FACE Analysis as a Fast and Reliable Methodology to Monitor the Sulfation and Total Amount of Chondroitin Sulfate in Biological Samples of Clinical Importance

Molecules

Evgenia Karousou³  Athanasia Asimakopoulou¹  Luca Monti²  Vassiliki Zafeiropoulou¹  Nikos Afratis¹  Panagiotis Gartaganis¹  Antonio Rossi²  Alberto Passi³ 

[1] Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece; E-Mails:;Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, 27100 Pavia, Italy; E-Mails:;Department of Surgical and Morphological Sciences, University of Insubria, 21100 Varese, Italy

关键词: glycosaminoglycans;    chondroitin sulfate;    hyaluronan;    fluorophore-assisted carbohydrate electrophoresis;    capillary electrophoresis;    high performance liquid chromatography;   

DOI  :  10.3390/molecules19067959

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.