title: Stereoselective analysis of carvedilol in human plasma using HPLC/MS/MS after chiral derivatization.
abstract: A relatively high-throughput high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) method using a chiral derivatization reagent was developed for the quantitative determination of carvedilol enantiomers in human plasma. S-carvedilol and R-carvedilol are extracted from human plasma by protein precipitation using acetonitrile containing racemic [(2)H(5)]-carvedilol as an internal standard. Extracts are then derivatized with 2,3,4,6-tetra-O-acetyl-beta-glucopyranosyl isothiocyanate (GITC) and analysed using HPLC-MS/MS with a TurboIonspray (TIS) interface and selected reaction monitoring. Using 150microL of plasma, the method was validated over a concentration range of 0.2-200ng/mL. The maximum within-run precision observed in a three run quality control was 8.2% for S-carvedilol and 6.7% for R-carvedilol, respectively. The maximum percentage bias observed at all quality control sample concentrations was 9.4% for S-carvedilol and 11.6% for R-carvedilol, respectively. The HPLC-MS/MS method was also compared with a previously developed high-performance LC/fluorescence method by analysing 25 samples containing racemic carvedilol. Based on results obtained, these two methods were found to be equivalent. However, compared with LC/fluorescence method, HPLC-MS/MS method is more sensitive, uses less plasma, and also employs a less time-consuming sample preparation process.
authors.affiliation: Worldwide Bioanalysis, Drug Metabolism and Pharmacokinetics, 709 Swedeland Rd., King of Prussia, PA 19406, USA. eric.y.yang@gsk.com
authors.forename: Eric
authors.initials: E
authors.lastname: Yang
authors.forename: Sherry
authors.initials: S
authors.lastname: Wang
authors.forename: John
authors.initials: J
authors.lastname: Kratz
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Cyronak
mesh_terms: D002227:Carbazoles / Q000097:blood* / Q000737:chemistry*; D000077261:Carvedilol; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D011412:Propanolamines / Q000097:blood* / Q000737:chemistry*; D013237:Stereoisomerism
chemical_list: D002227:Carbazoles; D011412:Propanolamines; D000077261:Carvedilol