Improvements in wheat protein analysis and quality prediction by reversed-phase high-performance liquid chromatography
1987
Huebner, F.R. | Bietz, J.A.
Specific problems in wheat gluten protein analysis by reversed-phase high-performance liquid chromatography (RP-HPLC) and in evaluating data for predicting quality were found. Procedures were developed to improve the reliability and reproducibility of the method. To obtain accurate integration, a flat baseline is desirable; this was achieved by reducing (15 to 18%) the concentration of trifluoroacetic acid in acetonitrile. The similarity of gliadins extracted with 50 to 70% ethanol or after pre-extraction with 0.1 M NaCl was demonstrated; adjustment of extractant hydrophobicity prevented early elution of gliadins during preparative RP-HPLC. Long-term storage of gliadins at various temperatures was studied; -70°C is recommended. Various dissociating solvents for extraction, reduction and alkylation of glutenin were compared, and the need for acidification after alkylation for long-term storage was demonstrated. It was also demonstrated that computer analyses of chromatographic data can identify proteins correlated with quality.
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