Premium
Separation and characterization of barley ( Hordeum vulgare L.) hordeins by free zone capillary electrophoresis
Author(s) -
Lookhart George L.,
Bean Scott R.,
Jones Berne L.
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990601)20:7<1605::aid-elps1605>3.0.co;2-j
Subject(s) - hordein , capillary electrophoresis , chromatography , chemistry , hordeum vulgare , repeatability , extraction (chemistry) , resolution (logic) , electrophoresis , capillary action , analytical chemistry (journal) , storage protein , materials science , biochemistry , botany , poaceae , artificial intelligence , biology , computer science , composite material , gene
Extraction conditions, separation conditions, and capillary rinsing protocols were optimized for the separation of barley hordeins by free zone capillary electrophoresis. Stable hordein extracts were obtained with a single 5 min extraction after the albumins and globulins were removed. Hordeins had to be reduced for optimal resolution. Optimum separation conditions for hordein separations were 100 m M phosphate‐glycine buffer containing 20% acetonitrile and 0.05% hydroxypropylmethylcellulose. The addition of zwitterionic sulfobetaine detergents containing hydrocarbon tails of eight and ten carbons slightly improved the resolution of the separations, but not enough to warrant their use on a routine basis. The migration positions of the hordein subclasses were determined by two‐ dimensional reversed‐phase high‐performance liquid chromatography x free zone capillary electrophoresis mapping. The hordein subclasses formed clusters similar to those of wheat gliadins. Separation‐to‐separation repeatability was good, with migration time relative standard deviations < 1% for a 15‐run period. For routine discrimination of cultivars, a 2 min post‐separation rinse with 500 m M acetic acid was necessary to prevent protein build‐up on the capillary walls. An example of successfully differentiating barley cultivars using this technique is shown.