Phytochemical analysis and biological activity of Ferulago macedonica Micevski et Mayer, Echinophora sibthorpiana Guss., Laserpitium ochridanum Micevski, Heracleum orphanidis Boiss. and Angelica pancicii Vandas (Apiaceae)

Osnovni cilj ove doktorske disertacije je karakterizacija hemijskog sastava etarskih ulja i ekstrakata retkih i endemičnih vrsta iz Republike Makedonije koje su delimično ili potpuno neistražene: Ferulago macedonica Micevski et Mayer, Echinophora sibthorpiana Guss., Laserpitium ochridanum Micevski, Heracleum orphanidis Boiss. i Angelica pancicii Vandas (Apiaceae). Drugi deo ovog istraživanja odnosi se na utvrđivanje in vitro biološkog potencijala etarskih ulja, metanolnih, etanolnih i vodenih ekstrakata navedenih vrsta i to njihovog antioksidativnog, antimikrobnog i anti-quorum sensing dejstva. U ovom radu su prvi put predstavljeni rezultati hemijske analize etarskih ulja vršnog i nadzemnog dela F. macedonica kao i metanolnih ekstrakata nadzemnog dela i korena vrsta H. orphanidis i A. pancicii. Do sada nije bilo ni literaturnih podataka o biološkim aktivnostima vrsta F. macedonica, H. orphanidis i A. pancicii. GC-FID i GC-MS analiza sastava etarskih ulja proučavanih vrsta pokazala je dominantno prisustvo monoterpenskih i seskviterpenskih ugljovodonika i njihovih oksidovanih formi. Ove analize su pokazale da su etarska ulja vršnog i nadzemnog dela F. macedonica bogata α-pinenom (43,10%; 22,78%) i sabinenom (26,73%; 15,46%). Za etarsko ulje E. sibthorpiana karakteristično je prisustvo velike koncentracije metil eugenola (60,40%), p-cimena (11,18%) i α-felandrena (10,23%), a za etarsko ulje L. ochridanum veliki sadržaj sabinena (22,8%), viridiflorola (14,7%) i α-pinena (11,4%). U etarskom ulju H. orphanidis kao glavna jedinjenja identifikovani su n-oktanol (39,57%), n-oktil heksanoat (17,62%) i n-oktil acetat (14,12%), dok su bornil acetat (8,08%), n-oktanol (5,82%), kesan (4,26%) i β-selinen (4,25%) najzastupljenije komponente etarskog ulja A. pancicii. Rezultati tentativne analize sastava metanolnih ekstrakata odabranih vrsta, dobijenih pomoću HPLC-DAD/ESI-ToF-MS sistema, prvi put su predstavljeni i opisani u ovoj disertaciji. Identifikacija sekundarnih metabolita ekstrakata urađena je na osnovu molekulske formule dobijene korišćenjem Mass Hunter Workstation softvera za prikupljanje i obradu podataka i na osnovu literaturnih podataka o hemijskom sastavu vrsta odgovarajućih rodova i predstavnika drugih rodova familije Apiaceae...The main goal of this doctoral dissertation was chemical characterization of essential oils and methanolic extracts of rare and endemic unexplored or partially investigated species originating from Republic of Macedonia: Ferulago macedonica Micevski & Mayer, Echinophora sibthorpiana Guss. Laserpitium ochridanum Micevski, Heracleum orphanidis Boiss. and Angelica pancicii Vandas (Apiaceae). Other part of this investigation was dedicated to determination of in vitro biological potential of essential oils, methanol, ethanol and aqueous extracts of selected species including antioxidant, antimicrobial and anti-quorum sensing effects. To date, no published results are available on phytochemical investigation of essential oils of F. macedonica and H. orphanidis and metanolic extracts of H. orphanidis and A. pancicii aerial parts and roots. Additionally, there is no literature data about biological potential of F. macedonica, H. orphanidis and A. pancicii species. GC-FID and GC-MS analysis showed a dominant presence of monotererpene and sesquiterpene hydrocarbons and their oxygenated forms in essential oils. This analisys showed that essential oils of F. macedonica apical and aerial parts were rich in α-pinene (43,10%; 22,78%) and sabinene (26,73%; 15,46%). E. sibthorpiana essential oil was characterised by dominant presence of methyl eugenol (60,40%), p-cymene (11,18%) i α-phellandrene (10,23%), while the most abundant compounds of L. ochridanum essential oil were sabinene (22,8%), viridiflorol (14,7%) and α-pinene (11,4%). As the main components of H. orphanidis essential oil were identified n-octanol (39,57%), n-octyl hexanoate (17,62%) and n-octyl acetate (14,12%), while bornyl acetate (8,08%), n-octanol (5,82%), kessane (4,26%) and β-selinene (4,25%) were present in the highest percentage in A. pancicii essential oil. The results of tentative analysis of methanol extracts of selected species, using HPLC-DAD/ESI-ToF-MS system, were described for the first time in this work. The identification of secondary metabolites of these samples was based on molecular formulas obtained by Mass Hunter Workstation software and based on literature data on the composition of the respective genera and other species from Apiaceae family..

