Cereal Chem 72:217-221  |  VIEW ARTICLE

Analysis of Free and Glycosylated Vitamin B6 in Wheat by High- Performance Liquid Chromatography.

D. A. Sampson, L. A. Eoff, X. L. Yan, and K. Lorenz. Copyright 1995 by the American Association of Cereal Chemists, Inc. 

We developed an high-performance liquid chromatography (HPLC) method for analysis of vitamin B6 in wheat, based on modifications of existing reverse-phase ion-pairing procedures. The method is much simpler than multistep HPLC-based procedures required for exhaustive analysis of all B6 forms in foods, including minor components, or methods based on microbiological analysis. The method is suitable for analysis of major B6 forms in wheat, including pyridoxine glucoside. The method entails extraction of wheat B6 using metaphosphoric acid followed by quantitation of B6 in a single binary-gradient HPLC separation. Analysis of three randomly selected wheat cultivars with this method revealed significant variation in content of pyridoxine (2.2-3.3 nmol/g) and pyridoxine glucoside (1.8-9.4 nmol/g). The method should facilitate analysis of different wheat cultivars for vitamin B6 content.