title: Development and validation of a highly sensitive liquid chromatography/mass spectrometry method for simultaneous quantification of lenalidomide and flavopiridol in human plasma.
abstract: Lenalidomide, an immunomodulatory agent, and flavopiridol, a broad cyclin-dependent kinase inhibitor, are active therapies for clinical use in genomic high-risk chronic lymphocytic leukemia. A high-performance liquid chromatographic assay with tandem mass spectrometric detection has been developed to simultaneously quantify lenalidomide and flavopiridol in human and mouse plasma to facilitate their combined clinical development. Samples were prepared by liquid-liquid extraction with acetonitrile (ACN)-containing internal standard, genistein, followed by evaporation of solvent and reconstitution in 95/5 H2O/ACN. Lenalidomide and internal standard were separated by reversed-phase liquid chromatography on a C-18 column using a gradient of H2O and ACN, each with 0.1% formic acid. Atmospheric pressure chemical ionization in positive ion mode with single reaction monitoring on a triple quadrupole mass spectrometer was applied to detect transitions of lenalidomide (260.06 > 149.10) and flavopiridol (402.09 > 341.02). Lower limits of quantification of lenalidomide and flavopiridol were 1 and 0.3 nM, respectively. Recoveries of lenalidomide and flavopiridol from human plasma ranged from 99% to 116% throughout their linear ranges. Within- and between-run precision and accuracy of replicate samples were all less than 15%. This is the most sensitive analytical method reported to date for both lenalidomide and flavopiridol. This sensitivity will enable late terminal phase concentration measurements and accurate pharmacokinetic parameter estimation in a planned clinical trial with lenalidomide and flavopiridol in patients with chronic lymphocytic leukemia.
authors: Division of Pharmaceutics, College of Pharmacy, Comprehensive Cancer Center,The Ohio State University, 500 West 12th Avenue, Columbus, OH 43210, USA.
authors: Qing
authors: Q
authors: Liu
authors: Katherine L
authors: KL
authors: Farley
authors: Amy J
authors: AJ
authors: Johnson
authors: Natarajan
authors: N
authors: Muthusamy
authors: Craig C
authors: CC
authors: Hofmeister
authors: Kristie A
authors: KA
authors: Blum
authors: Larry J
authors: LJ
authors: Schaaf
authors: Michael R
authors: MR
authors: Grever
authors: John C
authors: JC
authors: Byrd
authors: James T
authors: JT
authors: Dalton
authors: Mitch A
authors: MA
authors: Phelps
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods* / Q000592:standards; D005419:Flavonoids / Q000097:blood* / Q000737:chemistry; D006801:Humans; D000077269:Lenalidomide; D009101:Multiple Myeloma / Q000097:blood / Q000188:drug therapy; D010880:Piperidines / Q000097:blood* / Q000737:chemistry; D012008:Recurrence; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods / Q000592:standards*; D013792:Thalidomide / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D013997:Time Factors
chemical_list: D005419:Flavonoids; D010880:Piperidines; C077990:alvocidib; D013792:Thalidomide; D000077269:Lenalidomide