A rapid ultra HPLC–MS/MS method for the quantitation and pharmacokinetic analysis of 3-deazaneplanocin A in mice

► We developed and validated a novel uHPLC–MS/MS assay for the quantification of 3-deazaneplanocin A (DZNep). ► This method has a faster run time and is more sensitive than previous methods. ► This method is accurate, precise, and a simple liquid extraction procedure recovers 90% of drug from mouse plasma. ► Successfully applied to preclinical pharmacokinetic study in mice. 3-Deazaneplanocin A (DZNep) has been shown to have anti-cancer activity in numerous cancer types and its continued preclinical, and eventual clinical, drug development will require rapid and sensitive bioanalytical methods in order to quantitate this drug for pharmacokinetic analyses. The ultra HPLC with positive thermospray tandem mass spectrometric (LC–MS/MS) detection affords the most sensitive (limit of quantitation 5ng/mL) and rapid (3min run time) bioanalytical method to date for DZNep. Due to the polar nature of this drug and the internal standard (tubercidin), a hydrophilic-interaction column (HILIC) was used. The method was accurate, with less than 10% deviation from nominal values, as well as precise, where both within-day and between-day precisions were less than 15%. A liquid–liquid extraction procedure was able to recover ∼90% of drug from a small volume (50μL) of mouse plasma. This method was successfully applied to a pharmacokinetic study in mice intravenously injected with DZNep.