Determination of topiramate in human plasma using liquid chromatography tandem mass spectrometry

The LC-MS/MS method for determination of the anti-epileptic drug topiramate (TPM) in human plasma was developed and validated for pharmacokinetic and bioequivalence study purposes. For quantitative determination of TPM values the method with deuterated internal standard (topiramate-d12) and liquid chromatography with tandem mass spectrometry was used. TPM was extracted from the human plasma using the solid-phase extraction procedure on a Strata X extraction column. Negative ions were monitored in the selected reaction monitoring mode (SRM) and transitions m/z 338.2 > 78.2 and m/z 350.3 > 78.2 were used for the quantitative evaluation of TPM and the internal standard, respectively. The results obtained from validation were statistically evaluated according to the requirements of European Medicines Agency (EMA) and Food and Drug Administration (FDA) regulatory guidelines. The linearity of the method was checked within a concentration range from 10 to 2000 ng/mL. Successful validation confirmed that this method is precise, accurate, sensitive and therefore suitable for determination of topiramate plasma levels in pharmacokinetic and bioequivalence studies.Razvili in validirali smo LC-MS/MS metodo za določevanje antiepileptičnega zdravila topiramat (TPM) v humani plazmi za namene študij farmakokinetike in bioekvivalence. Za kvantitativno določitev smo uporabili metodo devteriranega internega standarda (topiramat-d12) in tekočinsko kromatografijo s tandemsko masno spektrometrijo. Učinkovino TPM smo ekstrahirali iz humane plazme z ekstrakcijo na trdnem nosilcu na Strata X ekstrakcijskih kolonah. Spojine smo ionizirali z elektrorazprševanjem in merjenjem negativnih ionov, spremljali smo SRM prehoda 338,2 > 78,2 za TPM in 350,3 > 78,2 za interni standard. Rezultate vsebnosti topiramata smo statistično ovrednotili v skladu z zahtevami EMA regulative in FDA smernicami. Linearnost metode smo preverili v koncentracijskem območju od 10 do 2000 ng/ml. Uspešni rezultati testov so pokazali, da je razvita metoda natančna, točna in občutljiva ter primerna za določanje topiramata v humani plazmi v študijah farmakokinetike in bioekvivalence