Hunor Farkaš, Jog Raj, and Marko Vasiljević
PATENT CO., Mišićevo, Serbia
What are mycotoxins and why should we control them?
Animal feed can be contaminated with microorganisms, mycotoxins, animal by-products, organic pollutants and toxic metals. This contamination has negative effects, both on animal and human health. Among these contaminants, mycotoxins are emerging as a major contaminant of feed and food.
Mycotoxins are produced as secondary metabolites by various fungi. The major fungus producing mycotoxins are:
- ⇰ Aspergillus
- ⇰ Fusarium
- ⇰ Penicillium
Mycotoxins cause mycotoxicosis and cause significant economic losses in the livestock industry due to reduced productivity, increased disease incidence and decreased reproductive performance. The mycotoxins that pose a higher threat due to their toxicity and occurrence are:
- ⇰ Aflatoxin B1 (AFB1)
- ⇰ Deoxynivalenol (DON)
- ⇰ Ochratoxin A (OTA)
- ⇰ Zearalenone (ZEA)
- ⇰ Fumonisins (FB1 and FB2)
- ⇰ T-2 toxins
How can we detect mycotoxins?
There are many methods available for the detection of mycotoxins. Conventional methods for mycotoxins analysis include:
- ⇰ ELISA
- ⇰ Thin-layer chromatography (TLC)
- ⇰ High-performance liquid chromatography (HPLC)
- ⇰ Gas chromatography (GC)
Most of these methods employ solid phase column clean-up of extracts and immunoaffinity techniques to remove interferences and improve the measurement of mycotoxins.
- ⇰ ELISA is a method of choice when rapid analysis is required but requires confirmatory analysis by LC-MS/MS.
- ⇰ LC-MS/MS is the most sensitive and preferred method of analysis for detecting and quantifying mycotoxins in food and feed samples.
A QUICK PEAK INTO THE LC-MS/MS BASED MULTI-MYCOTOXINS METHOD
Since it is necessary to test animal feeds for mycotoxin contamination, PATENT CO has developed and perfected a fast and simple LC-MS/MS based method for the determination and quantification of all mycotoxins (Aflatoxins B1, B2, G1, and G2; Deoxynivalenol, Zearalenone, Fumonisin B1 and B2, T-2, HT-2, and Ochratoxin A) regulated in feed (EU Directive 2002/32/ EC, 2006/576/EC and 2013/165/EU).
The method is based on “dilute and shoot” principle, which involves a two-step extraction phase and the centrifugation of the extracts.
To compensate the matrix effects* in electrospray ionization, the extracts are mixed with [13C] labelled internal standards for each group of mycotoxins (13C AB1, 13C DON, 13C ZON, 13C OTA, 13C FB1 and 13C T-2) before injection onto LC-MS/MS.
*Effect on an analytical method caused by all other components of the sample except the specific compound to be quantified.