title: Simultaneous quantification of ruxolitinib and nilotinib in rat plasma by LC-MS/MS: application to a pharmacokinetic study.
abstract: Efficacy assessments using a combination of ruxolitinib and nilotinib necessitate the development of a high precision analytical method for determination of both drugs in plasma. A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of ruxolitinib and nilotinib in rat plasma. Extraction of ruxolitinib, nilotinib and dasatinib (internal standard; IS) from 50μl rat plasma was carried out by protein precipitation with methanol. Chromatographic separation of analytes was performed on YMC pack ODS AM (150mm×4.6mm, 5μm) column under gradient conditions with acetonitrile:2.0mM ammonium acetate buffer as the mobile phase at a flow rate of 1ml/min. Precursor ion and product ion transition for both analytes and IS were monitored on a triple quadrupole mass spectrometer, operated in the selective reaction monitoring with positive ionization mode. Method was validated over a concentration range of 0.16-247ng/ml for ruxolitinib and 0.86-219ng/ml for nilotinib. Mean extraction recovery for ruxolitinib, nilotinib, and IS of 99.6%, 97.6% and 90.3% were consistent across low, medium, and high QC levels. Precision and accuracy at low, medium and high quality control levels were less than 15% across analytes. Bench top, wet, freeze-thaw and long term stability were evaluated for both analytes. The analytical method was applied to support a pharmacokinetic study of simultaneous estimation of ruxolitinib and nilotinib in Wistar rat. Assay reproducibility was demonstrated by re-analysis of 18 incurred samples.
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India; CRD, PRIST University, Vallam, Thanjavur 613403, Tamilnadu, India. Electronic address: sridharveeraraghavan@gmail.com.
authors.forename: Sridhar
authors.initials: S
authors.lastname: Veeraraghavan
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Satheeshmanikandan
authors.initials: S
authors.lastname: Thappali
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Srikant
authors.initials: S
authors.lastname: Viswanadha
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Sandhyarani
authors.initials: S
authors.lastname: Chennupati
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Santhoshkumar
authors.initials: S
authors.lastname: Nalla
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Manikantakumar
authors.initials: M
authors.lastname: Golla
authors.affiliation: Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
authors.forename: Swaroopkumar
authors.initials: S
authors.lastname: Vakkalanka
authors.affiliation: Department of Pharmaceutics, Annai JKK Sampoorani Ammal College of Pharmacy, Komarapalayam, Namakkal 638183, Tamilnadu, India.
authors.forename: Manivannan
authors.initials: M
authors.lastname: Rangasamy
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D009570:Nitriles; D011720:Pyrazoles / Q000097:blood* / Q000737:chemistry* / Q000493:pharmacokinetics; D011743:Pyrimidines / Q000097:blood* / Q000737:chemistry* / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009570:Nitriles; D011720:Pyrazoles; D011743:Pyrimidines; C540383:ruxolitinib; C498826:nilotinib
keywords: LC–MS/MS; Nilotinib; Plasma; Ruxolitinib