Acta Chimica Slovaca (ACS) publishes papers on fundamental and applied aspects of chemistry, biochemistry, chemical technology, chemical engineering and process control, biotechnology and food technology. Welcome are also topics which include chemical aspects of materials, physical chemistry and chemical physics, analytical chemistry, macromolecular chemistry and biomedical engineering.

Development and validation of HPLC-FLD method for determination of aflatoxin M1 content in Slovakian milk and dairy products

Lukáš Kolarič *, Peter Šimko

Slovak University of Technology in Bratislava, Faculty of Chemical and Food Technology, Institute of Food Science and Nutrition, Radlinského 9, 812 37 Bratislava, Slovak Republic

E-mail: * lukas.kolaric@stuba.sk

Abstract: Milk and dairy products are the most consumed foods in human diet, thus the safety of these products is in the attention centre of the control authorities. Aflatoxin M1 (AFM1) is a dangerous toxin that can occur in milk and dairy products as a metabolite formed from aflatoxin B1 contained in contaminated animal feed. Therefore, the aim of this study was to develop and validate a reliable method for the determination of the AFM1 content in milk and dairy products based on HPLC with fluorescence detection with previous extraction procedure using immunoaffinity columns (IAC). The optimal chromatographic separation of AFM1 was obtained using water/acetonitrile mixture (80/20, v/v) as a mobile phase, column with a stationary phase of C18 with temperature maintained at 25 °C, and fluorescence detection at excitation wavelengths at 360 nm and emission at 440 nm. As found, the efficacy of AFM1 extraction from the samples was influenced by the elution agent composition. The best results were noticed using 1.25 mL of acetonitrile/methanol (3/2, v/v) + 1.25 mL of water. The validation parameters of the proposed method passed the criteria set by European legislation with a limit of detection and quantification at 0.002 and 0.007 µg/kg, respectively. Also, the suitability of the method was confirmed by its application for determination of AFM1 content in certified reference material. Finally, the method was applied for the determination of the AFM1 content in 25 milk and dairy products collected in Slovakia and it was found that its content was below the limit of quantification. It was concluded that the method is suitable for monitoring of AFM1 content in milk and dairy products.

Keywords: aflatoxin M1, HPLC, immunoaffinity chromatography, milk, dairy products, validation, optimisation, food safety

Acta Chimica Slovaca, Vol. 16, No. 1, 2023, pp. 99—108, DOI: 10.2478/acs-2023-0010