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Analysis of cyclophosphamide and five metabolites from human plasma using liquid chromatography–mass spectrometry and gas chromatography–nitrogen–phosphorus detection |
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| Authors: | Thomas F. Kalhorn Song Ren William N. Howald Ross F. Lawrence John T. Slattery |
| Affiliation: | a Department of Pharmaceutics, University of Washington, Seattle, WA 98195, USA;b Department of Medicinal Chemistry, University of Washington, Seattle, WA 98195, USA;c Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N., AB-122, P.O. Box 19024, Seattle, WA 98104, USA |
| Abstract: | An assay method for the quantification of cyclophosphamide (CY) and five metabolites from human plasma is presented. The procedure is adapted to the chemical properties of the compounds of interest: non-polar compounds are extracted into methylene chloride, concentrated and analyzed by GC–NPD after derivatization, and the remaining aqueous fraction is deproteinated with acetonitrile–methanol prior to separation via reversed-phase HPLC and detection using atmospheric pressure ionization (API)-MS. Standard curves are linear over the required range and reproducible over five months. Plasma concentration–time profiles of CY and metabolites from a patient receiving CY by intravenous infusion (60 mg/kg, once a day for two days) are presented. |
| Keywords: | Cyclophosphamide Deschloroethylcyclophosphamide 4-Oxocyclophosphamide Carboxyethylphosphoramide mustard Phosphoramide mustard Hydroxypropylphosphoramide mustard |
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