High-Performance Liquid Chromatographic Determination of Chiral Amino Acids Using Pre-Column Derivatization with <i>o</i>-Phthalaldehyde and <i>N</i>-<i>tert</i>-Butyloxycarbonyl-D-cysteine and Application to Vinegar Samples

Aogu Furusho, Mintranee Obromsuk, Takeyuki Akita, Masashi Mita, Masanobu Nagano, Pornchai Rojsitthisak, Kenji Hamase

Research output: Contribution to journalArticlepeer-review

Abstract

<p>A reversed-phase high-performance liquid chromatographic (HPLC) method using pre-column derivatization with <i>o</i>-phthalaldehyde (OPA) plus <i>N</i>-<i>tert</i>-butyloxycarbonyl-D-cysteine (Boc-D-Cys) has been developed for the determination of aspartic acid (Asp), serine (Ser) and alanine (Ala) enantiomers. D-Amino acids in the real world samples are trace in most cases, and their small peaks should be eluted faster than the huge peaks of the L-forms in order to avoid overlapping. Amino acids were rapidly derivatized at room temperature with OPA plus Boc-D-Cys under simple conditions and were detected by their fluorescence. The target amino acid enantiomers were separated within 60 min on a reversed-phase column, CAPCELL PAK C18 MG II (4.6 x 200 mm), and their resolution values were higher than 2.14. The developed system was successfully validated using standard amino acids, and sufficient calibration lines (<i>r</i><sup>2</sup> > 0.9983) and precision (RSD < 5.29%) results were obtained. In a Japanese traditionally fermented amber rice vinegar, all of the target D-amino acids were observed, and their %D values were 21.5 for Asp, 6.8 for Ser and 22.9 for Ala.</p>
Original languageEnglish
Pages (from-to)147-151
Number of pages5
JournalCHROMATOGRAPHY
Volume41
Issue number3
DOIs
Publication statusPublished - 2020

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