Parameterization of contribution of sugar units to elution volumes in reverse-phase HPLC of 2-pyridylaminated oligosaccharides.

The reverse-phase high-performance liquid chromatography elution data, expressed in glucose units, of 2-pyridylaminated oligosaccharides (PA-oligosaccharides) originated from N-glycosides of glycoproteins have been analyzed with multiple regression to obtain parameters of contribution ascribable to each monosaccharide unit. For the best results, the known PA-oligosaccharides were classified into M-series (the "high mannose" type), X-series (those containing xylose), F-series (those containing L-Fuc alpha (1,6)GlcNAc-PA), and Z-series (all others). Within each series, a variable ("unit contribution") is assigned to every sugar component in the oligosaccharide, and the elution value of a given oligosaccharide was assumed to be the sum total of the unit contributions. The unit contribution obtained from such multiple regression calculations led to a set of elution values for oligosaccharides in each series, which showed very good agreement with the observed elution values. These unit contribution values are useful in predicting the elution value when the exact structure is known, and they also aid in predicting the structure from the known elution value.