DETERMINATION OF THIOPURINE NUCLEOTIDES IN HUMAN LUNG-TISSUE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A method for the analysis of thiopurine nucleotides in human transbron chial lung biopsy was developed. The sample treatment procedure is bas ed on perchloric acid homogenisation and deproteinisation with dithiot hreitol and hydrolysis of thiopurine nucleotides into their free bases by heating of the acid extract. Then, the free bases were analyzed in the gradient elution mode on a Hypersil ODS, 3-mu m column using dihy drogenphosphate buffer-methanol as eluent. Mean analytical recoveries for 6-thioguanosine monophosphate and 6-thioinosinic acid from lung ti ssue were 97.0 +/- 2.0 and 98.0 +/- 1.8% at a concentration of 3.0 nmo l/ml and the minimum detectable amounts were 3.5 and 2 pmol, respectiv ely. The procedure described is simple and represents a suitable metho d for the investigation of thiopurine nucleotides in tissues.