豌豆种子中α-半乳糖苷分析的改进毛细管区带电泳方法。与高效液相色谱-三重脉冲安培检测法的比较。

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.

作者信息

Frias J, Price K R, Fenwick G R, Hedley C L, Sørensen H, Vidal-VALverde C

机构信息

Department of Applied Genetics, John Innes Centre, Norwich, UK.

出版信息

J Chromatogr A. 1996 Jan 5;719(1):213-9. doi: 10.1016/0021-9673(95)00236-7.

DOI:10.1016/0021-9673(95)00236-7

PMID:8589830

Abstract

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.

摘要

本文描述了一种毛细管区带电泳（CZE）方法，用于分析豌豆种子中的棉子糖系列寡糖（RFO）。RFO的提取在80%乙醇中进行，提取物通过Sep-Pak C18柱。这被证明是对目前公认的纯化技术的一种改进。获得了高质量的电泳图，可用于蔗糖、棉子糖、水苏糖和毛蕊花糖的分离和定量。将CZE结果与通过阴离子交换高效液相色谱-三脉冲安培检测（HPAC-PAD）获得的结果进行了比较。样品取自四个豌豆品系，除了r和rb位点的基因外，它们是近等基因系。所有豌豆品系的RFO组成都获得了高度的精密度和重现性。使用配对学生t检验，在两种分析技术之间未发现统计学上的显著差异（p≤0.05）。