Strukturna karakterizacija febuksostata i njegovih onečišćenja HPLC-DAD-MSn metodom

master's thesis

• Title (english): Structural characterization of febuxostat and its impurities by HPLC-DAD-MSn method
• Author: Tomislav Kukor
• Mentor: Miranda Sertić (mentor)
• Committee member: Miranda Sertić (predsjednik povjerenstva)
• Committee member: Mirela Matić (član povjerenstva)
• Committee member: Biljana Nigović (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2017-07-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Giht je metabolička bolest karakterizirana ponavljanim napadima upalnog artritisa čiji je glavni uzrok stanje povećane koncentracije mokraćne kiseline u krvi, hiperuricemija. Srećom, postotak je populacije s dijagnozom gihta još uvijek nizak, međutim, incidencija je gihta u porastu. Ukoliko bolest nije dobro kontrolirana nefarmakološkim mjerama, potrebna je administracija lijekova, pri čemu glavnu ulogu imaju lijekovi za snižavanje razine mokraćne kiseline u krvi. Prva je linija liječenja alopurinol, ali, u slučaju neučinkovitosti ili nepodnošenja alopurinola, lijek je izbora febuksostat.
Febuksostat je relativno nov lijek – selektivan, nepurinski inhibitor ksantin oksidaze, enzima koji katalizira nastajanje mokraćne kiseline u organizmu. Učinkovit je u prevenciji i smanjenju učestalosti akutnih napada gihta. Febuksostat još uvijek nema svoju monografiju u Europskoj farmakopeji, stoga je potrebno razviti brze, selektivne, osjetljive i točne metode za potvrdu identiteta, ispitivanje čistoće te određivanje sadržaja febuksostata u aktivnim farmaceutskim supstancijama i gotovim ljekovitim oblicima.
Cilj je ovog diplomskog rada ispitivanje čistoće febuksostata. Korištena je visoko sofisticirana HPLC-DAD-MSn metoda te je postignuto odjeljivanje febuksostata i njegovih triju onečišćenja (febuksostat amid, febuksostat DEE, etil-febuksostat), a nakon toga masenom su spektrometrijom pretpostavljeni fragmentacijski putovi svakog analita. Predloženi fragmentacijski putovi mogu pomoći u razumijevanju profila onečišćenja febuksostata te olakšati i ubrzati ispitivanje njegove čistoće u gotovom ljekovitom obliku. S tim je ciljem pripremljen i analiziran uzorak febuksostata ekstrahiranog iz tablete, a uočeno je više kromatografskih pikova koji odgovaraju onečišćenjima. Od triju interesnih onečišćenja, u uzorku febuksostata iz tablete detektirana su dva, dok se prisutnost trećeg onečišćenja ne može sa sigurnošću potvrditi. Također, uočena su još četiri pika onečišćenja koje u nastavku rada treba strukturno karakterizirati i identificirati.

Abstract (english)

Gout is a metabolic disease characterized by recurrent attacks of inflammatory arthritis. It is mostly connected with the state of abnormally high blood level of uric acid called hyperuricemia, which is considered to be the main cause of gout. The percentage of the population diagnosed with this disease is quite small, but the incidence of gout tends to grow nowadays. If non-pharmacological measures are not effective in controlling the disease, drug administration is needed. Although there are a few pharmacological groups of drugs that can be used in gout treatment, the main role is played by drugs that lower the blood concentration of uric acid. Allopurinol represents the first line of treatment, but in the case of its ineffectiveness or intolerance, febuxostat can be used.
Febuxostat is a relatively new drug, which acts like selective, non-purine xanthine oxidase inhibitor. Xanthine oxidase is an enzyme that catalyzes the production of uric acid, therefore, its inhibition may help prevent and reduce the frequency of acute gout attacks. Febuxostat still does not have an official monograph in pharmacopoeia so it is highly necessary to develop fast, selective, sensitive and accurate analytical methods for the identification and determination of febuxostat and its impurities in active pharmaceutical ingredients and finished drug products.
The aim of this diploma thesis was to analyze impurities in febuxostat. In order do to so, highly sophisticated HPLC-DAD-MSn method was used to separate febuxostat and its three impurities (febuxostat amide, febuxostat DEE, ethyl-febuxostat). Then, fragmentation pathways for all analytes were determined using mass spectrometry. Suggested pathways of fragmentation can be helpful in understanding the impurity profile of febuxostat, and can also contribute to a fast and easy way of impurity profiling of finished drug product. Hence, the sample of febuxostat extracted from a tablet was taken and analyzed. Many chromatographic peaks that matched impurities were noticed. Two out of three impurities of interest were detected in this sample, while the presence of the third impurity could not be confirmed with certainty. Also, four more peaks were noticed but they will be structuraly characterized and identified in later research.

• Keywords: febuksostat; onečišćenja; HPLC; masena spektrometrija; putovi fragmentacije
• Keywords (english): febuxostat; impurities; HPLC; mass spectrometry; fragmentation pathways
• Language: croatian
• URN:NBN: urn:nbn:hr:163:547928
• 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: 2017-11-21 13:17:41