Quantitative gas-liquid chromatography of neutral sugars in human serum glycoproteins. Fucose, mannose, and galactose as predictors in ovarian and small cell lung carcinoma

Charles W. Gehrke, T. Phillip Waalkes, Ernest Borek, Wade F. Swartz, Thomas F. Cole, Kenneth C. Kuo, Martin Abeloff, David S. Ettinger, Neil Rosenshein, Robert C. Young

Research output: Contribution to journalArticle

Abstract

A precise and accurate gas-liquid chromatographic (GLC) method has been developed for the quantitative analysis of the neutral sugars l-fucose (6-deoxygalactose), mannose, galactose, and glucose in ethanol precipitates of human serum proteins. The chromatographic conditions and sample preparation resulted in short analysis times (20 min per run) and made routine analyses practicable (twelve samples per day). The alditol acetate derivatization yielded single derivatives for each sugar. Complete separation was achieved on a 2.0 m × 2 mm I.D. column with 2.0% Silar-7 CP on Chromosorb W AW 80-100 mesh. The results of hydrolysis showed that the release of fucose and galactose preceded the release of mannose. Hydrolysis with AG 50W-X8 (H+) ion-exchange resin in 0.5 N HCI at 100° for 7 h optimized glycosidic bond cleavage with only minimal destruction of fucose, mannose and galactose. A combination of strong cation- and anion-exchange resin columns was used to remove chromatographic background of peptides, amino acids, amino sugars, and inorganic ions. An average R.S.D. of less than 4% with recovery of <86% for the three sugars was achieved. The homogeneity of the chromatographic peaks for the neutral sugars of normal human serum glycoproteins was confirmed by GLC-mass spectrometry. Significantly elevated ratios of fucose, galactose, and mannose to serum protein were observed for patients with small cell lung and ovarian carcinomas.

Original languageEnglish (US)
Pages (from-to)507-528
Number of pages22
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume162
Issue number4
DOIs
StatePublished - Apr 11 1979

ASJC Scopus subject areas

  • Chemistry(all)

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