Simultaneous quantification of preactivated ifosfamide derivatives and of 4-hydroxyifosfamide by high performance liquid chromatography-tandem mass spectrometry in mouse plasma and its application to a pharmacokinetic study.
The antitumor drug, ifosfamide (IFO), requires activation by cytochrome P450 (CYP) to form the active metabolite, 4-hydroxyisfosfamide (4-OHIFO), leading to toxic by-products at high dose. In order to overcome these drawbacks, preactivated ifosfamide derivatives (RXIFO) were designed to release 4-OHIFO without CYP involvement. A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the simultaneous quantification of 4-OHIFO, IFO and four derivatives RXIFO in mouse plasma using multiple reaction monitoring. Because of its instability in plasma, 4-OHIFO was immediately converted to the semi-carbazone derivative, 4-OHIFO-SCZ. For the six analytes, the calibration curves were linear from 20 to 5000ng/mL in 50μL plasma and the lower limit of quantitation was determined at 20ng/mL with accuracies within ±10% of nominal and precisions less than 12%. Their recoveries ranged from 62 to 96% by using liquid-liquid extraction. With an improved assay sensitivity compared to analogues, the derivative 4-OHIFO-SCZ was stable in plasma at 4°C for 24h and at -20°C for three months. For all compounds, the assay was validated with accuracies within ±13% and precisions less than 15%. This method was applied to a comparative pharmacokinetic study of 4-OHIFO from IFO and three derivatives RXIFO in mice. This active metabolite was produced by some of the novel conjugates with good pharmacokinetic properties.
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France. Electronic address: alain.deroussent@gustaveroussy.fr.
Alain
A
Deroussent
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Charles
C
Skarbek
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Adeline
A
Maury
CNRS UMR 8612, Laboratory of New Anticancer Targeting Strategies, Paris-Sud University, Galien Institute, Châtenay-Malabry, France.
Hubert
H
Chapuis
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Estelle
E
Daudigeos-Dubus
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Ludivine
L
Le Dret
Mass Spectrometry Metabolomics Facility, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Sylvère
S
Durand
CNRS UMR 8612, Laboratory of New Anticancer Targeting Strategies, Paris-Sud University, Galien Institute, Châtenay-Malabry, France.
Patrick
P
Couvreur
CNRS UMR 8612, Laboratory of New Anticancer Targeting Strategies, Paris-Sud University, Galien Institute, Châtenay-Malabry, France.
Didier
D
Desmaële
CNRS UMR 8203, Laboratory of Vectorology and Anticancer Therapeutics, Paris-Sud University, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France; Pharmacology and Drug Analysis Department, Gustave Roussy Cancer Campus Grand Paris, Villejuif, France.
Angelo
A
Paci
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D007069:Ifosfamide / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D016014:Linear Models; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
C039771:4-hydroxyifosfamide; D007069:Ifosfamide
4-Hydroxyifosfamide; HPLC–MS/MS; Mouse plasma; Pharmacokinetics; Preactivated ifosfamide