Identification of 6-methylmercaptopurine derivative formed during acid hydrolysis of thiopurine nucleotides in erythrocytes, using liquid chromatography-mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance assay.
6-Thioguanine and 6-methylmercaptopurine (Me6-MP) nucleotides are the two major thiopurine metabolites of azathioprine found in erythrocytes. During the acid hydrolysis required for the conversion of thiopurine nucleotides into their free bases, Me6-MP was converted into a compound that could be analyzed on a Purospher RP18-e column with dihydrogen phosphate-methanol buffer as eluent. The pH of the acid extract strongly influenced the conversion of Me6-MP into its derivative. The Me6-MP derivative was identified using liquid chromatography-mass spectrometry and infrared and nuclear magnetic resonance spectrometric methods. During the acid hydrolysis of thiopurine nucleotides in erythrocytes, Me6-MP undergoes degradation, leading to 4-amino-5-(methylthio)carbonyl imidazole.
Dervieux
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T
Université Claude Bernard Lyon 1, Institut des Sciences Pharmaceutiques et Biologiques, Lyon Cedex, France.
Boulieu
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R
D001379:Azathioprine / Q000097:blood; D002853:Chromatography, Liquid; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D006868:Hydrolysis; D007093:Imidazoles / Q000097:blood*; D007166:Immunosuppressive Agents / Q000097:blood; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D015122:Mercaptopurine / Q000031:analogs & derivatives* / Q000097:blood; D010472:Perchlorates; D013055:Spectrophotometry, Infrared; D013873:Thionucleotides / Q000737:chemistry*
C116263:4-amino-5-(methylthio)carbonyl imidazole; D007093:Imidazoles; D007166:Immunosuppressive Agents; D010472:Perchlorates; D013873:Thionucleotides; C010240:6-methylthiopurine; D015122:Mercaptopurine; D001379:Azathioprine