title: Development of a rapid method for simultaneous separation of hyaluronic acid, chondroitin sulfate, dermatan sulfate and heparin by capillary electrophoresis.
abstract: This study reports the use of diethylenetriamine as background electrolyte for the simultaneous separation of hyaluronan acid, chondroitin sulfate, dermatan sulfate and heparin. The analytes were baseline separated by using an uncoated fused silica capillary at 37°C with a run time of 23min. The migration order, with hyaluronan acid at first and heparin at last, was related to the sulfation degree. The effect of salt concentration on resolution and migration order was also investigated. The developed method was applied to the simultaneous determination of hyaluronan acid and chondroitin sulfate in mouse plasma. Interferences in plasma were removed by protein precipitation and glycosaminoglycans were further purified by ethanol precipitation. The method was validated over the concentration range from 50 to 600μg/mL for hyaluronan acid and 500 to 6000μg/mL for chondroitin sulfate in mouse plasma. Results from assay validations showed that the method was selective and robust.
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Zhao
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Xinlei
authors.initials: X
authors.lastname: Song
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Tan
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Linghua
authors.initials: L
authors.lastname: Xu
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Mingxiu
authors.initials: M
authors.lastname: Yu
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Siyi
authors.initials: S
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China. Electronic address: liuxium@sdu.edu.cn.
authors.forename: Xiumei
authors.initials: X
authors.lastname: Liu
authors.affiliation: School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China; Shandong Provincial Key Laboratory of Carbohydrate Chemistry and Glycobiology, Jinan 250012, China. Electronic address: fswang@sdu.edu.cn.
authors.forename: Fengshan
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods; D002809:Chondroitin Sulfates / Q000032:analysis*; D003871:Dermatan Sulfate / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000032:analysis*; D006820:Hyaluronic Acid / Q000032:analysis*; D051379:Mice; D010949:Plasma / Q000737:chemistry*; D011073:Polyamines / Q000737:chemistry
chemical_list: D011073:Polyamines; C005391:diethylenetriamine; D003871:Dermatan Sulfate; D006820:Hyaluronic Acid; D006493:Heparin; D002809:Chondroitin Sulfates
keywords: Capillary electrophoresis; Glycosaminoglycans; Plasma; Simultaneous separation