A comprehensive HPLC analytical system for the identification and quantification of hexoses that employs 2-aminobenzamide coupling

Kayo Hasehira, Shin Ichi Nakakita, Nobumitsu Miyanishi, Wataru Sumiyoshi, Shie Hayashi, Kaoru Takegawa, Jun Hirabayashi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Rare sugars are defined as monosaccharides with extremely low natural abundances. Their natural distributions and biological functions remain to be clarified. To establish a robust analytical system that can separate, identify and quantify rare sugars, 12 d-hexoses - including five rare aldohexoses and three rare ketohexoses - were labelled with 2-aminobenzamide (2-AB), and the fluorescently tagged monosaccharides were separated by high-performance liquid chromatography (HPLC). Because the ketohexoses were much less reactive than were the aldohexoses, the reaction conditions were optimized to achieve the maximum yields (>75) for both aldohexoses and ketohexoses. The calibration curve determined for the rare ketohexose, d-psicose (Psi), was linear between 1 pmol and 1 mol and had a correlation coefficient of 0.9999. Using the developed method, the psicose content in a leaf of Itea virginica, in which the presence of psicose has previously been reported, was found to be 2.7 mg psicose/g leaf. The result proved feasibility of the method even for natural products. Because the system is a comprehensive tool, it should help answer questions concerning the biosyntheses and functions of rare hexose sugars.

Original languageEnglish
Pages (from-to)501-509
Number of pages9
JournalJournal of biochemistry
Volume147
Issue number4
DOIs
Publication statusPublished - Apr 1 2010

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Hexoses
High performance liquid chromatography
High Pressure Liquid Chromatography
Sugars
Monosaccharides
Biosynthesis
Biological Products
Calibration
anthranilamide
psicose

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

A comprehensive HPLC analytical system for the identification and quantification of hexoses that employs 2-aminobenzamide coupling. / Hasehira, Kayo; Nakakita, Shin Ichi; Miyanishi, Nobumitsu; Sumiyoshi, Wataru; Hayashi, Shie; Takegawa, Kaoru; Hirabayashi, Jun.

In: Journal of biochemistry, Vol. 147, No. 4, 01.04.2010, p. 501-509.

Research output: Contribution to journalArticle

Hasehira, Kayo ; Nakakita, Shin Ichi ; Miyanishi, Nobumitsu ; Sumiyoshi, Wataru ; Hayashi, Shie ; Takegawa, Kaoru ; Hirabayashi, Jun. / A comprehensive HPLC analytical system for the identification and quantification of hexoses that employs 2-aminobenzamide coupling. In: Journal of biochemistry. 2010 ; Vol. 147, No. 4. pp. 501-509.
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