Abstract

Abstract To support a phase 1 trial in patients with lymphomas, we developed a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for tazemetostat quantitation in 20 μL of human plasma. After protein precipitation, chromatographic separation employed a Kinetex C18 column and a gradient of 0.1% formic acid in both water and acetonitrile, during a 3‐min run time. Detection was achieved using a SCIEX 6500+ tandem mass spectrometer with electrospray positive‐mode ionization. Validation was based on the latest Food and Drug Administration guidance. With a stable isotopic internal standard, the assay was linear within the range of 10–5000 ng/mL and proved to be accurate (91.9%–103.7%) and precise (<4.4% imprecision). Recovery varied between 93.3% and 121.1%, and matrix effect ranged from −25.5% to −4.9%. Hemolysis, lipemia, and dilution did not impact quantitation. Plasma stability was confirmed after three freeze–thaw cycles, 24 h at room temperature, and 4 months at −80°C. Incurred sample reanalysis yielded 94.4% samples within 20% difference ( n = 36). External validation showed a mean bias of −11.1%. Pharmacokinetic (PK) data obtained from three patients suggested variable concentration time profiles, warranting collection of further data. The assay proved to be suitable for tazemetostat quantitation in human plasma and will support clinical studies by defining tazemetostat PKs.