title: Development and validation of sensitive liquid chromatography/tandem mass spectrometry method for quantification of bendamustine in mouse brain tissue.
abstract: A liquid chromatography-tandem mass spectrometry method for quantification of bendamustine in mouse brain tissue was developed and fully validated. Methanol was used to precipitate proteins in brain tissue. Bendamustine and internal standard (chlorambucil) were separated with reverse-phase chromatography on a C-18 column with a gradient of water and 95% methanol in 0.1% formic acid. Positive mode electrospray ionization was applied with selected reaction monitoring to achieve 5 ng/ml lower limits of quantitation in mouse brain tissue. The calibration curve for bendamustine in mouse brain was linear between 5 and 2000 ng/ml. The within- and between-batch accuracy and precision of the assay were within 15% at 10, 100 and 1000 ng/ml. The recovery and matrix effect of bendamustine in mouse brain tissue ranged from 41.1% to 51.6% and 107.4% to 110.3%, respectively. The validated method was then applied to quantitate bendamustine in an animal study. Results indicate the assay can be applied to evaluate bendamustine disposition in mouse brain tissue. This assay will be applied in the future to detect and quantify bendamustine in human brain tissue samples.
authors.affiliation: Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH 43210, United States.
authors.forename: Lei
authors.initials: L
authors.lastname: He
authors.forename: John C
authors.initials: JC
authors.lastname: Grecula
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Ling
authors.forename: Michael D
authors.initials: MD
authors.lastname: Enzerra
authors.forename: Mario
authors.initials: M
authors.lastname: Ammirati
authors.forename: Kari
authors.initials: K
authors.lastname: Kendra
authors.forename: Robert
authors.initials: R
authors.lastname: Cavaliere
authors.forename: Nina
authors.initials: N
authors.lastname: Mayr
authors.forename: John
authors.initials: J
authors.lastname: McGregor
authors.forename: Thomas
authors.initials: T
authors.lastname: Olencki
authors.forename: Ewa
authors.initials: E
authors.lastname: Mrozek
authors.forename: Mani
authors.initials: M
authors.lastname: Matharbootham
authors.forename: Chima
authors.initials: C
authors.lastname: Oluigbo
authors.forename: Mitch A
authors.initials: MA
authors.lastname: Phelps
mesh_terms: D000818:Animals; D000069461:Bendamustine Hydrochloride; D001923:Brain Chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D016018:Least-Squares Analysis; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009588:Nitrogen Mustard Compounds / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009588:Nitrogen Mustard Compounds; D000069461:Bendamustine Hydrochloride