20.500.12556/DKUM-76725 Antioksidativne lastnosti biološko aktivnih komponent v ekstraktih črne kumine in njihov vpliv na viabilnost humanih celic magistrsko delo Antioxidant properties of biologicallz active components in black cumin extracts and their effect on the viabilitz of human cells V magistrskem delu smo v ekstraktih iz semen črne kumine določevali vsebnost timola, timokinona, karvakrola in α-hederina z GC-MS in LC-MS/MS analizo. Razvijali smo HPTLC metodo za ločevanje in identificiranje teh komponent iz mešanice standardov in nekaterih ekstraktov. Ekstraktom smo določevali antioksidativno učinkovitost z DPPH in β-karotensko metodo ter jo primerjali z učinkovitostjo BHT in askorbinske kisline. Ugotavljali smo vpliv nekaterih ekstraktov in α-hederina na viabilnost človeških črevesnih celic (HUIEC) in črevesnih rakavih celic (CaCo-2) z MTT testom, z nanašanjem različnih koncentracij ekstraktov na celice. Ekstrakti so bili pripravljeni s konvencionalno Soxhletovo ekstrakcijo, ultrazvočno ekstrakcijo in superkritično ekstrakcijo. Ekstrakti so vsebovali največ timokinona (do 4000 µg/g). Metanolne raztopine ekstraktov so vsebovale okoli 700 µg/g karvakrola z izjemo enega SC ekstrakta in okoli 200 µg/g timola. Najvišjo vsebnost α-hederina je vsebovala metanolna raztopina ekstrakta Soxhletove ekstrakcije s heksanom (47 µg/g). S HPTLC metodo z mobilno fazo toluen, etil acetat, metanol v razmerju 9 : 1 : 0,5 (v/v) smo ločili timol, timokinon in karvakrol iz mešanice stnadardov, vendar z manj kot 20 % obnovljivostjo. Identificirali smo karvakrol v SC ekstraktu (40 °C in 100 barov). Najvišjo antioksidativno učinkovitost smo dosegli z SC ekstrakti v vodni raztopini. Večina ekstraktov z izjemo UZ ekstraktov je dosegla višjo antioksidativno učinkovitost kot sintetična antioksidanta. Ekstrakta SC ekstrakcije pri 60 °C in 300 barih in s kotopilom pri 40 °C in 200 barih ter standard α-hederin so pozitivno vplivali na viabilnost HUIEC celic pri nižjih koncentracijah. Z višanjem koncentracije smo opazili toksičnost za celice. Viabilnost celic CaCo-2 je padala pri višanju koncentracij ekstraktov. Contents of thymol, thymoquinone, carvacrol and α-hederine were determined in extracts of black cumin seeds with GC-MS and LC-MS/MS analysis. HPTLC method was developed for separation and identification of those components from mix of standards and some extracts. Antioxidant activity of extracts was determined with DPPH and β-carotene method and compared to antioxidant activity of BHT and ascorbic acid. Effect of some extracts and α-hederine on the viability of human intestinal epithelial cell line (HUIEC) and human colon carcinoma cell line (CaCo-2) was determined with MTT cell viability assay. Extracts were prepared with Soxhlet, ultrasound and supercritical extraction. Extracts contained the most timoquinone (up to 400 µg/g). Methanolic solutions of extracts contained around 70 µg/g of carvacrol with exception of SC extract (40°C, 100 bar) and around 25 µg/g of thymol. Methanolic solution of Soxhlet extract with hexsane contained the most α-hederine (47 µg/g). We succesfully separated thymol, thymoquinone and carvacrol from mix of standards with HPTLC method with toluene, ethyl acetate and metanol in ratio 9:1:0,5 (v/v) as the mobile phase but we managed to reach only 20% recovery. We identified carvacrol in SC extract (40 °C, 100 bar). The highest antioxidant efficacy was found in SC extracts dissolved in water. The majority of extracts, with exception of ultrasound extracts, reached higher antioxidant efficacy than BHT and ascorbic acid. SC extract (40 °C, 100 bar), SC extract (cosolvent, 40 °C, 200 bar) and standard α-hederine showed positive effect on HUIEC cell viability at lower concentrations. At higher concentrations we noticed toxicity for cells. Viability of CaCo-2 cells was decreasing with higer concentrations. eterična olja LC-MS/MS GC-MS HPTLC antioksidativna aktivnost viabilnost celic essential oils LC-MS/MS GC-MS HPTLC antioxidant activity cell viability true true false [Š. Lesičar] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2020-06-26 11:55:24 2020-07-30 14:37:15 2022-08-09 13:53:53 2020-08-07 03:09:50 2020 0 Maribor 0 Maribor X, 79 str. 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 66.094.3.097.8:665.52(043.2) 24459523 URN:SI:UM:DK:9UCVUJXO MAG_Lesicar_Spela_2020.pdf MAG_Lesicar_Spela_2020.pdf 1 C153504B5DE8DE35D95312F4C3E0E768 b6808dcbb2c58f3ea20015ee95939023c11937ea61db9c55f4fecf3f9862c483 6988723a-7c11-11eb-bb7a-00155d0001ca 20.500.12556/dkum/d97f99af-6159-4fa7-b09c-25567cddaa44 https://dk.um.si/Dokument.php?lang=slv&id=143607 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0