Sample preparation for the determination of purine nucleotide analogues in tissues

Journal of Chromatography. a
R Boulieu

Abstract

A sample treatment procedure for the determination of thiopurine and ganciclovir nucleotides in human tissues was developed. Owing to the lack of suitable standards for most of the active nucleotide analogues, the procedure was based on two steps: (1) perchloric acid homogenization and deproteinization of the tissue specimen and (2) conversion of purine nucleotides into parent drug or free bases by enzymatic or acid hydrolysis. The parent drug or purine bases formed were then analyzed on a Hypersil ODS column using isocratic elution with dihydrogenphosphate buffer for ganciclovir nucleotides or the gradient elution mode with dihydrogenphosphate buffermethanol for thiopurine nucleotides. The sample treatment procedure was evaluated using guanosine triphosphate (GTP), 6-thioinosinic acid (6TIMP) and 6-thioguanosine monophosphate (6TGMP) as standards. Mean analytical recoveries determined by adding known concentrations of standards to the tissue specimen before sampling processing were higher than 97%. The sample preparation described is simple and represents a suitable method for the investigation of active nucleotide pool in tissues.

References

Apr 26, 1991·Pharmaceutisch Weekblad. Scientific Edition·E H Wiltink, R Janknegt
Jan 1, 1989·Journal of Pharmaceutical and Biomedical Analysis·R D McDowallV S Picot
Jan 1, 1985·Drug Metabolism Reviews·K G Van ScoikW R Porter
Mar 10, 1995·Journal of Chromatography. B, Biomedical Applications·R Boulieu, A Lenoir
Sep 1, 1994·Journal of Pharmaceutical and Biomedical Analysis·R Boulieu, N Bleyzac
Aug 5, 1994·Journal of Chromatography. B, Biomedical Applications·N Bleyzac, R Boulieu

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