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A rapid and sensitive ultra-high-performance liquid chromatography–quadrupole-time-of-flight mass spectrometric (UHPLC–Q-TOF-MS) method was developed for quantification of imipramine, one of the most widely used tricyclic antidepressants, and desipramine, an active metabolite of imipramine, in mouse serum. The developed method included a simple protein precipitation with acetonitrile in 50 μL of serum and analyte separation on an Acquity UPLC BEH C18 column using a gradient elution of acetonitrile with 0.1% formic acid and 20 mM ammonium formate. As a result, the entire analysis time was &lt;20 min including the sample preparation and the LC–MS analysis. The limit of quantification was 5.0 ng mL −1  for both imipramine and desipramine, and calibration curves were linear over the concentration range of 5.0–1,000.0 and 5.0–250.0 ng mL −1  for imipramine and desipramine, respectively. Intraday precisions at three levels were 2.2–3.6 and 1.7–4.2% for imipramine and desipramine, respectively, whereas interday precisions were 2.6–5.0 and 2.0–8.4% for imipramine and desipramine, respectively. Accuracy ranged between 93.6 and 106.6% for imipramine and 94.1 and 106.4% for desipramine. Absolute recovery was 96.0–97.6% for imipramine and 87.0–99.5% for desipramine. Finally, the described method was applied to mice administered with imipramine, demonstrating the suitability for quantification of imipramine and desipramine for therapeutic drug monitoring or bioequivalence studies.

journal of chromatographic science

A Simple, Rapid and Reliable Method to Determine Imipramine and Desipramine in Mouse Serum Using Ultra-High-Performance Liquid Chromatography–Quadrupole-Time-of-Flight Mass Spectrometry