Razvoj i validacija UHPLC metode prema načelu kvalitete ugrađene u dizajn (QbD) za određivanje onečišćenja fosamprenavira

master's thesis

• Title (english): Development and validation of UHPLC method using quality by design (QbD) approach for fosamprenavir impurities determination
• Author: Sara Perko
• Mentor: Biljana Nigović (mentor)
• Committee member: Biljana Nigović (predsjednik povjerenstva)
• Committee member: Ana Mornar Turk (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2021-05-21, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Cilj ovog diplomskog rada bio je razviti brzu, selektivnu i osjetljivu metodu tekućinske kromatografije
ultravisoke djelotvornosti (UHPLC) za određivanje onečišćenja fosamprenavir kalcija, amprenavira i IMP
0,93 na. Primjenom načela kvalitete ugrađene u dizajn (QbD) znatno je skraćeno vrijeme razvoja metode te
je utrošeno manje resursa što je ekološki prihvatljivije od tradicionalnog razvoja metode koji zahtjeva veći
utrošak kemikalija. Prikladni pH za odjeljivanje sastavnica uspješno je predviđen pomoću programskog
paketa Marvin Beans. Sustavno je proveden razvoj s UV detektorom uparene UHPLC metode obrnutih faza
korištenjem DryLab® softvera za modeliranje. U programu ispitivani parametri bili su: stacionarna faza,
vrijeme eluacije, temperatura kolone i pH mobilne faze. Programski paket DryLab® uključuje i vizualni
prikaz robusnog područja metode čime omogućava izbor eksperimentalnog područja u kojem promjena
parametara metode neće značajno utjecati na rezultat analize. Na temelju dobivenih podataka metoda je
razvijena na koloni Agilent, Zorbax EclipsePlus C18 RRHD (1,8 μm 2,1 x 50 mm). Kao pokretne faze
korištene su amonijev acetat pH 4 i acetonitril. Optimizirana metoda može razdijeliti fosamprenavir kalcij
od odabranih onečišćenja u vremenu od 3 minute i pri temperaturi kolone 50°C. Nakon razvoja, metoda je
validirana prema ICH Q2 (R1) smjernicama. Dokazana je selektivnost, linearnost, točnost i preciznost
metode te stabilnost ispitivanih otopina. Utvrđene su granice dokazivanja i određivanja te radno područje
metode. Razvijena i validirana metoda može se koristiti u kontroli kvalitete za određivanje onečišćenja u
farmaceutskim oblicima koji sadrže fosamprenavir kalcij kao djelatnu tvar.

Abstract (english)

The main goal of this diploma thesis was the development of fast, selective and sensitive ultra-highperformance
liquid chromatography method (UHPLC) used for detection and quantification of impurities,
amprenavir and IMP 0.93, in the samples with fosamprenavir calcium as API. Employing Quality by
Design approach (QbD) the method development time was significantly reduced and the materials used
were saved, which is ecologically more acceptable than traditional way requiring higher consumption of
chemicals. The right pH value of the mobile phase was successfully predicted with Marvin Beans software.
A new reversed phase UHPLC method with UV detection development was systematically performed using
the DryLab® software modelling package where stationary phase, gradient time, column temperature and
the pH of the eluent were explored parameters. What DryLab® also includes is a visual presentation of
method robust space which allows choosing the right experimental domain where the parameter change
would not significantly affect analysis results. Based on the results of DryLab® experiment, the method
was developed on Agilent, Zorbax EclipsePlus C18 RRHD (1.8 μm 2.1 x 50 mm) column. Ammonium
acetate pH 4 and acetonitrile were used as mobile phases. Optimised gradient method was able to separate
fosamprenavir calcium and selected impurities in 3 minutes with the column temperature of 50°C. After the
development, validation was conducted according to Q2 (R1) guidelines of International Conference
Harmonization (ICH). The developed method was validated with respect to selectivity, linearity, accuracy
and precision as well as the solution stability. The limit of detection, limit of quantification and working
range were also established. Developed and validated method can be used for quality control of
pharmaceutical forms containing fosamprenavir calcium as an active substance.

• Keywords: fosamprenavir; UHPLC; onečišćenja; QbD
• Keywords (english): fosamprenavir; UHPLC; impurities; QbD
• Language: croatian
• URN:NBN: urn:nbn:hr:163:552163
• 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: 2022-02-03 09:36:13