MYCOTOXINS DETECTION 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.
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IMMUNOCHEMICAL METHODS Mycotoxins 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
ELISA TEST The enzyme-linked immunosorbent assay (ELISA) is an antibody-based assay that is commonly used for mycotoxins
immunosorbent assay)
detection.
RIA (radioimmunoassay)
There are several commercial
immunoaffinity columns
deoxynivalenol, Fumonisins,
ELISA kits available for aflatoxins, Ochratoxins and Zearalenone.
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Most of these methods are very
ELISA is one of the most
sensitive, specific and easy to
affordable methods for
use, which has given rise to a
mycotoxins detection, but the
new dimension in terms of
limit of detection for many
methodology for mycotoxins
mycotoxins often exceeds
analysis.
0.2 ppm.
INSTRUMENTAL METHODS Chromatography and Spectrometry
The use of chromatographic
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
GAS CHROMATOGRAPHY
techniques requires prior preparation of a sample
High-performance liquid
Gas chromatography (GC) is used
using different methods of
chromatography (HPLC) represents
less frequently since most of the
extraction and purification.
the technique of choice for the
mycotoxins are not sufficiently
Depending on the mycotoxins
analysis of mycotoxins or for
volatile and, therefore, must be
to be analysed and the
confirmation of positive results by
derivatized, which increases the time
feed matrix, different
ELISA, as it is sensitive, reproducible,
and cost of analysis.
types of techniques such
accurate and has a higher degree of
as immunoaffinity columns
automation.
based preparations or other procedures based on solidliquid (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
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methods for the detection and quantification of mycotoxins.
MASS SPECTROMETRY ABSORPTION AND EMISSION SPECTROMETRIC TECHNIQUES
Mass spectrometry (MS) is an analytical technique for determining the distribution of molecules of a
The most widely used absorption
substance based on its mass.
and emission spectrometric techniques are ultraviolet (UV)
HPLC and GC/MS, apart from
and fluorescence (FL) detection,
having detection limit of <0.05
the latter being preferred when
ppm for many mycotoxins, also
mycotoxins exhibit natural
require expensive equipment and
fluorescence.
technical support.
LIQUID CHROMATOGRAPHY COUPLED TO A TANDEM MASS SPECTROMETRY DETECTOR Liquid chromatography coupled to a
Liquid chromatography combined
tandem mass spectrometry detector
with tandem mass spectrometry
(LC-MS/MS) LC-MS/MS based highly
(LC-MS / MS) is nowadays the
sensitive and selective methods, are
gold standard for the evaluation of
used for the analysis of mycotxoins
mycotoxins and the most reliable
and their metabolites at very low
way of their quantification.
concentrations and from complex biological samples. LC-MS/MS can analyse most of the mycotoxins, with increased sensitivity, precision and better assay reproducibility.
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