Abstract
Presence of aflatoxin M1 (AFM1) in milk should be continuously controlled in order to protect the population from risks associated with its proven toxicity and carcinogenicity. During recent years, there has been an increase in demand for development of sensitive, accurate, simple and fast method which is reliable for detection of AFM1 at low concentrations found in milk samples. For that purpose, enzyme linked immunosorbent asssay (ELISA), high performance liquid chromatography with fluorescence detector (HPLC-FLD) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) were optimized and validated in order to apply them for AFM1 analysis in naturally contaminated milk samples, and to assess the closeness of agreement between results of three different methods. The obtained validation parameters indicate that all three methods are suitable for determination of AFM1 in milk samples. The statistical analysis of variance between the methods and the obtained correlation coefficients indicate that there is a strong correlation between methods. All three methods are satisfactory in meeting the requirements for official control purposes. To the best of author's knowledge, this study represents the first report of an investigation and comparison of ELISA, HPLC-FLD and HPLC-MS/MS methods for determination of AFM1 in naturally contaminated milk samples.
Highlights
Aflatoxins (AFs) are one of the main groups of mycotoxins due to their toxicity and prevalence in food and feed
limit of quantification (LOQ) for all three procedures were less than the proposed LOQ (0.02 μg/kg) for aflatoxin M1 (AFM1) according Technical Report CEN/TR 16059:2010.31 According to 2002/657/EC24 trueness of the methods were expressed after analysis of certified reference material (CRM)
It is acceptable that trueness of measurements is assessed through recovery of additions of known amounts of the analyte to the unknown samples it should be pointed that the added analyte is not chemically bound in the real matrix and that results obtained by this approach have lesser validity than those achieved through the use of CRM
Summary
Aflatoxins (AFs) are one of the main groups of mycotoxins due to their toxicity and prevalence in food and feed. Among eighteen identified AFs,[1] aflatoxin M1 (AFM1) has demonstrated the greatest potential for presence in the human diet since it is commonly found as contaminant of milk, which is consumed daily. There has been an increase in demand for finding of simple, accurate and fast method which is reliable for detection of AFM1 at concentrations found in milk samples
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