The evaluation of a novel approach for the profiling and identification of N-linked glycan with a procainamide tag by HPLC with fluorescent and mass spectrometric detection

J Pharm Biomed Anal. 2010 Nov 2;53(3):315-24. doi: 10.1016/j.jpba.2010.03.045. Epub 2010 Apr 7.

Abstract

Procainamide was investigated as a multifunctional oligosaccharide label for glycan profiling and identification in a HPLC-FL/ESI-QTOF system. Addition of this aromatic amine to glycans through reductive amination improves fluorescence detection and ESI ionization efficiency. Both procainamide and 2-AB derivatives of N-linked glycans released from three glycoproteins (Human IgG, Mouse IgG, and RNase B) were quantitatively profiled with HPLC-FL and identified with ESI-QTOF. The procainamide derivatives produced FL glycan profiles comparable to the 2-AB derivatives, but with a few extra minor peaks, which suggests better labeling efficiency for procainamide derivatives for minor peaks. The procainamide derivatives also improve ESI ionization efficiency by 10-50 times over the respective 2-AB derivatives and the ESI-QTOF method sensitivity is at the low picomole to high femtomole level. Using the procainamide tag, all N-linked glycans released from three tested glycoproteins can be quantitatively detected with HPLC-FL and identified with ESI-QTOF at the same time. Monosaccharide sequence confirmation was also demonstrated in this study.

MeSH terms

  • Animals
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Mice
  • Monosaccharides / chemistry
  • Polysaccharides / analysis*
  • Polysaccharides / chemistry
  • Procainamide / chemistry*
  • Spectrometry, Fluorescence / methods*
  • Spectrometry, Mass, Electrospray Ionization / methods*

Substances

  • Monosaccharides
  • Polysaccharides
  • Procainamide