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Clinical Chemistry 21: 1314-1322, 1975;
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Clinical Chemistry, Vol 21, 1314-1322, Copyright © 1975 by the American Association for Clinical Chemistry

Liquid-Chromatographic Analysis for Neutral Carbohydrates in Serum Glycoproteins

John E. Mrochek 1, Stanley R. Dinsmore 1, and T. Phillip Waalkes 2

1 Oak Ridge National Laboratory, Oak Ridge, Tenn. 37830 (Operated for the Energy Research and Development Administration by the Union Carbide Corp.).
2 Laboratory of Chemical Pharmacology, National Cancer Institute, NIH, Bethesda, Md. 20014.

We describe a sensitive, reproducible procedure of analysis for the six neutral carbohydrates in glycoproteins, by high-resolution anion-exchange chromatography. As many as 16 neutral carbohydrates can be separated by elution with a concentration gradient of boric acid (pH 7, 67 to 672 mmol/liter). The carbohydrates are detected with a cerate oxidimetric detector system, which monitors the fluorescence of Ce3+ produced by the reaction of the eluted constituents with Ce4+. Sensitivity to 1 nmol of fucose is demonstrated. Analytical methods and results are presented for mannose, fucose, and galactose in serum glycoproteins of both normal women and those with metastatic breast cancer. We briefly discuss the possibility of separating and analyzing for the three neutral carbohydrates in serum glycoproteins in 4 h by isocratic (constant eluent concentration) elution from a chromatographic column.


Key Words: cerate oxidimetric detector • metastatic breast cancer • anion-exchange chromatography in borate media • values for sera of normal women

Submitted on April 21, 1975
Accepted on May 12, 1975






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Copyright © 1975 by the American Association for Clinical Chemistry.