title: Dried blood spot UHPLC-MS/MS analysis of oseltamivir and oseltamivircarboxylate--a validated assay for the clinic.
abstract: The neuraminidase inhibitor oseltamivir (Tamiflu®) is currently the first-line therapy for patients with influenza virus infection. Common analysis of the prodrug and its active metabolite oseltamivircarboxylate is determined via extraction from plasma. Compared with these assays, dried blood spot (DBS) analysis provides several advantages, including a minimum sample volume required for the measurement of drugs in whole blood. Samples can easily be obtained via a simple, non-invasive finger or heel prick. Mainly, these characteristics make DBS an ideal tool for pediatrics and to measure multiple time points such as those needed in therapeutic drug monitoring or pharmacokinetic studies. Additionally, DBS sample preparation, stability, and storage are usually most convenient. In the present work, we developed and fully validated a DBS assay for the simultaneous determination of oseltamivir and oseltamivircarboxylate concentrations in human whole blood. We demonstrate the simplicity of DBS sample preparation, and a fast, accurate and reproducible analysis using ultra high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer. A thorough validation on the basis of the most recent FDA guidelines for bioanalytical method validation showed that the method is selective, precise, and accurate (≤15% RSD), and sensitive over the relevant clinical range of 5-1,500 ng/mL for oseltamivir and 20-1,500 ng/mL for the oseltamivircarboxylate metabolite. As a proof of concept, oseltamivir and oseltamivircarboxylate levels were determined in DBS obtained from healthy volunteers who received a single oral dose of Tamiflu®.
authors.affiliation: Department of Neurology, Laboratory of Neuro-Oncology and Clinical and Cancer Proteomics, University Medical Center Rotterdam (Erasmus MC), Rotterdam, The Netherlands. g.hooff@erasmusmc.nl
authors.forename: Gero P
authors.initials: GP
authors.lastname: Hooff
authors.forename: Roland J W
authors.initials: RJ
authors.lastname: Meesters
authors.forename: Jeroen J A
authors.initials: JJ
authors.lastname: van Kampen
authors.forename: Nick A
authors.initials: NA
authors.lastname: van Huizen
authors.forename: Birgit
authors.initials: B
authors.lastname: Koch
authors.forename: Asmar F Y
authors.initials: AF
authors.lastname: Al Hadithy
authors.forename: Teun
authors.initials: T
authors.lastname: van Gelder
authors.forename: Albert D M E
authors.initials: AD
authors.lastname: Osterhaus
authors.forename: Rob A
authors.initials: RA
authors.lastname: Gruters
authors.forename: Theo M
authors.initials: TM
authors.lastname: Luider
mesh_terms: D000998:Antiviral Agents; D001793:Blood Preservation; D001800:Blood Specimen Collection / Q000379:methods*; D002624:Chemistry, Clinical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D053139:Oseltamivir / Q000031:analogs & derivatives / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000998:Antiviral Agents; D053139:Oseltamivir