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Highly sensitive determination of N-acetyl- and N-glycolylneuraminic acids in human serum and urine and rat serum by reversed-phase liquid chromatography with fluorescence detection.

Abstract
A simple, rapid and highly sensitive high-performance liquid chromatographic method for the determination of N-acetyl- and N-glycolylneuraminic acids in serum and urine is described. The neuraminic acids, released by hydrolysis of serum and urine, are converted in dilute sulphuric acid with 1,2-diamino-4,5-dimethoxybenzene, a fluorogenic reagent for alpha-keto acids, to highly fluorescent derivatives. The derivatives are separated isocratically within 8 min by reversed-phase chromatography using a Radial-Pak cartridge C18 column and detected fluorimetrically. The limit of detection is 40 fmol (12 pg) for both neuraminic acids in 10-microliters injection volume [0.3 nmol (90 ng)/ml) of serum or urine]. This sensitivity permits the precise determination of the neuraminic acids in 5 microliters of serum or urine. The method was applied to the determination of the neuraminic acids in sera from normal subjects and cancer patients, normal urine and rat serum.
AuthorsS Hara, M Yamaguchi, Y Takemori, M Nakamura, Y Ohkura
JournalJournal of chromatography (J Chromatogr) Vol. 377 Pg. 111-9 (Apr 25 1986) Netherlands
PMID3711202 (Publication Type: Journal Article)
Chemical References
  • Neuraminic Acids
  • Sialic Acids
  • N-glycolylneuraminic acid
Topics
  • Animals
  • Chromatography, High Pressure Liquid
  • Humans
  • Liver Neoplasms (metabolism)
  • Neuraminic Acids (analysis, blood, urine)
  • Rats
  • Rats, Inbred Strains
  • Sialic Acids (analysis, blood, urine)
  • Spectrophotometry, Ultraviolet

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