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.
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.
Hara
S
S
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Takemori
Y
Y
Yamaguchi
M
M
Nakamura
M
M
Ohkura
Y
Y
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D005732:Gangliosides / Q000032:analysis; D006017:Glycolipids / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D006736:Horses; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D008832:Microchemistry; D009077:Mucins / Q000032:analysis; D009438:Neuraminic Acids / Q000032:analysis* / Q000097:blood; D009961:Orosomucoid / Q000032:analysis; D011786:Quality Control; D051381:Rats; D012756:Sheep; D012794:Sialic Acids / Q000032:analysis* / Q000097:blood; D013050:Spectrometry, Fluorescence; D013552:Swine
D005732:Gangliosides; D006017:Glycolipids; D006023:Glycoproteins; D009077:Mucins; D009438:Neuraminic Acids; D009961:Orosomucoid; D012794:Sialic Acids; C032592:N-glycolylneuraminic acid