Comparison of Thin-Layer Chromatography and an Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Deoxynivalenol in Corn and Wheat†

1996 ◽  
Vol 59 (4) ◽  
pp. 438-440 ◽  
Author(s):  
CHARLENE E. WOLF-HALL ◽  
LLOYD B. BULLERMAN

The trichothecene, deoxynivalenol (DON), is a common mycotoxin found in wheat and corn. Detection methods for DON have limitations in accuracy, sensitivity, ease of use, and turn-around time. Two methods, thin-layer chromatography (TLC) and an enzyme-linked immunosorbant assay (ELISA) for DON detection and quantification were compared. The methods varied considerably in the amounts of DON detected in 51 different grain samples, with the ELISA method showing higher concentrations than the TLC method. Much of the unaccounted-for DON may have been lost during sample preparation for the TLC method. Recovery rates for the TLC method at DON levels of 1 and 5 ppm were 46% and 25% in corn grits and 32% and 26% in ground wheat respectively. Recovery rates for the ELISA method at 1 and 5 ppm levels were both 96% in corn grits and respectively 83% and 69% in ground wheat. The ELISA method was much faster and less laborious than the TLC method and did not use organic solvents, which are required in the TLC method.

2020 ◽  
Vol 21 (5) ◽  
pp. 1670
Author(s):  
Akhikun Nahar ◽  
Anthony L. Baker ◽  
David S. Nichols ◽  
John P. Bowman ◽  
Margaret L. Britz

In addition to cell membrane phospholipids, Actinobacteria in the order Corynebacteriales possess a waxy cell envelope containing mycolic acids (MA). In optimized culture condition, some species can also accumulate high concentrations of intracellular triacylglycerols (TAG), which are a potential source of biodiesel. Bacterial lipid classes and composition alter in response to environmental stresses, including nutrient availability, thus understanding carbon flow into different lipid classes is important when optimizing TAG synthesis. Quantitative and qualitative analysis of lipid classes normally requires combinations of different extraction, derivatization, chromatographic and detection methods. In this study, a single-step thin-layer chromatography-flame ionization detection (TLC-FID) technique was applied to quantify lipid classes in six sub-Antarctic Corynebacteriales strains identified as Rhodococcus and Williamsia species. A hexane:diethyl-ether:acetic acid solvent system separated the total cellular lipids extracted from cells lysed by bead beating, which released more bound and unbound MA than sonication. Typical profiles included a major broad non-polar lipid peak, TAG and phospholipids, although trehalose dimycolates, when present, co-eluted with phospholipids. Ultra-performance liquid chromatography-tandem mass-spectrometry and nuclear magnetic resonance spectroscopy detected MA signatures in the non-polar lipid peak and indicated that these lipids were likely bound, at least in part, to sugars from cell wall arabinogalactan. Waxy esters were not detected. The single-solvent TLC-FID procedure provides a useful platform for the quantitation and preliminary screening of cellular lipid classes when testing the impacts of growth conditions on TAG synthesis.


2006 ◽  
Vol 78 (3) ◽  
pp. 905-911 ◽  
Author(s):  
Michael Lancaster ◽  
David M. Goodall ◽  
Edmund T. Bergström ◽  
Sean McCrossen ◽  
Peter Myers

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