS method for the simultaneous determination of a wide range of mycotoxins and plant toxins (110 toxins) in raw milk has been developed and validated according to the latest EU regulations for mycotoxins (EU 2023/2782) and plant toxins (EU 2023/2783) in terms of recovery, precision and limit of quantification (LOQ). The method involved QuEChERS-based extraction and LC-MS/MS analysis using triple quadrupole mass spectrometry.

The validated method was applied to the INTAQT project samples. Milk samples collected in summer and winter seasons from farms in 5 European countries (France, Germany, Ireland, Italy and Switzerland) were examined. More than 150 pooled raw cow milk samples were analyzed using the validated LC-MS/MS multi-method. The results revealed generally low levels of mycotoxins and plant toxins across the different regions.

The aim of this work was to investigate the levels of several mycotoxins and plant toxins and their co-occurrence in milk. The validated LC-MS/MS multi-method proved to be a reliable and efficient tool for the simultaneous determination of multiple mycotoxins and plant toxins in milk.

[1] Pietruszka K et al. (2023) Survey of the enniatins and beauvericin in raw and UHT cow's milk in Poland. J Vet Res 67:259-266.

[2] Leite M (2023) Regulated and Emerging Mycotoxins in Bulk Raw Milk: What Is the Human Risk?. Toxins 15:605.

[3] Klein LM et al. (2024) Pyrrolizidine alkaloids and tropane alkaloids in milk samples from individual dairy farms of the German federal states of Bavaria and Schleswig-Holstein. Food Additives & Contaminants: Part A 41:629-647.

[4] Engel AM et al. (2022) Investigations on the Transfer of Quinolizidine Alkaloids from Lupinus angustifolius into the Milk of Dairy Cows. J. Agric. Food Chem. 70:11749–11758.