Razvoj visoko zmogljivega preskusa sproščanja na avtomatskem pipetorju : [diplomska naloga]

Čuček, Evelin (avtor), Žakelj, Simon (mentor), Kristan, Katja (komentor)

Razvoj visoko zmogljive metode je za farmacevtsko industrijo pomemben, saj lahko tako istočasno analiziramo veliko več vzorcev, hkrati pa porabimo manjše količine medija za sproščanje (9 ml namesto 1 L). Proces je tudi avtomatiziran, s tem pa zmanjšamo še variabilnost zaradi same izvedbe poskusa. V farmacevtski industriji je preskus raztapljanja aktivne učinkovine iz končnih zdravilnih oblik pomembno orodje v procesu odkrivanja novih učinkovin in pri analizi kakovosti zdravil. Cilj diplomske naloge je bil, da za farmacevtsko obliko s prirejenim sproščanjem razvijemo test raztapljanja z visoko zmogljivo metodo na avtomatskem pipetorju. Uporabili smo različne vzorce, ki so vsebovali težko topno učinkovino ketoprofen v obliki pelet. Testirali smo parametre, ki bi lahko vplivali na sproščanje ketoprofena: število analiziranih pelet, hitrost stresanja, temperatura medija za sproščanje in predinkubacija v kislini. Ugotovili smo, da se sproščanje učinkovine pri večji količini pelet zmanjša, verjetno zato, ker raztapljanje omejuje topnost ketoprofena. Posledično smo naše nadaljnje analize izvajali na manjši količini pelet (25 % mase zatehte v kapsuli ali manj). Izkazalo se je, da so naše pelete zelo robustne in odporne, saj se sproščanje učinkovine pri najvišji hitrosti stresanja ni bistveno povečalo. Je pa sproščanje učinkovine močno odvisno od temperature, saj se je pri višji temperaturi, kot pričakovano, sprostilo več učinkovine. Dodatno smo še ugotovili, da se učinkovina iz obloženih pelet, ki so se inkubirale v 1 M HCL, ni sprostila. Da bi preverili vplive kombinacij različnih parametrov na sproščanje učinkovine (količina pelet 25, 17,5 in 10 % mase polnjenja v kapsule, temperatura 22, 37 in 50 °C in hitrost stresanja 0, 6 in 8 Hz), smo se poslužili statističnega načrtovanja eksperimentov in po tem načrtu izvajali testiranja. Za identifikacijo pogojev, pri katerih je raztapljanje pelet ketoprofena z visoko zmogljivo metodo najbolj primerljivo z raztapljanjem v USP aparaturi 1, smo za vsako kombinacijo pogojev izračunali korelacijo med obema metodama. Spremljali smo naklon korelacijske premice (k), standardni odklon od premice (Syx) in korelacijski koeficient (R²). Rezultate smo vstavili v program Modde, ki nam je podatke prilagodil na regresijski model. Najboljšo korelacijo med obema metodama za pelete s ketoprofenom smo dobili pri eksperimentu, kjer smo uporabili 25 % mase pelet, temperaturo 22 °C in je eksperiment potekal brez stresanja (0 Hz), saj so bile vrednosti R2 najvišje (0,9707) in vrednosti Syx najnižje (5,35). Izkazalo se je, da je za dobro korelacijo pomembno bolj počasno sproščanje pri nizki temperaturi in višja količina pelet, ki zmanjša variabilnost. Rezultati kažejo, da lahko s pravilno izbiro parametrov dosežemo dobro korelacijo med obema metodama.

• Jezik: Slovenski jezik
• Ključne besede: testi raztapljanja statistično načrtovanje eksperimentov temperatura pH pelete ketoprofen tresenje inkubatorjev
• Vrsta gradiva: Diplomsko delo
• Tipologija: 2.11 - Diplomsko delo
• Organizacija: FFA - Fakulteta za farmacijo
• Kraj izida: Ljubljana
• Založnik: [E. Čuček]
• Leto izida: 2014
• Št. strani: VI, 53 f., P1-P43
• PID: 20.500.12556/RUL-121068
• UDK: 615.45.014+66.061+661.12(043.2)
• COBISS.SI-ID: 3624305
• Datum objave v RUL: 29.09.2020

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Development of a high-throughput automated dissolution method on automated pipettor

Izvleček

In the pharmaceutical industry, dissolution testing of the active ingredient of the final dosage forms is an important tool in the process of drug discovery and analysing the quality of medicinal products. The aim of the thesis was to develop the high-performance dissolution method for a pharmaceutical form with modified release, utilizing an automatic pipettor. Pellet samples with different amount of controlled release coating and an insoluble substance ketoprofen were used. We tested the parameters that could affect the release of ketoprofen: the amount of pellets, agitation rate, temperature and the influence of pre-incubation in acid. We found that the release of the active ingredient from a larger quantity of pellets is reduced, probably because the solubility of ketoprofen limits the dissolution. As a result further analyses were carried out on a smaller amount of pellets (25% of the weight in the capsule or less). It was found that the tested pellets were very robust and resistant to agitation, because the agitation rate did not influence the release of the active ingredient significantly. However, the release of the active ingredient depends strongly on the temperature. As was expected, the release of the active ingredient was higher at higher temperature. Additionally, we observed that the active ingredient of the pellets, which were incubated in 1 M HCl, was not released. To achieve the best correlation to the classical method, various parameters, influencing the release of the active ingredient (amount of pellets: 25, 17,5 and 10 % of capsule filling mass, the temperature of 22, 37 and 50 ° C and the agitation rate of 0, 6 and 8 Hz) were evaluated by design of experiments (DoE). The responses studied were the correlation of the scaled-down dissolution method with the classical method (R2), the slope of the correlation curve (k), and the standard deviation between the methods (Syx). The responses were adjusted to the regression model. The results have revealed that low T (22 °C), no shaking and high amount of pellets (25% capsule filling mass) were parameters that resulted in the best correlation between both dissolution methods for ketoprofen controlled release pellets. At these conditions the value of R2 (0,9707) was the highest and value of SYX (5.35) the lowest. (It has thus been shown that for good correlation it is important to achieve a slow release at low temperature, while a higher amount of pellets reduces variability. These results show that with proper selection of parameters we can achieve a good correlation between the two methods. Since the rate and extent of samples analysed by classical dissolution testing is limited, it is important to develop alternative high-throughput methods for pharmaceutical industry.

• Ključne besede: dissolution high-throughput dissolution method design of experiments temperature pH