Mycotoxins

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.

Only few countries have recommended levels for these mycotoxins in food and animal feed but it is often tested to prevent them from entering the food chain and to prevent losses in animal production. Like with all mycotoxin tests, sampling is the most crucial step on the way to a reliable result. Our Romer® mills are especially 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 to get quantitative results in a short time with low investment costs is ELISA technology. We recommend using Agraquant® DON or Agraquant® T-2 Toxin if many samples need to be analyzed at once 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 clean up columns for sample preparation prior to TLC. Especially if an Autospotter is used to apply the samples, highly reproducible and reliable result can be achieved. 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 lately also in combination with Mass Spectrometry (MS). We offer a variety of clean up columns for the sample preparation and a vast variety of reference materials including isotope labled trichothecenes which can be used as internal standards in MS applications.

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

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® Fumonisin if many samples need to be analyzed at once, 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 a clean up column for the sample preparation prior to TLC. Especially if an Autospotter is used to apply the samples, highly reproducible and reliable result can be achieved. 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 clean up columns for the sample preparation prior to the injection of a sample into the HPLC system.

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 like 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 on the way to a reliable result. Romer® mills are especially designed for grinding large quantities of various commodities and to subsample 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 to receive quantitative results within a short time with low investment is ELISA technology. We recommend using Agraquant® Ochratoxin if a number of sample has to be analyzed at one shot.

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 clean up 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 clean up columns for clean up of Ochratoxin A and B prior to HPLC analysis.

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 the reproductive systems makes it a concern in animal husbandry.

Only 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 on the way to a reliable result. Our Romer® mills are especially designed for grinding large quantities of various commodities and to subsample the lot in one step. For the testing of cereals and of other possibly infected commodities, many methods and technologies are available.

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 at once. 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 clean up columns for the sample preparation necessary prior to TLC. Especially if an Autospotter is used to apply the samples, highly reproducible and reliable result can be achieved with TLC. 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 clean up columns for the sample preparation prior to the injection of a sample into the HPLC system.

Patulin is produced by Penicillium species and Aspergillus species The contamination with this mycotoxin mainly occurs on damaged or rotting fruits like 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 clean up columns for simple and rapid clean up of various commodities prior to the injection into HPLC. Our clean up columns shorten the 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 are mycotoxins produced by fungi of all members of the Claviceps species, most important in terms of frequency of occurrence 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 vasoconstriction that may progress into vaso-occlusion and gangrenous changes, but also abortions. The neurotoxic signs comprise feed refusal and dizziness but also convulsion. Typical dopaminergic effects are agalactia accompanied with insufficient nursing of suckling animals such as piglets and foals.

Romer Labs® offers special clean up columns as well as reference materials for ergot analysis.

For the chemical analysis of ergot alkaloids and for the direct monitoring of the occurrence of ergot alkaloids in food and feed mainly chromatographic methods are used. 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 previously common very extensive and time consuming cleanup step.

Other Mycotoxins

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. As manifold as their chemical structures are their effects on human and animals. Most countries worldwide have regulatory limits for some mycotoxins, the most often regulated one is aflatoxin B1. Usually the regulation also includes a detailed sampling procedure as this is the most crucial step on the way to a reliable result. Our Romer® mills are especially designed for grinding large quantities of various commodities and to subsample 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 good 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. For the clean up of the raw extract we offer a specialized one-step clean up column.

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

Cyclopiazonic acid
This mycotoxin is often co occurring 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.

For more information about mycotoxins visit www.mycotoxins.info