CARBOHYDRATE FRACTIONATION FACILITY

Project: Research project

Project Details

Description

The aim of this proposal is to establish a Carbohydrate Fractionation
Facility that uses the latest HPLC techniques for rapidly separating and
quantitating complex mixtures of monosaccharides, oligosaccharides and
glycopeptides with maximal resolution and sensitivity. The facility will
consist of three HPLC Systems designed to separate analytical or
quantitative amounts of carbohydrates on the basis of charge, size, lectin-
binding properties and anionic properties at high pH. A Rainin HPLC system
presently available to the group will be used for separations based on
charge and size. A Perkin Elmer HPLC system is requested to be dedicated
for use in lectin-affinity HPLC which separates carbohydrates according to
their abilities to bind different plant lectins. The third system in the
facility will be the Dionex Carbohydrate Gradient HPLC system, a novel
method developed recently, which allows rapid determination of as little as
0.1n mole complex carbohydrates and the separation of oligosaccharides with
identical composition that differ only in a single glycosidic linkage. The
Dionex system includes a pulsed amperometric detector (PAD) that is
extremely sensitive and will detect 10-100 pmoles carbohydrate. A second
PAD is requested for use with the Rainin HPLC system. This method of
detection cannot be applied to monitoring lectin-affinity HPLC since high
concentrations of sugars are used in the buffers. Therefore, a continuous
flow Berthold radiation detector is requested for efficient monitoring of
these columns. The combination of HPLC systems and detectors described
represents "state-of-the-art" technology in rapid and sensitive analysis of
complex mixtures of eukaryotic carbohydrates and would be enormously
helpful to the research programs described herein.
StatusFinished
Effective start/end date4/1/903/31/91

Funding

  • National Center for Research Resources

ASJC

  • Radiation

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