title: A sensitive LC-MS/MS method for quantifying clofarabine triphosphate concentrations in human peripheral blood mononuclear cells.
abstract: Clofarabine triphosphate is an intracellular active metabolite of clofarabine. In the present study, we developed and validated a rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for quantifying clofarabine triphosphate concentrations in human peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from blood using the Ficoll gradient centrifugation method. Chromatographic separation was performed on a CN column using an isocratic mobile phase comprising acetonitrile/5mM ammonium acetate with 0.001% ammonium hydroxide (20/80, v/v) at a flow rate of 0.60 mL/min. Detection was carried out by MS/MS in the multiple reaction monitoring mode using a negative electrospray ionization interface. The method was validated in concentration ranges of 1.25-100 ng/10(7) cells with acceptable accuracy and precision using 50 μL of cell extract. Clofarabine triphosphate was stable in a series of stability studies with bench-top, auto-sampler, and repeated freeze-thaw cycles. The validated method was successfully used to measure the concentrations of clofarabine triphosphate in PBMCs from cancer patients treated with clofarabine.
authors: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors: Xiaowei
authors: X
authors: Tu
authors: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors: Youming
authors: Y
authors: Lu
authors: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors: Dafang
authors: D
authors: Zhong
authors: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors: Yifan
authors: Y
authors: Zhang
authors: Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China. Electronic address: xychen@simm.ac.cn.
authors: Xiaoyan
authors: X
authors: Chen
mesh_terms: D000227:Adenine Nucleotides / Q000032:analysis* / Q000378:metabolism / Q000627:therapeutic use; D000964:Antimetabolites, Antineoplastic / Q000032:analysis* / Q000378:metabolism / Q000627:therapeutic use; D001087:Arabinonucleosides / Q000032:analysis* / Q000378:metabolism / Q000627:therapeutic use; D002853:Chromatography, Liquid / Q000191:economics / Q000379:methods*; D000077866:Clofarabine; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D057230:Limit of Detection; D009369:Neoplasms / Q000188:drug therapy; D011122:Polyphosphates / Q000032:analysis* / Q000378:metabolism / Q000627:therapeutic use; D053719:Tandem Mass Spectrometry / Q000191:economics / Q000379:methods*
chemical_list: D000227:Adenine Nucleotides; D000964:Antimetabolites, Antineoplastic; D001087:Arabinonucleosides; D011122:Polyphosphates; D000077866:Clofarabine; C005692:triphosphoric acid
keywords: Clofarabine; Clofarabine triphosphate; Liquid chromatography–tandem mass spectrometry; Peripheral blood mononuclear cells