title: Development and validation of dried blood spots technique for quantitative determination of topiramate using liquid chromatography-tandem mass spectrometry.
abstract: An LC-MS/MS method for determination of the anti-epileptic drug topiramate (TPM) in dried blood spot (DBS) samples was developed and validated. DBS samples were prepared by spotting 30 μL of spiked whole blood onto FTA(TM) DMPK-C cards and drying for at least 3 h. Six-millimetre punched spots were then extracted by using a mixture of methanol and water (90:10, v/v) with deuterated internal standard (topiramate-d12). The extracted samples were injected into a liquid chromatograph equipped with a tandem mass spectrometric detector. Negative ions were monitored in the selected reaction monitoring mode and transitions m/z 338.2 → 78.1 and m/z 350.3 → 78.1 were used for the quantitative evaluation of TPM and internal standard, respectively. The results obtained from validation were statistically evaluated according to the requirements of the European Medicines Agency and US Food and Drug Administration regulatory guidelines. The linearity of the method was checked within a concentration range from 10 to 2000 ng/mL. The validation results indicate that the method is accurate, precise, sensitive, selective and reproducible.
authors.affiliation: Krka, dd, Novo mesto, Šmarješka cesta 6, Novo mesto, Slovenia. tanja.vnucec-popov@krka.biz
authors.forename: Tanja Vnučec
authors.initials: TV
authors.lastname: Popov
authors.forename: Lea Cvitkovič
authors.initials: LC
authors.lastname: Maričič
authors.forename: Helena
authors.initials: H
authors.lastname: Prosen
authors.forename: Darinka Brodnjak
authors.initials: DB
authors.lastname: Vončina
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D059788:Dried Blood Spot Testing / Q000379:methods*; D004355:Drug Stability; D005632:Fructose / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000077236:Topiramate
chemical_list: D000077236:Topiramate; D005632:Fructose
keywords: dried blood spots; liquid chromatography tandem mass spectrometry; pharmacokinetic study; topiramate