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Title:Antioksidativne lastnosti biološko aktivnih komponent v ekstraktih črne kumine in njihov vpliv na viabilnost humanih celic : magistrsko delo
Authors:ID Lesičar, Špela (Author)
ID Kotnik, Petra (Mentor) More about this mentor... New window
ID Knez Marevci, Maša (Comentor)
ID Maver, Uroš (Comentor)
Files:.pdf MAG_Lesicar_Spela_2020.pdf (3,57 MB)
MD5: C153504B5DE8DE35D95312F4C3E0E768
PID: 20.500.12556/dkum/d97f99af-6159-4fa7-b09c-25567cddaa44
 
Language:Slovenian
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract: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.
Keywords:eterična olja, LC-MS/MS, GC-MS, HPTLC, antioksidativna aktivnost, viabilnost celic
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[Š. Lesičar]
Year of publishing:2020
Number of pages:X, 79 str.
PID:20.500.12556/DKUM-76725 New window
UDC:66.094.3.097.8:665.52(043.2)
COBISS.SI-ID:24459523 New window
NUK URN:URN:SI:UM:DK:9UCVUJXO
Publication date in DKUM:30.07.2020
Views:1463
Downloads:204
Metadata:XML DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:26.06.2020

Secondary language

Language:English
Title:Antioxidant properties of biologicallz active components in black cumin extracts and their effect on the viabilitz of human cells
Abstract: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.
Keywords:essential oils, LC-MS/MS, GC-MS, HPTLC, antioxidant activity, cell viability


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