Comparison of ELISA, HPLC-FLD and HPLC-MS/MS Methods for Determination of Aflatoxin M1 in Natural Contaminated Milk Samples

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.

High-performance liquid chromatography ultraviolet-photodiode array detection method for aflatoxin B1 in cattle feed supplements

“…Analysis of concentrations <0.2 ppm is recommended with other HPLC instruments’ different detectors, i.e., HPLC with fluorescence detector. Although the theoretical detection limit was 3.5 × 10 −6 ppm, it must be analyzed by liquid chromatography–electrospray ionization mass spectrometry to confirm [ 2 , 28 ]. As described on Protocol Cartagena that all about biologic products included mycotoxin must be clearly detected by instrument analysis with correlation between physic and chemistry of the compounds [ 29 ].…”

Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food

“…ELISA methods have been validated in a wide variety of food matrices [7,75]. While ELISA can be performed in several ways such as direct assay, competitive direct assay, and competitive indirect assay, a competitive direct assay is most commonly used [103,104]. The principle of ELISA is based on the competitive interactions between mycotoxins (acting as an antigen) and assigned antibodies labelled with toxin-enzyme conjugate for many binding sites [76,105].…”

Adverse Effects, Transformation and Channeling of Aflatoxins Into Food Raw Materials in Livestock

“…Furthermore, countermeasures may also include the application of various mycotoxin binding agents mixed with the feed (De Mil et al, 2015;Vila-Donat et al, 2018). Besides agricultural and technological approaches combating aflatoxins successfully, we also need to develop more sensitive and more reliable analytical methods (Kos et al, 2016).…”