Separation of anionic oligosaccharides by high-performance liquid chromatography

Journal Article · Wed Oct 01 00:00:00 EDT 1986 · Anal. Biochem.; (United States)

DOI:https://doi.org/10.1016/0003-2697(86)90585-3· OSTI ID:6943381

Green, E D; Baenziger, J U

The authors have developed methods for rapid fractionation of anionic oligosaccharides containing sulfate and/or sialic acid moieties by high-performance liquid chromatography (HPLC). Ion-exchange HPLC on amine-bearing columns (Micropak AX-10 and AX-5) at pH 4.0 is utilized to separate anionic oligosaccharides bearing zero, one, two, three, or four charges, independent of the identity of the anionic moieties (sulfate and/or sialic acid). Ion-exchange HPLC at pH 1.7 allows separation of neutral, mono-, di-, and tetrasialylated, monosulfated, and disulfated oligosaccharides. Oligosaccharides containing three sialic acid residues and those bearing one each of sulfate and sialic acid, however, coelute at pH 1.7. Since the latter two oligosaccharide species separate at pH 4.0, analysis at pH 4.0 followed by analysis at pH 1.7 can be utilized to completely fractionate complex mixtures of sulfated and sialylated oligosaccharides. Ion-suppression amine adsorption HPLC has previously been shown to separate anionic oligosaccharides on the basis of net carbohydrate content (size). In this study they demonstrate the utility of ion-suppression amine adsorption HPLC for resolving sialylated oligosaccharide isomers which differ only in the linkages of sialic acid residues (..cap alpha..2,3 vs ..cap alpha..2,6) and/or location of ..cap alpha..2,3- and ..cap alpha..2,6-linked sialic acid moieties on the peripheral branches of oligosaccharides. These two methods can be used in tandem to separate oligosaccharides, both analytically and preparatively, based on their number, types, and linkages of anionic moieties.

• Research Organization: Washington Univ. Medical School, St. Louis, MO
• OSTI ID: 6943381
• Journal Information: Anal. Biochem.; (United States), Vol. 158:1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
OLIGOSACCHARIDES
ION EXCHANGE CHROMATOGRAPHY
LIQUID COLUMN CHROMATOGRAPHY
TRITIUM COMPOUNDS
CLOSTRIDIUM PERFRINGENS
GLUCOPROTEINS
BACTERIA
CARBOHYDRATES
CHROMATOGRAPHY
CLOSTRIDIUM
LABELLED COMPOUNDS
MICROORGANISMS
ORGANIC COMPOUNDS
PROTEINS
SACCHARIDES
SEPARATION PROCESSES
550201* - Biochemistry- Tracer Techniques