Knowledge > Mycotoxins
 

Mycotoxins

Mycotoxins are secondary metabolites of moulds, contaminating a wide range of crop plants and fruits before or after harvest, the most important mycotoxins being: Aflatoxins, Deoxynivalenol, Ochratoxin A, Fumonisins, Zearalenone, Patulin and T-2 Toxin. The acute and chronic impact of mycotoxins on human and animal health is proven scientifically. Mycotoxin contamination is recognized as an unavoidable risk because the formation of fungal toxins is weather dependant and effective prevention is impossible.

According to the FAO, more than 25 % of the world's agricultural production is contaminated with mycotoxins. This equates to economic losses estimated at $923 million annually in the US grain industry alone. Most countries have adopted regulations to limit exposure to mycotoxins, which has strong impact on food and animal crop trade. The presence of mycotoxins is unavoidable and, therefore, testing of raw materials and products is required to keep our food and feed safe

Aflatoxins

Aflatoxin M1
Aflatoxin B1

Aflatoxins are a group of mycotoxins produced by some Aspergillus species like A. flavus or A. parasiticus. Aflatoxin B1, B2, G1 and G2 and the hydroxylated metabolite M1 are of primary interest, with aflatoxin B1 the most frequently occurring of these. Aflatoxins can be found on a wide range of commodities including cereals, nuts, spices, figs and dried fruit. Aflatoxin M1, the metabolite of Aflatoxin B1, is found in milk and dairy products. Aflatoxins are of major interest because of their impact on both human and animal health. Aflatoxin B1 is one of the most potent hepato-carcinogens known and hence levels of aflatoxins in the diet are an important consideration for human health.
Most countries have established regulatory limits for either aflatoxin B1 or for total aflatoxins, which includes the sum of aflatoxin B1, B2, G1, and G2, as well as regulatory limits for Aflatoxin M1. Very often regulations also include detailed sampling procedures, as this is one of the most crucial steps in ensuring reliable results.

 

Romer mills® are designed especially for grinding large quantities of various commodities and to subsample the lot within the same step. A variety of methods and technologies are available for the analysis of grains, cereals, nuts and other possibly infected commodities.

Qualitative Yes/No test
We recommend using the AgraStrip® Afla for testing raw commodities such as grain or groundnuts for the presence of aflatoxins with a certain “cut off level” (e.g. 4µg/kg, 10µg/kg, 20µg/kg). This lateral flow test with three different cut-off levels (4ppb, 10ppb, 20ppb) can be performed in less than five minutes and needs no additional laboratory equipment.

Low cost screening of cereals – sequential analysis of samples
If subsequent samples have to be tested and a quantitative result is required as soon as possible, we recommend using FluoroQuant® Afla. This test gives quantitative results within five minutes, needs very little equipment, and can be processed easily.

Low cost screening – testing many samples at once
Another possibility for getting quantitative results within a short time period and with low investment costs is ELISA technology. We recommend using AgraQuant®Afla if a large number of samples need to be analyzed at the same time, as this is the most efficient way to use this technology.

Cost efficient way to semiquantitative results
Thin Layer Chromatography (TLC) is a cost-efficient way to establish methods for the detection of mycotoxins, especially if laboratory facilities and trained personnel are available. We offer a variety of clean-up columns for the use prior to TLC. Highly reproducible and reliable results can be achieved, if an Autospotter is used to apply samples.

Reference method
A widely used reference method in case of regulatory issues is High Performance Liquid Chromatography (HPLC) with fluorescence detection (FLD). We offer a variety of cleanup columns, a post-column derivatization unit (either photochemical or electrochemical) and a vast variety of reference materials including isotope labled calibrants which can be used as internal standards in MS applications.

Trichothecenes

Deoxynivalenol
T-2 Toxin

Trichothecenes (T-2, DON and others) are a group of sesquiterpenes produced by various Fusarium species like F. graminearum, F. sporotrichioides, F. poae or F. equiseti. The most important structural features causing the biological activities of trichothecenes are: the 12,13-epoxy ring, the presence of hydroxyl or acetyl groups at appropriate positions on the trichothecene nucleus and the structure and position of the side-chain. They are produced on many different grains like wheat, oats or maize.

This group of structurally related mycotoxins has a strong impact on the health of animals and humans due to their immunosuppressive effects. Type A trichothecenes (e.g. T-2 toxin, HT-2 toxin, Diacetoxyscirpenol) are of special interest because they are even more toxic than the related type B trichothecenes (e.g. Deoxynivalenol, Nivalenol, 3- and 15-Acetyldeoxynivalenol). Their major effects – related to their concentration in the commodity – are reduced feed uptake, vomiting and immuno-suppression.

