May 28, 2013
Reversed-phase UHPLC/HPLC is routinely used for peptide mapping, but if the digest contains hydrophilic peptides, such as glycopeptides, valuable information can be missed. This note demonstrates peptide mapping of digested glycoprotein erythropoietin (EPO) protein using an Agilent ZORBAX Rapid Resolution High Definition 300-HILIC 1.8 ?m LC column and a TOF MS. Taking advantage of the high organic solvent system of the mobile phase for HILIC (hydrophilic interaction chromatography), the glycopeptides are effectively dissolved, retained and separated by the HILIC column. This app note includes the separation, sequence coverage and comparison of reversed-phase and HILIC data to demonstrate the utility of HILIC as an orthogonal and complementary approach to RPLC/MS for peptide analysis.