Analiza sekundarnih metabolita i ispitivanje farmakološke aktivnosti odabranih vrsta roda Laserpitium L. (Apiaceae)
Analysis of secondary metabolites and investigation of pharmacological activity of selected species of the genus Laserpitium L. (Apiaceae)
Докторанд
Popović, Višnja B.Ментор
Petrović, SilvanaЧланови комисије
Stepanović-Petrović, RadicaMilenković, Marina
Niketić, Marjan
Heyerick, Arne
Метаподаци
Приказ свих података о дисертацијиСажетак
Predmet doktorske disertacije je analiza sekundarnih metabolita i ispitivanje
farmakološke aktivnosti hloroformskih i metanolnih ekstrakata i etarskih ulja tri vrste
roda Laserpitium L. (Apiaceae): Laserpitium latifolium L., Laserpitium zernyi Hayek i
Laserpitium ochridanum Micevski.
Laserpitium latifolium je višegodišnja biljka, koja je široko rasprostranjena na
evropskom kontinentu. Laserpitium zernyi je skardo-pindska endemična vrsta, sa
prirodnim staništem na prostorima Srbije (Kosovo), Albanije, Makedonije i Grčke.
Ranije je smatrana podvrstom Laserpitium siler L., subsp. zernyi (Hayek) Tutin.
Laserpitium ochridanum je stenoendemit planine Galičice u Makedoniji.
Sekundarni metaboliti vrsta roda Laserpitium do sada su analizirani samo u
nekoliko šire distribuiranih predstavnika. Od biljaka koje predstavljaju predmet ove
doktorske disertacije, široko rasprostranjena vrsta L. latifolium je do sada samo
delimično ispitivana u pogledu hemijskog sastava. Druge dve, endemične biljne vrst...e -
L. zernyi i L. ochridanum, nisu hemijski i farmakološki proučavane.
Etarska ulja odabranih vrsta roda Laserpitium izolovana su destilacijom
vodenom parom i hemijski okarakterisana metodama GC-FID i GC-MS. U etarskim
uljima podzemnih organa sve tri ispitivane vrste roda Laserpitium, α-pinen je
predstavljao najzastupljeniju komponentu (44,4%, 33,2% i 31,6% u uljima
L. latifolium, L. ochridanum i L. zernyi, redom). U etarskim uljima podzemnih organa
L. zernyi i L. ochridanum utvrđen je i značajan sadržaj α-bisabolola (30,9% i 10,3%,
redom), a u etarskom ulju podzemnih organa L. ochridanum i hamazulena (14,9%). U
etarskim uljima plodova L. latifolium i L. ochridanum utvrđen je visok sadržaj
monoterpenskih ugljovodonika: u etarskom ulju L. latifolium α-pinena (44,0%), dok je u
etarskom ulju L. ochridanum dominanatan satojak limonen (57,7%), a u oba etarska ulja
detektovan je značajan procenat sabinena (26,8% i 14,4%, u etarskim uljima plodova
L. latifolium i L. ochridanum, redom). U etarskom ulju herbe L. ochridanum
najzastupljenija komponenta je sabinen (25,9%). U etarskom ulju cvasti L. zernyi
najzastupljenije komponente su sabinen, limonen i β-felandren (18,5%, 12,0% i 12,0%,
redom), a u etarskom ulju herbe L. zernyi β-pinen (20,0%). Ova dva ulja su, za razliku
od drugih ispitivanih ulja vrsta roda Laserpitium, imala relativno visok sadržaj
monoterpenskog alkohola terpinen-4-ola (10,6% i 12,0% u etarskom ulju herbe i cvasti
L. zernyi, redom).
Iz dela hloroformskog ekstrakta podzemnih organa L. latifolium rastvorljivog u
metanolu, kombinovanjem pogodnih hromatografskih tehnika (hromatografija na koloni
silikagela i semipreparativna HPLC) i instrumentalnih metoda za određivanje strukture
(1D i 2D NMR i IR spektroskopija i HR-MS ESI+ spektrometrija), kao glavni sastojci
izolovana su i okarakterisana dva daukanska estra: laserpicin (1) i
acetildezoksodehidrolaserpicin (2), i dva fenilpropana: laserin (3) i latifolon (4).
