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Home / Equine Research Bank / Anal Biochem / A microtiter plate assay for the determination of uronic aci...
Analytical biochemistry1998; 257(2); 107-111; doi: 10.1006/abio.1997.2538

A microtiter plate assay for the determination of uronic acids.

Abstract: The amount of uronic acid residues in samples containing glycosaminoglycans or pectin is an important parameter in the quantitative and structural analysis of these complex carbohydrates. This paper describes a method to determine the content of uronic acids in biological samples, using conventional polystyrene microtiter plates and microtiter plate-reading equipment with standard interference filters (i.e., 540 or 492 nm). This assay is a modification of a commonly used procedure, viz. hydrolysis of uronic acid containing carbohydrate polymers in 80% sulfuric acid containing tetraborate ions at 80 degrees C followed by a coloring step with an m-hydroxydiphenyl reagent at room temperature. The use of microtiter plates has several practical advantages: (i) less risk of handling hot, concentrated sulfuric acid is present; (ii) an accurate estimate of background absorbance by multiple reading of the plates is possible; and (iii) many samples can be assayed in one series without errors due to fading of the final color. The validity of the assay was checked for the quantification of hyaluronic acid in equine synovial fluid samples. We consider this the method of choice when a large number of samples must be analyzed for their content of uronic acid residues.
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Publication Date: 1998-03-26 PubMed ID: 9514779DOI: 10.1006/abio.1997.2538Google Scholar: Lookup
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  • 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.

This paper presents a method for determining uronic acid content in complex carbohydrates, using microtiter plates and standard equipment. This technique modifies an existing process and reduces the risks associated with handling sulfuric acid while also providing more accurate results.

Research Purpose and Methodology

  • The main aim of this study was to devise a safer and more effective way to add up the volume of uronic acid leftovers in natural samples, an important factor in analysing intricate carbohydrates like glycosaminoglycans or pectin. This was achieved by modifying an existing method and incorporating the use of polystyrene microtiter plates and universal microtiter plate-reading accessories.
  • The altered method involves the breakdown of carbohydrate polymers that contain uronic acid in a solution of 80% sulfuric acid with tetraborate ions, at a temperature of 80 degrees Celsius. This is succeeded by a colorization phase with an m-hydroxydiphenyl reagent at room temperature.

Advantages of the Proposed Method

  • Using these microtiter plates has several benefits over the traditional process:
    • It reduces the risks associated with manipulating hot, concentrated sulfuric acid, thereby improving safety conditions.
    • It enables a highly precise calculation of background absorbance through multiple readings of the plates, leading to more accurate results.
    • It permits the analysis of many samples in one series with no errors due to the final color fading, which increases the efficiency of the process.

Validation of the Method

  • The method’s authenticity and reliability were validated by applying it to measure the quantity of hyaluronic acid in equine synovial fluid samples. Since the method proved successful in this application, the researchers recommend it as a preferred technique when a considerable number of samples are required to be analysed for uronic acid content.

Cite This Article

APA
van den Hoogen BM, van Weeren PR, Lopes-Cardozo M, van Golde LM, Barneveld A, van de Lest CH. (1998). A microtiter plate assay for the determination of uronic acids. Anal Biochem, 257(2), 107-111. https://doi.org/10.1006/abio.1997.2538

Publication

Analytical biochemistry
ISSN: 0003-2697
NlmUniqueID: 0370535
Country: United States
Language: English
Volume: 257
Issue: 2
Pages: 107-111

Researcher Affiliations

van den Hoogen, B M
  • Department of Veterinary Basic Sciences, Graduate School of Animal Health, Utrecht University, The Netherlands.
van Weeren, P R
    Lopes-Cardozo, M
      van Golde, L M
        Barneveld, A
          van de Lest, C H

            MeSH Terms

            • Animals
            • Cattle
            • Colorimetry / instrumentation
            • Glucose / analysis
            • Horses
            • Hyaluronic Acid / analysis
            • Polystyrenes
            • Reproducibility of Results
            • Serum Albumin / analysis
            • Synovial Fluid / chemistry
            • Time Factors
            • Uronic Acids / analysis

            Citations

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