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Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 1989 Jul;48(7):565–570. doi: 10.1136/ard.48.7.565

Determination of synovial fluid hyaluronate concentration and polymerisation by high performance liquid chromatography.

H Saari 1, Y T Konttinen 1
PMCID: PMC1003817  PMID: 2774697

Abstract

High performance liquid chromatography (HPLC) with a size exclusion column and ultraviolet monitoring was used to study the effect of synovial fluid hyaluronate concentration and degree of polymerisation on viscosity and mucin clot formation. Rotational viscometry measurements showed an exponential relation between the synovial fluid hyaluronate concentration and relative viscosity, the viscosity increasing particularly steeply with hyaluronate concentrations exceeding 2-2.5 mg/ml. The scattering of individual values observed around the expected curve was eliminated when both the hyaluronate concentration and its degree of polymerisation were taken into account. Hyaluronate concentration and degree of polymerisation also correlated with the quality of mucin clot, though only HPLC provided more detailed quantitative information about this association. Because HPLC allows reproducible and rapid simultaneous analysis of the synovial fluid hyaluronate concentration and the degree of polymerisation in small volumes of unprocessed samples it can be used in well equipped rheumatological units to replace other methods used previously.

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Selected References

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

  1. Arnett F. C., Edworthy S. M., Bloch D. A., McShane D. J., Fries J. F., Cooper N. S., Healey L. A., Kaplan S. R., Liang M. H., Luthra H. S. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988 Mar;31(3):315–324. doi: 10.1002/art.1780310302. [DOI] [PubMed] [Google Scholar]
  2. BOLLET A. J. The intrinsic viscosity of synovial fluid hyaluronic acid. J Lab Clin Med. 1956 Nov;48(5):721–728. [PubMed] [Google Scholar]
  3. Bjelle A., Andersson T., Granath K. Molecular weight distribution of hyaluronic acid of human synovial fluid in rheumatic diseases. Scand J Rheumatol. 1983;12(2):133–138. doi: 10.3109/03009748309102899. [DOI] [PubMed] [Google Scholar]
  4. Cleland R. L., Wang J. L. Ionic polysaccharides. 3. Dilute solution properties of hyaluronic acid fractions. Biopolymers. 1970;9(7):799–810. doi: 10.1002/bip.1970.360090706. [DOI] [PubMed] [Google Scholar]
  5. Hutadilok N., Ghosh P., Brooks P. M. Binding of haptoglobin, inter-alpha-trypsin inhibitor, and alpha 1 proteinase inhibitor to synovial fluid hyaluronate and the influence of these proteins on its degradation by oxygen derived free radicals. Ann Rheum Dis. 1988 May;47(5):377–385. doi: 10.1136/ard.47.5.377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Scher I., Hamerman D. Isolation of human synovial-fluid hyaluronate by density-gradient ultracentrifugation and evaluation of its protein content. Biochem J. 1972 Mar;126(5):1073–1080. doi: 10.1042/bj1261073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Shimada E., Matsumura G. Viscosity and molecular weight of hyaluronic acids. J Biochem. 1975 Sep;78(3):513–517. doi: 10.1093/oxfordjournals.jbchem.a130935. [DOI] [PubMed] [Google Scholar]

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