Only a few countries have recommended levels for these mycotoxins in food and animal feed, but tests are often used to prevent them from entering the food chain and losses in animal production. As with all mycotoxin tests, sampling is the most crucial step in ensuring reliable results. Our Romer® mills are specially designed for grinding large quantities of various commodities and to subsample the lot in one step. For the testing of cereals and other relevant commodities, many methods and technologies are available:

Low cost screening – testing many samples at once
A possibility for getting quantitative results in a short time with low investment costs is ELISA technology. We recommend using Agraquant® DON or Agraquant® T-2 Toxin if a large number of samples need to be analyzed at the same time, as this is the most efficient way to use this technology.

Cost efficient way to semiquantitative results
Thin Layer Chromatography (TLC) is a cost efficient way to test trichothecenes. We offer a variety of cleanup columns for sample preparation prior to TLC. Highly reproducible and reliable result can be achieved, especially if an Autospotter is used to apply the samples. This technology can be recommended if a laboratory and trained personnel is available.

Reference method
The reference method is either Gas Chromatography (GC) combined with Electron capture detection (ECD) or with Mass Spectrometry (MS) or High Performance Liquid Chromatography (HPLC) with Variable Wavelength Detection (VWD) or, more recently, also in combination with Mass Spectrometry (MS). We offer a variety of cleanup columns for sample preparation and a vast variety of reference materials including isotope labled trichothecenes which can be used as internal standards in MS applications.

Fumonisins

Fumonisin B1
Fumonisins are a group of mycotoxins produced by Fusarium species such as F. moniliforme and F. proliferatum. Corn is the  commodity mainly affected. Conditions favoring Fumonisins formation are drought stress followed by warm, wet weather. Fumonisins have various effects on humans and animals, causing equine leukoencephalomalacia, pulmonary edema in swine, and they are suspected to influence the formation of esophageal cancer in humans.

Our Romer® mills are specially designed for the grinding of large quantities of various commodities and to subsample the lot in one step. Many methods and technologies are available for the testing of corn, cereals and other commodities.

Low cost screening – testing many samples at once
A possibility for getting quantitative results in a short time with low investment costs is ELISA technology. We recommend using AgraQuant® Fumonisin if a large number of samples need to be analyzed the same time, as this is the most efficient way to use this technology.

Cost efficient way to semiquantitative results
Thin Layer Chromatography (TLC) is a cost-efficient way to detect Fumonisins. We offer cleanup columns for sample preparation prior to TLC. Highly reproducible and reliable result can be achieved, especially if an Autospotter is used to apply the samples.This technology can be recommended if a laboratory and trained personnel is available.

Reference method
The reference method is High Performance Liquid Chromatography (HPLC) with fluorescence detection (FLD) after a derivatization step or the use of an LC-MS system (Liquid chromatography combined with mass spectrometry). We offer cleanup columns and reference materials, including stable isotope labeled calibrants for fumonisins.

Ochratoxin

Ochratoxin

Ochratoxin A, B, and C are mycotoxins produced by some Aspergillus species and Penicilium species, like A. ochraceus or P. viridicatum, with ochratoxin A as the most prevalent and relevant fungal toxin of this group.
Ochratoxin A is known to occur in commodities such as cereals, coffee, dried fruit and red wine. It is considered as a human carcinogen and is of special interest as it can be accumulated in the meat of animals. Thus meat and meat products can be contaminated with this toxin.

Regulatory limits have been established in many countries worldwide, also in the European Union. Very often regulations include detailed sampling procedures, as this is one of the most crucial steps in ensuring reliable results. Romer® mills are specially designed for grinding large quantities of various commodities and for subsampling the sample batch within one step.
For testing of cereals, wine, dried fruit and of other possibly infected commodities a variety of methods and technologies are available:

Low cost screening – testing many samples at once
A fast and cost-effective possibility for receiving quantitative results within a short time with low investment is ELISA technology. We recommend using AgraQuant® Ochratoxin if a number of samples have to be analyzed at one time.

Cost efficient way to semiquantitative results
Thin Layer Chromatography (TLC) is a cost-efficient way of establishing methods for the detection of mycotoxins, especially if laboratory facilities and trained personnel are available. We offer a variety of cleanup columns for the cleanup of Ochratoxin A and B prior to TLC. Highly reproducible and reliable results can be achieved, if an Autospotter is used to apply samples.

Reference method
A widely used and accepted reference method is High Performance Liquid Chromatography (HPLC) with Fluorescence Detection (FLD). We offer a variety of cleanup columns for the cleaning up of Ochratoxin A and B prior to HPLC analysis.

Zearalenone

Zearalenone

Zearalenone is a mycotoxin produced by Fusarium species like F. graminearum. It mainly occurs in grains and cereal products. Zearalenone is not acutely toxic but is a problem because of its estrogenic effects on mammals. The negative effects on reproductive systems makes it a concern in animal husbandry.
Only a few countries have imposed recommended levels for this mycotoxin in animal feed, but it is often tested to prevent losses in animal husbandry. As with all mycotoxin tests, sampling is the most crucial step ensuring reliable results.

