title: Quantitative analysis of glycosaminoglycans, chondroitin/dermatan sulfate, hyaluronic acid, heparan sulfate, and keratan sulfate by liquid chromatography-electrospray ionization-tandem mass spectrometry.
abstract: We developed a method using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) with a selected reaction monitoring (SRM) mode for simultaneous quantitative analysis of glycosaminoglycans (GAGs). Using one-shot analysis with our MS/MS method, we demonstrated the simultaneous quantification of a total of 23 variously sulfated disaccharides of four GAG classes (8 chondroitin/dermatan sulfates, 1 hyaluronic acid, 12 heparan sulfates, and 2 keratan sulfates) with a sensitivity of less than 0.5 pmol within 20 min. We showed the differences in the composition of GAG classes and the sulfation patterns between porcine articular cartilage and yellow ligament. In addition to the internal disaccharides described above, some saccharides derived from the nonreducing terminal were detected simultaneously. The simultaneous quantification of both internal and nonreducing terminal saccharides could be useful to estimate the chain length of GAGs. This method would help to establish comprehensive "GAGomic" analysis of biological tissues.
authors.affiliation: Department of Biochemistry, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan. Electronic address: biochem1@med.shimane-u.ac.jp.
authors.forename: Harumi
authors.initials: H
authors.lastname: Osago
authors.affiliation: Center for Integrated Research in Science, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Shibata
authors.affiliation: Department of Biochemistry, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Nobumasa
authors.initials: N
authors.lastname: Hara
authors.affiliation: Department of Orthopaedic Surgery, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Suguru
authors.initials: S
authors.lastname: Kuwata
authors.affiliation: Department of Orthopaedic Surgery, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Michihaya
authors.initials: M
authors.lastname: Kono
authors.affiliation: Department of Orthopaedic Surgery, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Uchio
authors.affiliation: Department of Biochemistry, Shimane University Faculty of Medicine, Izumo, Shimane 693-8501, Japan.
authors.forename: Mikako
authors.initials: M
authors.lastname: Tsuchiya
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D003871:Dermatan Sulfate / Q000031:analogs & derivatives / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis*; D006497:Heparan Sulfate / Q000032:analysis; D006820:Hyaluronic Acid / Q000032:analysis; D007632:Keratan Sulfate / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D007632:Keratan Sulfate; C069524:dermatan sulfate chondroitin sulfate
keywords: Electrospray ionization (ESI); Glycosaminoglycan (GAG); High-sulfated disaccharide; Selected reaction monitoring (SRM); Tandem mass spectrometry (MS/MS)