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Determination of individual vitamin B(6) compounds based on enzymatic conversion to 4-pyridoxolactone.

Saki S Nishimura, Sayaka S Nagano, Chan C A Crai, Nana N Yokochi, Yu Y Yoshikane, Fei F Ge, and Toshiharu T Yagi

J Nutr Sci Vitaminol (Tokyo) 54(1):18-24 (2008) PMID 18388403

A determination method for individual natural vitamin B(6) compounds was developed. The vitamin B(6) compounds were specifically converted into 4-pyridoxolactone (PAL), a highly fluorescent compound, through a combination of enzymatic reactions and HCl-hydrolysis. PAL was then determined by HPLC. Pyridoxal was completely oxidized to PAL with pyridoxal 4-dehydrogenase (PLDH). Pyridoxine and pyridoxamine were totally converted into PAL through a coupling reaction involving pyridoxine 4-oxidase and PLDH, and one involving pyridoxamine-pyruvate aminotransferase and PLDH, respectively. The 5'-phosphate forms and pyridoxine-beta-glucoside were hydrolyzed with HCl, and then determined as their free forms. Pyridoxine 5'-phosphate and pyridoxine-beta-glucoside were not separately determined here. Three food samples were analyzed by this method.

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