mycotoxins definitions

and analysis methods


Mycotoxins are secondary metabolites of low molecular weight and therefore test methods for analysis of mycotoxins face different problems ranging from the need to have a well-equipped laboratory, in terms of instrumentation and analytical capacity, to personnel with sufficient experience in handling of samples and reference standards.

Mycotoxins in feed are commonly detected and quantified using antibody-based assays and chromatography technique.

Immunochemical Methods in Mycotoxin detection

Mycotoxin are not antigenic. That is why one of the first developed studies were focused on how to achieve conjugation with proteins or polypeptides that could serve as transporters of antibody production in rabbits and other animals under the optimal conditions.

Different types of immunoassays were developed, among which the most important ones are:
  • ELISA test (enzyme-linked immunosorbent assay)
  • RIA (radioimmunoassay)
Most of these methods are very sensitive, specific and easy to use, which has given rise to a new dimension in terms of methodology for mycotoxins analysis.
ELISA test

The enzyme-linked immunosorbent assay (ELISA) is an antibody-based assay that is commonly used for mycotoxins detection.

Affordable methods

There are several commercial ELISA kits available for Aflatoxins, Deoxynivalenol, Fumonisins, Ochratoxins and Zearalenone.

ELISA is one of the most affordable methods for mycotoxins detection, but the limit of detection for many mycotoxins often exceeds 0.2 ppm.


Instrumental methods
Chromatography and Spectrometry

The use of chromatographic techniques requires prior preparation of a sample using different methods of extraction and purification. Depending on the mycotoxins to be analysed and the feed matrix, different types of techniques such as immunoaffinity columns based preparations or other procedures based on solid-liquid (Quechers, DMFS, etc.) or liquid-liquid (DLLME) extractions are used.

High performance liquid chromatography (HPLC) and gas chromatography/mass spectrometry (GC/MS) are two of the most commonly used methods for the detection and quantification of mycotoxins.

High-performance liquid chromatography
High-performance liquid chromatography (HPLC) represents the technique of choice for the analysis of mycotoxins or for confirmation of positive results by ELISA, as it is sensitive, reproducible, accurate and has a higher degree of automation.
Gas chromatography
Gas chromatography (GC) is used less frequently since most of the mycotoxins are not sufficiently volatile and, therefore, must be derivatized, which increases the time and cost of analysis.
Abssorption and emission spectrometric techniques
The most widely used absorption and emission spectrometric techniques are ultraviolet (UV) and fluorescence (FL) detection, the latter being preferred when mycotoxins exhibit natural fluorescence.
Mass spectrometry
Mass spectrometry (MS) is an analytical technique for determining the distribution of molecules of a substance based on its mass. HPLC and GC/MS, apart from having detection limit of 0.05 ppm for many mycotoxins, also require expensive equipment and technical support.
Liquid chromatography coupled to a tandem mass spectrometry detector
Liquid chromatography coupled to a tandem mass spectrometry detector (LC-MS/MS) LC-MS/MS based highly sensitive and selective methods, are used for the analysis of mycotxoins and their metabolites at very low concentrations and from complex biological samples. LC-MS/MS can analyse most of the mycotoxins, with increased sensitivity, precision and better assay reproducibility.
Liquid chromatography combined with tandem mass spectrometry (LC-MS / MS) is nowadays the gold standard for the evaluation of mycotoxins and the most reliable way of their quantification.
Liquid Chromatography
Micotoxicosis prevention
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