MALDI-FTMS characterization of oligosaccharides labeled with 9-aminofluorene

Abstract

9-Aminofluorene (9AmFL) was investigated as an oligosaccharide label. The label was amenable to high UV detectability but did not interfere with mass spectrometric analysis. The 9AmFL label has high molar absorptivity (ε = 1.4 X 10⁴ L cm⁻¹ mol⁻¹ at λ = 267 nm), is chemically stable, and adds easily in reductive amination to the aldehyde terminus of oligosaccharides. Various linear and branched oligosaccharides were labeled with 9AmFL and the products were purified by chromatography on porous graphitized carbon (PGC). The derivatization reaction gave excellent yields (>95%). Up to 100-fold increase in UV sensitivity at λ = 206 nm, compared to the corresponding alditol, was observed. Mass spectra were recorded for the labeled compounds. In the presence of sodium dopant, series of Y- and B-fragments were observed. Protonation of the labeled compounds prior to mass spectrometric analysis resulted in simplified spectra (Y-fragments only) and allowed for complete sequence analysis. The retention of the positive charge at the label in the protonated species was consistent with the basicity of the amine. The smallest amount of labeled sugar to be detected by photo-diode array (PDA) was 5 pmol (λ = 267 nm).