20.500.12556/DKUM-95239 Uporaba funkcionaliziranih MOFs v mikroreaktorju za remediacijo odpadnih vod magistrsko delo Application of functionalised MOFs in a microreactor for wastewater remediation Poraba vode in naraščajoče onesnaževanje te predstavljata resen globalni izziv. Odpadne vode vsebujejo številna onesnaževala, med katerimi so posebej problematična mikroonesnaževala, kot so farmacevtski izdelki. Med njimi izstopajo nesteroidna protivnetna zdravila, predvsem diklofenak (DCF). Zaradi svoje široke uporabe v humani in veterinarski medicini se pogosto pojavljajo v odpadnih vodah, kjer lahko škodljivo vplivajo na vodni ekosistem. Zato je nujen razvoj naprednih postopkov čiščenja odpadnih voda, ki omogočajo učinkovito odstranjevanje tovrstnih onesnaževal. V magistrskem delu je prikazana študija adsorpcije in razgradnje DCF v mikroreaktorskem sistemu. Kot adsorbent so bila uporabljena metalo-organska ogrodja (MOFs), natančneje zeolitno imidazolatno ogrodje-8 (ZIF-8), sintetizirano s precipitacijsko metodo. V nadaljevanju smo v ZIF-8 z metodo kovalentnega povezovanja imobilizirali encim lakazo in tako sintetizirali kompozit lakaza@ZIF-8. Lakaza je oksidoreduktazni encim, ki katalizira oksidacijo fenilnih obročev v molekuli DCF, zato je kompozit lakaza@ZIF-8 primeren za razgradnjo DCF v mikroreaktorskem sistemu. Učinkovitost poroznih materialov pri adsorpciji in razgradnji DCF pri različnih pretokih skozi mikroreaktor smo zasledovali z visoko zmogljivostno tekočinsko kromatografijo (HPLC). Funkcionaliziranemu nano ogrodju lakaza@ZIF-8 smo določili aktivnost imobilizirane lakaze in učinkovitost imobilizacije. Proučili smo vpliv deleža volumna mrežnega povezovalca gluteraldehida (GA) in epiklorohidrina (ECH) ter koncentracije lakaze na učinkovitost imobilizacije in preostalo aktivnost encima. Izvedli smo tudi študijo ponovne uporabe, termične stabilnosti in skladiščenja imobilizirane lakaze. Rezultati so pokazali, da je bil optimalen delež GA 10 (v/v) %, ECH pa 7,5 (v/v) %, ob optimalni koncentraciji lakaze 10 mg/mL. Preostala aktivnost lakaze po imobilizaciji je znašala 129,00 % s specifično aktivnostjo 4,48 U/mg (GA) in 118,13 % s specifično aktivnostjo 5,78 U/mg (ECH). Rezultati nakazujejo na hiperaktivacijo encima, ko smo ga imobilizirali na ZIF-8. Učinkovitost imobilizacije je znašala 95,60 % (GA) in 93,15 % (ECH). Imobilizirano lakazo smo lahko večkrat uporabili, izkazala se je za dobro termično stabilno z možnostjo daljšega skladiščenja pri 4 °C. Pri proučevanju adsorpcije in razgradnje DCF z ZIF-8 in lakazo@ZIF-8 v mikroreaktorju pri pretokih 125 µL/min in 250 µL/min smo ugotovili, da zmanjšanje začetnega pretoka na polovico nima bistvenega vpliva na končni delež adsorpcije oz. razgradnje DCF, vpliva le na čas, potreben za razgradnjo/odstranitev enake količine DCF. Ne glede na izbrani pretok se je izkazalo, da je imobilizirana lakaza na ZIF-8 učinkovitejša za odstranjevanje DCF, doseženi delež razgradnje je znašal okoli 90,00 %. Water consumption and increasing water pollution represent a serious global challenge. Wastewater contains numerous pollutants, among which micro-pollutants such as pharmaceutical products are particularly problematic. Non-steroidal anti-inflammatory drugs, especially diclofenac (DCF), are notable in this regard. Due to its widespread use in human and veterinary medicine, diclofenac frequently appears in wastewater, where it can harm the aquatic ecosystem. Therefore, it is necessary to develop advanced wastewater treatment processes that enable the effective removal of such pollutants. This master's thesis presents a study on the adsorption and degradation of DCF in a microreactor system. A metal-organic frameworks (MOFs), specifically zeolite imidazolate framework-8 (ZIF-8), synthesised by the precipitation method, was used as the adsorbent. The enzyme laccase was then immobilised in ZIF-8 using a covalent bonding method, resulting in the composite laccase@ZIF-8. Laccase is an oxidoreductase enzyme that catalyses the oxidation of phenyl rings in the DCF molecule, making the laccase@ZIF-8 composite suitable for DCF degradation in a microreactor system. The efficiency of the porous materials in the adsorption and degradation of DCF at different flow rates through the microreactor was monitored using high-performance liquid chromatography (HPLC). The activity of immobilised laccase and the efficiency of immobilisation were determined for the functionalised laccase@ZIF-8 nanostructure. The effects of the volume fraction of the crosslinkers glutaraldehyde (GA) and epichlorohydrin (ECH), as well as the concentration of laccase, on immobilisation efficiency and residual enzyme activity were investigated. Studies on the reuse, thermal stability, and storage of immobilised laccase were also conducted. The results showed that the optimal proportion of GA was 10 (v/v) %, ECH was 7.5 (v/v) %, and the optimal concentration of laccase was 10 mg/mL. The residual activity of laccase after immobilisation was 129.00 % with a specific activity of 4.48 U/mg (GA) and 118.13 % with a specific activity of 5.78 U/mg (ECH). These results indicate enzyme hyperactivation when immobilised on ZIF-8. The immobilisation efficiency was 95.60 % (GA) and 93.15 % (ECH). The immobilised laccase could be used multiple times and proved to be thermally stable, with the possibility of prolonged storage at 4 °C. When studying the adsorption and degradation of DCF with ZIF-8 and laccase@ZIF-8 in a microreactor at flow rates of 125 µL/min and 250 µL/min, we found that reducing the initial flow rate by half had no significant effect on the final percentage of DCF adsorption or degradation, but only affected the time required to degrade or remove the same amount of DCF. Regardless of the selected flow rate, immobilised laccase on ZIF-8 proved to be more effective for DCF removal, with a degradation rate of approximately 90.00 %. MOFs ZIF-8 lakaza GA ECH DCF mikroreaktor HPLC imobilizacija razgradnja adsorpcija MOFs ZIF-8 laccase GA ECH DCF microreactor HPLC immobilization degradation adsorption true true false [N. Savec] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2025-09-09 19:48:50 2025-10-28 13:54:17 2025-11-05 03:11:39 0000-00-00 00:00:00 2025 0 Maribor 0 Maribor 1 spletni vir (1 datoteka PDF (XII, 53 str.)) 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 66.067.8.081.3:577.152.1(043.2) 255748867 MAG_Savec_Neja_2025.pdf MAG_Savec_Neja_2025.pdf 1 EF49336DA5D147A8138D53860996C620 9e7c62e1d89cb530860613ea695090f66929798cee7d0c76b55561a2675f0c91 d6cbef94-b361-11f0-a3bd-00155d000105 https://dk.um.si/Dokument.php?lang=slv&id=200231 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0