Određivanje ostataka tetraciklinskih antibiotika u mlijeku LC-MS/MS metodom

master's thesis

• Title (english): Determination of tetracycline antibiotic residues in milk by LC-MS/MS method
• Author: Andreja Petek
• Mentor: Blaženka Kos (mentor)
• Mentor: Ivana Varenina (komentor)
• Committee member: Andreja Leboš-Pavunc (predsjednik povjerenstva)
• Committee member: Ivana Varenina (član povjerenstva)
• Committee member: Ksenija Durgo (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Antibiotic, Enzyme, Probiotic and Starter Cultures Technology); Zagreb
• Defense date and country: 2023-06-07, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Antibiotici se koriste u liječenju i prevenciji bolesti životinja, te kao promotori rasta što može dovesti do prisutnosti rezidua u hrani životinjskog podrijetla pa tako i u mlijeku. Prema regulativi Europske unije, NDK (najveća dopuštena koncentracija) za tetraciklinske antibiotike u mlijeku iznosi 100 μg/kg. Cilj ovog rada bio je primijeniti LC-MS/MS (tekućinska kromatografija-tandemska spektrometrija masa) metodu i ELISA (engl. enzyme-linked immunosorbent assay) test, usporediti korištene ekstrakcijske metode i detektirati prednosti i nedostatke istih. Optimizirani parametri LCMS/MS metode omogućili su bolju selektivnost, razdvajanje analita, te točnije rezultate analize. Usporedbom pročišćavanja uzoraka QuECHERS (engl. quick, easy, cheap, effective, rugged, and safe) i ekstrakcijom organskim otapalom dokazano je da je ekstrakcijski prinos veći, a matriks efekt manji, primjenom ekstrakcije organskim otapalom. Korištenjem natrijevog sulfata u odnosu na magnezijev sulfat u QuECHERS esktrakciji postiže se veća efikasnost ekstrakcije. LC-MS/MS metoda može se koristiti u određivanju rezidua tetraciklinskih antibiotika dok se ELISA test zbog neponovljivosti rezultata ne može koristiti u kvantitativne svrhe niti kvalitativne svrhe s obzirom da ne pokriva sve tetraciklinske antibiotike koji se mogu naći kao rezidue u mlijeku.

Abstract (english)

Antibiotics are used in the treatment and prevention of animal diseases, and as growth promoters, which can lead to the presence of residues in food of animal origin, which includes milk. According to European Union regulations, MRL (maximum residue limit) for tetracycline antibiotics in milk is 100 μg/kg. The aim of this work was to apply the LC-MS/MS (liquid chromatography tandem mass spectrometry) method and the ELISA (enzyme-linked immunosorbent assay) test, to compare the used extraction methods and to detect their advantages and disadvantages. The optimized parameters of the LC-MS/MS method enabled better selectivity, analyte separation, and more accurate analysis results. By comparing the purification of samples after QuECHERS (quick, easy, cheap, effective, rugged, and safe) and extraction with an organic solvent, it was proven that the extraction yield is higher, and the matrix effect is lower when using extraction with an organic solvent. By using sodium sulfate in relation to magnesium sulfate in QuECHERS extraction, a higher extraction efficiency is achieved. The LCMS/MS method can be used to determine residues of tetracycline antibiotics, while the ELISA test cannot be used for quantitative or qualitative purposes due to the unreproducibility of the results and considering that it does not cover all tetracycline antibiotics that can be found as residues in milk.

• Keywords: tetraciklinski antibiotici; LC-MS/MS; ELISA test; QuECHERS ekstrakcija; ekstrakcija organskim otapalom
• Keywords (english): tetracycline antibiotics; LC-MS/MS; ELISA test; QuECHERS extraction; organic solvent extraction
• Language: croatian
• URN:NBN: urn:nbn:hr:159:556242
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-05-15 11:52:15