title: Validated bioanalytical method for the quantification of RGB-286638, a novel multi-targeted protein kinase inhibitor, in human plasma and urine by liquid chromatography/tandem triple-quadrupole mass spectrometry.
abstract: A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantitative determination of RBG-286638, a novel multi-targeted protein kinase inhibitor, in 200 microl aliquots of human potassium EDTA plasma with deuterated RGB-286638 as internal standard. The sample extraction and cleaning-up involved a simple liquid-liquid extraction with 100 microl aliquots of acetonitrile and 1 ml aliquots of n-butylchloride. Urine was accurately 5- and 10-fold diluted in blank plasma prior to extraction. Chromatographic separations were achieved on a reversed phase C18 column eluted at a flow-rate of 0.250 ml/min on a gradient of 0.2 mM ammonium formate and acetonitrile both acidified with 0.1% formic acid. The overall cycle time of the method was 7 min, with RGB-286638 eluting at 1.9 min. The multiple reaction monitoring transitions were set at 546>402 (m/z), and 549>402 (m/z) for RGB-286638 and the internal standard, respectively. The calibration curves were linear over the range of 2.00 to 1000 ng/ml with the lower limit of quantitation validated at 2.00 ng/ml. The within-run and between-run precisions were within 7.90%, while the accuracy ranged from 92.2% to 99.7%. The method was successfully applied to samples derived from a clinical study.
authors.affiliation: Department of Medical Oncology, Erasmus MC-Daniel den Hoed Cancer Center, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: de Bruijn
authors.forename: Inge M Ghobadi
authors.initials: IM
authors.lastname: Moghaddam-Helmantel
authors.forename: Maja J A
authors.initials: MJ
authors.lastname: de Jonge
authors.forename: Thorsten
authors.initials: T
authors.lastname: Meyer
authors.forename: Mei-Ho
authors.initials: MH
authors.lastname: Lam
authors.forename: Jaap
authors.initials: J
authors.lastname: Verweij
authors.forename: Erik A C
authors.initials: EA
authors.lastname: Wiemer
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Loos
mesh_terms: D002138:Calibration; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D004492:Edetic Acid / Q000737:chemistry; D005561:Formates / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D015233:Models, Statistical; D047428:Protein Kinase Inhibitors / Q000097:blood* / Q000652:urine*; D011720:Pyrazoles / Q000097:blood* / Q000652:urine*; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D014508:Urea / Q000031:analogs & derivatives* / Q000097:blood / Q000652:urine
chemical_list: D005561:Formates; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; C546217:RGB 286638; C030544:formic acid; D014508:Urea; D004492:Edetic Acid