Development and validation of an HPLC method for the rapid and simultaneous determination of 6-mercaptopurine and four of its metabolites in plasma and red blood cells

Ahmed F. Hawwa, Jeff S. Millership, James C. McElnay, Paul S. Collier

Research output: Contribution to journalArticlepeer-review

Abstract

An HPLC method has been developed and validated for the rapid determination of mercaptopurine and four of its metabolites; thioguanine, thiouric acid, thioxanthine and methylmercaptopurine in plasma and red blood cells. The method involves a simple treatment procedure based on deproteinisation by perchloric acid followed by acid hydrolysis and heating for 45min at 100 degrees C. The developed method was linear over the concentration range studied with a correlation coefficient >0.994 for all compounds in both plasma and erythrocytes. The lower limits of quantification were 13, 14, 3, 2, 95pmol/8 x 10(8) RBCs and 2, 5, 2, 3, 20ng/ml plasma for thioguanine, thiouric acid, mercaptopurine, thioxanthine and methylmercaptopurine, respectively. The method described is selective and sensitive enough to analyse the different metabolites in a single run under isocratic conditions. Furthermore, it has been shown to be applicable for monitoring these metabolites in paediatric patients due to the low volume requirement (200microl of plasma or erythrocytes) and has been successfully applied for investigating population pharmacokinetics, pharmacogenetics and non-adherence to therapy in these patients.
Original languageEnglish
Pages (from-to)401-409
Number of pages9
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume49
Issue number2
DOIs
Publication statusPublished - 20 Feb 2009

Keywords

  • 6-mercaptopurine
  • 6-thioguanine nucleotides
  • paediatrics
  • leukaemia
  • Drug Inflammatory bowel disease
  • HPLC

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