Fluorometric high-performance liquid chromatography of N-acetyl- and N-glycolylneuraminic acids and its application to their microdetermination in human and animal sera, glycoproteins, and glycolipids

Anal Biochem. 1987 Jul;164(1):138-45. doi: 10.1016/0003-2697(87)90377-0.

Abstract

A simple, rapid, and highly sensitive high-performance liquid chromatographic method is described for the determination of N-acetyl- and N-glycolylneuraminic acids in human and animal sera, glycoproteins, and glycolipids. The neuraminic acids, released by acid hydrolysis of these biological samples, are converted in dilute sulfuric acid with 1,2-diamino-4,5-methylene-dioxybenzene, a fluorogenic reagent for alpha-keto acids, to highly fluorescent derivatives. The derivatives are separated within 12 min on a reversed-phase column (Radial-Pak cartridge C18) with an isocratic elution and detected fluorometrically. The detection limits are 25 fmol (7.7 pg) for N-acetylneuraminic acid and 23 fmol (7.5 pg) for N-glycolylneuraminic acid in a 10-microliter injection volume at a signal-to-noise ratio of 2. This method permits precise determination of the neuraminic acids in 5 microliter of human and animal sera or in 0.25-2.5 micrograms of glycoproteins and glycolipids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid*
  • Gangliosides / analysis
  • Glycolipids / analysis*
  • Glycoproteins / analysis*
  • Horses
  • Hot Temperature
  • Humans
  • Mice
  • Microchemistry
  • Mucins / analysis
  • Neuraminic Acids / analysis*
  • Neuraminic Acids / blood
  • Orosomucoid / analysis
  • Quality Control
  • Rats
  • Sheep
  • Sialic Acids / analysis*
  • Sialic Acids / blood
  • Spectrometry, Fluorescence
  • Swine

Substances

  • Gangliosides
  • Glycolipids
  • Glycoproteins
  • Mucins
  • Neuraminic Acids
  • Orosomucoid
  • Sialic Acids
  • N-glycolylneuraminic acid