Extraction and separation of water-soluble proteins from different wheat species by acidic capillary electrophoresis.
Optimization of protein extraction and a capillary zone electrophoresis method for water-soluble protein analysis in wheat is described. The optimal separation was obtained with a 50 microm i.d. x 27 cm (20 cm to detector) uncoated capillary filled with 0.1 M phosphoric acid/beta-alanine, pH 2.5, buffer containing urea (1 M), 0.05% (w/v) hydroxypropylmethylcellulose, and 20% (v/v) acetonitrile. Separation was carried out at 15 kV and 35 degrees C for 9 min. Extract stability was also investigated within 2 h from the extraction. Good visual peak parameters and a higher sensitivity can be obtained when 30% ethanol is used as an extraction medium. The method was successfully used to analyze extracts obtained from whole and refined meals of six Triticum spp. Moreover, the described methodology could be applied to the discrimination of species with different ploidy levels and to the detection of durum wheat adulteration, as well as to screen wheat collections for enzymes involved with the quality of wheat derivatives.
Piergiovanni
Angela R
AR
Istituto di Genetica Vegetale-CNR, via Amendola 165/a, 70126 Bari, Italy. angelarosa.piergiovanni@igv.cnr.it
D000418:Albumins / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D010940:Plant Proteins / Q000302:isolation & purification*; D012639:Seeds / Q000737:chemistry; D012995:Solubility; D014908:Triticum / Q000737:chemistry*; D014867:Water
D000418:Albumins; D010940:Plant Proteins; D014867:Water