title: Determination of the unsaturated disaccharides of hyaluronic acid in equine synovial fluid by high-performance liquid chromatography and fluorescence detection.
abstract: BACKGROUND
  The purpose of this study was to develop and validate an analytical method to determine the presence of hyaluronic acid derived disaccharides in equine synovial fluid.
  
  
  RESULTS
  A high-performance liquid chromatography method for the determination of hyaluronic acid derived unsaturated disaccharides in equine synovial fluid was developed and validated. The method is based on the measurement of unsaturated disaccharides released by digestion of linear hyaluronic acid molecules. The method showed linearity (r(2) = 0.996) over the full working concentration range 0.89-30 mg/l. Relative standard deviation of intra- and inter-day precision ranged from of 4.3-6.7% and 7.1-7.8% respectively. The detection limit was 0.3 mg/l corresponding to 20 mg/l in synovial fluid. Accuracy of the assay was 97-103%. This method was evaluated by determining the concentration of unsaturated disaccharides from hyaluronic acid in synovial fluid of horses with lameness in the metacarpo-/metatarsophalangeal joint localized with positive response to intra-articular anesthesia.
  
  
  CONCLUSIONS
  The described method is valid for determination of hyaluronic acid derived disaccharides in equine synovial fluid. This method was applied to a larger research project dealing with a new form of intra-articular therapy in horses with arthritic diseases.
authors.affiliation: Department of Equine and Small Animal Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland. kaisa.aaltonen@helsinki.fi.
authors.forename: Kaisa
authors.initials: K
authors.lastname: Aaltonen
authors.affiliation: Department of Equine and Small Animal Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland. tytti.niemela@helsinki.fi.
authors.forename: Tytti
authors.initials: T
authors.lastname: Niemelä
authors.affiliation: Department of Equine and Small Animal Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland. satu.sankari@helsinki.fi.
authors.forename: Satu
authors.initials: S
authors.lastname: Sankari
authors.affiliation: Department of Equine and Small Animal Medicine, University of Helsinki, P.O. Box 57, 00014, Helsinki, Finland. riitta.tulamo@helsinki.fi.
authors.forename: Riitta-Mari
authors.initials: RM
authors.lastname: Tulamo
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary*; D004187:Disaccharides / Q000378:metabolism*; D006736:Horses / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000378:metabolism*; D013582:Synovial Fluid / Q000737:chemistry*
chemical_list: D004187:Disaccharides; D006820:Hyaluronic Acid