Radical scavenging and antimicrobial activity of essential oil and extracts of Echinophora sibthorpiana Guss. from Macedonia

This study was undertaken to determine the antioxidant and antimicrobial effect of essential oil and extracts of Echinophora sibthorpiana Guss. (fam. Apiaceae) collected in Macedonia. The chemical composition of E. sibthorpiana essential oil was characterized by the presence of methyl eugenol (60.40%), p-cymene (11.18%) and α-phellandrene (10.23%). The free radical scavenging activity of extracts and essential oil was evaluated by DPPH and ABTS assays. The aqueous extract of aerial parts exhibited the strongest scavenging activity (IC50=1.67 mg/ml); results of the ABTS test showed that the most effective was the ethanol extract of aerial parts (1.11 mg vit. C/g). The essential oil showed stronger antioxidant activity compared to hydroxyanisole, ascorbic acid and quercetin that were used in the DPPH and ABTS tests, respectively. The total phenolic and flavonoid concentrations in the extracts ranged between 38.65-60.72 mg GA/g, and 3.15-19.00 mg Qu/g, respectively. The antimicrobial properties of the extracts and essential oil were investigated using a micro-well dilution technique against human pathogenic strains. The results were comparable with the effects of the positive controls, streptomycin and fluconazole. These findings indicate that E. sibthorpiana extracts and oil can be used in preventive treatments and as an alternative for synthetic preservatives. [Projekat Ministarstva nauke Republike Srbije, br. 173029 i br. 173021

Supplementary data for article: Mileski, K. S.; Trifunović, S. S.; Ćirić, A. D.; Šakić, Ž. M.; Ristić, M. S.; Todorović, N. M.; Matevski, V. S.; Marin, P. D.; Tešević, V. V.; Džamić, A. M. Research on Chemical Composition and Biological Properties Including Antiquorum Sensing Activity of Angelica Pancicii Vandas Aerial Parts and Roots. Journal of Agricultural and Food Chemistry 2017, 65 (50), 10933–10949. https://doi.org/10.1021/acs.jafc.7b04202

Supporting information for: [https://doi.org/10.1021/acs.jafc.7b04202]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2572

Laserpitium ochridanum: antioxidant, antimicrobial and anti-quorum sensing activities against Pseudomonas aeruginosa

