Determination of sialic acids in biological fluids using reversed-phase ion-pair high-performance liquid chromatography

Citation
Pa. Siskos et Mhe. Spyridaki, Determination of sialic acids in biological fluids using reversed-phase ion-pair high-performance liquid chromatography, J CHROMAT B, 724(2), 1999, pp. 205-212
Citations number
31
Categorie Soggetti
Chemistry & Analysis
Journal title
JOURNAL OF CHROMATOGRAPHY B
ISSN journal
13872273 → ACNP
Volume
724
Issue
2
Year of publication
1999
Pages
205 - 212
Database
ISI
SICI code
1387-2273(19990319)724:2<205:DOSAIB>2.0.ZU;2-A
Abstract
A simple, rapid and sensitive reversed-phase ion-pair high-performance liqu id chromatographic method for the determination of N-acetylneuraminic acid and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in biological fluids is des cribed. Determination of N-acetylneuraminic acid released by acidic hydroly sis, in serum, urine and saliva, and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid in urine, without hydrolysis, was accomplished by injecting the sampl e without derivatization, into the chromatograph. Measurements were carried out isocratically within 6 min using a C-18 column and a mobile phase of a queous solution of triisopropanolamine, as ion-pair reagent, 60 mM, pH 3.5 at room temperature with UV absorbance detection. The present method is rep orted for the first time for the determination of sialic acids in biologica l fluids. Recoveries in serum, urine and saliva ranged from 90 to 102% and the limits of detection were 60 nM and 20 nM for the two sialic acids, resp ectively. The method has been applied to normal and pathological sera from patients with breast, stomach, colon, ovarian and cervix cancers, to normal urine and urine from patient with sialuria and to normal saliva. (C) 1999 Elsevier Science B.V. All rights reserved.