Nucleotides, nucleosides, and oxypurines in human kidneys measured by use of reversed-phase high-performance liquid chromatography.

An HPLC technique is presented for determining adenine nucleotides and related substances in renal cortical tissue. Nineteen metabolic substances can be resolved in a single 25-min run, with use of a gradient-elution system. The mean intra-assay CV is 2.4%, the interassay CV 5%. The lower detection limit for substances commonly present in kidney tissue--such as ATP, ADP, AMP, GTP, GDP, GMP, IMP, inosine, adenosine, hypoxanthine, and xanthine--ranges from 0.6 to 3.6 mumol/L, corresponding to 18 and 107 pmol applied to the column. For reliable analysis, a specimen of renal cortex weighing at least 5 mg (wet weight), taken during donor nephrectomy, during cold storage of the kidney, and 1 h after the onset of reperfusion, can be used. The method presented provides a rapid, reproducible diagnostic tool for assessing the chemical energy status of human kidneys in renal surgery and transplantation.