20.500.12556/RUL-117403 Določanje fenolnih spojin v invazivnih tujerodnih dresnikih (Fallopia spp.) Determination of phenolic compounds in invasive alien knotweeds (Fallopia spp.) Japonski dresnik, sahalinski dresnik in češki dresnik so zelo razširjene invazivne tujerodne rastlinske vrste, ki tako v Evropi kot Severni Ameriki predstavljajo ekološki in ekonomski problem. Z namenom izkoriščanja te biomase, ki vsebuje različne bioaktivne spojine, smo v listih in korenikah teh dresnikov s tankoplastno kromatografijo visoke ločljivosti (HPTLC) sklopljeno z denzitometrijo, analizo slik in masno spektrometrijo (HPTLC–MS/MS) proučevali naslednje fenolne spojine: flavan-3-ole in proantocianidine. Z analizo slik HPTLC silikagelnih plošč smo na osnovi celokupnih ploščin določili najvišjo vsebnost proantocianidinov v korenikah japonskega dresnika. Z uporabo standardnih raztopin(–)-epikatehina in procianidina B2 smo določili vsebnosti monomerov in dimerov v listih in korenikah vseh treh dresnikov. Vsebnosti monomerov v listih japonskega, češkega in sahalinskega dresnika so bile 0,84, 1,39 in 2,36 kg/t suhega materiala, vsebnosti dimerov pa 0,99, 1,40 in 2,06 kg/t suhega materiala. Vsebnosti monomerov v korenikah japonskega, češkega in sahalinskega dresnika so bile 2,99, 1,52 in 2,36 kg/t suhega materiala, medtem ko so bile vsebnosti dimerov 2,81, 1,09 in 2,17 kg/t suhega materiala. S HPTLC v kombinaciji z denzitometrijo in analizo slik smo ugotovili, da so kromatografski profili proantocianidinov korenik japonskega, sahalinskega in češkega dresnika kvalitativno enaki, kar pa ne velja za profile listov teh dresnikov, saj ima sahalinski dresnik v profilu tri dodatne vrhove. Znotraj iste vrste pa ima le sahalinski dresnik enak profil za oba rastlinska dela, medtem ko imata japonski in češki dresnik v primerjavi z listi dva dodatna vrhova v profilih korenik. V listih vseh dresnikov in v korenikah češkega in sahalinskega dresnika smo s HPTLC-MS/MS prvi identificirali vse proantocianidine tipa B od monomerov do dekamerov, v korenikah češkega in sahalinskega dresnika ter v listih sahalinskega dresnika pa tudi nekatere njihove galate (od monomerov do heksamerov) in dimer digalate. Kot prvi smo identificirali proantocianidine v listih češkega dresnika. Med listi vseh treh dresnikov smo največjo pestrost v kvalitativnem profilu galatov odkrili v listih sahalinskega dresnika. V listih japonskega dresnika smo potrdili le prisotnost monomer galatov in dimer galatov, medtem ko smo v listih češkega dresnika poleg teh potrdili tudi dimer digalate. Torej lahko povzamemo, da imajo listi japonskega, češkega in sahalinskega dresnika enake kemijske profile proantocianidinov glede na stopnjo polimerizacije, medtem ko se njihovi kemijski profili glede na galate razlikujejo. Korenike češkega in sahalinskega dresnika pa imajo enake kemijske profile proantocianidinov glede na stopnjo polimerizacije in tudi glede na galate. Znotraj posamezne vrste ima le sahalinski dresnik v listih in korenikah enak kemijski profil proantocianidinov glede na stopnjo polimerizacije in glede na galate. Japanese knotweed, giant knotweed and Bohemian knotweed are three widely spread, invasive alien plants that represent an ecological and economic problem in Europe and North America. In order to exploit this biomass containing various bioactive compounds, high performance thin-layer chromatography (HPTLC) coupled with densitometry, image analysis and mass spectrometry (HPTLC-MS/MS) was used to study phenolic compounds flavan-3-ols and proanthocyanidins in leaves and rhizomes of these knotweeds. Image analysis of the HPTLC silica gel plates showed the highest proanthocyanidins content (based on the total peak areas) in rhizomes of Japanese knotweed. Using (-)-epicatechin and procyanidin B2 standards, the contents of monomers and dimers in leaves and rhizomes of all three knotweeds were determined. In leaves of Japanese, Bohemian and giant knotweed the contents of monomers were 0.84, 1.39, 2.36 kg/t of dry material, respectively, while the contents of dimers were 0.99, 1.40, 2.06 kg/t of dry material, respectively. In rhizomes of Japanese, Bohemian and giant knotweed the contents of monomers were 2.99, 1.52, 2.36 kg/t of dry material, respectively and the contents of dimers were 2.81, 1.09, 2.17 kg/t of dry material, respectively. With HPTLC combined with densitometry and image analysis we found qualitatively equal chromatographic profiles of proanthocyanidins in rhizomes of Japanese, Bohemian and giant knotweed, which was not the case with profiles of leaves, where giant knotweed's profile has three additional peaks. Within the same species, only giant knotweed has the same profile for both plant organs, while Japanese and Bohemian knotweed have two additional peaks in the profiles of rhizomes compared to leaves. Using HPTLC-MS/MS we were the first to identify all B-type proanthocyanidins from monomers to decamers in leaves of all knotweeds and in rhizomes of Bohemian and giant knotweed, as well as some of their gallates (from monomers to hexamers) and dimer digallates in rhizomes of Bohemian and giant knotweed. We were the first to identify proanthocyanidins in leaves of Bohemian knotweed. Among the leaves of all three knotweeds, the highest diversity in the qualitative profile of gallates was detected in leaves of giant knotweed. Monomer gallates and dimer gallates were identified in leaves of Japanese knotweed, while in addition to these digallate dimer was also confirmed in leaves of Bohemian knotweed. Leaves of Japanese, Bohemian and giant knotweed have the same chemical profiles of proanthocyanidins with respect to the degree of polymerization, although their chemical profiles with respect to gallates differ. Rhizomes of Bohemian and giant knotweed have the same chemical profiles of proanthocyanidins in terms of degree of polymerization and also in terms of gallates. Within each of the three individual species, only giant knotweed has the same chemical profile of proanthocyanidins in leaves and rhizomes, both with respect to the degree of polymerization and with respect to gallates. invazivni tujerodni dresniki proantocianidini flavan-3-oli HPTLC HPTLC-MS/MS invasive alien knotweeds proanthocyanidins flavan-3-ols HPTLC HPTLC-MS/MS true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2020-07-09 10:35:01 2020-07-09 10:35:02 2022-08-29 03:43:12 0000-00-00 00:00:00 2020 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 6681 23546371 Bensa_Maja_-_Dolocanje_fenolnih_spojin_v_invazivnih_tujerodnih_dresnikih_(Fallopia_spp.).pdf Bensa_Maja_-_Dolocanje_fenolnih_spojin_v_invazivnih_tujerodnih_dresnikih_(Fallopia_spp.).pdf 1 831444242628E14677F32B4A0BB111E4 b31bdc8069af9e53425d8484048e1bcefb1d7e039c66acf4f0628ebda335ee76 2cc6637e-a1b9-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=131266 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0