Uporednom HPLC analizom, u uslovima optimizovanim za razdvajanje terpenoida,
uočena je velika kvalitativna sličnost hloroformskih ekstrakata podzemnih organa i herbe L. latifolium. Kvantifikacija ekstrakata izvršena je pod istim uslovima,
korišćenjem izolovanih jedinjenja kao eksternih standarda. Konstatovano je da je
sadržaj sva četiri jedinjenja viši u ekstraktu podzemnih organa u odnosu na ekstrakt
herbe L. latifolium...
In the scope of the doctoral dissertation are analysis of secondary metabolites
and testing of pharmacological activity of chloroform and methanol extracts and
essential oils of three species of the genus Laserpitium L. (Apiaceae): Laserpitium
latifolium L., Laserpitium zernyi Hayek and Laserpitium ochridanum Micevski.
Laserpitium latifolium is perenial species, widely distributed throughout
European continent. Laserpitium zernyi is scardo-pindic endemic species, that is
distributed in Serbia (Kosovo), Albania, Former Yugoslav Republic of Macedonia
(FYROM) and Greece. Previously, it was treated as a subspecies of colsely related
Laserpitium siler L., subsp. zernyi (Hayek) Tutin. Laserpitium ochridanum is endemic
species at Galičica Mountain, FYROM.
Secondary metabolites of Laserpitium species have been so far analyzed just in
several widely distributed plants. Considering investigation of the species that are in the
scope of this dissertation, only widely distributed L. latifolium has ...been partially
chemically examined. The other two endemic species - L. zernyi and L. ochridanum
have not been either chemically or pharmacologically investigated before.
Essential oils of selected Laserpitium species are isolated by a steam distillation
and chemically characterised by GC-FID and GC-MS. In the essential oils of the
underground parts of three Laserpitium species, the main constituent was α-pinene
(44.4%, 33.2% and 31.6% in essential oils of L. latifolium, L. ochridanum and L. zernyi,
respectively). In the essential oils of underground parts of L. zernyi and L. ochridanum
certain content of α-bisabolol was found (30.9% and 10.3%), and in essential oil of
underground parts of L. ochridanum was also detected chamazulene (14.9%). The fruit
oils of L. latifolium and L. ochridanum were rich is monoterpene hydrocarbons:
essential oil of L. latifolium was rich in α-pinene (44.0%), whilst in essential oil of
L. ochridanum dominant constituent was limonene (57.7%), and in both oils certain
percent of sabinene was detected (26.8% and 14.4%, in fruit oil of L. latifolium and
L. ochridanum, respectively). In herb essential oil of L. ochridanum the most abundand
constituent was sabinene (25.9%). In the essential oil flower of L. zernyi sabinene,
limonene and β-phellandrene were detected in higher percent (18.5%, 12.0% and
12.0%, respectively), and in oil of L. zernyi herb higher content of β-pinene (20.0%) was
detected. Two latter oils, unlike the other oils, were characterised with relatively high
content of terpinene-4-ol (12.0% and 10.6% in oils of L. zernyi flower and herb, respectively).
From the methanol-soluble part of chloroform extract of the underground parts
of L. latifolium, by combination of adequate chromatographic techniques (FC and
semi-preparative HPLC) and instrumental methods for structure elucidation (1D and
2D NMR and IR spectroscopy and HR-MS (ESI+) spectrometry), as the main
constituents were isolated and elucidated the two daucane esters: laserpitin (1) and
acetyldesoxodehydrolaserpitin (2), as well as the two phenylpropanes: laserin (3) and
latifolon (4). By comparative HPLC analysis in a system optimised for terpenoid separation, a huge qualitative similarity of chloroform extracts of the underground
parts and herb of L. latifolium was observed. Quantitative analyses was conducted in
the same system, using isolated compounds as external standards. It was demonstrated
that all compounds were present in higher concentrations in the extract of the
underground parts than in the herb extract of L. latifolium...
Факултет:
Универзитет у Београду, Фармацеутски факултетДатум одбране:
30-09-2013Пројекти:
- Испитивање лековитог потенцијала биљака: морфолошка, хемијска и фармаколошка карактеризација (RS-173021)
- Испитивање механизма дејства, интеракција и токсичних ефеката адјувантних аналгетика (RS-175045)