title: Simultaneous quantification of lenalidomide, ibrutinib and its active metabolite PCI-45227 in rat plasma by LC-MS/MS: application to a pharmacokinetic study.
abstract: Efficacy assessments using a combination of ibrutinib and lenalidomide necessitate the development of an analytical method for determination of both drugs in plasma with precision. A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of lenalidomide, ibrutinib, and its active metabolite PCI45227 in rat plasma. Extraction of lenalidomide, ibrutinib, PCI45227 and tolbutamide (internal standard; IS) from 50 μl rat plasma was carried out by liquid-liquid extraction with ethyl acetate:dichloromethane (90:10) ratio. Chromatographic separation of analytes was performed on YMC pack ODS AM (150 mm × 4.6 mm, 5 μm) column under gradient conditions with acetonitrile:0.1% formic acid buffer as the mobile phases at a flow rate of 1 ml/min. Precursor ion and product ion transition for 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 0.72-183.20 ng/ml for ibrutinib, 0.76-194.33 ng/ml for PCI-45227 and 1.87-479.16 ng/ml for lenalidomide. Mean extraction recovery for ibrutinib, PCI-45227, lenalidomide and IS of 75.2%, 84.5%, 97.3% and 92.3% 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 was evaluated for all the analytes. The analytical method was applied to support a pharmacokinetic study of simultaneous estimation of lenalidomide, ibrutinib, and its active metabolite PCI-45227 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: sridhar.veeraraghavan@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: 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: 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: Babu
authors.initials: B
authors.lastname: Govindarajulu
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: Dept. of Pharmaceutics, Annai JKK Sampoorani Ammal College of Pharmacy, Komarapalayam, Namakkal 638 183, Tamilnadu, India.
authors.forename: Manivannan
authors.initials: M
authors.lastname: Rangasamy
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004355:Drug Stability; D000077269:Lenalidomide; D059625:Liquid-Liquid Extraction / Q000379:methods; D008297:Male; D010880:Piperidines; D010949:Plasma / Q000737:chemistry*; D011720:Pyrazoles / Q000097:blood* / Q000737:chemistry*; D011743:Pyrimidines / Q000097:blood* / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods; D013792:Thalidomide / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D014044:Tolbutamide / Q000097:blood / Q000737:chemistry
chemical_list: C000602566:PCI-45227; D010880:Piperidines; D011720:Pyrazoles; D011743:Pyrimidines; C551803:ibrutinib; D013792:Thalidomide; D014044:Tolbutamide; D000077269:Lenalidomide; D000225:Adenine
keywords: Ibrutinib; LC–MS/MS; Lenalidomide; PCI-45227; Plasma