Validated liquid chromatography-tandem mass spectrometry method for simultaneous quantitation of bendamustine and copanlisib in mouse plasma: Application to a pharmacokinetic study in mice.
Harsha K TripathyS V Nair ManjuRama Murthy BesthaVinay KiranSreekanth DittakaviRamesh MullangiPublished in: Biomedical chromatography : BMC (2022)
In this study, we report the development and validation of an LC-tandem mass spectrometry method for the simultaneous quantitation of bendamustine and copanlisib in mouse plasma as per the US FDA regulatory guidelines. The sample processing involves extraction of bendamustine and copanlisib along with internal standard (IS; warfarin) from 50 μL mouse plasma using a liquid-liquid extraction method. The chromatographic separation of bendamustine, copanlisib and the IS was achieved on an Atlantis dC 18 column using an isocratic mobile phase (5 mM ammonium acetate:methanol, 20:80 v/v). Bendamustine, copanlisib and the IS eluted at 0.88, 1.39 and 0.74 min, respectively, with a total run time of 2.5 min. The calibration curve ranged from 3.99-2996 and 4.33-3248 ng/mL for bendamustine and copanlisib, respectively. Inter- and intra-day precision and accuracy, stability in processed samples and upon storage, dilution integrity and incurred sample reanalysis were investigated for both the analytes. The intra- and inter-day precisions were in the ranges of 2.01%-5.05% and 2.74%-6.13% and 1.98%-7.64 and 8.62%-9.04% for bendamustine and copanlisib, respectively. Stability studies showed that both analytes were stable on bench top for 6 h, in auto-sampler for 24 and at -80°C for 30 days. The validated method was successfully applied to a pharmacokinetic study in mice.
Keyphrases
- liquid chromatography tandem mass spectrometry
- simultaneous determination
- liquid chromatography
- tandem mass spectrometry
- solid phase extraction
- ultra high performance liquid chromatography
- high performance liquid chromatography
- chronic lymphocytic leukemia
- mass spectrometry
- high resolution mass spectrometry
- gas chromatography
- ms ms
- venous thromboembolism
- high resolution
- atrial fibrillation
- high fat diet induced
- direct oral anticoagulants
- transcription factor
- metabolic syndrome
- type diabetes
- clinical practice
- dendritic cells