The Importance of mycotoxins analysis

The

IMPORTANCE of

MYCOTOXINS ANALYSIS Jog Raj, Hunor Farkaš y Marko Vasiljević PATENT CO., Serbia

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Analysing incoming feed materials for mycotoxin contamination is crucially important as feed can be affected by climate, agricultural practices and local legislation on mycotoxins.

In a survey of the 2017 corn harvest in Serbia and Bosnia and Herzegovina, high levels of Aflatoxin B1, Fumonisin B1, Fumonisin B2 were discovered.

The global use of feed materials in the production of animal feed is increasing the risk of chemical and microbiological contaminants in foodproducing animals.

The feed can be contaminated

This contamination of animal feeds

Variations in mycotoxins are

with:

has a negative effect on both animal

increasing the importance of

and human health. Among these,

analysing raw materials before they

mycotoxins are emerging as a major

enter the feed chain.

Micro-organisms, Mycotoxins, Animal by-products,

contaminant of feed and food.

Organic pollutants Toxic metals.

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MYCOTOXINS Mycotoxins are produced as secondary metabolites by various fungi. The major fungus producing mycotoxins are aspergillus, fusarium and penicillium.

Aflatoxins, ochratoxin A, fumonisins, deoxynivalenol, T-2 toxin and zearalenone are the most common mycotoxins found in food and feed samples.

Many food and feed samples can become contaminated with mycotoxins before harvest, during transport and during their storage.

COMMODITIES AND PRODUCTS FREQUENTLY CONTAMINATED Commodities and products frequently contaminated with mycotoxins and used in animal feed include: Corn Wheat Barley

ECONOMIC IMPACT

Mycotoxins produce a wide range of adverse and toxic effects in animals affecting their overall health and productivity. Mycotoxins cause mycotoxicosis and cause significant economic losses in animals due to:

Rice

Reduced productivity,

Oats

Increased disease incidence

Nuts

Decreased reproductive performance.

Milk

MYCOTOXINS TOXICIT Y The mycotoxins of most concern due to their toxicity and occurrence are: Aflatoxin (AFB1) Deoxynivalenol (DON) Ochratoxin (Ochra A) Zearalenone (ZEA) Fumonisin (FB1 y FB2) T-2 toxins

Cheese Peanuts Cottonseed, etc.

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Regulations for major mycotoxins in the food and feed commodities exist in at least 100 countries.

Most of these regulations are related to aflatoxins and the maximum tolerated levels differs greatly among countries.

Analysis of mycotoxins To determine whether feed material and other commodities are contaminated with mycotoxins, it must be tested for mycotoxins. Proper sampling procedures are a pre-requisite for obtaining reliable results because of the heterogeneous distribution of mycotoxins in grains and other commodities.

These variations in tolerated levels of mycotoxins and non-regulation of other mycotoxins in other countries are posing a big challenge for the animal feed industry and these variations are increasing the importance of mycotoxin analysis of incoming raw materials before they enter the food/feed chain.

METHODS FOR DETECTION OF MYCOTOXINS There are many methods available for detection of mycotoxins.

Conventional methods for mycotoxins include:

Most of these methods employ a solid phase column clean-up of extracts and immunoaffinity techniques to remove

ELISA

interferences to improve the

Thin-Layer

measurement of mycotoxins.

Chromatography (TLC) High-performance Liquid Chromatography (HPLC) Gas Chromatography (GC).

ELISA is a method of choice where 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 mycotoxins in food and feed samples.

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Multi-mycotoxins methods Since it is necessary to test animal feeds for mycotoxin contamination, PATENT CO. is using a fast and simple UHPLC-based multi-mycotoxin method for the determination and accurate quantitation of all mycotoxins (Aflatoxin B1, B2, G1, G2, Deoxynivalenol, Zearalenone, Fumonisin B1 and B2, T-2, HT-2, Ochratoxin A) regulated in feed, by

The method is based on a ‘dilute and shoot’ principle. It involves two step extraction and centrifugation of the extracts.

liquid chromatography coupled with tandem mass spectrometry.

To compensate the matrix effects in electrospray ionization, the extracts are mixed with [13C] labelled internal standards for each

RESULTS

group of mycotoxins (13C AB1, 13C DON, 13C ZON, 13C OTA, 13C FB1 and 13C T-2) before injection onto LCMS/MS. The method was successfully validated on:

The blank samples were spiked with a mixture of 11 mycotoxin standards on two levels (LOQ and 10xLOQ) in 12 replicates. The RSDr of the method were between 2.5% and 13.4% and the apparent recoveries were between 62% and 115% for all analytes.

Corn Compound feed Wheat Barley Soya meal Wheat bran

CONCLUSION

Sunflower meal TMR It was therefore concluded that the ‘dilute and shoot’ method with

PARÁMETROS Method performance

addition of [13C] labelled internal standard is capable for determining all EU regulated mycotoxins in animal feed and compound feed.

parameters were obtained by in-house validation.

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Balkan corn survey in 2017 In a recent study, corn samples

A total of 113 samples were

Aflatoxin B1 (AB1), Fumonisin B1

received from Serbia and

received for analysis. Out of these,

(FB1), Fumonisin B2 (FB2) and HT-2

Bosnia and Herzegovina

53% of the samples were found to

were detected in 13%, 44%, 24%

during August to November

be contaminated with mycotoxins

and 8% of the samples (Figure 1).

2017 were screened.

and 28% of these contaminated samples were found to contain more than one mycotoxin.

Figure 1.

13%

Aflatoxin B1 (AB1)

HT-2

8%

Fumonisin B2 (FB2)

24%

44%

Fumonisin B1 (FB1)

CONCLUSIONES Los niveles de estas micotoxinas detectadas en las muestras de maĂ­z fueron:

Aflatoxin B1: 0,59-5.644 ppb

The summer of 2017 was very hot and dry in the Balkans.

Climate conditions have a huge effect on mycotoxin production as this may have a impact on plant–pathogen interactions.

Toxin HT-2: 9-66 ppb

Temperature, humidity, insect attack, and stress conditions in

Fumonisin B2: 53-2.540 ppb

the plants affects the ability of fungi to produce mycotoxins.

Fumonisin B1: 48-8.623

Therefore, changes in temperature would directly affect the growth of mould and hence, the mycotoxin production capacity.

High temperatures and drought stress can increase the risk of aflatoxin and fumonisin contamination in the corn. Similar effects were seen in this survey as corn was found to be contaminated with aflatoxins and fumonisin B1 and B2.

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