Masnospektrometrična kvantifikacija polifenolnih spojin v ekstraktih invazivnih dresnikov

Pepelko, Sara (author), Pogačnik da Silva, Lea (mentor), Kosjek, Tina (comentor)

Japonski dresnik (Fallopia japonica), sahalinski dresnik (Fallopia sachalinensis) in njun medvrstni križanec, češki dresnik (Fallopia × bohemica) so invazivne tujerodne rastline rodu Fallopia in družine Polygonaceae. Japonski in sahalinski dresnik sta bila priljubljena kot okrasni rastlini, zato sta se hitro razširila po Evropi in v zadnjih dveh stoletjih postala ekološki in ekonomski problem zaradi ogrožanja biodiverzitete in infrastrukture. V dresnikih so določili bioaktivne sekundarne metabolite iz različnih skupin, kot so stilbeni, flavonoidi, fenolne kisline, karotenoidi, klorofili in triterpenske kisline. Kvantitativni podatki o sekundarnih metabolitih so izhodišče za načrtovanje uporabe dresnikov kot novega vira biološko aktivnih spojin. Iz liofiliziranih vzorcev poganjkov in olupkov korenin treh vrst dresnikov smo pripravili etanolne ekstrakte. Določili smo skupne fenolne spojine po metodi Folin-Ciocalteu in ekstrakte analizirali še z metodo tekočinske kromatografije ultra visoke ločljivosti, sklopljene z masnim spektrometrom tipa kvadrupol z detektorjem na čas preleta ionov (UHPLC-MS/MS). Ločbo fenolnih spojin smo dosegli z reverzno fazno kromatografijo, kvantificirali pa smo jih po ionizaciji z razprševanjem v električnem polju z visokoločljivostno in tandemsko masno spektrometrijo. Kvantitativno določitev polifenolnih spojin smo dosegli z uporabo metode standardnega dodatka raztopin referenčnih standardov polifenolnih spojin. Kvantitativno smo v vsaj enem vzorcu dresnikov določili 7 polifenolnih spojin. Po metodi Folin-Ciocalteu smo določili največ skupnih polifenolnih spojin v olupkih korenin sahalinskega dresnika, z metodo UHPLC-MS/MS pa smo največ polifenolnih spojin določili v poganjkih češkega dresnika. Potrdili smo, da sta metodi Folin-Ciocalteu in UHPLC-MS/MS primerni za kvantitativno določitev polifenolnih spojin v dresnikih, njuni rezultati pa se ne ujemajo vedno.

• Language: Slovenian
• Keywords: dresnik, Fallopia, polifenolne spojine, kvantifikacija, tekočinska kromatografija ultra visoke ločljivosti sklopljena s tandemsko masno spektrometrijo, Folin-Ciocalteu
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [S. Pepelko]
• Year: 2024
• PID: 20.500.12556/RUL-158540
• UDC: 547.56:582.665:543.51
• COBISS.SI-ID: 199056387
• Publication date in RUL: 15.06.2024

PEPELKO, Sara, 2024, Masnospektrometrična kvantifikacija polifenolnih spojin v ekstraktih invazivnih dresnikov [online]. Master’s thesis. S. Pepelko. [Accessed 31 May 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=158540

Secondary language

• Language: English
• Title: Mass spectrometric quantification of polyphenol compounds in the extracts of invasive knotweed species

Abstract

Japanese knotweed (Fallopia japonica), Giant knotweed (Fallopia sachalinensis) and their hybrid, Bohemian knotweed (Fallopia × bohemica), are invasive plant species from the genus Fallopia and family Polygonaceae. The Japanese and Giant knotweeds were popular as ornamental garden plants and spreaded rapidly across Europe, becoming an ecological and economic problem in the last two centuries due to threats to biodiversity and infrastructure. Bioactive secondary metabolites from different compounds such as stilbenes, flavonoids, phenolic acids, carotenoids, chlorophylls and triterpenic acids have been identified in Japanese, Giant and Bohemian knotweed. Ethanolic extracts were prepared from freeze-dried shoots and root peels samples of three species of knotweeds. Total phenolic compounds were determined by the Folin-Ciocalteu method. Ethanolic extracts were further analysed by ultra-high-performance liquid chromatography coupled to a hybrid quadrupole-ion trap mass detector (UHPLC-MS/MS). The separation of phenolic compounds was achieved by reversed-phase chromatography and quantified after electrospray ionisation by tandem mass spectrometry. Seven polyphenolic compounds were quantified in at least one sample. This confirmed that the two methods used are suitable for the quantitative determination of polyphenolic compounds in knotweed extracts. The highest total polyphenolic compounds was with Folin-Ciocalteu method determined in the peels of Giant knotweed roots, while with UHPLC-MS/MS method we determined the highest amount of polyphenolic compounds in the shoots of Bohemian knotweed. We confirmed that the Folin-Ciocalteu and UHPLC-MS/MS methods are suitable for the quantitative determination of polyphenolic compounds in knotweeds and that they may give incomparable results.

• Keywords: knotweed species, Fallopia, polyphenolic compounds, ultra-high performance liquid chromatography coupled with tandem mass spectrometry, Folin-Ciocalteu