title: A comprehensive compositional analysis of heparin/heparan sulfate-derived disaccharides from human serum.
abstract: The analysis of heparan sulfate glycosaminoglycans (HSGAGs) variations in human serum at the disaccharide level has a great potential for disease diagnosis and prognosis. However, the lack of available analytical methodology for the compositional analysis of HSGAGs in human serum remains to be addressed to delineate the possible role of HSGAGs on the onset and/or progression of a disease. In this study, we have developed a method for the in-depth compositional analysis of the 12 heparin/HS-derived disaccharides from human serum using a combination of technologies--fractionation, exhaustive digestion, solid phase extraction, and LC-MS/MS. The method exhibits high recovery (72-110%) and good reproducibility (standard deviation of less than 5%) with a low limit of detection and quantification. Errors from the method validation were within 1.1%. Nondetectable non- or low-sulfated disaccharides in human serum were also detected using the optimized protocol. Further applying this method, the comprehensive analysis of HSGAGs compositions in human sera from female donors showed considerable variations in disaccharide patterns and compositions.
authors.affiliation: Department of Chemistry, University of California, Davis, California 95616, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Milady R
authors.initials: MR
authors.lastname: Niñonuevo
authors.forename: Anish
authors.initials: A
authors.lastname: Sharma
authors.forename: Lieza M
authors.initials: LM
authors.lastname: Danan-Leon
authors.forename: Julie A
authors.initials: JA
authors.lastname: Leary
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004187:Disaccharides / Q000097:blood* / Q000302:isolation & purification; D005260:Female; D006493:Heparin / Q000097:blood / Q000737:chemistry*; D006497:Heparan Sulfate / Q000097:blood / Q000737:chemistry*; D006801:Humans; D008297:Male; D052616:Solid Phase Extraction / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D004187:Disaccharides; D006493:Heparin; D006497:Heparan Sulfate