title: Simultaneous determination of fludarabine and clofarabine in human plasma by LC-MS/MS.
abstract: A method for quantification of fludarabine (FDB) and clofarabine (CFB) in human plasma was developed with an API5000 LC-MS/MS system. FDB and CFB were extracted from EDTA plasma samples by protein precipitation with trichloroacetic acid. Briefly, 50 μL plasma sample was mixed with 25 μL internal standard (50 ng/mL aqueous 2-Cl-adensosine) and 25 μL 20% trichloroacetic acid, centrifuged at 25,000 × g (20,000 rpm) for 3 min, and then transfered to an autosampler vial. The extracted sample was injected onto an Eclipse extend C18 column (2.1 mm×150 mm, 5 μm) and eluted with 1mM NH4OH (pH 9.6) - acetonitrile in a gradient mode. Electrospray ionization in positive mode (ESI(+)) and multiple reaction monitoring (MRM) were used, and ion pairs 286/134 for FDB, 304/170 for CFB and 302/134 for the internal standard were selected for quantification. The retention times were typically 3.72 min for FDB, 4.34 min for the internal standard, 4.79 min for CFB. Total run time was 10 min per sample. Calibration range was 0.5-80 ng/mL for CFB and 2-800 ng/mL for FDB. The method was applied to a clinical pharmacokinetic study in pediatric patients.
authors.affiliation: Department of Clinical Pharmacy, University of California at San Francisco, San Francisco, CA 94143, USA. Electronic address: huangl@sfghsom.ucsf.edu.
authors.forename: Liusheng
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Clinical Pharmacy, University of California at San Francisco, San Francisco, CA 94143, USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Lizak
authors.affiliation: Department of Pediatrics, University of California at San Francisco, San Francisco, USA.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Dvorak
authors.affiliation: Department of Clinical Pharmacy, University of California at San Francisco, San Francisco, CA 94143, USA.
authors.forename: Francesca
authors.initials: F
authors.lastname: Aweeka
authors.affiliation: Department of Clinical Pharmacy, University of California at San Francisco, San Francisco, CA 94143, USA.
authors.forename: Janel
authors.initials: J
authors.lastname: Long-Boyle
mesh_terms: D000227:Adenine Nucleotides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D001087:Arabinonucleosides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D000077866:Clofarabine; D006801:Humans; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014740:Vidarabine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D000227:Adenine Nucleotides; D001087:Arabinonucleosides; D000077866:Clofarabine; D014740:Vidarabine; C024352:fludarabine
keywords: Clofarabine; Fludarabine; LC–MS/MS; Plasma