Determination of Aflatoxin M1 in Raw Milk Using an HPLC-FL Method in Comparison with Commercial ELISA Kits-Application in Raw Milk Samples from Various Regions of Greece.

Martha Maggira,Ioannis Sakaridis,Georgios Samouris,Maria Ioannidou

doi:10.3390/vetsci8030046

Abstract

The highly toxic Aflatoxin M1 (AFM1) is most often detected in milk using an Enzyme-Linked-Immunosorbent Assay (ELISA) for screening purposes, while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-FL) is the reference method used for confirmation. The aim of the present study was the comparison between three commercially available ELISA kits and a newly developed HPLC-FL method for the determination of the AFM1 in milk samples. The developed HPLC-FL method was validated for the AFM1 and Aflatoxin M2 (AFM2), determining the accuracy, precision, linearity, decision limit, and detection capability with fairly good results. All three ELISA kits were also validated and showed equally good performance with high recovery rates. Moreover, the Limit Of Detection (LOD) and Limit Of Quantification (LOQ) values were found to be significantly lower than the Maximum Residue Limit (MRL) (50 ng kg−1). After the evaluation of all three commercial kits, the ELISA kit with the optimum performance along with the HPLC method was used for the determination of AFM1 in raw cow’s, goat’s, and sheep’s milk samples (396) obtained from producers in different regions of Greece. The evaluation of both methods showed that this ELISA kit could be considered as a faster and equally reliable alternative method to HPLC in routine analysis for the determination of AFM1 in milk.

Highlights

Aflatoxins (AFs) are one of the most widely known and investigated groups of mycotoxins, which can be found as contaminants in food commodities worldwide [1]
Aflatoxin M1 (AFM1) for screening purposes is EnzymeLinked-Immunosorbent Assay (ELISA), while High-Performance Liquid Chromatography with Fluorescence Detector (HPLC-fluorescence detection (FL)) is still the reference method used for confirmation
The present study shows that the newly developed high-performance liquid chromatography (HPLC)-FL method of HPLC was found to be accurate, sensitive, and repeatable, and it is suitable for the determination of AFM1 and Aflatoxin M2 (AFM2) in raw milk samples

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Aflatoxins (AFs) are one of the most widely known and investigated groups of mycotoxins, which can be found as contaminants in food commodities worldwide [1]. They are toxic substances produced by the secondary metabolism of several fungal species, such as Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius, which grow in major food crops under a wide range of climatic conditions [2]. Aflatoxins are chemical substances that cannot be perceived by smell or taste, are fluorescent in ultraviolet light, and are resistant to high temperatures as they withstand exposure above

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Conclusion