Utilizing ¹³C-Labeled internal standards to advance the analysis of heparan sulfate

Heparan sulfate (HS) is a highly sulfated and structurally heterogeneous polysaccharide that plays key roles in numerous biological processes. Due to its complex structure and variable sulfation patterns, accurately characterizing and quantifying HS in biological samples poses significant analytical challenges. This review presents an advanced high-performance liquid chromatography-tandem mass spectroscopy (LC-MS/MS) methodology that utilizes isotope-labeled internal standards for the precise quantification of HS disaccharides and rare 3-O-sulfated tetrasaccharides, alongside monitoring 6-O-endosulfatase enzyme activity and the metabolism of synthetic HS oligosaccharides in biological systems. The combination of isotope-labeled standards with LC-MS/MS technology provides a powerful and sensitive approach for comprehensive analysis of HS modifications, offering valuable insights into HS metabolism and its alterations across various biological contexts.