HPLC METHOD FOR MEASUREMENT OF PURINE NUCLEOTIDE DEGRADATION PRODUCTSIN CEREBROSPINAL-FLUID

We describe a convenient method for the separation and quantification of xanthine, hypoxanthine, and uric acid in 20 mu L of cerebrospinal f luid (CSF) with use of HPLC and ultraviolet detection, The analysis is performed on a Sepharon SGX C-18 column and the elution system consis ts of potassium phosphate buffer, pH 5.1, with 20 mL/L methanol. The l ower limit of detection was 4 pmol for hypoxanthine and xanthine and 6 pmol for uric acid. Analytical recoveries of purine metabolites range d from 98.6% to 102.9%. The intra- and interassay CVs were <3%. The ap plicability of the method is illustrated with the determination of mic romolar concentrations of xanthine, hypoxanthine, and uric acid in CSF samples obtained from 113 patients with various neurological disorder s.