Razvoj kapilarnoelektroforetske metode za istovremeno određivanje gemcitabina i njegovog onečišćenja

master's thesis

• Title (english): Development of a capillary electrophoretic method for simultaneous determination of gemcitabine and its impurity
• Author: Marija Hranilović
• Mentor: Miranda Sertić (mentor)
• Committee member: Miranda Sertić (predsjednik povjerenstva)
• Committee member: Biljana Nigović (član povjerenstva)
• Committee member: Zrinka Rajić Džolić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2017-06-13, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Gemcitabin, 2',2'-difluoro-2'-deoksicitidin, antimetabolit pirimidina, je antineoplastični lijek koji se koristi u liječenju brojnih uznapredovalih ili metastatskih karcinoma kao monoterapija ili u kombinaciji s drugim citostaticima, ovisno o indikaciji.
Cilj je ovog istraživanja bio razviti novu, brzu, jednostavnu i ekološki prihvatljivu kapilarnoelektroforetsku metodu za istovremenu analizu gemcitabina i njegovog onečišćenja cizotina. Kao prikladna tehnika odabrana je kapilarna elektroforeza kao "zelena" alternativa već razvijenoj HPLC metodi za određivanje sadržaja gemcitabina i određivanje onečišćenja uvrštenoj u 8. izdanje Europske farmakopeje.
Analiza je vršena u kapilari duljine 35 cm (27 cm) pri temperaturi od 25 °C, na valnoj duljini detekcije od 275 i 237 nm. Tijekom razvoja nove metode ispitane su različite vrste radnih pufera: boratni (pH 9,3), fosfatni (pH 7,0) i acetatni (pH 3,0). Na temelju rezultata eksperimenata najbolji puferom pokazao se fosfatni pufer (pH 7,0), dok je bolje razdvajanje i oblik pikova postignut dodatkom veće koncentracije SDS-a. Optimalnim se pokazala 50 mM koncentracija SDS-a. Ispitan je i utjecaj napona na vrijeme analize te razlučivanje između analita. Kod napona od 10 kV postignuto je bolje razdvajanje gemcitabina i citozina. Međutim, primjenom nižeg napona produljuje se vrijeme putovanja analita kroz kapilaru pa je potrebno voditi računa o tome da je razlučivanje između analita što bolje, no istovremeno uz što kraće vrijeme analize.
Predlaže se daljnja optimizacija metode kako bi se postiglo prikladno razlučivanje pikova te njihov odgovarajući oblik i simetrija uz što kraće vrijeme analize.

Abstract (english)

Gemcitabine, 2', 2'-difluoro-2'-deoxycytidine, pyrimidine antimetabolite, is an antineoplastic drug used to treat many advanced or metastatic cancers as monotherapy or in combination with other cytostatics, depending on the indication.
The aim of this study was to develop a new, fast, simple and environmentally friendly capillary electrophoretic method for simultaneous identification and determination of gemcitabine and its impurity cytosine. Capillary electrophoresis was chosen as a suitable "green" alternative to an already developed HPLC method for the determination of gemcitabine and its impurities included in the 8th edition of European Pharmacopoeia.
The analysis was carried out in 35 cm (27 cm) capillary at a temperature of 25 ° C and detection was at 275 i 237 nm. Three different buffers were studied while developing the method: borate (pH 9.3), phosphate (pH 7.0) and acetate (pH 3.0). Based on the experimental results, best buffer was shown to be the phosphate buffer (pH 7.0), while a better separation and peak shape were achieved by adding a higher concentration of SDS. 50 mM SDS was chosen as the optimal concentration. The effect of voltage on the analysis time and resolution between the analytes was investigated. It has been shown that a better separation of gemcitabine and cytosine was achieved at 10 kV. However, applying a lower voltage is prolonging the migration time of the analytes through the capillary. Therefore, it is necessary to take into account that a good resolution is achieved but that the analysis time is not too long.
Further method optimization is required to achieve the appropriate peak separation, shape and symmetry with as short analysis time as possible.

• Keywords: gemcitabin; onečišćenja u lijekovima; kapilarna elektroforeza; micelarna elektrokinetička kromatografija
• Keywords (english): gemcitabine; drug impurities; capillary electrophoresis; micellar electrokinetic chromatography
• Language: croatian
• URN:NBN: urn:nbn:hr:163:839213
• 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
• Created on: 2018-10-03 15:27:44