Razvoj in validacija analizne metode za določanje plazemskih koncentracij antidepresivov desvenlafaksina, duloksetina, mirtazapina, sertralina in venlafaksina

Agović, Lejla (avtor), Vovk, Tomaž (mentor)

Terapevtsko spremljanje koncentracij zdravil (angl. Therapeutic Drug Monitoring – TDM) omogoča oblikovanje personaliziranega režima zdravljenja za posameznika z določanjem koncentracij učinkovin v bioloških tekočinah. Tak režim upošteva farmakokinetične in farmakodinamične variabilnosti posameznika. S tem zagotavlja najvišjo učinkovitost terapije in najmanj možnosti za pojav toksičnih ali neželenih učinkov, ter za pojav recidiva. V okviru magistrske naloge smo razvili metodo za sočasno določanje antidepresivov desvenlafaksina, duloksetina, mirtazapina, sertralina in venlafaksina. Vzorce obogatene plazme smo naalkalili in dodali paroksetin kot interni standard. Metoda je temeljila na ekstrakciji tekoče-tekoče s topilom etil acetat:cikloheksan 1:9 (v/v), sušenjem s prepihovanjem z dušikom in raztapljanjem suhega preostanka s topilom acetonitril:fosfatni pufer 1:9 (v/v). Kromatografsko metodo smo razvili na reverzno fazni koloni. Uporabili smo gradientno elucijo, s pretokom mobilne faze 1,2 ml/min, ki je trajala 20 minut. Desvenlafaksin in venlafaksin smo detektirali z UV-VIS detektorjem pri valovni dolžini 226 nm, duloksetin pri 230 nm, sertralin pri 220 nm, paroksetin pri 290 nm, mirtazapin pa s fluorescenčnim detektorjem pri ekscitacijski valovni dolžini 290 nm in emisijski valovni dolžini 370 nm. Razvito metodo za določanje plazemskih koncentracij desvenlafaksina, duloksetina in mirtazapina smo uspešno validirali po smernicah za validacijo bioanaliznih metod EMA. Dosegli smo območja metode med 10 in 300 ng/ml za desvenlafaksin, 5 in 200 ng/ml za duloksetin ter 10 in 200 ng/ml za mirtazapin. Metoda je selektivna, točna in natančna znotraj in med dnevi za te tri analite. Stabilnost vzorcev v avtomatskem vzorčevalniku pri 10 °C je za desvenlafaksin manj kot 6 ur, za duloksetin vsaj 24 ur in za mirtazapin 6 ur. Metode za določanje sertralina in venlafaksina nismo uspešno validirali. Na osnovi uspešnosti ekstrakcije razvite metode, uspešnosti ločbe analitov in validacijskih parametrov lahko ocenimo, da je razvita metoda primerna za določanje desvenlafaksina, duloksetina in mirtazapina pri terapevtskem spremljanju koncentracij zdravil.

• Jezik: Slovenski jezik
• Ključne besede: antidepresivi, TDM, desvenlafaksin, duloksetin, mirtazapin, paroksetin, sertralin, venlafaksin
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2021
• PID: 20.500.12556/RUL-128127
• Datum objave v RUL: 03.07.2021

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Development and validation of analytical method for plasma concentration determination of antidepressants desvenlafaxine, duloxetine, mirtazapine, sertraline and venlafaxine

Izvleček

Therapeutic Drug Monitoring enables determination of personalized treatment regimen created for individuals by determination of plasma concentrations of drugs in bodily fluids. Such regimens take into account pharmacokinetic and pharmacodynamic variability, thus ensuring the highest effectiveness of therapy and the least chance of toxic or side effects, as well as relapse. In master thesis we developed the analytical method for determination of antidepressants desvenlafaxine, duloxetine, mirtazapine, sertraline and venlafaxine. Spiked plasma samples were alkanized and paroxetine was added as internal standard. Analytes were extracted with liquid-liquid extraciton with solvent ethyl acetate:cyclohexane 1:9 (v/v), evaporated and reconstituted with acetonitrile:phosphate buffer 1:9 (v/v). Chromatographic separation was achieved using reversed phase column. Mobile phase was delivered at 1,2 ml/min with gradient program, with run time of developed method 20 minutes. UV-VIS detection was performed at wavelenght 226 nm for desvenlafaxine and venlafaxine, 230 nm for duloxetine, 290 for paroxetine and at 220 nm for sertraline. Mirtazapine was detected with fluorescence detector (λex 290 nm / λem 370 nm). Developed analytical method was successfully validated for determination of desvenlafaxine, duloxetine and mirtazapine, according to EMA guideline on bioanalytical method validation. Ranges were 10 – 300 ng/ml for desvenlafaxine, 5 – 200 ng/ml for duloxetine and 10 – 200 ng/ml for mirtazapine. Method is selective, accurate and precise within-run and between-run for these three analytes. Stability in autosampler at 10°C was less than 6 hours for desvenlafaxine, at least 24 hours for duloxetine and 6 hours for mirtazapine. Method was not successfully validated for determination of sertraline and venlafaxine. Based on successful recoveries of extraction, separation on the column and validation we can conclude that the developed method for plasma concentration determination of desvenlafaxine, duloxetine and mirtazapine is suitable for use in TDM.

• Ključne besede: antidepressants, TDM, desvenlafaxine, duloxetine, mirtazapine, paroxetine, sertraline, venlafaxine