title: FACE analysis as a fast and reliable methodology to monitor the sulfation and total amount of chondroitin sulfate in biological samples of clinical importance.
abstract: Glycosaminoglycans (GAGs) due to their hydrophilic character and high anionic charge densities play important roles in various (patho)physiological processes. The identification and quantification of GAGs in biological samples and tissues could be useful prognostic and diagnostic tools in pathological conditions. Despite the noteworthy progress in the development of sensitive and accurate methodologies for the determination of GAGs, there is a significant lack in methodologies regarding sample preparation and reliable fast analysis methods enabling the simultaneous analysis of several biological samples. In this report, developed protocols for the isolation of GAGs in biological samples were applied to analyze various sulfated chondroitin sulfate- and hyaluronan-derived disaccharides using fluorophore-assisted carbohydrate electrophoresis (FACE). Applications to biologic samples of clinical importance include blood serum, lens capsule tissue and urine. The sample preparation protocol followed by FACE analysis allows quantification with an optimal linearity over the concentration range 1.0-220.0 µg/mL, affording a limit of quantitation of 50 ng of disaccharides. Validation of FACE results was performed by capillary electrophoresis and high performance liquid chromatography techniques.
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, 21100 Varese, Italy. jenny.karousou@uninsubria.it.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Karousou
authors.affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece. athanasia_asimakopoulou@yahoo.gr.
authors.forename: Athanasia
authors.initials: A
authors.lastname: Asimakopoulou
authors.affiliation: Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, 27100 Pavia, Italy. luca.monti02@universitadipavia.it.
authors.forename: Luca
authors.initials: L
authors.lastname: Monti
authors.affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece. vickzaf@yahoo.gr.
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Zafeiropoulou
authors.affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece. nafratis@upatras.gr.
authors.forename: Nikos
authors.initials: N
authors.lastname: Afratis
authors.affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece. pgartaganis@yahoo.gr.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Gartaganis
authors.affiliation: Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, 27100 Pavia, Italy. antrossi@unipv.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rossi
authors.affiliation: Department of Surgical and Morphological Sciences, University of Insubria, 21100 Varese, Italy. alberto.passi@uninsubria.it.
authors.forename: Alberto
authors.initials: A
authors.lastname: Passi
authors.affiliation: Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece. n.k.karamanos@upatras.gr.
authors.forename: Nikos K
authors.initials: NK
authors.lastname: Karamanos
mesh_terms: D002809:Chondroitin Sulfates / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004586:Electrophoresis / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods; D006025:Glycosaminoglycans / Q000032:analysis
chemical_list: D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates