Validacija kapilarnoelektroforetske metode za određivanje pirfenidona u ljekovitom obliku

master's thesis

• Title (english): Validation of capillary electrophoretic method for determination of pirfenidone in dosage form
• Author: Lana Bratelj
• Mentor: Miranda Sertić (mentor)
• Committee member: Miranda Sertić (predsjednik povjerenstva)
• Committee member: Ivana Perković (član povjerenstva)
• Committee member: Kristina Pavić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical analysis); Zagreb
• Defense date and country: 2020-07-20, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Idiopatska plućna fibroza rijetka je i smrtonosna bolest. Terapijskih opcija u liječenju idiopatske plućne fibroze je malo, a jedna od njih je liječenje pirfenidonom. Pirfenidon djeluje antifibrotički, protuupalno i antioksidativno, a u Europi je registriran u obliku Esbriet kapsule koja sadrži 267 mg pirfenidona.
Cilj ovog diplomskog rada je validirati metodu, odnosno ispitati prikladnost primjene novorazvijene metode za određivanje pirfenidona u Esbriet kapsuli tehnikom kapilarne elektroforeze. Validacijski parametri ispitani u ovom radu jesu: linearnost, preciznost, robustnost i točnost. Kao unutarnji standard koristila se salicilna kiselina.
Analiza je provedena u kapilari od izvučenog kvarca ukupne duljine 36 cm, udaljenosti do detektora 27cm, unutrašnjeg promjera 50 μm, pri temperaturi od 25 °C i naponu od 25 kV. Kao radni pufer koristila se smjesa 20 mM borata pH 9,1 i 70 mM SDS-a. Dobivena linearnost je zadovoljavajuća za koncentracijski raspon od 10 do 300 μg/mL, pri čemu jednadžba dobivenog regresijskog pravca glasi y = 0,0185x + 0,023, s koeficijentom korelacije R = 0,999. Ponovljivost predložene analitičke metode je također zadovoljavajuća, pri čemu su sve RSD vrijednosti bile manje od 2%. Za ispitivanje robustnosti provedena su ispitivanja na različitoj temperaturi i naponu. Utjecaj napona na vrijeme zadržavanja i površinu pikova nije velik, a temperatura ne utječe značajno na vrijeme zadržavanja i površinu pikova. Ispitivanje točnosti nije bilo u potpunosti zadovoljavajuće; recovery vrijednosti za 3 ispitivane koncentracije su nešto niže od 90%. Primjećeno je da se struja tijekom mjerenja mijenjala i vjerojatno treba češće mijenjati pufere ili promijeniti ekstrakcijski postupak ukoliko nije ekstrahiran sav pirfenidon iz ljekovitog oblika.

Abstract (english)

Idiophatic pulmonary fibrosis is a rare and deadly disease. There are not many therapeutic options in the treatment of idiophatic pulmonary fibrosis, and one of the options is treatment with pirfenidone. Pirfenidone has antifibrotic, anti-inflammatory and antioxidant effect and it is registered in Europe in the form of an Esbriet capsule which contains 267 mg of pirfenidone.
The aim of this thesis is to validate and examine the suitability of the application of the newly developed method for the determination of pirfenidone in Esbriet capsule with capillary electrophoresis technique. Validation parameters examined in this thesis are: linearity, precision, robustness and accuracy. Salicylic acid was used as an internal standard.
The analysis was performed in an fused silica capillary with a total lenght of 36 cm, a detector lengt of 27 cm, an inner diameter of 50 μm, at a temperature of 25 °C and a voltage of 25 kV. A mixture of 20 mM borate pH 9,1 and 70 mM SDS was used as a running buffer. The obtained linearity is satisfactory for the concentration range from 10 to 300 μg/mL, with belonging equation y = 0,0185x + 0,023, with a correlation coefficient R=0,999. The repetability of the analytical method is also satisfactory, with all RSD values being less than 2%. To test robustness, tests were preformed at different temperatures and voltages. The influence of voltage on retention time and peak area is not large, and temperature does not have a significant affect on retention time and peak area. The accuracy test were less satisfactory; recovery values for the 3 tested concentrations are little under than 90%. It has been observed that the current changed during the measurement and it is likely that the buffer should be changed more frequently or that the extraction procedure should be changed if not all pirfenidone has been extracted from the capsule.

• Keywords: pirfenidon; kapilarna elektroforeza; validacija; idiopatska plućna fibroza
• Keywords (english): pirfenidone; capillary electroforesis; bioanalytics; validation; idiophatic pulmonary fibrosis
• Language: croatian
• URN:NBN: urn:nbn:hr:163:023003
• 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: 2022-08-26 13:40:00