alexa A sensitive LC-MS MS method for quantifying clofarabine triphosphate concentrations in human peripheral blood mononuclear cells.
Chemical Engineering

Chemical Engineering

Journal of Chromatography & Separation Techniques

Author(s): Tu X, Lu Y, Zhong D, Zhang Y, Chen X

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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. Copyright © 2014 Elsevier B.V. All rights reserved. This article was published in J Chromatogr B Analyt Technol Biomed Life Sci and referenced in Journal of Chromatography & Separation Techniques

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