Validacija LC-MS/MS metode za određivanje N-nitrozo-hidroklorotiazida u hidroklorotiazid aktivnoj supstanci

master's thesis

• Title (english): Validation of LC-MS/MS method for the determination of N-nitroso-hydrochlorothiazide in hydrochlorothiazide active substance
• Author: Marino Tropčić
• Mentor: Sandra Babić (mentor)
• Committee member: Sandra Babić (predsjednik povjerenstva)
• Committee member: Dragana Mutavdžić Pavlović (član povjerenstva)
• Committee member: Irena Škorić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Chemical Engineering and Technology; Zagreb
• Defense date and country: 2024-02-21, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Validacija je važan korak u stvaranju analitičke metode nakon njenog razvitka, te se validacija definira kao postupak dokazivanja je li metoda prikladna za namjeravanu upotrebu. Validacija analitičke metode izvodi se u laboratoriju provođenjem ispitivanja izvedbenih karakteristika metode kao što su selektivnost/specifičnost, linearnost, preciznost, točnost, granica detekcije i kvantifikacije, radno područje te robusnost. Ovisno o namjeni analitičke metode ne moraju se nužno ispitivati sve izvedbene karakteristike već samo odabrane. Glavni cilj ovog rada bio je potvrditi prikladnost LC-MS/MS metode za određivanje N-nitrozo-hidroklorotiazida u hidroklorotiazid aktivnoj supstanci. Eksperimentalni dio rada izveden je u laboratoriju Odjela Kontrole kvalitete, Pliva Hrvatske d.o.o. ispitivanjem svih potrebnih izvedbenih karakteristika. Postignuta je dobra linearnost (R^2 = 1,000) te su također zadovoljeni kriteriji za preciznost (RSD ≤ 20 %) i točnost (RSD ≤ 25 %). Definirana je granica kvantifikacije (LOQ = 0,60 ng/ml) te radno područje metode (0,60 ng/ml – 50 ng/ml). Dodatno je ispitana stabilnost otopina standarda N-nitrozo-hidroklorotiazida i hidroklorotiazid aktivne supstance i te robusnost metode. Rezultatima validacije potvrđena je prihvatljivost metode za određivanje N-nitrozo-hidroklorotiazida u hidroklorotiazid aktivnoj supstanci što omogućuje njenu primjenu u svakodnevnom rutinskom radu laboratorija.

Abstract (english)

Validation is a key step in the creation of an analytical method after its development, and it is defined as the process of proving whether the method is suitable for the intended purpose. Analytical method validation is conducted in the laboratory by testing performance characteristics of the method, such as selectivity, specificity, linearity, working range, precision, trueness, limit of detection and quantification, and robustness. Depending on the purpose of the analytical method, it is not necessary to examine all performance features, but only selected ones. The main goal of this work is to validate the LC-MS/MS method for the determination of N-nitroso-hydrochlorothiazide in hydrochlorothiazide active substance. The experimental part of the work covered determination of all performance characteristics and was conducted in the Quality Control Department, Pliva Croatia d.o.o. The method achieved good linearity (R^2=1,000) and met the criteria for precision (RSD ≤ 20%) and accuracy (RSD ≤ 25%). The limit of quantification (LOQ = 0,60 ng/ml) as well as the working range of the method (0,60 ng/ml – 50 ng/ml) are defined. In addition, the stability of the solutions of N-nitroso-hydrochlorothiazide standard and hydrochlorothiazide active substance and the robustness of the method were evaluated. The validation results confirmed the acceptability of the method for the determination of N-nitroso-hydrochlorothiazide in hydrochlorothiazide active substance was confirmed, given that the obtained values of performance characteristics are within the given limits of acceptability. This enables the application of the method in the daily routine work of the laboratory.

• Keywords: validacija; LC-MS/MS; hidroklorotiazid; nitrozaminiska onečišćenja
• Keywords (english): validation; LC-MS/MS; hydrochlorotiazide; nitrosamine impurities
• Language: croatian
• URN:NBN: urn:nbn:hr:149:635663
• Study programme: Title: Materials Science and Engineering - Graduate study; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra inženjer/inženjerka kemijskog inženjerstva (sveučilišni/a magistar/magistra inženjer/inženjerka kemijskog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2034-02-21
• Created on: 2024-02-22 11:14:34