Saturday, July 05, 2008

Methods of analysis for aflatoxins

Recent Methods of Analysis for Aflatoxins in Foods and Feeds

Sampling and Sample Preparation :

Sampling and sample preparation remain a considerable source of error in the analytical identification of aflatoxins. Thus, systematic approaches to sampling, sample preparation, and analysis are absolutely necessary to determine aflatoxins at the parts-per-billion level . In this regard, specific plans have been developed and tested rigorously for some commodities such as corn, peanuts, and tree nuts; sampling plans for some other commodities have been modeled after them. A common feature of all sampling plans is that the entire primary sample must be ground and mixed so that the analytical test portion has the same concentration of toxin as the original sample.

Solid-Phase Extraction :

All analytical procedures include three steps: extraction, purification, and determination. The most significant recent improvement in the purification step is the use of solid-phase extraction.Test extracts are cleaned up before instrumental analysis(thin layer or liquid chromatography) to remove coextracted materials that often interfere with the determination of target analytes.

Thin-Layer Chromatography :

Thin layer chromatography (TLC) , also known as flat bed chromatography or planar chromatography is one of the most widely used separation techniques in aflatoxin analysis. Since 1990, 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. The TLC method is also used to verify findings by newer, more rapid techniques .

Liquid Chromatography :

Liquid chromatography (LC) is similar to TLC in many respects, including analyte application, stationary phase, and mobile phase. Liguid chromatography and TLC complement each other. For an analyst to use TLC for preliminary work to optimize LC separation conditions is not unusual. Liquid chromatography methods for the determination of aflatoxins in foods include normal-phase LC (NPLC), reversed-phase LC (RPLC) with pre- or before-column derivatization (BCD), RPLC followed by postcolumn derivatization (PCD), and RPLC with electrochemical detection.

Immunochemical Methods :

Thin layer chromatography and LC methods for determining aflatoxins in food are laborious and time consuming . Often, these techniques require knowledge and experience of chromatographic techniques to solve sepatation and and interference problems. Through advances in biotechnology, highly specific antibody-based tests are now commercially available that can identify and measure aflatoxins in food in less than 10 minutes. These tests are based on the affinities of the monoclonal or polyclonal antibodies for aflatoxins. The three types of immunochemical methods are radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), and immunoaffinity column assay (ICA).

Confirmation of Identities of the Aflatoxins :

Although analytical methods might consist of different extraction, clean-up, and quantitation steps, the results of the analyses by such methods should be similar when the methods are applied properly. Since the reliability of the quantitative data is not in question, the problem still to be solved is the confirmation of identity of the aflatoxins. The confirmation techniques used involve either chemical derivatization or mass spectrometry (MS).


Nabil Saad. (2004). Aflatoxins: Occurrence and Health Risks. Retrieved July 4, 2008 from ansci.cornell.edu Website: http://www.ansci.cornell.edu/plants/toxicagents/aflatoxin/aflatoxin.html

Aflatoxins in human health

Aflatoxins and Human Health

Humans are exposed to aflatoxins by consuming foods contaminated with products of fungal growth . Such exposure is difficult to avoid because fungal growth in foods is not easy to prevent . Even though heavily contaminated food supplies are not permitted in the market place in developed countries, concern still remains for the possible adverse effects resulting from long-term exposure to low levels of aflatoxins in the food supply . Evidence of acute aflatoxicosis in humans has been reported from many parts of the world , namely the Third World Countries, like Taiwan, Ouganda, India, and many others . The syndrome is characterized by vomiting, abdominal pain, pulmonary edema, convulsions, coma, and death with cerebral edema and fatty involvment of the liver , kidneys , and heart.

Conditions increasing the likelihood of acute aflatoxicosis in humans include limited availability of food, environmental conditions that favor fungal development in crops and commodities, and lack of regulatory systems for aflatoxin monitoring and control.Because aflatoxins, especially aflatoxin B1, are potent carcinogens in some animals, there is interest in the effects of long-term exposure to low levels of these important mycotoxins on humans . In 1988, the IARC placed aflatoxin B1 on the list of human carcinogens. This is supported by a number of epidemiological studies done in Asia and Africa that have demonstrated a positive association between dietary aflatoxins and Liver Cell Cancer (LCC) . Additionally , the expression of aflatoxin-related diseases in humans may be influenced by factors such as age, sex, nutritional status, and/or concurrent exposure to other causative agents such as viral hepatitis (HBV) or parasite infestation.


Nabil Saad. (2004). Aflatoxins: Occurrence and Health Risks. Retrieved July 4, 2008 from ansci.cornell.edu Website: http://www.ansci.cornell.edu/plants/toxicagents/aflatoxin/aflatoxin.html

Factors affecting the growth of aflatoxins and the occurance

From my research, i had found that:

Aflatoxins are found in:

In Raw Agricultural Products:

Aflatoxins often occur in plants in the field prior to harvest. Postharvest contamination can occur if crop drying is delayed and during storage of the crop if water is allowed to exceed critical values for the mold growth. Insect or rodent infestations facilitate mold invasion of some stored commodities.Aflatoxins are detected occasionally in milk, cheese, corn, peanuts, cottonseed, nuts, almonds, figs, spices, and a variety of other foods and feeds. Milk, eggs, and meat products are sometimes contaminated because of the animal consumption of aflatoxin-contaminated feed. However, the commodities with the highest risk of aflatoxin contamination are corn, peanuts, and cottonseed.


Factors Favourizing Aflatoxin Production

Fungal growth and aflatoxin contamination are the consequence of interactions among the fungus, the host and the environment. The appropriate combination of these factors determine the infestation and colonization of the substrate, and the type and amount of aflatoxin produced. However, a suitable substrate is required for fungal growth and subsequent toxin production , although the precise factor(s) that initiates toxin formation is not well understood . Water stress, high-temperature stress, and insect damage of the host plant are major determining factors in mold infestation and toxin production.

Similarly, specific crop growth stages, poor fertility, high crop densities, and weed competition have been associated with increased mold growth and toxin production. Aflatoxin formation is also affected by associated growth of other molds or microbes. For example, preharvest aflatoxin contamination of peanuts and corn is favored by high temperatures, prolonged drought conditions, and high insect activity; while postharvest production of aflatoxins on corn and peanuts is favored by warm temperatures and high humidity.


Nabil Saad. (2004). Aflatoxins: Occurrence and Health Risks. Retrieved July 4, 2008 from ansci.cornell.edu Website: http://www.ansci.cornell.edu/plants/toxicagents/aflatoxin/aflatoxin.html