TY - JOUR
T1 - Quantitative gas-liquid chromatography of neutral sugars in human serum glycoproteins. Fucose, mannose, and galactose as predictors in ovarian and small cell lung carcinoma
AU - Gehrke, Charles W.
AU - Waalkes, T. Phillip
AU - Borek, Ernest
AU - Swartz, Wade F.
AU - Cole, Thomas F.
AU - Kuo, Kenneth C.
AU - Abeloff, Martin
AU - Ettinger, David S.
AU - Rosenshein, Neil
AU - Young, Robert C.
N1 - Funding Information:
The authors wish to thank Dr. James Hueser, Medical Oncologist, for obtaining the normal female control samples through the local Chapter of the American Cancer Society. Special thanks are extended to Linda F’riedrich and Debi Whisenand for their expert technical assistance. Sincere appreciation is also expressed to the professional staff and graduate students of the Experiment Station Chemical Laboratories for their help and encouragement. The work was supported in part by a contract from the National Cancer Institute, Bethesda, Md., U.S.A. (Contract No. NIH NO1 CM 12323).
PY - 1979/4/11
Y1 - 1979/4/11
N2 - 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.
AB - 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.
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U2 - 10.1016/S0378-4347(00)81831-9
DO - 10.1016/S0378-4347(00)81831-9
M3 - Article
C2 - 528665
AN - SCOPUS:0018411768
SN - 0378-4347
VL - 162
SP - 507
EP - 528
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - 4
ER -