An ultrafiltration technique coupled with liquid chromatography and spectrofluorimetric (λ(ex) = 373 nm, λ(em) = 448 nm) detection is described for rapidly assaying free N-acetylneuraminic acid (Neu5Ac) in the serum of normal and cancer patients. Free Neu5Ac was obtained from serum after ultrafiltration hy centrifugation through an Amicon Centrifree micropartition filter which has a relative molecular mass cut-off of 1000 daltons. Aspects of the ultrafiltration procedure include the extent of non-specific binding of Neu5Ac to the ultrafilters, precision of recovery and the effect of temperature. Non-specific losses of Neu5Ac in serum to the ultrafilter system were low (recovery > 91.2%). The detection limit (S/N = 3) = 2 ng/ml. For free Neu5Ac at 180-1421 ng ml-1, the intra- and inter-day C.V.s for the ultrafiltration method were ≤ 2.8% and ≤ 3.9%, respectively. Free Neu5Ac concentrations measured by this method were compared to those measured by the acetonitrile procedure. The results showed an average value of 245 ± 90 ng ml-1for 30 serum samples of normal subjects. The average value for 20 lung cancer patients was found to be 996 ± 310 ng ml-1.

Determination of free N-acetylneuraminic acid by ultrafiltration and liquid chromatography in the serum of normal and cancer patients / Lagana', Aldo; Marino, Aldo; Fago, Giovanna; B., Pardo Martinez. - In: ANALYTICA CHIMICA ACTA. - ISSN 0003-2670. - STAMPA. - 306:1(1995), pp. 65-71. [10.1016/0003-2670(94)00577-9]

Determination of free N-acetylneuraminic acid by ultrafiltration and liquid chromatography in the serum of normal and cancer patients

LAGANA', Aldo;MARINO, Aldo;FAGO, Giovanna;
1995

Abstract

An ultrafiltration technique coupled with liquid chromatography and spectrofluorimetric (λ(ex) = 373 nm, λ(em) = 448 nm) detection is described for rapidly assaying free N-acetylneuraminic acid (Neu5Ac) in the serum of normal and cancer patients. Free Neu5Ac was obtained from serum after ultrafiltration hy centrifugation through an Amicon Centrifree micropartition filter which has a relative molecular mass cut-off of 1000 daltons. Aspects of the ultrafiltration procedure include the extent of non-specific binding of Neu5Ac to the ultrafilters, precision of recovery and the effect of temperature. Non-specific losses of Neu5Ac in serum to the ultrafilter system were low (recovery > 91.2%). The detection limit (S/N = 3) = 2 ng/ml. For free Neu5Ac at 180-1421 ng ml-1, the intra- and inter-day C.V.s for the ultrafiltration method were ≤ 2.8% and ≤ 3.9%, respectively. Free Neu5Ac concentrations measured by this method were compared to those measured by the acetonitrile procedure. The results showed an average value of 245 ± 90 ng ml-1for 30 serum samples of normal subjects. The average value for 20 lung cancer patients was found to be 996 ± 310 ng ml-1.
1995
Cancer patients; Fluorimetry; Liquid chromatography; N-acetylneuramic acid; Neu5Ac; Serum
01 Pubblicazione su rivista::01a Articolo in rivista
Determination of free N-acetylneuraminic acid by ultrafiltration and liquid chromatography in the serum of normal and cancer patients / Lagana', Aldo; Marino, Aldo; Fago, Giovanna; B., Pardo Martinez. - In: ANALYTICA CHIMICA ACTA. - ISSN 0003-2670. - STAMPA. - 306:1(1995), pp. 65-71. [10.1016/0003-2670(94)00577-9]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/455804
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