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

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 derlvatized with 4'-hydrazino-2-stilbazole, a fluorogenic reagent for a-keto acid. The derivatives were separated by reversed phase chromatography on a Radial PAK Cartridge Cig 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 nl. This method was applied to the determination of NANA in human blood and urine. © 1985, The Japan Society for Analytical Chemistry. All rights reserved.