Our Romer® mills are specially designed for grinding large quantities of various commodities and subsampling the lot in one step. Many methods and technologies are available for the testing of cereals and of other possibly infected commodities.

Low cost screening – testing many samples at once
A possibility to get quantitative results in a short time with low investment costs is ELISA technology. We recommend using AgraQuant® Zearalenone if a number of samples need to be analyzed the same time. This is the most cost-efficient way to use this technology.

Cost efficient way to semiquantitative results
Thin Layer Chromatography (TLC) is a cost-efficient way to detect mycotoxins. We offer a variety of cleanup columns for  sample preparation necessary prior to TLC. Highly reproducible and reliable result can be achieved with TLC, especially if an Autospotter is used to apply the samples. This technology can be recommended if a small laboratory and trained personnel is available.

Reference method
The reference method is High Performance Liquid Chromatography (HPLC) with Fluorescence Detection (FLD). We offer a variety of cleanup columns for sample preparation prior to the injection of a sample into the HPLC system.

Patulin

Patulin

Patulin is produced by Penicillium species and Aspergillus species Contamination with this mycotoxin mainly occurs on damaged or rotting fruits such as apples, pears, peaches and grapes. Patulin is suspected to be carcinogen.
As Patulin occurs mainly in fruits and fruit juices which are often used as baby or infant food, some countries have established regulatory limits. For testing possibly affected commodities like fruits and fruit juices we recommend using HPLC-UV.

We offer cleanup columns for simple and rapid cleanup of various commodities prior to the injection into HPLC. Our cleanup columns shorten sample preparation time significantly compared to the usually used liquid liquid partition. Also the reduction in working steps and the easier handling helps to reduce errors.

Ergot Alkaloids

Ergotamine

Ergot alkaloids are mycotoxins produced by fungi of all members of the Claviceps species, most important in terms of frequency being C. purpurea. These fungi infect the seed heads of plants during the flowering period and produce a wintering body, also known as sclerotium. Such structures are mainly found on rye, wheat and triticale, but also on other cereals and grasses. Sclerotia contain different classes of alkaloids, the most prominent being ergometrine, ergotamine, ergosine, ergocristine, ergocryptine and ergocornine.

Ergot alkaloids exert toxic effects in all animal species, and the most prominent toxic signs can be attributed to the interaction of ergot alkaloids with adrenergic, serotinergic and dopaminergic receptors. Typical clinical symptoms are vasoconstrictions that may progress into vaso-occlusion and gangrenous changes, but also abortions. The neurotoxic signs comprise feed refusal and dizziness but also convulsions. Typical dopaminergic effects are agalactia accompanied with insufficient nursing of suckling animals such as piglets and foals.
Romer Labs® offers special cleanup columns as well as reference materials for ergot analysis. Chromatographic methods are mainly used for the chemical analysis of ergot alkaloids and for the direct monitoring of the occurrence of ergot alkaloids in food and feed. Nowadays, the most important analytical technique for the analysis of ergot alkaloids is reversed phase HPLC-FLD. We now offer a cleanup column for sample preparation before HPLC analysis. This column eliminates the very extensive and time consuming cleanup step previously in many instances.

Other Mycotoxins

Citrinin
Mycotoxins are secondary metabolites of fungi and occur on a wide range of commodities. There are more than 200 currently known mycotoxins belonging to different groups of chemical structures. Their effects on human and animals are as manifold as their chemical structures. Most countries worldwide have regulatory limits for some mycotoxins, the most frequently regulated one is aflatoxin B1. Regulation usually also include a detailed sampling procedure as this is the most crucial step in ensuring reliable results. Our Romer® mills are especially designed for grinding large quantities of various commodities and for subsampling the lot in one step as well.

Citrinin
Citrinin can be detected using Thin Layer Chromatography (TLC) or High Performance Liquid Chromatography. In both cases, food a clean-up of the raw extract is extremely important to obtain reliable results.

Moniliformin
Moniliformin is a very polar molecule that can be detected by Thin Layer Chromatography (TLC) or High Performance Liquid Chromatography. We offer a specialized one-step clean up column for the cleanup of the raw extract.

Sterigmatocystin
A mycotoxin structurally related to the family of the aflatoxins. It can be detected by the use of Thin Layer Chromatography after the use of a simple one-step clean up column.

Cyclopiazonic acid
This mycotoxin often co-occurrs with aflatoxins. A method for the detection of this toxin is Thin Layer Chromatography. The clean up can be done using a Romer® clean-up column.
 

References: ROMER LABS GUIDE TO MYCOTOXINS (4th Edition), Oct. 2012