Improved method for the analysis of alpha-galactosides in pea seeds by capillary zone electrophoresis. Comparison with high-performance liquid chromatography-triple-pulsed amperometric detection.
A capillary zone electrophoretic (CZE) method is described for the analysis of the raffinose family of oligosaccharides (RFO) in pea seeds. Extraction of RFO was carried out in 80% ethanol and the extract was passed through a Sep-Pak C18 cartridge. This proved to be an improvement on the currently accepted purification technique. High-quality electrophoregrams were obtained which allowed the separation and quantitation of sucrose, raffinose, stachyose and verbascose The CZE results were compared with those obtained by anion-exchange high-performance liquid chromatography coupled to a triple-pulsed amperometric detection (HPAC-PAD). The samples were obtained from four pea strains which were near-isogenic except for genes at the r and rb loci. A high degree of precision and reproducibility was obtained for the RFO compositions of all the pea strains. No statistically significant differences (p < or equal to 0.05) were found between the two analytical techniques using paired Student-t tests.
Frias
J
J
Department of Applied Genetics, John Innes Centre, Norwich, UK.
Price
K R
KR
Fenwick
G R
GR
Hedley
C L
CL
Sørensen
H
H
Vidal-VALverde
C
C
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007887:Fabaceae / Q000737:chemistry*; D005697:Galactosides / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis; D010946:Plants, Medicinal; D011887:Raffinose / Q000032:analysis; D012639:Seeds / Q000737:chemistry*; D013395:Sucrose / Q000032:analysis
D005697:Galactosides; D009844:Oligosaccharides; C005695:stachyose; D013395:Sucrose; D011887:Raffinose