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Composition of essential oils from ripe dried fruits of Peucedanum longifolium Waldst. & Kit. (PL) and P. aegopodioides (Boiss.) Vandas (PA) was investigated for the first time. Both taxa are distributed on the Balkan Peninsula, Asia Minor and Transcaucasia, and some authors include the last species in a separate genus, Rhizomatophora Pimenov [1]. Plant material was collected in Serbia, PL on Vis hill in Sićevo Gorge, PA in vicinity of Pirot, near village Basara. Oils were obtained by hydrodistillation in Clevenger-type apparatus; yields 0.91% (PL) and 0.02% (PA), w/w. GC-FID-MS analysis revealed presence of 46 components in PL oil and 48 in PA oil, accounting for 98.0% and 90.0% of total oils. PL oil was dominated by monoterpene hydrocarbons (74.4%), mainly α-phellandrene (26.2%), β-phellandrene + limonene (21.0%) and myrcene (9.5%), followed by sesquiterpene hydrocarbons (17.9%), mainly germacrene B (9.5%). On the other hand, most abundant in PA oil were non-terpenic aliphatic hydrocarbons (46.1%), mainly n-undecane (16.5%) and n-nonane (11.3%). However, this oil also contained significant amounts of both non-oxygenated and oxygenated sesquiterpenes (11.4% and 13.6%), with (E)-sesquilavandulol being most prominent (10.0%). Previously, composition of essential oils from some other plant parts of PL was investigated. For example, essential oil from fresh leaves and young stems of this plant was dominated by sesquiterpene β-elemene (24.7%) and monoterpene (E)-β-ocimene (11.7%) [2], which were present in small amounts in dried fruit oil investigated in current study (0.5% and 0.6%). 5th International Conference on Natural Products Utilization: from Plants to Pharmacy Shelf - ICNPU-2023, Sts. Constantine & Helena, Bulgaria, 30th May to 2nd June 2023

Composition of dichloromethane and methanol extracts from roots and fruits of Prangos trifida (Mill.) Herrnst. & Heyn s.l. (collected in Sićevo Gorge, Serbia in 2020) was investigated. Extracts were obtained by bimaceration, firstly with dichloromethane, then with methanol; solvents were removed under reduced pressure. GC-FID-MS analysis, conducted after saponification and methylation of samples, showed that palmitic, oleic and linoleic (14.2-51.8%) were dominant fatty acids in root dichloromethane extract, and oleic, linoleic and petroselinic in oily supernatant of fruit dichloromethane extract (10.2-49.9%); in both samples 16 fatty acids were identified. Dominant phytosterols, investigated by GC-FID-MS after unsaponifiable fractions silanization, were β-sitosterol (48.2 and 25.1%) and stigmasterol (14.9 and 12.8%). LC-DAD-MS analysis of dichloromethane and methanol extracts revealed presence of 7-11 furanocoumarins, including oxypeucedanin and imperatorin (up to 105.9 and 49.2 mg/g in fruit dichloromethane extract crystalline precipitate), followed by smaller amounts of oxypeucedanin hydrate, isoimperatorin, heraclenol, heraclenin and/or xanthotoxin. All extracts were rich in one 2',3'-dihydrofuranocoumarin derivative. Previously, such derivative, prantschimgin, as well as imperatorin and isoimperatorin were isolated from this plant and their in vitro anti-inflammatory activity was demonstrated [1]. In current investigation, in root methanol extract, chlorogenic (31.6 mg/g), two dicaffeoylquinic (12.8 and 48.1 mg/g), and smaller amounts of one caffeoylquinic and one feruloylquinic acids were also determined. Furthermore, low quantities of chlorogenic and one dicaffeoylquinic acid and two quercetin-type flavonoids were detected in fruit methanol extract, and one methylluteolin-type flavonoid in both methanol extracts. 5th International Conference on Natural Products Utilization: from Plants to Pharmacy Shelf - ICNPU-2023, Sts. Constantine & Helena, Bulgaria, 30th May to 2nd June 2023

