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OF AFLATOXIN M1
Mycotoxins may be defined as metabolites of fungi, which could incur diseases in man and animals. It causes mycotoxicoses or toxicity syndromes due to the intake of mycotoxins by man and animals, typically through ingestion, inhalation or skin contact. The toxic factor is produced by two fungi, Aspergillus parasiticus and Aspergillus flavus, thus the name "aflatoxin" was given to it derived from the name of Aspergillus flavus. Aflatoxins are toxic metabolites produced by certain fungi in/on foods and feeds/feedstuffs. They are probably the best known and most intensively researched mycotoxins in the world. Aflatoxins have been associated with various diseases, such as aflatoxicosis in livestock, domestic animals and humans throughout the world. The occurrence of aflatoxins is influenced by certain environmental factors; hence the extent of contamination will vary with geographic location, agricultural and agronomic practices, and the susceptibility of commodities to fungal invasion during pre-harvest, storage, and/or processing periods. Aflatoxins have received greater attention than any other mycotoxins because of their carcinogenic effect in susceptible laboratory animals and their acute toxicological effects in humans. Among the mycotoxins, the most significant in milk and milk products is aflatoxin M1, the 4-hydroxy derivative of aflatoxin B1. The contamination of animal products caused by caused by feeds/feedstuffs given to animals contaminated with aflatoxin B1.
This makes them very applicable and useful for scientists and technicians in developing countries. After that. a. it is possible to judge whether the sample contains more or less aflatoxins than the standard. depending on their physicochemical properties. The minicolumn is embedded with successive zones of absorbents including alumina.Methods of Analysis for Aflatoxins Chromatographic Procedures Chromatography involves solute partitioning between two phases. A special design. there are also limitations within this method. In contast to thin layer chromatography (TLC). A chloroform extract is placed on the top of the column. can be used for detection of some mycotoxins in certain products. where they can be detected by their blue fluorescence under the ultraviolet (UV) light. false positive results may occur. column chromatography does not distinguish the different aflatoxins. Hence. containing fluorescent compounds other than the toxin of interest are applied to the column. The stationary phase slows down more or less the progress of substances through the bed. Column Chromatography Column chromatography is a technique often used in clean-up procedures. hence. descending chromatography having a mixture of chloform and acetone is applied thus trapping the aflatoxins as a tight band at the top of the FlorisilR layer. a sample column with a column containing a known amount of aflatoxins. and drained by gravity. Silica gel has been extensively used in column chromatography clean-up. This method requires little time and no costly equipment. a stationary phase (the chromatoghraphic bed) and a mobile phase (liquid or gas). For mycotoxin assays. In comparison. However. the glass minicolumn with an intenal diameter of 5 mm. The method is best when used semi-quantitatively and has a higher limit of . separation into components is achieved. The interpretation of the picture on the mincolumn needs some expertise and it may be quite difficult when dirty extracts. column chromatography and thin layer chromatography can be used. FlorisilR and silica gel with calcium phosphate drier at both ends and held in place with glass wool. that carries substances to be separated through the chromatographic bed.
Silica gel TLC plates are most often used as this type of adsorbent generally offers the best possibility of separating the toxin of interest from matrix components. It has been considered the AOAC official method and the method of choice to identify and quantitate aflatoxins at levels as low as 1 ng/g dated 1991. it became a powerful separation technique in which second development is carried out in a direction at right angles to the first one. the sample is spotted at a corner of the TLC plate and two developments are carried out successively parallel to the two sides of the plate using two different solvents. The precision syringes allow the intermittent application of larger volumes under inactive atmosphere by using them together with a repeating dispenser eventually incorporated in a spotting device. Calcium sulphate can be added to act as a binder of the silica gel to the glass plate. Thin Layer Chromatography (TLC) Thin layer chromatography (TLC) likewise known as flat bed chromatography or planar chromatography is one of the most widely used separation techniques in aflatoxin analysis being simple and cost-effective. This enables much better separation than one-dimensional TLC especially when Aflatoxin M1 in milk has to be detected. Intilaly. using a different developing solvent.determination and less sensitivity and selectivity than what is obtained in TLC and other chromatographic procedures. The TLC method is also used to verify findings by newer. Different applicator types are used. EDTA can also be used as a complexing agent for contaminants in the silica gel to prevent streaking of citrinin spots. 5-15 micro liter of extract is applied to the plate. more rapid techniques. In TLC. . disposable capillary pipettes or precision syringes are used. Later on. the stationary phase consists of a thin layer of adsorbent particles within a plate and the mobile phase runs through this layer by capllary forces. In a two-dimensional TLC. Silica gel can be used as an adsorbent in TLC. b. The solvents must have compatibility and independence or else the spots would agglomerate along the bisector of the plate. two-dimensional approach to TLC was introduced to mycotoxin research. For screening. In the TLC determination of mycotoxins. separations were carried out in one dimension using a single developing solvent.
pp.cornell.V. pp. A. Betina (Ed. Food and Agricultural Organization (FAO) of the United Nations (UN).Conclusion: Determination of mycotoxins particularly the aflatoxin M1 in milk. and quantization steps that are helpful to properly attain reliability of results. (1993). September 2010. V.E. 4-5. and Gibbs. (2009).html.). R. J. Coker. Aflatoxins: Ocurence and Health Risks. 21-24. Elsevier Science Publishers B. milk products and feeds/feedstuffs is important because its presence in such products bring out various bad effects to humans and animals. References: Cornell Univeristy's Departement of Animal Science. Available Retrieved at 02 http://www.P.: Amsterdam. Chromatography of Mycotoxins: Techniques and Applications. clean-up. .edu/plants/toxicagents/aflatoxin/aflatoxin. John..ansci. (1990). Manuals of Food Quality Control: Training in Mycotoxins Analysis. Analytical methods like column chromatography and thin layer chromatography involves a sequence of discrete operations such as extraction. FAO-UN: Rome.A. 12-13.