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
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.