In this study composition and antimicrobial activity of essential oils of roots, leaves, stems and fruits of Prangos trifida (Mill.) Herrnst. & Heyn s.l. (Apiaceae) were investigated. Plant material was collected in Sićevo Gorge (Kusača) in 2020. Essential oils were isolated by hydrodistillation in Clevenger-type apparatus and analyzed by GC-FID-MS. Antimicrobial activity was tested by microdilution method against food contaminants: Gram-positive (Staphylococcus aureus, Bacillus cereus and Listeria monocytogenes), Gram-negative bacteria (Escherichia coli, Enterobacter cloacae and Salmonella Typhimurium), and molds (Aspergillus fumigatus, A. niger, A. versicolor, Penicillium funiculosum, P. verrucosum var. cyclopium and Trichoderma harzianum); artificial food preservatives E211 and E224 were used as controls. Dominant constituents of essential oils were monoterpenes (66.2-87.2%). Terpinolene (36.2%) and p-cymene (11.5%) prevailed in root oil, (E)-β-ocimen (23.2%) and terpinolene (18.1%) in leaf oil, p-cimen-8-ol (21.8%) and p-cimen (14.1%) in stem oil, and p-cimen (25.4%) and limonene (14.4%) in fruit oil. Essential oils showed significant antifungal activity (MIC=0.10-0.78 mg/mL; MFC=0.20-1.56 mg/mL), better compared to both tested commercial preservatives, and interesting antibacterial activity (MIC=0.20-6.25 mg/mL; MBC=0.39-12.5 mg/mL), which was in the case of root and stem oils better than activity of both preservatives against S. aureus and B. cereus. It should be noted that composition of tested oils was in accordance with composition of the oils of this plant from vicinity of Madrid (1) and from Crimea (2), while their antimicrobial activity was tested in this work for the first time. Obtained results justify further research of P. trifida essential oils as new natural food preservatives. Cilj ovog rada bio je da se ispitaju hemijski sastav i antimikrobna aktivnost etarskih ulja korena, listova, stabljika i plodova Prangos trifida (Mill.) Herrnst. & Heyn s.l. (Apiaceae). Biljni materijal je prikupljen na području Sićevačke klisure (Kusača) 2020. god. Etarska ulja su dobijena destilacijom vodenom parom u aparaturi po Klevendžeru i analizirana metodom GC-FID-MS. Antimikrobna aktivnost testirana je mikrodilucionom metodom na kontaminantima hrane: Gram-pozitivnim (Staphylococcus aureus, Bacillus cereus i Listeria monocytogenes), Gram-negativnim bakterijama (Escherichia coli, Enterobacter cloacae i Salmonella Typhimurium), i plesnima (Aspergillus fumigatus, A. niger, A. versicolor, Penicillium funiculosum, P. verrucosum var. cyclopium i Trichoderma harzianum); kao kontrole korišćeni su sintetski konzervansi E211 i E224. Dominantni sastojci etarskih ulja bili su monoterpeni (66,2-87,2%). Najzastupljeniji u ulju korena bili su terpinolen (36,2%) i p-cimen (11,5%), u ulju listova (E)-β-ocimen (23,2%) i terpinolen (18,1%), u ulju stabljika p-cimen- 8-ol (21,8%) i p-cimen (14,1%), a u ulju plodova p-cimen (25,4%) i limonen (14,4%). Etarska ulja su pokazala značajnu antifungalnu aktivnost (MIK=0,10-0,78 mg/mL; MFK=0,20-1,56 mg/mL), bolju u poređenju sa oba testirana komercijalna konzervansa, i interesantnu antibakterijsku aktivnost (MIK=0,20-6,25 mg/mL; MBK=0,39-12,5 mg/mL), koja je u slučaju ulja korena i stabljika prema S. aureus i B. cereus bila bolja u odnosu na oba konzervansa. Treba istaći da je sastav ispitivanih etarskih ulja bio u skladu sa sastavom ulja ove biljke iz okoline Madrida (1) i sa Krima (2), dok je njihova antimikrobna aktivnost ispitivana u ovom radu po prvi put. Dobijeni rezultati predstavljaju dobru osnovu za dalja istraživanja etarskih ulja P. trifida kao novih prirodnih konzervanasa. VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beograd

The genus Hieracium L. s. str. (Cichorieae, Asteraceae) is one of the most taxonomically complex genera of flowering plants. Chemosystematic significance of triterpenes especially for Asteraceae genera is scarcely assessed. The subject of this research are 28 Hieracium species from Balkan Peninsula (mostly collected at Mt Durmitor, Montenegro): 12 principal belonging to sections Pannosa, Naegeliana, Drepanoidea and Villosa, and 16 hybridogenous originated from the species from those four sections and Glauciformia, Hieracium, Prenanthoidea and/or Italica. Previously, in dried dichloromethane extracts of flowering aerial plant parts, α-amyrin and α-amyrin acetate, as well as β-amyrin, β-amyrin acetate and/or lupeol acetate were identified and quantified using GC-FID-MS. The aim was to evaluate chemosystematic significance of these five triterpenes using multivariate statistical methods (principal components analysis-PCA and non-metric multidimensional scaling-nMDS). Statistical analysis supported the current taxonomical classification of the investigated species, despite overlapping of certain groups. In PCA, all five triterpenes significantly contributed to the differences between the species. Moreover, lupeol acetate could be a significant chemosystematic marker for the most of principal species belonging to the section Pannosa, as well as for some hybridogenous species between H. gymnocephalum Gris. ex Pant. and the members from the other sections. 14th Symposium on the Flora of Southeastern Serbia and Neighboring Regions, Kladovo, 26th to 29th June 2022

The wood-sage (woodland germander) Teucrium scorodonia L. is widespread in European Atlantic coast and in the western part of Central Europe, with some isolated populations in Mediterranean. Recently discovered population on the territory of Serbia, in the valley of Štavica river (vicinity of Loznica), represents the most eastern enclave in the species range. Preliminary chemical analysis of aerial flowering parts and its dried ethanol and hydroethanol (70% v/v) extracts, and assessment of antioxidant potential of selected (hydroethanol) extract were performed. Using appropriate spectrophotometrical methods, total polyphenols in herb (3.48%, expressed as pyrogallol), ethanol and hydroethanol extracts (100.05 and 96.65 mg gallic acid equivalents - GAE/g), tannins in herb (0.96%, expressed as pyrogallol), ethanol and hydroethanol extracts (11.86 and 12.31 mg GAE/g), flavonoids in herb, ethanol and hydroethanol extracts (0.19%, 0.83% and 0.74%, expressed as hyperoside) were quantified. Also, in ethanol and hydroethanol extracts dihydroxycinnamic acid derivatives were determined (8.96% and 8.37%, expressed as acteoside). By LC-MS, in selected (hydroethanol) extract, four flavonoid and five phenylethanoid glycosides were detected; verbascoside (acteoside) was identified and quantified (4.42%) using external standard. This extract exhibited significant total antioxidant activity (1.37 mmol Fe2+/g) and moderate anti-DPPH potential (SC50=51.28 μg/mL). 14th Symposium on the Flora of Southeastern Serbia and Neighboring Regions, Kladovo, 26th to 29th June 2022

Significance of pharmacognostic studies on previously untested or scarcely tested wild plants is constantly rising, considering growing demand of pharmaceutical industry for new bioactive isolates, including essential oils and extracts. Accordingly, comprehensive research that included all nine taxa of genus Heracleum L. (Apiaceae) (three of which are endemic) from Serbia, Montenegro, North Macedonia and Slovenia was performed (60 samples of various plant organs collected from 14 localities in period 2009-2016). Plants of this genus (cow parsnips) got their Latin name after ancient hero Heracles, because of their habitus and use in folk medicine in treatment of various diseases. Also, some of them were the main ingredient of borscht, a traditional dish in Eastern European countries. They are interesting because of wide range of metabolites: from essential oil components, coumarins, flavonoids, phenolic acids, triterpenes and phytosterols, to some specific fatty acids. Analysis of these secondary and primary metabolites was performed using modern chromatographic and spectroscopic techniques (GC-FID-MS, LC-MS and NMR). In the case of 35 selected essential oils, antimicrobial, cytotoxic and/or antioxidant activities were also tested. It was revealed that studied plants represent potential new sources of compounds