LC–MS/MS-ESI method for simultaneous quantification of darolutamide and its active metabolite, ORM-15341 in mice plasma and its application to a pharmacokinetic study

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 145; pp. 454 - 461
• Main Authors: Dittakavi, Sreekanth, Nagasuri, Pavan Kumar V.S.P., Sulochana, Suresh P., Saim, Syed Mohd, Mallurwar, Sadanand R., Zainuddin, Mohd, Dewang, Purushottam, Rajagopal, Sriram, Mullangi, Ramesh
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 25.10.2017
• Subjects: Animals; Calibration; Chromatography, Liquid; Darolutamide; LC–MS/MS; Liquid-Liquid Extraction; Method validation; Mice; Mice plasma; ORM-15341; Pharmacokinetics; Pyrazoles - blood; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Mice plasma; Darolutamide; ORM-15341; Method validation; Pharmacokinetics; LC–MS/MS
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2017.06.074

[Display omitted] •First LC–MS/MS method being reported for the simultaneous quantification of darolutamide and ORM-15341 in mice plasma.•Simple sample processing method and shorter run time (2.5min) enables this method as a high throughput assay.•Method was validated as per regulatory guidelines.•The method is specific, precise, accurate and no matrix effect was observed.•Method was used in a mice pharmacokinetic study to derive the pharmacokinetic parameters for analytes. A sensitive and rapid LC–MS/MS method was developed and validated for the simultaneous quantitation of darolutamide and its active metabolite i.e. ORM-15341 in 50μL mice plasma using bicalutamide as an internal standard (I.S.) as per regulatory guidelines. Sample processing was accomplished through liquid-liquid extraction. Chromatographic separation was achieved using an Atlantis C18 column with an isocratic mobile phase comprising 0.2% formic acid:acetonitrile (35:65, v/v) at a flow rate of 0.8mL/min within 2.5min. Detection and quantitation were done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 397→202, 395→202 and 429→255 for darolutamide, ORM-15341 and I.S, respectively in the negative ionization mode. The calibration curve was linear from 0.61–1097ng/mL for both darolutamide and ORM-15341. The intra- and inter-day precisions were in the range of 1.34–13.8 and 4.85–12.9 and 3.91–13.7 and 6.54–14.2%, for darolutamide and ORM-15341, respectively. Darolutamide and ORM-15341 were found to be stable under different stability conditions. The validated method was applied to a pharmacokinetic study in mice.