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.
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.
Hara
S
S
Yamaguchi
M
M
Takemori
Y
Y
Nakamura
M
M
Ohkura
Y
Y
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D009438:Neuraminic Acids / Q000032:analysis* / Q000097:blood / Q000652:urine; D051381:Rats; D011919:Rats, Inbred Strains; D012794:Sialic Acids / Q000032:analysis* / Q000097:blood / Q000652:urine; D013056:Spectrophotometry, Ultraviolet
D009438:Neuraminic Acids; D012794:Sialic Acids; C032592:N-glycolylneuraminic acid