Reliability of 1,9-dimethylmethylene blue tests in comparison to agarose gel electrophoresis for quantification of urinary glycosaminoglycans

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dc.contributor.author Lima, Cilene R. de
dc.contributor.author Baccarin, Raquel Yvonne Arantes
dc.contributor.author Michelacci, Yara M.
dc.date.accessioned 2016-01-24T12:41:55Z
dc.date.available 2016-01-24T12:41:55Z
dc.date.issued 2007-03-01
dc.identifier http://dx.doi.org/10.1016/j.cca.2006.11.025
dc.identifier.citation Clinica Chimica Acta. Amsterdam: Elsevier B.V., v. 378, n. 1-2, p. 206-215, 2007.
dc.identifier.issn 0009-8981
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/29542
dc.description.abstract Background: the relevance of glycosaminoglycan determination in biological fluids is gradually gaining importance in the literature. Nevertheless, the results obtained by different methods vary widely. We evaluated 1,9-dimethylmethylene blue (DMB) dye-binding assays for quantification of urinary glycosaminoglycans, in comparison to densitometry after agarose gel electrophoresis.Methods: Urinary glycosaminoglycans from different mammalian species were quantified by 3 different DMB dye-binding assays. the results were compared to those obtained by densitometry after agarose gel electrophoresis of glycosaminoglycans isolated from urine samples by ion exchange chromatography.Results: Densitometry after agarose gel electrophoresis showed glycosaminoglycan urinary concentrations of 1-20 mg/l, and glycosaminoglycan/creatinine ratios of 2-25 x 10(-3), for all the mammalian species here studied. A decrease with age was observed for humans, cats and horses. in comparison, DMB assays gave much higher results - up to 200 mg/l and 500 x 10(-3) glycosaminoglycan/creatinine ratios. These values were greatly reduced after 4-h dialysis, suggesting that low molecular weight compounds do interfere. Furthermore, urinary anions such as sulfate, phosphate and citrate, react with metachromatic dyes, such as Toluidine Blue and DMB.Conclusion: DMB assays, although rapid and simple, are not appropriate to quantify urinary glycosaminoglycans in normal mammalians, since other urinary components interfere with the reactions. (c) 2006 Elsevier B.V. All rights reserved. en
dc.format.extent 206-215
dc.language.iso eng
dc.publisher Elsevier B.V.
dc.relation.ispartof Clinica Chimica Acta
dc.rights Acesso restrito
dc.subject glycosaminoglycan en
dc.subject urine en
dc.subject agarose gel electrophoresis en
dc.subject DMB assay en
dc.title Reliability of 1,9-dimethylmethylene blue tests in comparison to agarose gel electrophoresis for quantification of urinary glycosaminoglycans en
dc.type Artigo
dc.rights.license http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.contributor.institution Universidade de São Paulo (USP)
dc.description.affiliation Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, SP, Brazil
dc.description.affiliation Univ São Paulo, Fac Med Vet & Zootecn, Dept Clin Med, São Paulo, SP, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, SP, Brazil
dc.identifier.doi 10.1016/j.cca.2006.11.025
dc.description.source Web of Science
dc.identifier.wos WOS:000244997600033



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