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Fluorometric Determination of N-Acetyl and N-Glycolyl Neuraminic Acids by High Performance Liquid Chromatography as Their 4′-Hydrazino-2-stilbazole Derivatives

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Abstract

A sensitive and specific method for the determination of N-acetylneuraminic acid and N-glycolylneuraminic acid was developed by using high performance liquid chromatography with fluorometric detection. Both neuraminic acids were derivatized with 4′-hydrazino-2-stilbazole, a fluorogenic reagent for α-keto acid. The derivatives were separated by reversed phase chromatography on a Radial PAK Cartridge C18 Column (Waters) with methanol-aqueous 1.5% triethylamine solution adjusted the pH to 2.3 with phosphoric acid (35:65 v/v) as a mobile phase at a flow rate of 1 ml/min. The detection limits of both derivatives were 0.5 ng in an injection volume of 5 μl. This method was applied to the determination of NANA in human blood and urine.

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Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, Chiba University, Yayoi-cho, Chiba, 260, Japan

    Kazunori Kobayashi, Yumi Akiyama, Kayo Kawaguchi, Shinzo Tanabe & Toshio Imanari

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  1. Kazunori Kobayashi
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  2. Yumi Akiyama
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  3. Kayo Kawaguchi
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  4. Shinzo Tanabe
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  5. Toshio Imanari
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Kobayashi, K., Akiyama, Y., Kawaguchi, K. et al. Fluorometric Determination of N-Acetyl and N-Glycolyl Neuraminic Acids by High Performance Liquid Chromatography as Their 4′-Hydrazino-2-stilbazole Derivatives. ANAL. SCI. 1, 81–84 (1985). https://doi.org/10.2116/analsci.1.81

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  • DOI: https://doi.org/10.2116/analsci.1.81

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