Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1973 Feb;131(2):343–350. doi: 10.1042/bj1310343

The quantitative measurement of Alcian Blue–glycosaminoglycan complexes

Paul Whiteman 1
PMCID: PMC1177474  PMID: 4269149

Abstract

1. A simple new quantitative micro method was developed to study the interaction of the cationic dye Alcian Blue 8GX and acid glycosaminoglycans under different conditions. After washing with ethanol the precipitated Alcian Blue–glycosaminoglycan complex was dissociated in Manoxol IB solution and the amount of bound dye measured spectrophotometrically. 2. Reaction profiles of complex-formation were determined in the presence of different concentrations of MgCl2 at pH5.8, and could be used to study the critical electrolyte concentrations of glycosaminoglycans. At least 50mm-MgCl2 was required to produce maximum precipitation of, and maximum uptake of, Alcian Blue by standard glycosaminoglycans. Maximum uptake of Alcian Blue by glycosaminoglycans in the urine of a patient with Hurler's syndrome required the presence of 25–50mm-MgCl2. 3. Under standard conditions of maximum interaction, calibration curves for the quantitative determination of a series of standard glycosaminoglycans in 20μl volumes were nearly linear over the range 1–10μg. 4. The technique was used to determine the molecular binding ratios of Alcian Blue to glycosaminoglycans under controlled conditions.

Full text

PDF
343

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  2. Bolton P. G., Dean A. C. Phosphatase synthesis in Klebsiella (aerobacter) aerogenes growing in continuous culture. Biochem J. 1972 Mar;127(1):87–96. doi: 10.1042/bj1270087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Calatroni A., Donnelly P. V., Di Ferrante N. The glycosaminoglycans of human plasma. J Clin Invest. 1969 Feb;48(2):332–343. doi: 10.1172/JCI105989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dean M. F., Muir H., Ewins R. J. Hurler's, Hunter's and Morquio's syndromes. A biochemical study in the light of current views of the underlying defects. Biochem J. 1971 Aug;123(5):883–894. doi: 10.1042/bj1230883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Di Ferrante N. M. The measurement of urinary mucopolysaccharides. Anal Biochem. 1967 Oct;21(1):98–106. doi: 10.1016/0003-2697(67)90087-5. [DOI] [PubMed] [Google Scholar]
  6. Kaplan D. Classification of the mucopolysaccharidoses based on the pattern of mucopolysacchariduria. Am J Med. 1969 Nov;47(5):721–729. doi: 10.1016/0002-9343(69)90166-1. [DOI] [PubMed] [Google Scholar]
  7. Kaplan D., McKusick V., Trebach S., Lazarus R. Keratosulfate-chondroitin sulfate peptide from normal urine and from urine of patients with Morquio syndrome (mucopolysaccharidosis IV). J Lab Clin Med. 1968 Jan;71(1):48–55. [PubMed] [Google Scholar]
  8. LAURENT T. C., SCOTT J. E. MOLECULAR WEIGHT FRACTIONATION OF POLYANIONS BY CETYLPYRIDINIUM CHLORIDE IN SALT SOLUTIONS. Nature. 1964 May 16;202:661–662. doi: 10.1038/202661a0. [DOI] [PubMed] [Google Scholar]
  9. Manley G., Williams U. Urinary excretion of glycosaminoglycans in the various forms of gargoylism. J Clin Pathol. 1969 Jan;22(1):67–75. doi: 10.1136/jcp.22.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Neri G., Di Ferrante N., Hrgovcic R. Measurement of plasma glycosaminoglycans: an extension of the method. Clin Chim Acta. 1971 Dec;35(2):359–362. doi: 10.1016/0009-8981(71)90206-3. [DOI] [PubMed] [Google Scholar]
  11. SCOTT J. E. The solubility of cetylpyridinium complexes of biological polyanions in solution of salts. Biochim Biophys Acta. 1955 Nov;18(3):428–429. doi: 10.1016/0006-3002(55)90108-6. [DOI] [PubMed] [Google Scholar]
  12. Scott J. E., Dorling J. Differential staining of acid glycosaminoglycans (mucopolysaccharides) by alcian blue in salt solutions. Histochemie. 1965 Oct 1;5(3):221–233. doi: 10.1007/BF00306130. [DOI] [PubMed] [Google Scholar]
  13. Scott J. E. Histochemistry of Alcian blue. I. Metachromasia of Alcian blue, Astrablau and other cationic phthalocyanin dyes. Histochemie. 1970;21(3):277–285. doi: 10.1007/BF00304219. [DOI] [PubMed] [Google Scholar]
  14. Scott J. E. Histochemistry of alcian blue. II. The structure of alcian blue 8GX. Histochemie. 1972;30(3):215–234. [PubMed] [Google Scholar]
  15. Scott J. E., Quintarelli G., Dellovo M. C. The chemical and histochemical properties of Alcian Blue. I. The mechanism of Alcian Blue staining. Histochemie. 1964 Jul 17;4(2):73–85. doi: 10.1007/BF00306149. [DOI] [PubMed] [Google Scholar]
  16. Scott J. E. The histochemistry of Alcian blue. Note on the presence and removal of boric acid as the major diluent in alcian blue 8GX. Histochemie. 1972;29(2):129–133. doi: 10.1007/BF00277280. [DOI] [PubMed] [Google Scholar]
  17. Scott J. E., Willett I. H. Binding of cationic dyes to nucleic acids and ther biological polyanions. Nature. 1966 Mar 5;209(5027):985–987. doi: 10.1038/209985a0. [DOI] [PubMed] [Google Scholar]
  18. Whiteman P. The quantitative determination of glycosaminoglycans in urine with Alcian Blue 8GX. Biochem J. 1973 Feb;131(2):351–357. doi: 10.1042/bj1310351. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES