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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 ek, 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 w

A series of 18 novel N-Mannich bases derived from 5-adamantyl-1,2,4-triazole-3-th ione was synthesized and characterized using NMR spectroscopy and X-ray diffraction technique. All derivatives were evaluated for their anticancer potential against four human cancer cell lines. Several tested compounds exerted good cytotoxic activities on K562 and HL-60 cell lines, along with pronounced selectivity, showing lower cytotoxicity against normal fibroblasts MRC-5 compared to cancer cells. The effects of compounds 5b,5e, and 5j on the cell cycle were investigated by flow cytometric analysis. It was found that these compounds cause the accumulation of cells in the subG1 and G1 phases of the cell cycle and induce caspase-dependent apoptosis, while the anti-angiogenic effects of 5b, 5e, and 5j have been confirmed in EA. hy926 cells using a tube formation assay. Further, the interaction of Bax protein with compound 5b was investigated by means of molecular modeling, applying the combined molecular docking/molecular dynamics approach.

The essential oil, different extracts, and isolated compounds of Angelica pancicii Vandas (Apiaceae) were investigated for the first time. The GC-FID and GC-MS analyses revealed sesquiterpenoids as the main constituents of A. pancicii essential oil of aerial parts with bornyl acetate (8.08%), n-octanol (5.82%), kessane (4.26%), and beta-selinene (4.26%) as the main constituents. Analysis of methanol extracts, using an HPLC-DAD/ESI-ToF-MS system, showed a total of 52 compounds in the aerial parts and 53 in the roots, indicating coumarins as the main constituents. In addition, new chromone (1) and six known furanocoumarins (2-7) were isolated from the roots and structurally elucidated by combined spectroscopic methods. The aerial part extracts exhibited higher polyphenolic contents and antioxidant activity evaluated by three radical scavenging assays. Using a microwell dilution method, the strongest antibacterial activity profiles were determined for ethanol and methanol root extracts (minimum bactericidal concentrations (MBCs) = 0.25-3.00 mg/mL), which were comparable to the activity of streptomycin (MBCs = 0.34-1.24 mg/mL), while the strongest antibacterial compound of A. pancicii was oxypeucedanin hydrate (MBCs = 0.50-8.00 mg/mL). Antifungal potential was in moderate extent, and the highest activity was obtained for root methanol extract (minimum fungicidal concentrations (MFCs) = 4.00-14.00 mg/mL). Tested sub-minimum inhibitory concentrations (subMICs) of the extracts and isolated compounds inhibited selected Pseudomonasaeruginosa PAO1 virulence determinants. The most reduced growth of P. aeruginosa colony was in the presence of isolated oxypeucedanin. Ethanol (17.36-46.98%) and methanol (34.54-52.43%) root extracts showed higher anti-biofilm activity compared to streptomycin (49.40-88.36%) and ampicillin (56.46-92.16%).

Eight 1,3,4-oxadiazole derivatives containing phenolic acid moieties (7a-h) and eight of their diacylhydrazine precursors (6a-h) were synthesized, characterized using spectroscopic methods and examined by scavenging of stable DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals. The most potent phenolic 1,3,4-oxadiazoles showed better DPPH scavenging activity in comparison with their corresponding diacylhydrazine precursors as a result of participation of both aromatic rings and a 1,3,4-oxadiazole moiety in resonance stabilization of the formed phenoxyl radical. Four diacylhydrazines (6d, 6e, 6g, and 6h) and four 1,3,4-oxadiazoles (7d, 7e, 7g and 7h) with the best DPPH scavenging activity, were chosen for further evaluation of their antioxidant potential through various assays. The investigated compounds exerted pronounced ABTS radical scavenging capacity, moderate to good H2O2 scavenging properties and strong ferric ion reducing capacity. Further in vitro evaluation of the antioxidant properties of the most active compounds demonstrated their protective effects in normal lung fibroblasts MRC-5 against hydrogen peroxide induced oxidative stress. Diacylhydrazine 6h increased two times the activity of glutathione peroxidase in treated cells in comparison with a control sample and did not affect the superoxide dismutase activity.

Background Short ragweed (Ambrosia artemisiifolia) allergies affect more than 36 million people annually. Ragweed pollen grains release subpollen particles (SPP) of respirable size upon hydration or a change in air electrical conditions. The aim of this study was to characterize the proteomes and allergomes of short ragweed SPP and total pollen protein extract (TOT), and compare their effects with those of standard aqueous pollen protein extract (APE) using sera from short ragweed pollen-sensitized patients. Methods Quantitative 2D gel-based and shotgun proteomics, 1D and 2D immunoblotting, and quantitative ELISA were applied. Novel SPP extraction and preparation protocols enabled appropriate sample preparation and further downstream analysis by quantitative proteomics. Results The SPP fraction contained the highest proportion (94%) of the allergome, with the largest quantities of the minor Amb a 4 and major Amb a 1 allergens, and as unique, NADH dehydrogenases. APE was the richest in Amb a 6, Amb a 5 and Amb a 3, and TOT fraction was the richest in the Amb a 8 allergens (89% and 83% of allergome, respectively). Allergenic potency correlated well among the three fractions tested, with 1D immunoblots demonstrating a slight predominance of IgE reactivity to SPP compared to TOT and APE. However, the strongest IgE binding in ELISA was noted against APE. New allergenic candidates, phosphoglycerate mutase and phosphoglucomutase, were identified in all the three pollen fractions. Enolase, UTP-glucose-1-phosphate uridylyltransferase and polygalacturonase were observed in SPP and TOT fractions as novel allergens of the short ragweed pollen, as previously described. Conclusion and Clinical Relevance We demonstrated that the complete major (Amb a 1 and 11) and almost all minor (Amb a 3, 4, 5, 6, 8 and 9) short ragweed pollen allergen repertoire as well as NADH oxidases are present in SPP, highlighting an important role for SPP in allergic sensitization to short ragweed.

The essential oil, different extracts, and isolated compounds of Angelica pancicii Vandas (Apiaceae) were investigated for the first time. The GC-FID and GC-MS analyses revealed sesquiterpenoids as the main constituents of A. pancicii essential oil of aerial parts with bornyl acetate (8.08%), n-octanol (5.82%), kessane (4.26%), and beta-selinene (4.26%) as the main constituents. Analysis of methanol extracts, using an HPLC-DAD/ESI-ToF-MS system, showed a total of 52 compounds in the aerial parts and 53 in the roots, indicating coumarins as the main constituents. In addition, new chromone (1) and six known furanocoumarins (2-7) were isolated from the roots and structurally elucidated by combined spectroscopic methods. The aerial part extracts exhibited higher polyphenolic contents and antioxidant activity evaluated by three radical scavenging assays. Using a microwell dilution method, the strongest antibacterial activity profiles were determined for ethanol and methanol root extracts (minimum bactericidal concentrations (MBCs) = 0.25-3.00 mg/mL), which were comparable to the activity of streptomycin (MBCs = 0.34-1.24 mg/mL), while the strongest antibacterial compound of A. pancicii was oxypeucedanin hydrate (MBCs = 0.50-8.00 mg/mL). Antifungal potential was in moderate extent, and the highest activity was obtained for root methanol extract (minimum fungicidal concentrations (MFCs) = 4.00-14.00 mg/mL). Tested sub-minimum inhibitory concentrations (subMICs) of the extracts and isolated compounds inhibited selected Pseudomonasaeruginosa PAO1 virulence determinants. The most reduced growth of P. aeruginosa colony was in the presence of isolated oxypeucedanin. Ethanol (17.36-46.98%) and methanol (34.54-52.43%) root extracts showed higher anti-biofilm activity compared to streptomycin (49.40-88.36%) and ampicillin (56.46-92.16%).

This investigation examines surface sediment samples from rivers and artificial lakes (Serbia) to quantify levels of potentially harmful elements (As, Hg, and Se) and several ancillary elements (Al, Fe, Mn, Ca, Mg, C, H, N, and S). These data provide the first global analysis of Hg, As and Se levels in a well-studied system. Combination of different methods was applied for estimation of the environmental status of sediments and to determine the potential risk of ecological damage: digestion with aqua regia and determination of element content, determination of elements mobile forms by ammonium-acetate extraction, calculation of contamination indexes - index of geoaccumulation and enrichment factor, comparison with sediment quality guidelines and other literature data, and use of statistical methods. Results indicate that the sediments are slightly enriched with As and Hg, but not with Se. Fact that values for Se contents was less than the international threshold levels, suggesting the very probable absence of risk of contamination over the catchment with this element. The average pollution levels in Serbian river and artificial lakes expressed in terms of geoaccumulation indexes of As and Hg indicate that the environment is uncontaminated to moderately contaminated. All values of EF for Hg were below 2, indicating deficiency to minimal enrichment with Hg on studied localities. The EF values of As indicate deficiency to moderate enrichment (EF ranged from 0.11 to 3.54). According to the comparison to quality standard, the As in sediments of the studied area have potential risk. High content of arsenic in river and lake sediments indicate that arsenic pollution problem that lasts longer period of time.

Species within the genus Centaurium readily hybridize and polyploid complexes are often seen in natural populations. We describe phytochemical profiles of newly discovered allohexaploid hybrid, here named Centaurium pannonicum, and its parental tetraploid species C. erythraea and rare C. littorale ssp. compressum. Our aim was to examine chemodiversity of these taxa in the area of Vojvodina (North Serbia) and to perform metabolomics search for chemical classifiers which would provide high resolution discrimination of parental and hybrid individuals. In sum, UHPLC-MS/MS Orbitrap metabolomics fingerprinting revealed seventy compounds in methanol extracts. Despite the lack of qualitative chemical novelty in hybrid plants, UHPLC-qqqMS targeted metabolomics approach, aimed at three secoiridoid compounds and seventeen phenolics, pointed to considerable differences in quantitative composition of these dominant compounds among the plant taxa studied. In addition to the difference in the ploidy levels, the hybrid taxon was well distinguished from both parental species based on metabolite profiles, and, for most individuals, positioned intermediately to the parental taxa in both PCA and hierarchical clustering. After optimizing and comparing several statistical learning methods, it was possible to narrow the number of taxonomic classifiers to five (three xanthones, one secoiridoid glycoside, and one phenolic acid), while increasing the differentiation resolution. The presented metabolomics approach will certainly, along with morphometrics and molecular genetics studies, have high impact on further elucidation of complex relationships among taxa within the genus Centaurium.

This investigation examines surface sediment samples from rivers and artificial lakes (Serbia) to quantify levels of potentially harmful elements (As, Hg, and Se) and several ancillary elements (Al, Fe, Mn, Ca, Mg, C, H, N, and S). These data provide the first global analysis of Hg, As and Se levels in a well-studied system. Combination of different methods was applied for estimation of the environmental status of sediments and to determine the potential risk of ecological damage: digestion with aqua regia and determination of element content, determination of elements mobile forms by ammonium-acetate extraction, calculation of contamination indexes - index of geoaccumulation and enrichment factor, comparison with sediment quality guidelines and other literature data, and use of statistical methods. Results indicate that the sediments are slightly enriched with As and Hg, but not with Se. Fact that values for Se contents was less than the international threshold levels, suggesting the very probable absence of risk of contamination over the catchment with this element. The average pollution levels in Serbian river and artificial lakes expressed in terms of geoaccumulation indexes of As and Hg indicate that the environment is uncontaminated to moderately contaminated. All values of EF for Hg were below 2, indicating deficiency to minimal enrichment with Hg on studied localities. The EF values of As indicate deficiency to moderate enrichment (EF ranged from 0.11 to 3.54). According to the comparison to quality standard, the As in sediments of the studied area have potential risk. High content of arsenic in river and lake sediments indicate that arsenic pollution problem that lasts longer period of time.

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.

According to scientific community and professional specialists, urging necessity to increase capacity of sources of energy can be partially achieved by using biomass as a raw material. This approach is justified by both economic and ecological reasons. In this paper the results of chemical study of the biomass Miscanthus × giganteus Greef et Deu. cultivated in the Republic of Serbia in the experimental field of the Institute for Application of Nuclear Energy in Zemun were presented. Study of an amount of ashes, elemental analysis of the biomass, and calorific value of the biomass (both higher and lower), which are critical parameters when estimating the quality of a biofuel, points out the possibility of utilization of the miscantus biomass as a fuel. Samples of the plant material for this study were collected during two harvesting periods: in the fall, when the biological yield is the highest, and at the end of the winter, when a highest quality biomass with respect to the combustion is obtained do to a decrease in moisture content and remobilization of the macro- and micronutrients from the above-ground parts of the plants to the rhizomes. Values of the studied parameters indicate that biomass meets the 'green' energy requirements because the amounts of oxides of nitrogen and sulphur in the combustion products would not threaten current quality of the environment and the amount of carbon dioxide produced would be consumed in the process of photosynthesis. Results of the chemical analyses strongly suggest to a high potential of utilization of the miscantus biomass as a biofuel (renewable source of energy) to generate thermal energy., Delimičan odgovor stručne i naučne javnosti na pitanje kako povećati kapacitete energetskih izvora je upotreba biomase kao sirovine, kako iz ekonomskih, tako i iz ekoloških razloga. U ovom radu su prikazani rezultati fizičko-hemijskih ispitivanja biomase miskantusa (Miscanthus × giganteus Greef et Deu.), gajenog na teritoriji Republike Srbije, oglednom polju Instituta za primenu nuklearne energije, Zemun. Ispitivanjem količine pepela, elementarnog sastava biomase i kalorične vrednosti biomase (donje i gornje), koji su presudni za procenu kvaliteta biogoriva, ukazano je na mogućnost eksploatacije biomase miskantusa kao biogoriva. Prikupljanje uzoraka biljnog materijala za ovo istraživanje izvršeno je u dva žetvena roka: u jesen - vreme maksimuma biološkog prinosa i krajem zime, kada se dobija kvalitetnija biomasa za sagorevanje usled smanjenja sadržaja vlage i remobilizacije makro- i mikrohraniva iz nadzemnih delova rizoma. Vrednosti ispitivanih parametara pokazuju da biomasa ispunjava zahteve 'zelene' energije, jer produkti sagorevanja biomase svojom količinom (oksidi N i S) ne bi ugrožavali postojeći kvalitet životne sredine, a količina nastalog ugljenik (IV) - oksida ne bi predstavljala ekološki problem, jer bi se dalje uključila u proces fotosinteze. Rezultati hemijskih analiza argumentovano ukazuju na veliki potencijal upotrebe biomase miskantusa kao biogoriva (obnovljivog izvora energije) u proizvodnji toplotne energije.

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.

This study evaluated the chemical composition, antioxidant and antimicrobial activities of the essential oils of twelve Ocimum basilicum L. cultivars grown in Serbia. The essential oils were obtained by hydrodistillation and analyzed by gas chromatography-flame ionization detection and GC-mass spectrometry. The oil yields ranged from 0.65 to 1.90 %. A total of 75 compounds were identified as constituents of analyzed essential oils. GC/MS analyses revealed that a majority of the examined basil cultivars belonged to the "linalool chemotype". The results of the DPPH assay showed a very high antioxidant capacity of the basil oils, which was especially high for Blue Spice cultivar (IC50 = 0.03 mu g/mL). Significant antimicrobial activity was shown for all the tested oils. Essential oil of Compact cultivar was the most active against the bacterium Micrococcus flavus, with an MIC value of 0.009 mu g/mL while Osmin cultivar exhibited the strongest antifungal activity with MIC values ranging from 0.08-1.07 mu g/mL. All the examined essential oils showed a 10-to 100-fold greater ability to inhibit fungal growth compared to commercial antifungal agents.

Oxidative stress is directly related to several diseases and symptoms, where antioxidant compounds, such as xanthenes, may become important in prevention and/or treatmant. Ten biologically active 9-aryl substituted 2,6,7-trihydroxyxanthen-3-one derivatives were synthesized using reliable one-pot synthesis and their structures were confirmed by IR, H-1 and C-13 NMR spectroscopy and mass spectrometry. Some of the synthesized compounds were scanned for their antioxidant potency using electrochemical method cyclic voltammetry of immobilized microparticles. Substitution of hydrogen at the phenyl ring of 2,6,7-trihydroxy-9-phenylxanthen-3-one with an electron-donating group affected the reducing power of the compounds by lowering the biological oxidation potential. These results signify the importance of xanthen-3-one derivatives as antioxidant agents and their further biological evaluation.

The increasing use of sun-creams containing organic UV-filters has led to increased concentration of these compounds in aquatic environment. Chlorinated water can convert these chemicals into chlorinated products whose toxic effects are of primary concern. The new compound may be more toxic than the starting primary compound. Many studies have shown that UV filters absorb UV light and decompose under solar irradiation, due to their unstable properties. This may lead to formation of certain by-products with harmful effects. Their decomposition products can cause allergic and toxic reactions to the human skin. This study follows the stability of most commonly used UV filters, homosalate, in conditions that include those existent in swimming pools. Stability of the homosalate in chlorinated water was studied in simulated swimming pool water samples. UV spectroscopy was used to follow the reaction of homosalate in presence of free chlorine. Water samples were filtered, acidified, and extracted by use of solid-phase extraction. Gas chromatography with mass spectrometry was used to identify the major transformation by-products. Under the experimental conditions, homosalate reacted with chlorine following zero order reaction. The chemical transformation of the homosalate in chlorinated water led to formation of chlorinated by-products that was identified as: monochloro-homosalate, dichloro-homosalate and two diastereoisomers monochloro-homosalate.

Experimental details relating to the article: Mandić, B., Vlajic, M. D., Trifunović, S., Simić, M. R., Vujisić, L. V., Vučković, I., Novaković, M., Nikolic-Mandic, S. D., Tešević, V., Vajs, V.,& Milosavljević, S. (2015). Optimisation of isolation procedure for pyrrolizidine alkaloids from Rindera umbellata Bunge. Natural Product Research, 29(9), 887-890. https://doi.org/10.1080/14786419.2014.991929. Figure S1. Structures of monitored PAs from R. umbellata (1 - 6). Figure S2. Yields of PAs in relation to days of maceration in the methanol. Figure S3. Yields of PAs extracted with methanol, ethanol and sulphuric acid during four days at dark place and room temperature. Figure S4. Yields of PAs (1-6) in relation to molarity of sulphuric acid used for their extraction. Figure S5. Yields of PAs (1-6) in relation to days of maceration with 1 M sulphuric acid, at room temperature. Figure S6. Yields of PAs (1-6) in relation to days of ultrasonic assisted extraction with 1 M sulphuric acid. Figure S7. Yields of PAs (1-6) in relation to days of extraction with 1 M sulphuric acid by overhead rotary mixer. Figure S8. Comparison of yields of PAs (1-6) accomplished after four days of maceration, three days of extraction by ultrasonic bath and overhead rotary mixer assisted extractions of plant material with 1M sulphuric acid. Figure S9. Yields of PAs (1-6) isolated from 1.00 g of the plant material (with 1M sulphuric acid during the extraction by overhead rotary mixer for three days) in relation to the volume (ml) of sulphuric acid. Figure S10. Yields of isolated PAs (1-6) in relation to pH values of aqueous solutions for alkaline extraction of PAs with CH2Cl2. Figure S11. Yields of isolated PAs in relation to the number of continuous alkaline re-extractions with CH2Cl2 at pH 9.

Procedure for isolation of pyrrolizidine alkaloids (PAs) from Rindera umbellata Bunge plant species was optimised. Different extraction media (methanol, ethanol and sulphuric acid), concentration and volume of sulphuric acid, pH of PA solution for alkaline extraction, extraction time and techniques (maceration, ultrasonic and overhead rotary mixer assisted extraction) were investigated. The yields of six PAs (7-angeloyl heliotridane, 7-angeloyl heliotridine, lindelofine, 7-angeloyl rinderine, punctanecine and heliosupine) were monitored by GC-MS/FID. The best results for the isolation all of six PAs were obtained when the extraction was performed with 1M sulphuric acid (30mL per 1.00g of dried sample) by overhead rotary mixer during three days. Optimal pH value for alkaline extraction of PAs with CH2Cl2 was 9, and the extraction should be performed with four portions of 30mL of CH2Cl2. This procedure could be also useful for a plant sample preparation for GC and LC analyses of PAs.

Procedure for isolation of pyrrolizidine alkaloids (PAs) from Rindera umbellata Bunge plant species was optimised. Different extraction media (methanol, ethanol and sulphuric acid), concentration and volume of sulphuric acid, pH of PA solution for alkaline extraction, extraction time and techniques (maceration, ultrasonic and overhead rotary mixer assisted extraction) were investigated. The yields of six PAs (7-angeloyl heliotridane, 7-angeloyl heliotridine, lindelofine, 7-angeloyl rinderine, punctanecine and heliosupine) were monitored by GC-MS/FID. The best results for the isolation all of six PAs were obtained when the extraction was performed with 1M sulphuric acid (30mL per 1.00g of dried sample) by overhead rotary mixer during three days. Optimal pH value for alkaline extraction of PAs with CH2Cl2 was 9, and the extraction should be performed with four portions of 30mL of CH2Cl2. This procedure could be also useful for a plant sample preparation for GC and LC analyses of PAs.

Further phytochemical investigation into the aerial parts of Achillea clavennae has resulted in the 3 isolation of three new sesquiterpene lactones: two highly oxygenated germacranolides (1, 2) and 4 the iso-seco-guaianolide, 9(R)-acetoxy-3-O-methyl-iso-seco-tanapartholide (3). Eight known 5 compounds were also found in this plant species, of which 9 α -acetoxycanin (5), sintenin (6) and 6 oleanolic acid (7) were detected for the first time. The structures of the isolated compounds were 7 elucidated by combined spectroscopic methods (1D and 2D NMR, HRESIMS, CIMS, FTIR). 8 While the predominant metabolite germacranolide sintenin (6) was not cytotoxic, the new iso-9 seco-guaianolide (3) displayed cytotoxicity comparable to that of cisplatin and the lactone 10 apressin (4), inducing partly apoptotic death in human U251 and rat C6 glioma cell lines.

Rod Alnus Mill. (jove), sa više od 30 vrsta, pripada familiji Betulaceae. Alnus vrste koje rastu u Srbiji su A. glutinosa (crna jova), A. viridis subsp. viridis (zelena jova) i A. incana (siva jova). U ovom radu izolovani su i okarakterisani diarilheptanoidi iz kore crne jove i zelene jove i određena je njihova biološka aktivnost. Biljke ovog roda su u tradicionalnoj medicini korišćene u brojne svrhe, kao alterativi, astringenti, purgativi, emetici, antipiretici, hemostatici, antiparazitici i antihelmintici, za lečenje različitih bolesti, uključujući tumor. Jedinjenja izolovana iz jova poseduju širok spektar bioloških aktivnosti, a najvažnija su citotoksična, antitumorska, antivirusna, anti-inflamatorna, antimikrobna i antiparazitska. Vrste roda Alnus su bogat izvor diarilheptanoida. Oni se u biljci prvenstveno nalaze u kori i lišću, kao prosti, glikozidi ili složeni - diarilheptanoid glikozidi esterifikovani različitim fenolnim kiselinama, najčešće cinamičnim. U okviru ove disertacije iz kora zelene i crne jove izolovano je i okarakterisano 39 jedinjenja, od kojih je 19 novih prirodnih proizvoda. Iz zelene jove izolovano je 13 jedinjenja (1-13), a iz crne 28 (1, 5, 14-39), pri čemu su diarilheptanoidi platifilozid (1) i acerozid VII (5) izolovani iz obe biljne vrste. Od 13 jedinjenja izolovanih iz zelene jove, 11 pripada grupi diarilheptanoida (1-11), od kojih je 6 novih prirodnih proizvoda (3, 7, 8, 9, 11 i 13). Svih 28 jedinjenja izolovanih iz crne jove su diarilheptanoidi, a 13 (19, 27, 29-39) je novih. Strukturne karakteristike diarilheptanoida zelene jove su da isključivo imaju phidroksifenil grupe kao aromatične sisteme i da se, pored prostih, sreću glikozidi koji se sastoje iz mono- i disaharida. Složeni diarilheptanoidi su retki, izolovan je samo jedan sa galnom kiselinom (3). Za većinu diarilheptanoida crne jove karakteristično je da imaju bar jednu kateholsku grupu. Od glikozida su zastupljeni isključivo oni sa monosaharidnom jedinicom, prvenstveno glukoz, The genus Alnus Mill. (Alder), with more than 30 species, belongs to the Betulaceae family. Alnus species growing in Serbia are A. glutinosa (black alder), A. viridis subsp. viridis (green alder) and A. incana (gray alder). In this work, diarylheptanoids from the bark of black alder and green alder were isolated and characterized, and their biological activity was determined. Plants of this genus have been traditionally used for numerous purposes, e.g. as an alterative, astringent, cathartic, emetic, febrifuge, hemostatic, parasiticide, vermifuge, in the treatment of various diseases, including tumors. Compounds isolated from the Alder species possess a wide spectrum of biological activities and the most important are cytotoxic, antitumoral, antiviral, anti-inflammatory, antimicrobial and antiparasitic. Alnus species are a rich source of diarylheptanoids. These are predominantly found in the bark and leaves as simple diarylheptanoids, glycosides and complex diarylheptanoids – diarylheptanoid glycosides esterified with some phenolic acid, most commonly with some cinnamic acid. In this work, 39 compounds were isolated and characterized from the barks of green and black alder, of which 19 are new natural products. Thirteen compounds were isolated from green alder (1-13), and 28 from black alder (1, 5, 14-39), of which diarylheptanoids platyphylloside (1) and aceroside VII (5) were isolated from both species. Of the 13 compounds isolated from green alder, 11 belong to diarylheptanoids (1-11) from which 6 are new natural products (3, 7, 8, 9, 11, and 13). All 28 compounds isolated from black alder are diarylheptanoids and 13 (19, 27, 29-39) are new. The structural characteristics of the diarylheptanoids isolated from green alder are that they have only p-hydroxyphenyl groups as aromatic systems and that besides simple diarylheptanoids, diarylheptanoid glycosides are found which consist of mono- and disaccharides. Complex diarylheptanoids are rare and only one was isolate

