USE OF A POROUS GRAPHITIZED CARBON COLUMN FOR THE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF OLIGOSACCHARIDES, ALDITOLS AND GLYCOPEPTIDES WITH SUBSEQUENT MASS-SPECTROMETRY ANALYSIS

HPLC using a porous graphitised carbon (PGC) column eluted in acetonit rile-aqueous trifluoroacetic acid has been shown to give complementary chromatography to reversed-phase (ODS) HPLC for separation of peptide s and glycopeptides. The PGC column can also be used for separation of oligosaccharides and oligosaccharide alditols released from protein b y enzymes (N-linked chains) or base-borohydride degradation (0-linked chains). The advantages are that peptides, glycopeptides, reducing oli gosaccharides, sialylated oligosaccharides and oligosaccharide alditol s can be chromatographed under the same conditions. The samples can be readily recovered by evaporation for sensitive liquid secondary ion m ass spectrometric (LSI-MS) analysis and there is no contamination or d eterioration of chromatography from column leakage. LSI-MS analysis re vealed that complete peak separation of all of the possible oligosacch aride components of the standard glycoproteins fetuin and bovine subma xillary mucin was not achieved. However, PGC remains as a useful adjun ct to other HPLC profiling and separation techniques in particular whe re subsequent MS analysis is desired.