HPLC determination of adenosine in human synovial fluid

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 24; no. 5; pp. 1143 - 1146
• Main Authors: Sottofattori, Enzo, Anzaldi, Maria, Ottonello, Luciano
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 01-03-2001; Elsevier Science
• Subjects: Adenosine; Adenosine - analysis; Analysis; Arthritis, Rheumatoid - metabolism; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; General pharmacology; HPLC; Humans; Medical sciences; Pharmacology. Drug treatments; Reproducibility of Results; Rheumatoid arthritis; Sensitivity and Specificity; Spectrophotometry, Ultraviolet; Synovial fluid; Synovial Fluid - metabolism; Synovial fluid; Adenosine; Rheumatoid arthritis; HPLC; Human; Validation; Immunopathology; Biological fluid; Diseases of the osteoarticular system; HPLC chromatography; Biological marker; Autoimmune disease; Inflammatory joint disease; Isocratic condition; Chronic; Ultraviolet detector; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(00)00574-4

A high-performance liquid chromatographic method has been developed for the quantitative determination of adenosine in human synovial fluid. The method is simple, rapid and, overall, selective. No interference with the components of the biological matrix was observed in these chromatographic conditions. An ODS (250×4.6 mm) 5 μm column was used with an isocratic elution of a phosphate buffer–acetonitrile mobile phase. Detection was carried out on a UV detector at 260 nm. Calibration curve was found to be linear in the 0.7–70 μg ml
−1 range. Linear regression analysis of the data demonstrates the efficacy of the method in terms of precision and accuracy. The precision of this method, calculated as the relative standard deviation (RSD) of the recoveries (1.57–2.21%), was excellent. The limits of quantitation (LOQ) and detection (LOD) were respectively 0.7 and 0.2 μg ml
−1. The method was applied to some samples of synovial effusion from patients affected by rheumatoid arthritis. The concentrations of adenosine which were found were included in the range of the calibration curve.