of importance not only for pharmaceutical, but also for cosmetic, food and chemical industries. For components of essential oils and furanocoumarins, by multivariate statistical analysis, chemosystematic significance was also established. Results of pharmacological investigations of essential oils indicated their importance as potential new medicinal plant raw materials, under condition of respecting established maximum daily intakes for those oils in which furanocoumarins were detected. Značaj farmakognozijskih studija na do sada neispitivanim ili nedovoljno ispitivanim samoniklim biljkama konstantno raste s obzirom na sve veće potrebe farmaceutske industrije za novim bioaktivnim izolatima, uključujući i etarska ulja i ekstrakte. U skladu sa tim, sprovedeno je opsežno istraživanje svih devet taksona roda Heracleum L. (Apiaceae) (od kojih su tri endemična), sa područja Srbije, Crne Gore, Severne Makedonije i Slovenije (60 uzoraka različitih biljnih organa sakupljanih sa 14 lokaliteta u periodu od 2009-2016. god.). Biljke ovog roda (mečje šape) dobile su latinski naziv po antičkom heroju Heraklu, kako zbog svog habitusa, tako i zbog primene u narodnoj medicini u lečenju različitih bolesti. Takođe, neke od njih bile su glavni sastojak boršča, tradicionalnog jela u zemljama istočne Evrope. Interesantne su zbog širokog spektra metabolita: od komponenti etarskih ulja, kumarina, flavonoida, fenolkarboksilnih kiselina, triterpena i fitosterola, sve do nekih specifičnih masnih kiselina. Ispitivanje navedenih sekundarnih i primarnih metabolita ovih taksona sprovedeno je uz korišćenje savremenih hromatografskih i spektroskopskih tehnika (GC- FID-MS, LC-MS i NMR). U slučaju 35 odabranih etarskih ulja ispitivane su i antimikrobna, citotoksična i/ili antioksidantna aktivnost. Utvrđeno je da su proučavane biljke potencijalno novi izvori jedinjenja od značaja ne samo za farmaceutsku, već i za kozmetičku, prehrambenu i hemijsku industriju. Za komponente etarskih ulja i furanokumarine ustanovljeno je multivarijantnom statističkom analizom da imaju i hemosistematski značaj. Rezultati ispitivanja farmakoloških aktivnosti etarskih ulja ukazali su na njihovu važnost kao potencijalno novih lekovitih biljnih sirovina, uz uslov poštovanja ustanovljenih maksimalnih dozvoljenih dnevnih unosa za ona etarska ulja u kojima su detektovani furanokumarini. VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beograd

In this work, composition and antioxidant activity of dry dichloromethane and methanol extracts of Heracleum ternatum Velen. (Apiaceae) leaves and flowers from Mt Durmitor (Montenegro) were investigated. Using LC-MS, five furanocoumarins were identified in leaf dichloromethane extract (accounting for 23.00 mg/g of dry extract) and eight in flower dichloromethane extract (60.47 mg/g). Dominant in leaf extract was heraclenin (11.64 mg/g), followed by imperatorin (5.90 mg/g), and in flower extract heraclenol (17.55 mg/g), followed by heraclenin (15.04 mg/g). LC-MS analysis revealed eight flavonol glycosides in leaf methanol extract (accounting for 16.18 mg/g of dry extract) and 11 in flower methanol extract (65.82 mg/g), as well as chlorogenic acid in both extracts (4.72 mg/g and trace, respectively). Among detected flavonoids, dominant was quercetin 7-O-rhamnosyl 3-O-glucoside (vincetoxicoside A; 7.95 and 16.77 mg/g), followed by biosides and triosides of kaempferol and methylquercetin. Content of total polyphenols, determined spectrophotometrically using Folin-Ciocalteu reagent, was 87.29 and 98.12 mg of gallic acid equivalents/g of dry methanol extracts. Among methanol extracts, polyphenol richer flower extract showed higher, spectrophotometrically determined, total antioxidant and anti-DPPH activities compared to leaf extract (FRAP=0.80 and 0.65 mmol Fe2+/g; DPPH SC50=83.12 and 90.95 μg/mL). Dichloromethane extracts showed lower anti-DPPH activity (SC50=860.58 and 515.76 μg/mL). 14th Symposium on the Flora of Southeastern Serbia and Neighboring Regions, Kladovo, 26th to 29th June 2022

