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A capillary zone electrophoresis method for determining N-acetylneuraminic acid in glycoproteins and blood sera
dc.creator | Strousopoulou, K. | en |
dc.creator | Militsopoulou, M. | en |
dc.creator | Lamari, F. N. | en |
dc.creator | Karamanos, N. K. | en |
dc.date.accessioned | 2015-11-23T10:49:03Z | |
dc.date.available | 2015-11-23T10:49:03Z | |
dc.date.issued | 2002 | |
dc.identifier | 10.1002/bmc.143 | |
dc.identifier.issn | 2693879 | |
dc.identifier.uri | http://hdl.handle.net/11615/33470 | |
dc.description.abstract | A simple capillary zone electrophoresis (CZE) method for the determination of the content of the major sialic acid form N-acetylneuraminic acid (Neu5Ac) in glycoproteins was established. The present method utilizes a simplified hydrolysis-purification procedure consisting of mild acid hydrolysis (25 mM trifluoroacetic acid for 2 h at 80°C) to release Neu5Ac and ultrafiltration on Centricon-3 membrane to remove the obtained asialoglycoproteins and other macromolecules present in biologic samples. Derivatization with benzoic anhydride at 80°C for 20 min resulted in complete conversion of Neu5Ac to per-O-benzoylated Neu5Ac. CZE analysis was performed using the operating buffer 25 mM phosphate, pH 3.5, containing 50% (v/v) acetonitrile as organic modifier at 30 kV, and detection of the per-O-benzoylated Neu5Ac at 231 nm. The method showed excellent repeatability (RDS < 1.98%) and a linearity range from 5 μg/mL to 5 mg/mL with a detection limit of 2 μM. Application of the method to microanalysis of human α1-acid glycoprotein and blood serum samples showed excellent agreement with previously published values, suggesting a high precision for the developed CZE method. Copyright © 2002 John Wiley & Sons, Ltd. | en |
dc.source.uri | http://www.scopus.com/inward/record.url?eid=2-s2.0-0036194627&partnerID=40&md5=0ca06a5a43ed8a1aee128c46747c0500 | |
dc.subject | acetonitrile | en |
dc.subject | acid anhydride | en |
dc.subject | asialoglycoprotein | en |
dc.subject | benzene derivative | en |
dc.subject | benzoic acid | en |
dc.subject | buffer | en |
dc.subject | glycoprotein | en |
dc.subject | n acetylneuraminic acid | en |
dc.subject | organic compound | en |
dc.subject | orosomucoid | en |
dc.subject | phosphate | en |
dc.subject | sialic acid | en |
dc.subject | trifluoroacetic acid | en |
dc.subject | accuracy | en |
dc.subject | analytic method | en |
dc.subject | article | en |
dc.subject | blood analysis | en |
dc.subject | capillary zone electrophoresis | en |
dc.subject | carbohydrate analysis | en |
dc.subject | controlled study | en |
dc.subject | derivatization | en |
dc.subject | human | en |
dc.subject | hydrolysis | en |
dc.subject | macromolecule | en |
dc.subject | microanalysis | en |
dc.subject | pH | en |
dc.subject | protein analysis | en |
dc.subject | purification | en |
dc.subject | reproducibility | en |
dc.subject | separation technique | en |
dc.subject | ultrafiltration | en |
dc.subject | Electrophoresis, Capillary | en |
dc.subject | Female | en |
dc.subject | Glycoproteins | en |
dc.subject | Humans | en |
dc.subject | N-Acetylneuraminic Acid | en |
dc.subject | Reproducibility of Results | en |
dc.subject | Sensitivity and Specificity | en |
dc.title | A capillary zone electrophoresis method for determining N-acetylneuraminic acid in glycoproteins and blood sera | en |
dc.type | journalArticle | en |
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