Liquid chromatographic method for the determination of uridine in human serum

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2004-04, Vol.803 (2), p.345-351
• Main Authors: Zilly, M, Langmann, P, Winzer, R, Benesic, A, Schirmer, D, Walker, U.A, Klinker, H
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; HIV Infections - blood; Humans; Medical sciences; Pharmacology. Drug treatments; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Uridine; Uridine - blood
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2004.01.018

To evaluate uridine levels in humans we developed a very sensitive and specific high-performance liquid chromatographic method for the determination of uridine in serum. We use techniques which are available in a standard analytical laboratory. Chromatographic analysis was carried out on a Phenomenex Aqua C18 5 μ 125A column protected by a guard cartridge system. Potassium dihydrogen phosphate buffer–acetonitrile was used as an eluent and oxypurinol as the internal standard. All sample preparation steps were done at 4 °C and the autosampler was cooled down to 4 °C. The calibration curve was linear throughout the calibration range from 0.25 to 100 μmol/l. This method was primarily established to evaluate uridine serum levels in patients with HIV infection since patients on highly active antiretroviral therapy (HAART) might develop metabolic disturbances that could lead to severe and fatal lactic acidosis due to mitochondrial toxicity. It is suggested that a limited or inadequate uridine supply is at least in part responsible for the onset of such deterioration.