Razvoj analizne metode in vrednotenje sproščanja cinarizina iz tablet v medijih, ki ponazarjajo prisotnost lipidov v hrani

Šifrer, Nina (avtor), Trontelj, Jurij (mentor), Felicijan, Tjaša (komentor)

Cinarizin je lipofilna učinkovina s slabo vodotopnostjo, in vivo podatki pa kažejo velik vpliv hrane na njegovo biološko uporabnost. V sklopu magistrske naloge smo razvili analizno metodo na osnovi tekočinske kromatografije visoke ločljivosti (HPLC) in vrednotili sproščanje cinarizina iz tablet v medijih, ki ponazarjajo prisotnost lipidov v hrani. Razvito analizno metodo smo ovrednotili po smernicah za validacijo analiznih metod ICH Q2(R2). Razvili smo ustrezen postopek priprave vzorcev za analizo s HPLC. Sproščanje cinarizina smo testirali v 0,01 M HCl, fosfatnem pufru pH 6,8 in zmeseh SMOFlipid®-a z 0,01 M HCl oziroma s fosfatnim pufrom pH 6,8. Redčitve smo izbrali na podlagi literature, kjer so podoben lipidni medij uporabili pri pripravi biorelevantnega medija za ponazoritev stanja v želodcu po obroku. Teste sproščanja smo izvajali na napravi z vesli pri hitrosti vrtenja mešal 75 obratov/min. Koncentracijo sproščenega cinarizina smo izračunali s pomočjo umeritvenih premic, ki smo jih pripravili za vsak medij, v katerem smo testirali sproščanje. Analizirali smo delež sproščenega cinarizina v vodni (spodnji) in maščobni (zgornji) fazi vzorcev sproščanja. S preverjanjem masne bilance pri izbranih poskusih sproščanja in poskusih s standardi smo potrdili, da se s seštevkom določenih deležev cinarizina v posameznih fazah približamo 100 % celotnemu deležu v vsaki časovni točki sproščanja. Iz rezultatov poskusov sproščanja cinarizina smo ugotovili, da je sproščanje v zmeseh SMOFlipid®-a z 0,01 M HCl hitrejše od sproščanja v zmeseh s fosfatnim pufrom pH 6,8. Razlog je v dobri topnosti cinarizina v mediju s HCl in slabi topnosti v fosfatnem pufru. Lipidi, ki so prisotni v SMOFlipid®-u, vplivajo na sproščanje cinarizina iz tablet ter na porazdelitev cinarizina med maščobno in vodno fazo. Pri manj redčenih zmeseh SMOFlipid®-a s HCl je prisotnih več lipidov, ki sproščanje nekoliko upočasnijo. Več cinarizina se porazdeli v maščobno fazo, se pa zaradi dobre topnosti v mediju s HCl veliko cinarizina porazdeli tudi v vodno fazo. Manj kot so redčene zmesi SMOFlipid®-a s fosfatnim pufrom, hitrejše je sproščanje in večji je delež sproščenega cinarizina. V vseh zmeseh s fosfatnim pufrom se praktično ves cinarizin porazdeli v maščobno fazo. Dokazali smo, da lahko z različnimi deleži lipidov in pH vrednostmi medijev vplivamo na sam potek sproščanja cinarizina. Na podlagi tega sklepamo, da lahko z uporabo preizkušenih medijev dosežemo in vitro profil sproščanja cinarizina, ki je lahko reprezentativen za in vivo stanje po obroku.

• Jezik: Slovenski jezik
• Ključne besede: cinarizin, SMOFlipid®, HPLC, razvoj metode, sproščanje, masna bilanca
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2023
• PID: 20.500.12556/RUL-146722
• Datum objave v RUL: 09.06.2023

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Development of an analytical method and evaluation of cinnarizine release from tablets in media, simulating the presence of lipids in food

Izvleček

Cinnarizine is a lipophilic active ingredient with poor aqueous solubility. In vivo data show a significant impact of food on its bioavailability. We developed an analytical method based on high performance liquid chromatography (HPLC) and evaluated the release of cinnarizine from tablets in media, simulating the presence of lipids in food. The developed analytical method was evaluated according to the Guidelines for validation of analytical methods ICH Q2(R2). We developed an appropriate sample preparation process for subsequent HPLC analysis. The release of cinnarizine was tested in 0.01 M HCl, phosphate buffer pH 6.8, and in SMOFlipid® mixtures with 0.01 M HCl, and with phosphate buffer pH 6.8. Dilutions were selected based on the literature, where a similar lipid medium was used in the preparation of biorelevant media to simulate the gastric fed state conditions. Dissolution tests were performed on an apparatus II at 75 rotations per minute. The concentration of the released cinnarizine was calculated using the calibration curves, prepared in each of the tested media. We analyzed the percentage of cinnarizine released in the upper lipid and lower aqueous phase of release samples. With a mass balance analysis in selected release samples, and in standard solutions, we have confirmed that the sum of percentages of cinnarizine in individual phases, approaches 100 % of the total amount of cinnarizine at each release time-point. We concluded that the release of cinnarizine in SMOFlipid® mixtures with 0.01 M HCl is faster than in mixtures with phosphate buffer pH 6.8, due to the good solubility of cinnarizine in HCl and the poor solubility in phosphate buffer. The lipids present in SMOFlipid® affect the release of cinnarizne from tablets and the distribution of cinnarizine between the aqueous and lipid phase. In less diluted SMOFlipid® mixtures with HCl, more lipids are present which slow down the release slightly. More cinnarizine is distributed into the lipid phase, but due to its good solubility in media containing HCl, a significant amount of cinnarizine is also distributed into the aqueous phase. The less diluted are SMOFlipid® mixtures with phosphate buffer, the faster is the observed release, and the higher is the percentage of cinnarizine released. In all mixtures with phosphate buffer, practically all of the cinnarizine is distributed into the lipid phase. We have proven that with different proportions of lipids and pH values of the media, we can influence the release of cinnarizine. This leads us to conclusion that by the use of the tested media, it is possible to achieve an in vitro release profile of cinnarizine, that may be representative for the fed state in vivo.

• Ključne besede: cinnarizine, SMOFlipid®, HPLC, method developement, drug release, mass balance