Deuterated hydrazino-s-triazine as highly-efficient labelling reagent for glycan relative quantification analysis using electrospray ionization mass spectrometry

Herein, an innovative stable-isotope relative quantification strategy for N-glycans was achieved using a self-designed non-reductive hydrazino-s-triazine deuterated derivative as a labelling reagent combined with mass spectrometry. As much as a 20 Da mass shift could effectively distinguish different forms of N-glycans labelled with normal and heavy hydrazino-s-triazine at the reducing end. Especially for glycans with high molecular weight, qualitative identification could be significantly simplified on account of avoiding isotopic distributions overlapping in the mass spectra. Meanwhile, a deuterium derivative with high purity guaranteed the accuracy of the relative quantification. The obviously undifferentiated response toward normal and heavy hydrazino-s-triazine labelled glycans in electrospray ionization mass spectrometry confirmed the feasibility and reliability of our proposed strategy, which was further demonstrated by relative quantification of the mixtures of labelled N-glycans cleaved from ovalbumin as a model sample. Finally, we adopted human serum as a complex sample and successfully achieved highly accurate detection of 48 N-glycans. The deuterated hydrazino-s-triazine labelling reagent exhibited a great potential to promote the development of highly-efficient, accurate and reliable N-glycan relative quantification in pharmacy and diagnosis research.