Chiral amino acid analysis of Japanese traditional Kurozu and the developmental changes during earthenware jar fermentation processes

•Enantiomers of all proteinogenic amino acids plus allo forms in the Japanese traditional Kurozu vinegar were determined.•In addition to the widely observed d-Ala, d-Asp, d-Glu and d-Ser, considerable amounts of d-Leu and d-allo-Ile were found.•Developmental changes of these d-amino acids during fermentation and aging processes have also been clarified.•The present results are useful to design a highly functional natural beverage.

Enantioselective amino acid metabolome analysis of the Japanese traditional black vinegars (amber rice vinegar, Kurozu) was performed using two-dimensional high-performance liquid chromatography combining a microbore-monolithic ODS column and narrowbore-enantioselective columns. d-Amino acids, the enantiomers of widely observed l-amino acids, are currently paid attention as novel physiologically active substances, and the foodstuffs and beverages containing high amounts of d-amino acids are the subjects of interest. In the present study, the amino acid enantiomers were determined by two-dimensional HPLC techniques after pre-column fluorescence derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole. In the first dimension, the amino acid enantiomers are separated as their d plus l mixtures by the reversed-phase mode, then the d-amino acids and their l-counterparts are separately determined in the second dimension by the enantioselective columns. As a result, large amounts of d-Ala (800–4000 nmol/mL), d-Asp (200–400 nmol/mL) and d-Glu (150–500 nmol/mL) were observed in some of the traditionally produced Kurozu vinegars. Relatively large or small amounts of d-Ser (50–100 nmol/mL), d-Leu (10–50 nmol/mL) and d-allo-Ile (less than 20 nmol/mL) were also present in these samples. Developmental changes in the d-amino acid amounts during the fermentation and aging processes have also been investigated