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

J Nutr Sci Vitaminol (Tokyo). 2008 Feb;54(1):18-24. doi: 10.3177/jnsv.54.18.

Abstract

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.

MeSH terms

  • Alcohol Oxidoreductases / chemistry*
  • Animals
  • Capsicum
  • Chickens
  • Chromatography, High Pressure Liquid / methods
  • Food Analysis / methods*
  • Garlic
  • Glucosides / analysis
  • Glucosides / chemistry
  • Hydrochloric Acid / chemistry
  • Hydrolysis
  • Pyridoxic Acid / analogs & derivatives*
  • Pyridoxic Acid / analysis
  • Pyridoxic Acid / chemical synthesis
  • Pyridoxine / analogs & derivatives
  • Pyridoxine / analysis
  • Pyridoxine / chemistry
  • Time Factors
  • Transaminases / chemistry*
  • Vitamin B 6 / analysis
  • Vitamin B 6 / chemistry
  • Vitamin B Complex / analysis*
  • Vitamin B Complex / chemistry

Substances

  • Glucosides
  • Vitamin B Complex
  • 5'-O-(glucopyranosyl)pyridoxine
  • 4-pyridoxic acid lactone
  • Pyridoxic Acid
  • Vitamin B 6
  • Alcohol Oxidoreductases
  • pyridoxal dehydrogenase
  • pyridoxine 4-oxidase
  • Transaminases
  • pyridoxamine-pyruvate transaminase
  • Pyridoxine
  • Hydrochloric Acid