Relative molecular weight and concentration determination of sodium hyaluronate solutions by gel-exclusion high-performance liquid chromatography

Beaty, Narlin; Tew, William P.; Mello, Robert J.

doi:10.1016/0003-2697(85)90287-8

Cited by 39 publications

(12 citation statements)

References 13 publications

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“…0.1 pg/mL and shows an inter-assay coefficient of variation = 4.5 %. HPLC combined with refractometry has been used [19] in analysis for hyaluronan and requires ca. 20 pL of specimen with hyaluronan concentration > 100 pg/mL.…”

Section: Other Methods For Hyaluronan Determinationmentioning

confidence: 99%

Quantitative analysis of hyaluronan in vitreous humor using capillary electrophoresis

et al. 1994

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Capillary electrophoresis was used for the separation and quantitative analysis of the glycosaminoglycan hyaluronan in human and bovine vitreous with detection by UV absorbance at 200 nm. Calibration was carried out using standards made up from known concentrations of hyaluronan of umbilical cord origin. The purity of the standard was examined by 1H NMR (nuclear magnetic resonance). Concentrations as low as 25 micrograms/mL could be detected by capillary electrophoresis. Confirmation that the signal was due to hyaluronan was obtained by depolymerization of the native mucopolysaccharide by hyaluronidase. This resulted in loss of the hyaluronan peak and appearance of several new peaks corresponding to the oligomeric fragments which had shorter migration times. Capillary electrophoresis is a reproducible and sensitive technique for the quantification and characterisation of hyaluronan in vitreous samples.

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Section: Other Methods For Hyaluronan Determinationmentioning

confidence: 99%

Quantitative analysis of hyaluronan in vitreous humor using capillary electrophoresis

et al. 1994

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“…Concentrations were determined by HPLC (Waters Ltd., Watford, UK), using a size exclusion TosoHaas TSK G6000 PWXL column (Anachem Ltd., Luton, UK) (7.83300 mm) and absorbance at 206 nm [15]. The mobile phase was 50 mM phosphate buffer, adjusted to pH 5.55.…”

Section: Analyses By Hplcmentioning

confidence: 99%

Comparative study of hyaluronic derivatives: rheological behaviour, mechanical and chemical degradation

Prieto

Pulido

Zapico

et al. 2005

International Journal of Biological Macromolecules

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“…However, the sensitivity of this determination is restricted by protein absorption and the detection limit was found to be in units of micrograms. Beaty et al (1985) using HPGPC for the determination of HA in solution estimated the range of detectable concentrations as 100-300 pg/mL. Moreover, the determination of HA concentration in complex mixtures by these methods is impractical because impurities or other components absorb light at the adjusted wavelengths (280 nm) as has been shown by Hardingham and Muir (1974).…”

Section: ~~mentioning

confidence: 95%

Specific high performance liquid chromatographic determination of the molecular weight and concentration of hyaluronic acid in complex mixtures by labelled hyaluronate binding proteins

Orvisky

Kéry

Stancíková

1991

Biomedical Chromatography

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A simple High performance liquid chromatographic (HPLC) method for the specific determination of the molecular weight and concentration of hyaluronic acid (HA) in complex mixtures has been developed. Hyaluronate-binding proteins isolated from bovine cartilage labelled by 125I or fluoresceinisothiocyanate were used as specific markers. The specific binding affinities of the markers were compared and were found to have association constants of 1.6 x 10(7) M-1 and 1.2 x 10(7) M-1 respectively. The HA levels and molecular weight distributions can be easily determined in the range 10-500 ng/mL in complex mixtures by the use of markers, molecular sieving HPLC columns and appropriate detectors. It has been demonstrated clearly that the method is useful for the highly specific determination of the parameters in complex biological samples such as serum and synovial fluids and is recommended for clinical applications.

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Relative molecular weight and concentration determination of sodium hyaluronate solutions by gel-exclusion high-performance liquid chromatography

Cited by 39 publications

References 13 publications

Quantitative analysis of hyaluronan in vitreous humor using capillary electrophoresis

Quantitative analysis of hyaluronan in vitreous humor using capillary electrophoresis

Comparative study of hyaluronic derivatives: rheological behaviour, mechanical and chemical degradation

Specific high performance liquid chromatographic determination of the molecular weight and concentration of hyaluronic acid in complex mixtures by labelled hyaluronate binding proteins

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