Wheat cultivar discrimination by capillary electrophoresis of gliadins in isoelectric buffers

Electrophoresis
L CapelliP G Righetti

Abstract

A modified method is reported for screening of wheat cultivars: capillary zone electrophoresis of gliadins in isoelectric buffers. Previously published procedures recommended a 100 mM phosphate buffer, supplemented with 0.05% hydroxypropylmethylcellulose and 20% acetonitrile, in uncoated capillaries. Due to the very high conductivity of such a buffer (4.7 mmhos at 25 degrees C) high speed separations (10-12 min analysis time at 800 V/cm) could only be elicited in 20 microm internal diameter (ID) capillaries, at the expense of sensitivity. In the present report, we optimized the background electrolyte as follows: 40 mM aspartic acid (pH=pI=2.77) in the presence of 7 M urea and 0.5% short-chain hydroxyethylcellulose (Mn 27000 Da; apparent pH 3.9 in 7 M urea). As an alternative recipe, the same isoelectric buffer can be supplemented with a mixed organic solvent composed of 4 M urea and 20% acetonitrile (apparent pH 3.66). Due to the much lower conductivity (0.7 mmhos), separations can be carried out at 1000 V/cm in only 10 min, but in larger bore capillaries (50 microm ID), ensuring a five-times higher sensitivity. The gliadin patterns thus obtained are species-specific and allow easy identification of all cultivars tested of both...Continue Reading

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Citations

Feb 5, 2002·Electrophoresis·S R Bean, G L Lookhart
Jun 19, 2012·Analytical and Bioanalytical Chemistry·Angela R Piergiovanni
Oct 25, 2000·Journal of Chromatography. a·A BossiA P Turner
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Sep 21, 2010·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Luca ElliMaria Teresa Bardella
Jul 15, 2005·Journal of Separation Science·Frantisek Kvasnicka
Jul 22, 2009·Electrophoresis·Youssef AlahmadMyriam Taverna
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