Rapid Detection of Mycotoxins

Non-Technical Summary:<br/>
The risk of contamination of grains and other field crops by mycotoxins, a toxic byproduct of mold infestations affecting as much as one-quarter of global food and feed crop output, is a very important food safety concern. The economic effects attributed to mycotoxin infection are widely felt in all sectors of the production and consumption of grain products. Grain producers are affected by limited yields, restricted end markets, and price discounts. Grain handlers are affected by restricted storage options, cost of testing grain lots, and loss of end markets. The estimated cost related to mycotoxins in U.S. and Canada is $5 billion/year. Food contaminated with mycotoxins can sometimes cause fatal acute illness and is associated with increased cancer risk from long-term exposure. Among the mycotoxins, trichothecene mycotoxins are extremely hazardous to humans. Exposure to even nano gram quantities of trichothecene mycotoxins, such as T-2 toxin, can cause severe reactions. Central nervous systems injury, as well as gastrointestinal and hematological toxicity can occur from ingestion or inhalation of very small quantities of these toxins. To protect consumers from these health risks, many countries, including the United States, have adopted regulations to limit exposure to mycotoxins, often taking the form of product standards. However, currently available enzyme based immuno assays (ELISA) kits for detection of such toxins requires use of antibodies, enzymes and incubation for the signal enhancement and hence take relatively long time, and are also expensive. The ELISA kits usually need to be stored under controlled environment and also involves sample preparation and use of several reagents, and are generally not suitable for field applications. Thus, there is a great need for a simple, convenient, cost effective and field deployable mycotoxin detection system. To address the need we are proposing to develop a simple battery operated handheld system that utilizes disposable micro-cartridges without any bio-molecules like antibodies or enzymes for rapid detection of tricothecene mycotoxins in grains, other field crops and water. The proposed hand-held rapid mycotoxin detection system has a broad potential commercial application in detection of trichothecene mycotoxins in food, feed and water under various settings because of its ease of use, speed, portability and competitive cost. In addition, the proposed technology also addresses a critical need of first responders, fire departments and the military to field-test for possible mycotoxin contaminations.
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Approach:<br/>
The overall goal of this Phase I project is to demonstrate the feasibility of using a colorimetric indicator based cartridges to indicate the presence of trace quantities of trichothecene mycotoxins in grains and in aqueous solutions. The Phase I research will consist of characterization of the optical properties of the colorimetric indicator and its trichothecene mycotoxin complexes, selection of a suitable LED light source, selection of a suitable solid support for the colorimetric indicator and fabrication of test cartridges, evaluation and testing of the cartridges with trichothecene mycotoxins, and design of a prototype to be developed in the Phase II. This testing will include preparation of various trichothecene mycotoxin test samples in grain extract solution or in water in the range of 50 ng/mL - 5 ug/mL. T-2, DON, acetylated DON derivatives and nivalenol test solutions will be prepared by diluting respective standard solutions. All tests will be validated using commercially available detection kits, such as Charm ROSA mycotoxin detection kit, AgraQuant Mycotoxin ELISA Kit or VICAM detection kit.
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Progress:<br/>
2012/05 TO 2013/03<br/>
OUTPUTS: During the Phase I project, Lynntech investigated the detection of trichothecene via complexation with solvatochromic dyes in the DIPAIN family. The toxins and dyes were thoroughly investigated to isolate false positives and define the measurement parameters that were necessary to detect the interactions between the analytes. Matrix effects on the dye were also characterized to ensure that the proper environment was provided for the coordination. Areas investigated include solid phase extraction, support induced interaction, and solvent assisted coordination. Significant effort was also expended to determine the repeatability.
<br/>PARTICIPANTS: Bikas Vaidya; Justin Weeks; Marc Penny; Tim Bosser.
<br/>TARGET AUDIENCES: Nothing significant to report during this reporting period.
<br/>PROJECT MODIFICATIONS: Not relevant to this project.
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IMPACT: However, significant difficulties were encountered with uncontrolled environmental effects on the dye fluorescence intensity and peak shift during the Phase I research. Particularly strong changes were observed from dye mobility during solvent evaporation, and the peak shift caused by the presence of certain organic solvents in both liquid and gas phase. Interaction with vapor phase chloroform was particularly distinct. Lynntech has accomplished the characterization of the optical properties of DIPAIN derivative and its trichothecene mycotoxin complexes, selection of a suitable LED light source, selection of a suitable solid support for DIPAIN derivatives and fabrication of test cartridges, evaluation and testing of the cartridges with trichothecene mycotoxins.