Reversed-phase high-performance liquid chromatographic determination of tricyclic nucleoside and tricyclic nucleoside 5'-phosphate in biological specimens.
An isocratic, sensitive, high-performance liquid chromatographic assay was developed for the determination of the tricyclic nucleoside 1,4,5,6,8-pentaazaacenaphthylene -3-amino-1,5-dihydro-5-methyl-1-beta-D-ribofuranosyl 5'-monophosphate (TCN-P; NSC 280594) and its dephosphorylated metabolite TCN (NSC 154020). Separation was obtained using a C18 Sep-Pak precolumn, a reversed-phase column, and a mobile phase of phosphate buffer--methanol (87.5:12.5, v/v) containing 0.0025 M tetrabutylammonium hydroxide. The absorbance of both TCN and TCN-P was monitored at 280 nm with a sensitivity limit of 10 ng/ml. The recovery was 54 +/- 6% and 51 +/- 8% (mean +/- S.D.) from plasma for TCN and TCN-P, respectively. Rapid enzymatic dephosphorylation of TCN-P in plasma and tissue samples was prevented by adding a large excess of adenosine 5'-monophosphate. The assay was used to determine plasma and tissue concentrations of TCN-P and TCN after administration of TCN-P to cancer patients in a Phase I clinical study.
D000073:Acenaphthenes; D000818:Animals; D001826:Body Fluids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004355:Drug Stability; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D008297:Male; D010766:Phosphorylation; D051381:Rats; D011919:Rats, Inbred Strains; D012263:Ribonucleosides / Q000008:administration & dosage / Q000032:analysis*; D012265:Ribonucleotides / Q000008:administration & dosage / Q000032:analysis*; D014018:Tissue Distribution
D000073:Acenaphthenes; D012263:Ribonucleosides; D012265:Ribonucleotides; C023764:triciribine; C014205:triciribine phosphate