Method for determining the structure of oligosaccharide linkages and automated instrumentation thereof

An apparatus for use in determining the structure of a carbohydrate, comprising:

• at least one reaction vessel for reacting a carbohydrate sample with reagents to cleave the carbohydrates into subunits and introduce chromophoric entities onto the subunits;
• means for introducing the reagents into the reaction vessel;
• means for removing a sample from the reaction vessel;
• means for separating benzoates resulting from introduction of chromophoric entities onto the subunits;
• means for delivering a sample of the separated benzoates to each of a plurality of spectrophotometric instruments;
• means for determining the mass spectrum of a sample of the resulting benzoates;
• means for determining the ultraviolet spectrum of a sample of the resulting benzoates;
• means for determining the circular dichroic spectrum of a sample of the resulting benzoates;
• and
• means for comparing the mass, ultraviolet and circular dichroic spectra to reference spectra or calculated values in order to determine the structure of the sample carbohydrate.

A method for cleaving a carbohydrate comprising

• (a) perbenzoylating the carbohydrate with a perbenzoylating agent to protect free hydroxyl groups in the carbohydrate;
• (b) contacting the perbenzoylated carbohydrate with an amount of BrCH.sub.2 COBr/H.sub.2 O effective to cleave the carbohydrate;
• (c) treating the product of step (b) with AgOAc and methanol or AgOTf/TMU and methanol to effect glycosildation;
• and
• (d) treating the product of step (c) with thiourea to remove bromoacetate groups.