This study shows Laserpitium ochridanum essential oil composition, its antifungal potency, and possible antioxidant, antimicrobial and anti-quorum sensing activities of different extracts. Monoterpene hydrocarbons (40.9%) found to be the most abundant group of constituents in the oil. Sabinene (22.8%), viridiflorol (14.7%) and α-pinene (11.40%) were the main compounds of the oil. Ethanolic extract had the highest antioxidant capacity in DPPH and ABTS assays and was the richest in polyphenols. Microdilution method revealed the strongest antibacterial activity of ethanolic extracts in comparison to other tested extracts and streptomycin. L. ochridanum oil evidenced the best antifungal potential against used micromycetes. Results of anti-quorum sensing activity assay indicated high affection of aqueous extract in reduction of PAO1 pyocianin production (18.07%).Used samples possessed slight reduction of twitching and swimming mobility. Excellent antifungal oil activity, good antioxidant, antimicrobial and anti-quorum activity of L. ochridanum extracts was found. This study shows for the first time anti-guorum sensing activity against Pseudomonas aeruginosa of L. ochridanum, as well as its significant antioxidant potential. This study was conducted due to adventives of usage of natural products as efficient substitution for synthetic agents in treatment of various medical conditions

Inhibition of quorum sensing virulente factors of Pseudomonas aeruginosa PAO1 by Ferulago macedonica and Echinophora sibthorpiana extracts and essential oils

This study was conducted to analyse extracts and essential oils of Ferulago macedonica and Echinophora sibthorpiana (Apiaceae) for anti–quorum sensing potential using Pseudomonas aerugonisa PAO1 as biomonitoring system. Anti–quorum sensing screening was carried out by tree different tests which showed that examined samples possessed significant effects in suppressing of some virulent determinants of PAO1. The essential oils of F. macedonica and E. sibthorpiana aerial parts reduced the growth and modified the appearance of P. aeruginosa colonies and flagella to a large extent in twitching and motility assay. In the presence of essential oil of E. sibthorpiana, diameter of the colony was 6.67 mm, the flagella were absent and the color of the colony was changed from green to white. In anti–biofilm assay, the inhibitory activity of E. sibthorpiana was more notable than of F. macedonica. Applied colorimetric test revealed that examined samples strongly reduced the production of pigment pyocyanin where the highest inhibition of its synthesis showed F. macedonica oil (5.22%), followed by E. sibthorpiana oil (10.69%), which was singnificantly bigger potential in comparison to used antibiotics (141.55%)

Essential Oil Composition, Antioxidant and Antimicrobial Properties of Essential Oil and Deodorized Extracts of Helichrysum italicum (Roth) G. Don

The main aim of this work was to determine the chemical composition of essential oil from aerial parts and to evaluate potential antimicrobial and antioxidant activities of essential oil and deodorized extracts of Helichrysum italicum. The gas chromatography-flame ionization detector (GC/FID) and gas chromatography-mass spectrometry (GC/MS) techniques revealed the highest amount of sesquiterpene hydrocarbons (37.88 %), with neryl-acetate (20.44 %), γ-curcumene (14.08 %) and trans-α-bergamotene (6.99 %) as the main components. Total phenolic contents of tested extracts were determined by Folin-Ciocalteau assay and gallic acid was used as a standard. The highest phenolic concentration was measured in deodorised aqueous extract. The radical scavenging activity of the samples was evaluated by means of DPPH radicals. In this test, deodorised aqueous extract possessed the highest antioxidant effect (IC50 = 0.35 mg/ml), followed by deodorized methanol extract, while essential oil had the lowest scavengin..

Reactivity of particles under electron microscopy.