Further phytochemical investigation into the aerial parts of Achillea clavennae has resulted in the 3 isolation of three new sesquiterpene lactones: two highly oxygenated germacranolides (1, 2) and 4 the iso-seco-guaianolide, 9(R)-acetoxy-3-O-methyl-iso-seco-tanapartholide (3). Eight known 5 compounds were also found in this plant species, of which 9 α -acetoxycanin (5), sintenin (6) and 6 oleanolic acid (7) were detected for the first time. The structures of the isolated compounds were 7 elucidated by combined spectroscopic methods (1D and 2D NMR, HRESIMS, CIMS, FTIR). 8 While the predominant metabolite germacranolide sintenin (6) was not cytotoxic, the new iso-9 seco-guaianolide (3) displayed cytotoxicity comparable to that of cisplatin and the lactone 10 apressin (4), inducing partly apoptotic death in human U251 and rat C6 glioma cell lines.

A complex mixture of compounds was identified from the secretion of specimens of Unciger transsilvanicus. Phenol and p-cresol were detected for the first time in the family Julidae, and for the second time in the order Julida. Thirteen quinones were identified, with a great relative abundance of toloquinone and 2-methoxy-3-methyl- 1,4-benzoquinone. Hydroquinone was detected for the first time in the order Julida. Besides these compounds, isopentyl hexacosatetraenoate and isopentyl esters of saturated and unsaturated fatty acids with chain lengths from C-14 to C(20)were identified. The most abundant non-quinone compound was isopentyl eicosenoate. The relative abundance of quinone and non-quinone in the defensive fluid of U transsilvanicus was 77% and 23%, respectively. The phylogenetic importance of the registered compounds is briefly discussed.

Nine diarylheptanoids, 1-9, catechin (11), and a phenolic glucoside, 10, were isolated from the bark of green alder (Alnus viridis). Four of the isolated compounds, i.e., 2, 5, 8, 10, are new. The structures of 1-11 were determined on the basis of spectroscopic data. All isolated compounds were evaluated for their in vitro protective effects on chromosome aberrations in peripheral human lymphocytes using cytokinesis-block micronucleus (CBMN) assay. Almost all of them exerted a pronounced effect of decreasing DNA damage of human lymphocytes, acting stronger than the known synthetic protector amifostine.