1. Academic Validation
  2. Simultaneous quantification of lenalidomide, ibrutinib and its active metabolite PCI-45227 in rat plasma by LC-MS/MS: application to a pharmacokinetic study

Simultaneous quantification of lenalidomide, ibrutinib and its active metabolite PCI-45227 in rat plasma by LC-MS/MS: application to a pharmacokinetic study

  • J Pharm Biomed Anal. 2015 Mar 25;107:151-8. doi: 10.1016/j.jpba.2014.11.041.
Sridhar Veeraraghavan 1 Srikant Viswanadha 2 Satheeshmanikandan Thappali 2 Babu Govindarajulu 2 Swaroopkumar Vakkalanka 2 Manivannan Rangasamy 3
Affiliations

Affiliations

  • 1 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.
  • 2 Incozen Therapeutics Private Limited, 450, Alexandira Knowledge Park, Turkapplly, Hyderabad 500078, Andhra Pradesh, India.
  • 3 Dept. of Pharmaceutics, Annai JKK Sampoorani Ammal College of Pharmacy, Komarapalayam, Namakkal 638 183, Tamilnadu, India.
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.

Keywords

Ibrutinib; LC–MS/MS; Lenalidomide; PCI-45227; Plasma.

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