Ferulago macedonica Micevski & E. Mayer (Apiaceae) , an endemic species from Macedonia, was evaluated for its essential oil composition and biological activity for the first time. The essential oil composition from inflorescence and aerial parts, analyzed by gas chromatography and gas chromatography–mass spectrometry (GC and GC-MS), indicated α-pinene (43.10% and 22.78%) and sabinene (26.73% and 15.46%) as the main components. The antioxidant activities of extracts was determinated spectrophotometrically using 2,2-diphenyl-1-picrylhydrazyl ( DPPH). The strongest activity had methanol extracts of inflorescence and aerial parts (IC 50=490 mg/mL and IC 50=630 mg/mL of solution, respectively). According to 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) test, the most effective were ethanol and methanol extracts of F. macedonica inflorescence (1.69 and 1.63 mg Vit.C/g dw). Total phenolic and flavonoid contents evaluation indicated that their amounts were ranged between 34.12-59.68 mg gallic acid (GA)/g dw for phenols and 9.08 - 23.73 mg quercetin hydrate (QE)/g dw for flavonoids. Antimicrobial activity of extracts and essential oils was investigated by micro-well dilution assay. Minimum inhibitory (MIC) and minimum bactericidal/fungicidal concentrations (MBC/MFC) were defined. The methanol extract of inflorescence has shown to be the strongest in both, antibacterial ( MICs=4-8 mg/mL; MBCs=9-13 mg/mL ) and antifungal activity ( MICs=2-12 mg/mL; MFCs=6-18 mg/mL). Essential oil from inflorescence and aerial parts of F. macedonica manifested stronger inhibition on growth of pathogenic bacteria and fungy compared to tested extracts and positive standards

Heracleum orphanidis: chemical characterisation, and comparative evaluation of antioxidant and antimicrobial activities with specific interest in the influence on Pseudomonas aeruginosa PAO1

GC-FID and GC-MS were used to determine the chemical composition of the essential oil of the aerial parts of Heracleum orphanidis Boiss. Also, the HPLC-DAD/ESI-ToF-MS profile of the methanol extracts of aerial parts and roots was determined. The main components of the essential oil were n-octanol, octyl hexanoate and n-octyl acetate, while coumarins were the most prevalent compounds in methanol extracts. An evaluation of antioxidant activity showed that the methanol and aqueous extracts of the aerial parts had the highest potential. In terms of antimicrobial activity determined by microdilution assays, the essential oil and methanol extract of the roots showed the greatest effectiveness. The colonies of Pseudomonas aeruginosa PAO1 treated with H. orphanidis samples produced less toxic pyocyanin, showed lower twitching and flagella mobility and biofilm formatting was reduced. The analyses in this study showed the considerable biological potential of H. orphanidis considering free radicals and various pathogenic strains, including the wild type of P. aeruginosa.The peer-reviewed version: [http://cer.ihtm.bg.ac.rs/handle/123456789/3130

Areca catechu - from farm to food and biomedical applications

The family Arecaceae includes 181 genera and 2,600 species with a high diversity in physical characteristics. Areca plants, commonly palms, which are able to grow in nearly every type of habitat, prefer tropical and subtropical climates. The most studied species Areca catechu L. contains phytochemicals as phenolics and alkaloids with biological properties. The phenolics are mainly distributed in roots followed by fresh unripe fruits, leaves, spikes, and veins, while the contents of alkaloids are in the order of roots, fresh unripe fruits, spikes, leaves, and veins. This species has been reputed to provide health effects on the cardiovascular, respiratory, nervous, metabolic, gastrointestinal, and reproductive systems. However, in many developing countries, quid from this species has been associated with side effects, which include the destruction of the teeth, impairment of oral hygiene, bronchial asthma, or oral cancer. Despite these side effects, which are also mentioned in this work, the present review collects the main results of biological properties of the phytochemicals in A. catechu. This study emphasizes the in vitro and in vivo antioxidant, antimicrobial, anticancer, and clinical effectiveness in humans. In this sense, A. catechu have demonstrated effectiveness in several reports through in vitro and in vivo experiments on disorders such as antimicrobial, antioxidant, or anticancer. Moreover, our findings demonstrate that this species presents clinical effectiveness on neurological disorders. Hence, A. catechu extracts could be used as a bioactive ingredient for functional food, nutraceuticals, or cosmeceuticals. However, further studies, especially extensive and comprehensive clinical trials, are recommended for the use of Areca in the treatment of diseases.info:eu-repo/semantics/publishedVersio