Razvoj HPLC metode za analizu antibiotika doksiciklin hiklata i razgradnih produkata

master's thesis

• Title (english): Development of HPLC method for analysis of antibiotic doxycycline hyclate and its degradation products
• Author: Lana Štanfel
• Mentor: Antonija Trontel (mentor)
• Mentor: Anita Šporec (pomoć pri izradi, neposredni voditelj)
• Committee member: Mario Novak (predsjednik povjerenstva)
• Committee member: Andreja Leboš-Pavunc (član povjerenstva)
• Committee member: Blaženka Kos (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology); Zagreb
• Defense date and country: 2024-09-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Doksiciklin hiklat je jedan od često upotrebljavanih antibiotika za liječenje različitih bolesti kod ljudi i životinja. Stoga je neophodno ispuniti zahtjeve o kvaliteti lijeka kao što je mikrobiološka čistoća u cilju postizanja sigurnosti primjene lijeka. Uobičajeni pristupi inaktivacije antibiotika uključuju razrjeđivanje do niskih koncentracija ili kemijsku razgradnju, dok se u ovom radu istražuje enzimska razgradnja kao ekološki prihvatljivija alternativa. Odabran je enzim lakaza koji oksidacijom fenolnih skupina razgrađuje doksiciklin hiklat prisutan u farmaceutskom proizvodu odnosno tableti. Razvijena je gradijentna HPLC metoda za praćenje promjene koncentracije doksiciklin hiklata i srodnih spojeva. Primjenom Luna Omega Polar C18 kolone (150 x 3 mm, 3 µm) s mobilnom fazom koja se sastoji od acetatnog pufera pH=8,8 i acetonitrila, uz protok 0,8 mL/min i pri 274 nm, eluira se doksiciklin hiklat i njegovi razgradni spojevi unutar 25 minuta. Metodom se može učinkovito pratiti enzimska razgradnja, što je ključno za procjenu djelotvornosti lakaze u razgradnji antibiotika.

Abstract (english)

Doxycycline hyclate is one of the most commonly used antibiotics for treating various diseases in humans and animals. Therefore, it is essential to meet the quality requirements of the drug, such as microbiological purity, to ensure the safety of its application. Traditional approaches to antibiotic inactivation include dilution to low concentrations or chemical degradation, while this study investigates enzymatic degradation as a more environmentally friendly alternative. The enzyme laccase, which oxidizes phenolic groups, was selected to degrade doxycycline hyclate present in a pharmaceutical product (tablet). A gradient HPLC method was developed to monitor changes in the concentration of doxycycline hyclate and related compounds. Using a Luna Omega Polar C18 column (150 x 3 mm, 3 µm) with a mobile phase consisting of acetate buffer pH=8.8 and acetonitrile, at a flow rate of 0.8 mL/min and detection at 274 nm, doxycycline hyclate and its degradation products elute within 25 minutes. This method can effectively monitor the enzymatic degradation, which is crucial for assessing the effectiveness of laccase in the degradation of the antibiotic.

• Keywords: doksiciklin hiklat; lakaza; HPLC; razgradnja
• Keywords (english): doxycycline hyclate; laccase; HPLC; degradation
• Language: croatian
• URN:NBN: urn:nbn:hr:159:332334
• 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: 2024-10-17 12:13:46