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ARS Home » Southeast Area » Mississippi State, Mississippi » Crop Science Research Laboratory » Corn Host Plant Resistance Research » Research » Publications at this Location » Publication #328878

Title: Single corn kernel aflatoxin B1 extraction and analysis method

item REID, CEDRIC - Mississippi State University
item SPARKS, DARREL - Mississippi State University
item Williams, William
item BROWN, ASHLI - Mississippi State University

Submitted to: Natural Resources
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/2/2016
Publication Date: 7/5/2016
Citation: Reid, C., Sparks, D., Williams, W.P., Brown, A. 2016. Single corn kernel aflatoxin B1 extraction and analysis method. Natural Resources. 7:405-410.

Interpretive Summary: Aflatoxin, produced by the fungus Aspergillus flavus, occurs naturally in corn. It is the most potent carcinogen found in nature and is highly toxic to livestock, pets, and wildlife. Planting corn hybrids with resistance to aflatoxin accumulation is generally considered a desirable way to reduce losses to aflatoxin contamination. To produce such hybrids, corn germplasm lines with genetic resistance to aflatoxin accumulation must be identified. Understanding the underlying genetic basis of resistance is an important step in producing corn hybrids that are not contaminated by aflatoxin. Methods for extracting and quantifying aflatoxin from a single corn kernel were developed. This protocol will facilitate comparisons of aflatoxin in developing ears of corn with different levels of resistance to aflatoxin accumulation over time. This will aid in determining which genes are associated with resistance and when and how these genes are expressed. This will enhance development and deployment of corn hybrids with resistance to aflatoxin accumulation into farmers’ fields.

Technical Abstract: Aflatoxins are highly carcinogenic compounds produced by the fungus Aspergillus flavus. Aspergillus flavus is a phytopathogenic fungus that commonly infects crops such as cotton, peanuts, and maize. The goal was to design an effective sample preparation method and analysis for the extraction of aflatoxin B1 from Aspergillus flavus infected maize kernels. The method involves liquid extraction with a methanol/water solution. The samples were analyzed by liquid chromatography coupled to an electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) operating in positive ion multiple reaction monitoring (MRM) mode. The spiked levels for the recovery experiment were 4 ppb and 20ppb. Recoveries ranged between 72% and 113% (90.8% average), with RSD below 15% (10.6% on average).