Hemijska i farmakološka karakterizacija odabranih taksona roda Heracleum L. (Apiaceae), autohtonih za jugoistočnu Evropu

Abstract

Predmet istraņivanja doktorske disertacije je devet taksona roda Heracleum (Apiaceae): H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii i H. verticillatum iz grupe H. sphondylium, i H. orphanidis, svi iz (tipske) sect. Heracleum, kao i H. austriacum subsp. siifolium iz sect. Wendia.Gasnom hromatografijom sa FID i MS detekcijom analiziran je sastav etarskih ulja izolovanih destilacijom vodenom parom iz podzemnih organa, listova, cvasti i plodova. U uljima podzemnih organa dominirali su monoterpeni i/ili fenilpropani (taksoni grupe H. sphondylium), ili (Z)-falkarinol (H. orphanidis i H. austriacum); u uljima plodova alifatiĉni estri oktilacetat (sect. Heracleum) ili oktilheksanoat (H. austriacum), a u uljima listova i cvasti alifatiĉni estri (H. orphanidis) ili seskviterpeni i/ili fenilpropani. S obzirom da je u pojedinim uljima utvrĊeno prisustvo furanokumarina koji mogu delovati fototoksiĉno, definisan je bezbednosni profil ulja, tj. utvrĊen njihov maksimalni dozvoljeni dnevni unos.U dihlormetanskim ekstraktima podzemnih organa i plodova svih taksona identifikovano je ukupno 12 furanokumarina (LC-MS metodom pomoću standardnih supstanci ili na osnovu UV, MS, 1H i ROESY NMR spektara), od kojih su dominantni bili pimpinelin, bergapten, biakangelikol, heraklenin i/ili imperatorin.Hemosistematski znaĉaj komponenti etarskih ulja (u sluĉaju podzemnih organa i plodova i komponenti njihovih headspace frakcija) i furanokumarina utvrĊen je multivarijantnom statistiĉkom analizom (PCA, nMDS i UPGMA).U uljastim supernatantima dihlormetanskih ekstrakata plodova gasnom hromatografijom (GC-FID i GC-MS) analizirane su masne kiseline (dominantna je bila petroselinska), steroli (najzastupljeniji je bio β-sitosterol) i triterpeni.Antimikrobna aktivnost etarskih ulja pokazana je mikrodilucionom metodom na po osam sojeva bakterija i mikromiceta (u nekim sluĉajevima MIK su bile niņe ili bliske 0,1 mg/mL); selektivna citotoksiĉnost MTT testom na tumorske HeLa, LS174 i A549 ćelije (u nekim sluĉajevima IC50 su bile niņe ili bliske 30 μg/mL; ulja uglavnom nisuinhibirala rast normalnih MRC-5 ćelija)...

This doctoral dissertation focuses on nineHeracleumtaxa (Apiaceae): H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicumsubsp. pollinianum, H. pyrenaicumsubsp. orsiniiandH. verticillatum, belonging togroupH. sphondylium, andH. orphanidis, all from the (type)sect. Heracleum, as well asH.austriacum subsp.siifoliumfromsect. Wendia. By GC-FID and GC-MS, the composition of essential oils isolated by hydrodistillation fromunderground parts, leaves, flowers and fruits was analysed. Underground parts oils were dominated bymonoterpenesand/orphenylpropanoids (H. sphondyliumgroup), or(Z)-falcarinol (H. orphanidisandH. austriacum); fruit oils byaliphatic estersoctylacetate(sect. Heracleum) oroctylhexanoate(H. austriacum), andleaf and flower oilsby aliphatic esters(H. orphanidis) or sesquiterpenesand/orphenylpropanoids. Considering that in some of these oils potentially phototoxic furanocoumarins were detected, safety profileof the oils, i.e. their maximum daily intake wasestimated.In dichloromethane extracts of underground parts and fruits of investigated taxa,atotal of 12 furanocoumarins were identified(by LC-MS using standard compoundsorbased onUV, MS, 1H andROESY NMR spectra), among which pimpinellin, bergapten, byakangelicol, heraclenin and/orimperatorinwere dominant. Chemosystematicsignificance ofessential oil components(in the case ofunderground parts and fruits, of their headspacefractioncomponents as well) and furanocoumarinswas demonstrated usingmultivariatestatistics (PCA, nMDS andUPGMA).In oily supernatants of fruit dichloromethaneextracts, by GC-FID and GC-MS,fatty acids (petroselinic acid was dominant), sterols (β-sitosterolwas the most abundant) and triterpeneswere analysed. Antimicrobialactivity of the essential oilswas showedbymicrodilutionmethodoneightbacteria and eightfungi(in some cases MICs werelower or around0.1 mg/mL); selective cytotoxicityby MTT teston cancerHeLa, LS174 andA549 cells(in some cases IC50werelower or around 30 μg/mL; oils mostly did not inhibit growth ofnormalMRC-5 cells). Essential oils showed weakanti-DPPH potential. The composition of their active zones in TLC-DPPH test was analysed byGC-FID and GC-MS(oxygenated terpenes and phenylpropanoidswere dominant).