基于酶促转化为4-吡哆醇内酯对个体维生素B6化合物的测定

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

作者信息

Nishimura Saki, Nagano Sayaka, A Crai Chan, Yokochi Nana, Yoshikane Yu, Ge Fei, Yagi Toshiharu

机构信息

Department of Bioresources Science, Faculty of Agriculture, Kochi University, Kochi, Japan.

出版信息

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

DOI:10.3177/jnsv.54.18

PMID:18388403

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.

摘要

开发了一种用于测定个体天然维生素B(6)化合物的方法。通过酶促反应和盐酸水解相结合，将维生素B(6)化合物特异性地转化为4-吡哆醇内酯（PAL），一种高荧光化合物。然后通过高效液相色谱法测定PAL。吡哆醛用吡哆醛4-脱氢酶（PLDH）完全氧化为PAL。吡哆醇和吡哆胺分别通过涉及吡哆醇4-氧化酶和PLDH的偶联反应以及涉及吡哆胺-丙酮酸转氨酶和PLDH的偶联反应完全转化为PAL。5'-磷酸形式和吡哆醇-β-葡萄糖苷用盐酸水解，然后以其游离形式进行测定。此处未分别测定5'-磷酸吡哆醇和吡哆醇-β-葡萄糖苷。用该方法分析了三个食品样品。