Comparison of procainamide and 2-aminobenzamide labeling for profiling and identification of glycans by liquid chromatography with fluorescence detection coupled to electrospray ionization-mass spectrometry

Anal Biochem. 2015 Oct 1;486:38-40. doi: 10.1016/j.ab.2015.06.006. Epub 2015 Jun 12.

Abstract

One of the most widely used methods for glycan analysis is fluorescent labeling of released glycans followed by hydrophilic interaction chromatography-(ultra-)high-performance liquid chromatography [HILIC-(U)HPLC]. Here, we compare the data obtained by (U)HPLC-fluorescence (FLR) coupled to electrospray ionization-mass spectrometry (ESI-MS) for procainamide and 2-aminobenzamide (2-AB)-labeled N-glycans released from human immunoglobulin G (IgG). Fluorescence profiles from procainamide show comparable chromatographic separation to those obtained for 2-AB but gave higher fluorescence intensity as well as significantly improved ESI efficiency (up to 30 times that of 2-AB). Thus, labeling with procainamide increases the ability to identify minor glycan species that may have significant biological activity.

Keywords: ESI–MS; Fluorescent labeling; Glycan derivatization; HILIC–(U)HPLC; Procainamide.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Fluorescent Dyes / chemistry*
  • Humans
  • Polysaccharides / analysis*
  • Polysaccharides / chemistry*
  • Procainamide / chemistry*
  • Spectrometry, Fluorescence
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Staining and Labeling
  • ortho-Aminobenzoates / chemistry*

Substances

  • Fluorescent Dyes
  • Polysaccharides
  • ortho-Aminobenzoates
  • Procainamide
  • anthranilamide