A Method to Quantitate Total Sialic Acid, Glucosamine, and Galactosamine in Blood Serum and Glycoconjugates by HPLC

Makatsori, E.; Karamanos, Nikos K.; Anastassiou, Evangelos D.; Hjerpe, Anders; Tsegenidis, T.

doi:10.1080/10826079808006884

Cited by 9 publications

(3 citation statements)

References 42 publications

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“…Reduced disaccharides were separated and determined by elution of the column with 50 mm sodium dihydrogen orthophosphate, pH 2.50 at a flow rate of 0.7 mL/min. Quantitative determinations of glucuronic acid and iduronic acid as well as glucosamine and galactosamine were performed by HPLC (Karamanos et al, 1988;Makatsori et al, 1998).…”

Section: Analysis Of Versican Bound Glycosaminoglycansmentioning

confidence: 99%

Versican undergoes speciﬁc alterations in the ﬁne molecular structure and organization in human aneurysmal abdominal aortas

Theocharis

Tsolakis

Hjerpe

et al. 2003

Biomedical Chromatography

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Versican is the major matrix proteoglycan in aortic wall and participates in various biological functions of the tissue. In the present study the molecular characteristics of versican isolated from normal human aorta as well as those of versican expressed in aneurysmal aortic tissue were examined. Versican was isolated by combined anion-exchange and gel permeation chromatography and was further characterized by high-performance liquid chromatography, polyacrylamide gel electrophoresis and immunoblotting. In both tissues versican is exclusively substituted with chondroitin sulfate chains, in contrast to other human tissues where both chondroitin and dermatan sulfate chains are attached onto versican core proteins. Except for the significant decrease in the concentration of versican in the aneurysmal tissue, this PG undergoes specific alterations in the aneurysmal tissue. The molecular size of versican isolated from diseased tissue is decreased with a simultaneous increase in the ratio of glycosaminoglycan to protein in this tissue. The latter reflect the extensive fragmentation of versican in the diseased tissue and most probably the generation of shorter peptides enriched to glycosaminoglycan chains. Although the size of chondroitin sulfate chains is identical in both versican preparations, a significant increase in the percentage of 6-sulfated disaccharides is observed in chondroitin sulfate chains of versican in aneurysmal aortas, which is accompanied by decrease in 4-sulfated and non-sulfated units.

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Section: Analysis Of Versican Bound Glycosaminoglycansmentioning

confidence: 99%

Versican undergoes speciﬁc alterations in the ﬁne molecular structure and organization in human aneurysmal abdominal aortas

Theocharis

Tsolakis

Hjerpe

et al. 2003

Biomedical Chromatography

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“…Sulfuric acid diluted at concentrations of 25-100 mM is the most commonly used acid [33][34][35][36][37][38]. Others authors have used different solvents for hydrolysis: diluted HCl [32,39], microwave hydrolysis with acetic acid [9,40] that decreases the time of hydrolysis, sodium hydrosulfite [41], or trifluoroacetic acid [42,43].…”

Section: Release Of Sialic Acidsmentioning

confidence: 99%

Determination of sialic acid and gangliosides in biological samples and dairy products: A review

Lacomba

salcedo

Alegrı́a

et al. 2010

Journal of Pharmaceutical and Biomedical Analysis

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“…Five phenyl groups are introduced to the hydroxyl groups of Neu5Ac to give stable and strongly UV-absorbing per-O-benzoylated derivatives. This scheme of derivatization has been successfully used to determine both neutral monosaccharides and sialic acid in glycoproteins and blood sera using reversed-phase HPLC (Karamanos et al, 1990;Tsegenidis and Karamanos, 1998;Makatsori et al, 1998bMakatsori et al, , 1999. In this article we extend the utilization of this derivatization reaction to develop a simple and accurate CZE method for determining the levels of Neu5Ac in glycoproteins and blood serum samples.…”

Section: Introductionmentioning

confidence: 95%

A capillary zone electrophoresis method for determining N‐acetylneuraminic acid in glycoproteins and blood sera

Strousopoulou

Militsopoulou

Stagiannis

et al. 2002

Biomedical Chromatography

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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 2h at 80 degrees 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 degrees C for 20 min resulted in complete conversion of Neu5Ac to per-O-benzoylated Neu5Ac. CZE analysis was performed using the operating buffer 25mM 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 microg/mL to 5mg/mL with a detection limit of 2 microM. Application of the method to microanalysis of human alpha(1)-acid glycoprotein and blood serum samples showed excellent agreement with previously published values, suggesting a high precision for the developed CZE method.

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A Method to Quantitate Total Sialic Acid, Glucosamine, and Galactosamine in Blood Serum and Glycoconjugates by HPLC

Cited by 9 publications

References 42 publications

Versican undergoes speciﬁc alterations in the ﬁne molecular structure and organization in human aneurysmal abdominal aortas

Versican undergoes speciﬁc alterations in the ﬁne molecular structure and organization in human aneurysmal abdominal aortas

Determination of sialic acid and gangliosides in biological samples and dairy products: A review

A capillary zone electrophoresis method for determining N‐acetylneuraminic acid in glycoproteins and blood sera

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