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.
Maessen
J G
JG
Department of Physiology, University of Limburg, Maastricht, The Netherlands.
van der Vusse
G J
GJ
Vork
M
M
Kootstra
G
G
D000227:Adenine Nucleotides / Q000032:analysis; D000241:Adenosine / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003065:Coenzyme A / Q000032:analysis; D006150:Guanine Nucleotides / Q000032:analysis; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000032:analysis; D007288:Inosine / Q000032:analysis; D007291:Inosine Monophosphate / Q000032:analysis; D007672:Kidney Cortex / Q000032:analysis*; D016030:Kidney Transplantation; D009243:NAD / Q000032:analysis; D009249:NADP / Q000032:analysis; D009705:Nucleosides / Q000032:analysis*; D009711:Nucleotides / Q000032:analysis*; D011786:Quality Control; D019820:Xanthine; D014970:Xanthines / Q000032:analysis
D000227:Adenine Nucleotides; D006150:Guanine Nucleotides; D007042:Hypoxanthines; D009705:Nucleosides; D009711:Nucleotides; D014970:Xanthines; D009243:NAD; D007291:Inosine Monophosphate; D019820:Xanthine; D019271:Hypoxanthine; D009249:NADP; D007288:Inosine; D000241:Adenosine; D003065:Coenzyme A