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Home / Equine Research Bank / Hoppe Seylers Z Physiol Chem / Fluorimetric determination of unsubstituted and 9(8)-O-acety...
Hoppe-Seyler's Zeitschrift fur physiologische Chemie1982; 363(3); 255-262; doi: 10.1515/bchm2.1982.363.1.255

Fluorimetric determination of unsubstituted and 9(8)-O-acetylated sialic acids in erythrocyte membranes.

Abstract: A method is described for all quantitative determination of free or glycosidically bound sialic acids with special reference to erythrocyte membranes. Sialic acids, unsubstituted in their side chains, quantitatively yield formaldehyde after mild periodate oxidation (1 mM NaIO4, 15 min, 4 degrees C, in the dark). The formaldehyde is determined by the reaction with acetylacetone and ammonium acetate which leads to a sensitive fluorogen (F 410/510 nm). Sialic acids O-acetylated at C-9 or C-8 are not oxidized under these conditions. Therefore, they can be determined quantitatively by measuring the increase of fluorogen after O-deacetylation by 0.1 M NaOH. A minimum amount of 100 ng of sialic acids can be determined. Total sialic acids (microgram/ml packed erythrocytes) and the relative amount of 9(8)-mono-O-acetylated sialic acids were determined by this procedure in erythrocytes from the following mammals (data in brackets): donkey (360; 8%), horse (350; 4%), mouse (300; 60%), rat (280; 40%), cow (200; 0%), cat (190; 25%), man (180; 0%), guinea pig (160; 22%), pig (130; 12%) and rabbit (70; 20%).
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Publication Date: 1982-03-01 PubMed ID: 7076126DOI: 10.1515/bchm2.1982.363.1.255Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research describes a method for accurately determining the different types of sialic acids present in erythrocyte (red blood cell) membranes. The method uses mild oxidation to detect the presence of these molecules, which can give insights into different biological properties of species.

Objective of the Research

  • The aim of the study is to present a method to correctly determine the amount of free or bound sialic acids – specifically those that are unsubstituted or 9(8)-O-acetylated. The research uses this method to measure the presence of these acids in erythrocyte membranes of various mammals.

Methodology

  • Unsubstituted sialic acids are processed under mild periodate oxidation. This treatment induces a chemical reaction that results in the production of formaldehyde.
  • The formed formaldehyde is then measured with acetylacetone and ammonium acetate. This reaction forms fluorogen, a fluorescent compound, which can be detected and measured.
  • Sialic acids that are O-acetylated at either C-9 or C-8 positions are not oxidized under these conditions. This allows for their direct quantification by measuring the increase in fluorogen after the deacetylation process.
  • A minimum of 100 ng of sialic acids could be effectively determined with this technique.

Results and Discussion

  • The method was used to quantify total sialic acids (measured in micrograms per milliliter of packed erythrocytes) and the relative presence of 9(8)-mono-O-acetylated sialic acids in erythrocytes from various mammals.
  • The subjects studied include donkeys, horses, mice, rats, cows, cats, humans, guinea pigs, pigs, and rabbits.
  • The results varied according to the type of mammal. For example, mice were found to have 300 µg/mL packed erythrocytes and 60% of them were 9(8)-mono-O-acetylated sialic acids, while humans had 180 µg/mL packed erythrocytes with no 9(8)-mono-O-acetylated sialic acids.

Conclusion

  • The presented method was successful in allowing the quantification of different types of sialic acids in the erythrocyte membranes of a variety of mammals. This could provide a useful tool for further investigations in this field.

Cite This Article

APA
Shukla AK, Schauer R. (1982). Fluorimetric determination of unsubstituted and 9(8)-O-acetylated sialic acids in erythrocyte membranes. Hoppe Seylers Z Physiol Chem, 363(3), 255-262. https://doi.org/10.1515/bchm2.1982.363.1.255

Publication

Hoppe-Seyler's Zeitschrift fur physiologische Chemie
ISSN: 0018-4888
NlmUniqueID: 2985060R
Country: Germany
Language: English
Volume: 363
Issue: 3
Pages: 255-262

Researcher Affiliations

Shukla, A K
    Schauer, R

      MeSH Terms

      • Animals
      • Cats
      • Cattle
      • Erythrocyte Membrane / analysis
      • Erythrocytes / analysis
      • Guinea Pigs
      • Horses
      • Humans
      • Mice
      • Perissodactyla
      • Rabbits
      • Rats
      • Sialic Acids / blood
      • Species Specificity
      • Spectrometry, Fluorescence / methods
      • Structure-Activity Relationship
      • Swine

      Citations

      This article has been cited 24 times.
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