Determination of Individual Vitamin B<sub>6</sub> Compounds Based on Enzymatic Conversion to 4-Pyridoxolactone

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  • NISHIMURA Saki
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • NAGANO Sayaka
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • A. CRAI Chan
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • YOKOCHI Nana
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • YOSHIKANE Yu
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • GE Fei
    Department of Bioresources Science, Faculty of Agriculture, Kochi University
  • YAGI Toshiharu
    Department of Bioresources Science, Faculty of Agriculture, Kochi University

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  • Determination of individual vitamin B6 compounds based on enzymatic conversion to 4-pyridoxolactone

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A determination method for individual natural vitamin B6 compounds was developed. The vitamin B6 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-β-glucoside were hydrolyzed with HCl, and then determined as their free forms. Pyridoxine 5'-phosphate and pyridoxine-β-glucoside were not separately determined here. Three food samples were analyzed by this method.

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