Quantification of dimethyl-ifosfamide and its N-deschloropropylated metabolites in mouse plasma by liquid chromatography-tandem mass spectrometry

Alain Deroussent, Stéphanie Rodriguez, Sonia Martelli, Atmane Seck, Estelle Dubus-Daudigeos, Didier Desmaële, Gilles Vassal, Angelo Paci

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Résumé

Among antitumor oxazaphosphorine drugs, the prodrug ifosfamide (IFO) and its analogs require metabolic activation by specific liver cytochrome P450 (CYP) enzymes to become therapeutically active. New 7,9-dimethyl-ifosfamide analogs have shown greater cytotoxic activity than IFO, whereas side-chain oxidation still occurred leading to monochloroacetone after N-dechloropropylation. A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay was developed and validated for the simultaneous quantitation of the prodrug 7S,9S-dimethyl-ifosfamide (diMeIFO) and its two inactive metabolites, N2- and N3-deschloropropyl-dimethylifosfamide (N2-DCP-diMeIFO and N3-DCP-diMeIFO) in mouse plasma. After protein precipitation with methanol, the analytes were separated by isocratic reversed-phase chromatography with (methanol/ammonium formate pH 5.5, 60:40, v/v) and detected by tandem mass spectrometry using multiple reaction monitoring of transitions ions m/z 289→168 for diMeIFO, m/z 213→168 for N2-DCP-diMeIFO, m/z 213→92 for N3-DCP-diMeIFO and m/z 261→154 for IFO (internal standard). The calibration curves were linear over the concentration range of 20-10,000ng/mL for the three analytes. Mean extraction recoveries from mouse plasma were 99, 96, 99 and 100% for diMeIFO, N2-DCP-diMeIFO, N3-DCP-diMeIFO and IFO, respectively. The lower limit of quantitation for diMeIFO and its metabolites was 20ng/mL in 50μL plasma. The method was accurate with calculated bias from -5.8 to 4.0% for diMeIFO, from -1.1 to 10.6% for N2-DCP-diMeIFO and from -6.9 to 9.8% for N3-DCP-diMeIFO, and precise with coefficients of variation lower than 6.8%, 7.8% and 14.3%, respectively. The assay was successfully applied to a preliminary pharmacokinetic study of diMeIFO and of its metabolites in mice.

langue originaleAnglais
Pages (de - à)743-750
Nombre de pages8
journalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume879
Numéro de publication11-12
Les DOIs
étatPublié - 1 avr. 2011
Modification externeOui

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