Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5136360 | Journal of Chromatography B | 2017 | 10 Pages |
Abstract
Purine nucleoside analogues are widely used in the treatment of haematological malignancies, and their biological activity is dependent on the intracellular accumulation of their triphosphorylated metabolites. In this context, we developed and validated a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to study the formation of 5â²-triphosphorylated derivatives of cladribine, fludarabine, clofarabine and 2â²-deoxyadenosine in human cancer cells. Br-ATP was used as internal standard. Separation was achieved on a hypercarb column. Analytes were eluted with a mixture of hexylamine (5 mM), DEA (0.4%, v/v, pH 10.5) and acetonitrile, in a gradient mode at a flow rate of 0.3Â mLÂ minâ1. Multiple reactions monitoring (MRM) and electrospray ionization in negative mode (ESI-) were used for detection. The application of this method to the quantification of these phosphorylated cytotoxic compounds in a human follicular lymphoma cell line, showed that it was suitable for the study of relevant biological samples.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Jean-Yves Puy, Lars Petter Jordheim, Emeline Cros-Perrial, Charles Dumontet, Suzanne Peyrottes, Isabelle Lefebvre-Tournier,