Use of SDS to extract sorghum and maize proteins for free zone capillary electrophoresis (FZCE) analysis

Two different extraction methods for extracting sorghum (Sorghum bicolor L. Moench.) storage proteins for free zone capillary electrophoresis (FZCE) analysis were compared. A traditional solvent based on 60% t-butanol was compared with a pH 10 berate buffer containing the anionic detergent SDS followed by precipitation of nonkafirins using 60% t-butanol. FZCE analysis of both types of extracts showed identical patterns, despite the fact that the SDS should have given all proteins equal charge-to-mass ratios. This methodology was also successfully applied to maize proteins. The use of t-butanol to precipitate nonkafirins, combined with electrophoresis at low pH, is thought to have removed the SDS from the storage proteins. The SDS extraction procedure produced more stable extracts for FZCE analysis. These extracts could also be used directly for SDS capillary electrophoresis (SDS-CE) separations. Kafirins from 15 genotypes were extracted with this procedure and analyzed by FZCE and SDS-CE. Resolution of the kafirins by FZCE was much higher than the SDS-CE, demonstrating that the kafirin proteins possessed a high level of charge density variability within a relatively small molecular size distribution. Two distinct groups of alpha -kafirins could be seen in the FZCE electropherograms.