Comparison of high performance liquid chromatographic, gas liquid chromatographic, and Saccharomyces uvarum methods for the determination of B{u2086} compounds

A high performance liquid chromatographic (HPLC) system consisting of an acetonitrile/phosphate buffer, a Spherisorb ODS column, and UV detector was used to separate the B₆ compounds pyridoxol, pyridoxal, pyridoxamine, and 4'-deoxypyridoxine. A gas chromatography (GC) equipped with a ⁶³Ni electron capture (EC) detector was used to separate the N-methyl-bis-trif luoroacetamide derivatives of the B₆ compounds on a l.54m x 2mm i.d. glass column packed with 10% SP2100 on Supelcoport 90-100 mesh at 125°C and an inlet pressure of 40 psig. Clean and successful separations of all the B₆ forms were obtained by HPLC and GC-EC. Total B₆ values as determined by HPLC and GC-EC for all 3 foods were higher than the corresponding total B₆ values as determined by S. uvarum assay. Several of the B₆ vitamer values for foods obtained by GC-EC agree with corresponding values obtained by HPLC. The HPLC method seemed to be the most satisfactory of the 3 methods for the quantitation of B₆ vitamers in foods and has the following advantages: increased sensitivity, method simplicity, and good precision.