Sample Clean-Up and Analysis of N-Acetyl and N-Glycolylneuraminic Acids in Blood Serum and Tissue Specimen by HPLC
作者:
T. Tsegenidis,
N.K. Karamanos,
期刊:
Journal of Liquid Chromatography & Related Technologies
(Taylor Available online 1998)
卷期:
Volume 21,
issue 6
页码: 793-802
ISSN:1082-6076
年代: 1998
DOI:10.1080/10826079808000509
出版商: Taylor & Francis Group
数据来源: Taylor
摘要:
A procedure for sample clean-up, isolation of glycoconjugates, and analysis of the two major sialic acids, Neu5Ac and Neu5Gc, as per-O-benzoylated derivatives in biologic samples is described. In blood sera, the bulk of low molecular weight molecules was efficiently removed by precipitation of sialic acids containing glycoconjugates with saturated ammonium sulphate, pH 10, at 0°C, centrifugation, and gel chromatography on a Sephadex G-25. The tissue specimen was homogenized with water at 0°C and following centrifugation the water-soluble sialic acids containing glycoconjugates were recovered in the supernatant. Sialic acids were liberated from their isolated macromolecules by acidic hydrolysis with 25 mM trifluoroacetic acid at 80°C for 2 h. Constituents with positive charged groups and neutral monosaccharides were removed by ion-exchange chromatographies on Dowex 50X8 and Dowex 1X8, whereas sialic acids were eluted with 2M formic acid.
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