title: Analysis of cyclophosphamide and carboxyethylphosphoramide mustard enantiomers in human plasma and application to clinical pharmacokinetics.
abstract: This study describes for the first time a method for the sequential analysis of the enantiomers of cyclophosphamide (CY) and its metabolite carboxyethylphosphoramide mustard (CEPM) in human plasma. The CY and CEPM enantiomers were extracted from plasma using only ethyl acetate and separated on a Chiralpak(®) AD-RH column using a mixture of water:acetonitrile:ethanol (45:30:25, v/v/v) plus 0.1% trifluoroacetic acid as the mobile phase at a flow rate of 0.5mL/min. No matrix effect was observed in the analysis of the enantiomers of both analytes and the analytical method was linear in the range of 0.05-25.0μg and 250-1000ng of each enantiomer/mL plasma. The coefficients of variation and relative errors obtained for the assessment of intra- and interassay precision and accuracy were less than 15%. CY and CEPM were found to be stable in human plasma after three successive freeze/thaw cycles, during storage for 4h at room temperature, and after 24h inside the autosampler at 4°C, with deviations less than 15%. The method was applied to the study of the pharmacokinetics of CY and its metabolite CEPM in patients with multiple sclerosis (n=10) who received a CY pretransplant conditioning regimen for hematopoietic stem cell transplantation. The pharmacokinetic parameters showed plasma accumulation of the (S)-(-)-CY enantiomer (S/R ratio=1.3) and lack of enantioselective exposure to the CEPM metabolite (S/R ratio=1.0).
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Avenida do Café s.n., Campus da USP, 14040-903 Ribeirão Preto, SP, Brazil.
authors.forename: Francine
authors.initials: F
authors.lastname: Attié de Castro
authors.affiliation: Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Gabriel dos Santos
authors.initials: Gdos S
authors.lastname: Scatena
authors.affiliation: Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Quézia Bezerra
authors.initials: QB
authors.lastname: Cass
authors.affiliation: Departamento de Química, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
authors.forename: Belinda Pinto
authors.initials: BP
authors.lastname: Simões
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Avenida do Café s.n., Campus da USP, 14040-903 Ribeirão Preto, SP, Brazil. Electronic address: lanchote@fcfrp.usp.br.
authors.forename: Vera Lucia
authors.initials: VL
authors.lastname: Lanchote
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D003520:Cyclophosphamide / Q000097:blood* / Q000493:pharmacokinetics*; D004355:Drug Stability; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006801:Humans; D009103:Multiple Sclerosis / Q000097:blood / Q000188:drug therapy; D010752:Phosphoramide Mustards / Q000097:blood* / Q000493:pharmacokinetics*; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods; D019172:Transplantation Conditioning
chemical_list: D010752:Phosphoramide Mustards; C445000:carboxyethylphosphoramide mustard; D003520:Cyclophosphamide
keywords: Carboxyethylphosphoramide mustard; Cyclophosphamide; Enantiomers; LC-MS/MS; Pharmacokinetics; Plasma