creators_name: Yang, Yifang creators_name: Breadmore, MC creators_name: Thormann, W creators_id: creators_id: mcb@utas.edu.au creators_id: type: article datestamp: 2008-06-05 12:49:59 lastmod: 2008-07-18 11:01:26 metadata_visibility: show title: Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking ispublished: pub subjects: 250000 subjects: 250401 subjects: 250400 full_text_status: restricted keywords: Chondroitin sulfate • Electrokinetic injection • Hyaluronic acid • Serum • Stacking note: Copyright © 2005 John Wiley and Sons abstract: CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di-HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 {micro}g/mL (0.2-1.0 {micro}g/mL for di-CS), with correlation coefficients being >0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis. date: 2005-11 date_type: published publication: Journal of Separation Science volume: 28 number: 17 pagerange: 2381-2389 id_number: 10.1002/jssc.200500181 refereed: TRUE issn: Print 1615-9306 official_url: http://dx.doi.org/10.1002/jssc.200500181 related_url_url: http://www3.interscience.wiley.com/journal/76510662/home related_url_type: pub referencetext: [1] Kakehi, K., Kinoshita, M., Yasueda, S., J. Chromatogr. B 2003, 797, 347-355. [2] Vynios, D. H., Karamanos, N. K., Tsiganos, C. P., J. Chromatogr. 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