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Comparison of high-performance liquid chromatography with a radiometric assay for determination of the effect of intra-articular administration of corticosteroid and saline solution on synovial fluid hyaluronate concentration in horses.
Abstract: Two recently developed direct methods, radioassay-125I-labeled hyaluronic acid binding protein (125I-HABP)- and high-performance liquid chromatography (HPLC), were used to assess and compare the concentration of hyaluronate (HA) in synovial fluid of horses. Also determined were changes in the HA concentration in an experimental treatment model involving physiologic saline solution (PSS)-irrigated or methylprednisolone acetate-injected tarsocrural joints of clinically normal horses. Serum HA concentration was determined simultaneously, using the 125I-HABP assay. Synovial fluid HA concentration values obtained by use of the HPLC method were approximately double the values obtained by use of 125I-HABP assay. Correlation (r = 0.819) between the 2 methods was highly significant (P less than 0.001; linear regression analysis) for all samples studied and for various experimental subgroups. When pure HA standards were used, correlation between the 2 methods was close to 1 (r = 0.965; P less than 0.001), with higher values obtained by use of the 125I-HABP assay. It is suggested that the HA binding protein derived from endogenous cartilage proteoglycan interferes with the 125I-HABP assay on synovial fluid, resulting in excessively low values, compared with those obtained using the HPLC procedure. Intra-articular injection of methylprednisolone acetate significantly (P less than 0.01) increased synovial fluid HA concentration at 24 hours after injection. Increase was also detected after PSS irrigation, but owing to wide intersubject variation, this increase was not significant. The HPLC procedure, which provides simultaneous information about the concentration and degree of polymerization of HA, is recommended for the study of synovial fluid, whereas the 125I-HABP assay is more suitable for serum HA analysis.
Publication Date: 1991-12-01 PubMed ID: 1789505 The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Biochemistry
- Clinical Study
- Comparative Study
- Correlation Analysis
- Corticosteroids
- Disease Treatment
- Equine Health
- Experimental Methods
- High-performance Liquid Chromatography (HPLC)
- Horses
- Hyaluronic Acid
- In Vivo
- Intra-Articular Injection
- Laboratory Methods
- Pharmacology
- Physiology
- Radiology
- Regression Analysis
- Synovial Fluid
- Veterinary Medicine
Summary
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The research article discusses the use of two methods, high-performance liquid chromatography (HPLC) and a radiometric assay, to analyze the concentration of hyaluronate in horses’ synovial fluid. It was observed that the HPLC method yielded higher values and the radiometric assay was more suitable for serum analysis. The study also observed the effect of corticosteroid and saline solution on the hyaluronate concentration in horses.
Comparison of Methods
- The researchers used two direct methods; a radioassay with 125I-labeled hyaluronic acid binding protein (125I-HABP) and high-performance liquid chromatography (HPLC), to measure the concentration of hyaluronate (HA) in synovial fluid of horses.
- It was found that the values obtained through HPLC were approximately double the values from the use of 125I-HABP assay.
- Upon analysis, a significant correlation (r = 0.819, P less than 0.001) was found between the results obtained from both methods.
- It was discovered that when pure HA standards were used, the correlation was near perfect (r = 0.965, P less than 0.001), with 125I-HABP yielding higher values.
Issue with 125I-HABP Method
- The researchers suggest that interference may have occurred with the 125I-HABP assay from endogenous cartilage proteoglycan in the synovial fluid. This may have led to disproportionately low values compared to those obtained with HPLC.
Effects of Corticosteroid and Saline Solution
- The researchers also examined the effect of corticosteroid (methylprednisolone acetate) and saline solution on HA concentration in an experimental treatment model.
- It was found that intra-articular injection of methylprednisolone acetate significantly increased synovial fluid HA concentration 24 hours after injection.
- Although an increase was also detected after irrigation with a physiologic saline solution (PSS), due to wide intersubject variation, this increase was not statistically significant.
Recommended Method for Analysis
- The researchers recommend the use of HPLC for the study of synovial fluid as it provides simultaneous information about the concentration and degree of polymerization of HA.
- On the other hand, they suggest that the 125I-HABP assay is better suited for serum HA analysis.
Cite This Article
APA
Tulamo RM.
(1991).
Comparison of high-performance liquid chromatography with a radiometric assay for determination of the effect of intra-articular administration of corticosteroid and saline solution on synovial fluid hyaluronate concentration in horses.
Am J Vet Res, 52(12), 1940-1944.
Publication
Researcher Affiliations
- College of Veterinary Medicine, Department of Clinical Sciences, Helsinki, Finland.
MeSH Terms
- Animals
- Chromatography, High Pressure Liquid
- Horses / metabolism
- Hyaluronic Acid / analysis
- Methylprednisolone / pharmacology
- Radiometry
- Regression Analysis
- Sodium Chloride / pharmacology
- Synovial Fluid / chemistry
- Synovial Fluid / drug effects
Citations
This article has been cited 3 times.- Niemelä TM, Tulamo RM, Hielm-Björkman AK. A randomised, double-blinded, placebo-controlled clinical study on intra-articular hyaluronan treatment in equine lameness originating from the metacarpophalangeal joint. BMC Vet Res 2016 Mar 23;12:60.
- Wong BL, Kim SH, Antonacci JM, McIlwraith CW, Sah RL. Cartilage shear dynamics during tibio-femoral articulation: effect of acute joint injury and tribosupplementation on synovial fluid lubrication. Osteoarthritis Cartilage 2010 Mar;18(3):464-71.
- Palmer JL, Bertone AL, McClain H. Assessment of glycosaminoglycan concentration in equine synovial fluid as a marker of joint disease. Can J Vet Res 1995 Jul;59(3):205-12.
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