Procjena prikladnosti odabranih ciklodekstrina kao solubilizatora u in vitro ispitivanju permeabilnosti flutikazonpropionata

master's thesis

• Title (english): Suitability assessment of selected cyclodextrins as solubilizers in fluticasone propionate in vitro permeation studies
• Author: Mia Kobaš
• Mentor: Mario Jug (mentor)
• Mentor: Daniela Amidžić Klarić (mentor)
• Committee member: Mario Jug (predsjednik povjerenstva)
• Committee member: Anita Hafner (član povjerenstva)
• Committee member: Jasna Jablan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical technology); Zagreb
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2020-09-29, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Loša topljivost flutikazonpropionata u vodi predstavlja veliki izazov u razvoju i karakterizaciji novih formulacija
lijeka. U in vitro ispitivanjima oslobađanja i permeabilnosti nužno je uspostaviti uvjete osigurane topljivosti. U odnosu
na organska suotapala i površinski aktivne tvari, ciklodekstrini pružaju prednost izrazite biokompatibilnosti sa staničnim
modelima, što ih čini prikladnim solubilizatorima za tu namjenu.
Cilj ovog rada bio je razviti metodu tekućinske kromatografije visoke djelotvornosti za pouzdano određivanje
sadržaja flutikazonpropionata u vodenim otopinama α- i hidroksipropil-α-ciklodekstrina, te potom ispitati učinak
ciklodekstrina na povećanje topljivosti lijeka pri uvjetima definiranim potrebama provođenja ispitivanja permeabilnosti
na Calu-3 stanicama.
U ovom diplomskom radu uspješno je razvijena metoda obrnutofazne razdjelne kromatografije za kvantitativnu
analizu flutikazonpropionata. Analiza je provedena uz izokratnu eluaciju pri stalnom omjeru acetonitrila, metanola i
fosfatnog pufera pH 3,5 u mobilnoj fazi (50:15:35, V/V/V). Volumen injektiranja bio je 50 μl, temperatura kolone 40⁰C,
a brzina protoka mobilne faze iznosila je 1 ml/min. Pri navedenim uvjetima, područje linearnosti metode potvrđeno je
za koncentracijski raspon lijeka od 1 μg/ml do 30 μg/ml (koeficijent korelacije 0,9998). Granice dokazivanja i
određivanja metode iznosile su 0,24 μg/ml te 0,74 μg/ml. Uz ovu metodu uspješno su analizirani uzorci ispitivanja fazne
topljivosti kompleksa flutikazonpropionata s α- te hidroksipropil-α-ciklodekstrinom provedenih pri 37⁰C u Hankovoj
otopini soli koja sadrži ione kalcija (pH 7,4). Primjenom α- te hidroksipropil-α-ciklodekstrina pri koncentracijama od
50 mM, povećana je topljivost flutikazonpropionata za 19,5, odnosno 27,3 puta u odnosu na intrinzičnu topljivost lijeka.
Uzevši u obzir strukturne značajke ciklodekstrina i lijeka te profil izotermi topljivosti, zaključeno je da je povećanje
topljivosti flutikazonpropionata posljedica uklapanja lijeka u agregate kompleksa lijek/ciklodekstrin i/ili samih
ciklodekstrina neinkluzijskim mehanizmima.
Temeljem dobivenih rezultata, procijenjeno je kako solubilizacijski učinak α- te hidroksipropil-α-ciklodekstrina nije
dostatan za primjenu u in vitro ispitivanju permeabilnosti flutikazonpropionata na staničnim modelima. Rezultati ovog
rada mogu poslužiti kao temelj za daljnja istraživanja s drugim vrstama ciklodekstrina koji bi mogli imati bolji
solubilizacijski učinak na flutikazonpropionat.

Abstract (english)

Low aqueous solubility of fluticasone propionate poses a great challenge in new formulation development and
characterization. When conducting in vitro dissolution and permeation studies it is important to ensure that sink
conditions are met. Compared to organic cosolvents and surfactants, cyclodextrins offer the advantage of distinct
biocompatibility with cell lines, what makes them a suitable choice for solubilization purposes.
The aim of this study was to develop a reliable high performance liquid chromatography method for quantification
of fluticasone propionate in aqueous solutions containing α- and hydroxypropyl-α-cyclodextrin, as well as to investigate
the solubilization potential of these cyclodextrins at conditions defined by requirements of permeability tests on Calu-3
cell lines.
In this diploma thesis, a reverse-phase chromatographic method for quantitative analysis of fluticasone propionate
was successfully developed. The analysis was carried out with isocratic flow, at constant acetonitrile, methanol and
phosphate buffer pH 3.5 ratio within the mobile phase (50:15:35, V/V/V). The injection volume was 50 μl, column
temperature 40⁰C and flow rate 1 ml/min. At these conditions, the linearity range was confirmed within drug
concentration range of 1 μg/ml to 30 μg/ml (correlation coefficient 0.9998). The limit of detection and quantification
were found to be 0.24 μg/ml and 0.74 μg/ml, respectively. Using this method, samples obtained in phase solubility
studies of fluticasone propionate with α- and hydroxypropyl-α-cyclodextrin carried out at 37⁰C in Hank's balanced salt
solution containing calcium ions (pH 7.4), were successfully analysed. Using α- and hydroxypropyl-α-cyclodextrin at
50 mM concentration, the intrinsic solubility of fluticasone propionate was increased by 19.5 and 27.3 times,
respectively. Considering the structural characteristics of cyclodextrins and the drug, as well as the obtained phase
solubility diagram profiles, it was concluded that the enhancement in fluticasone propionate's solubility was the product
of its entrapment inside cyclodextrin and/or drug/cyclodextrin complex aggregates, through non-inclusion mechanisms.
The results obtained showed that the solubilization effect of α- and hydroxypropyl-α-cyclodextrin was not sufficient
enough to be applied for in vitro permeation studies of fluticasone propionate using cell models. The results of this study
could serve as a basis for further research of other cyclodextrins with better solubilization effect on fluticasone
propionate.

• Keywords: flutikazonpropionat; topljivost; α-ciklodekstrin; hidroksipropil-α-ciklodekstrin; HPLC analiza
• Keywords (english): fluticasone propionate; solubility; α-cyclodextrin; hydroxypropyl-α-cyclodextrin; HPLC analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:163:673303
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2022-09-29
• Created on: 2022-08-26 12:36:23