A Comprehensive Compositional Analysis of Heparin/Heparan Sulfate-Derived Disaccharides from Human Serum

摘要

ABSTRACT: The analysis of heparan sulfate glycosaminoglycans (HSGAGs)nvariations in human serum at the disaccharide level has a great potential for diseasendiagnosis and prognosis. However, the lack of available analytical methodology fornthe compositional analysis of HSGAGs in human serum remains to be addressed tondelineate the possible role of HSGAGs on the onset and/or progression of a disease.nIn this study, we have developed amethod for the in-depth compositional analysis ofnthe 12 heparin/HS-derived disaccharides fromhuman serumusing a combination ofntechnologies—fractionation, exhaustive digestion, solid phase extraction, and LCu0001MS/MS. The method exhibits high recoveryn(72u0001110%) and good reproducibility (standard deviation of less than 5%) with a low limit of detection and quantification. Errorsnfromthemethod validation were within 1.1%.Nondetectable non- or low-sulfated disaccharides in human serumwere also detectednusing the optimized protocol. Further applying this method, the comprehensive analysis of HSGAGs compositions in human seranfrom female donors showed considerable variations in disaccharide patterns and compositions.