INTRODUCTION: Cultivated parsnip (Pastinaca sativa subsp. sativa L., Apiaceae) root is a well-known vegetable, common ingredient of soups, stews, salads, casseroles etc. Besides, its leaves and young shoots can be added to soups and fruits are used as a condiment. Furthermore, young shoots of wild-growing parsnips, e.g. P. sativa subsp. urens (Req. ex Godr.) Čelak., are consumed pickled or in salads or soups and it is considered that their essential oil acts as a natural preservative. OBJECTIVES: To investigate and compare the antimicrobial activity of the essential oils obtained from the roots, leaves, stems, flowers and fruits of cultivated P. sativa subsp. sativa and wild-growing P. sativa subsp. urens collected in Serbia. Furthermore, the most active essential oil constituents (against the most susceptible microorganisms) were predicted in silico. METHOD / DESIGN: Minimum inhibitory concentrations (MICs) and minimum bactericidal/fungicidal concentrations (MBCs/MFCs) of the essential oils (isolated by hydrodistillation using Clevenger-type apparatus) were determined by microdilution method against three Gram-positive bacteria: Staphylococcus aureus ATCC 11632, Bacillus cereus clinical isolate and Listeria monocytogenes NCTC 7973, three Gram-negative bacteria: Escherichia coli ATCC 25922, Salmonella Typhimurium ATCC 13311 and Enterobacter cloacae ATCC 35030, three Candida standard strains: C. albicans ATCC 10231, C. tropicalis ATCC 750 and C. parapsilosis ATCC 22019, and three Candida isolates from oral cavity: C. albicans 475/15, C. krusei H1/16 and C. glabrata 4/6/15. Pharmacokinetic properties of the compounds present in at least one oil in the quantity ≥ 1% (determined by GC-FID and GC-MS) were initially evaluated using SwissADME web tool and molecular docking was performed using AutoDock Vina 1.1.2 (interactions were visualized using Discovery Studio Visualizer 2019). RESULTS: All the investigated essential oils of the two Pastinaca sativa subspecies were able to reduce the growth of different tested Candida strains (MIC range 0.25-2 mg/mL; MFC range 0.5-4 mg/mL). The most promising activity was observed for both root oils (MIC range 0.25-1 mg/mL; MFC range 0.5-2 mg/mL). Among investigated Candida strains, C. parapsilosis strain was the most sensitive to these essential oils (MIC range 0.25-1 mg/mL; MFC range 0.5-2 mg/mL). The antibacterial activity of the tested essential oils was lower compared to their anticandidal potential (MIC range 1-4 mg/mL; MBC range 2-8 mg/mL). Thirty compounds were present in at least one oil in the quantity ≥ 1%. Estimation of pharmacokinetic properties using SwissADME tool suggested that 23 of these compounds are inhibitors of some of the cytochrome P450 system isoenzymes. This fact led to assumption that they could also act against fungal sterol 14α-demethylase (CYP51), which is a common target of antifungal drugs (e.g., ketoconazole). Thus, the compounds (3D structures downloaded from PubChem) were docked to the active site of this enzyme (downloaded from Protein Data Bank, PDB code 5TZ1). The highest affinities were predicted for sesquiterpenes caryophyllene oxide, (E)-caryophyllene, germacrene D, α-copaene, β-bourbonene and δ-cadinene (free binding energies from -9.4 to -8.7 kcal/mol; ketoconazole -11.6 kcal/mol). These compounds were present in somewhat lower quantities in the essential oils (≤ 9.9%). For dominant compounds of the tested essential oils, e.g. myristicin, γ-palmitolactone and octyl butanoate a bit lower affinities were predicted (free binding energies from -7.3 to -5.8 kcal/mol). Tested compounds mostly docked near the heme of the enzyme and formed hydrophobic interactions with the amino acid residues of the active site. According to SwissADME tool, four of five most active compounds have low absorption from gastrointestinal tract and higher skin permeation value, while caryophyllene oxide and three dominant compounds have high absorption and lower skin permeation value (similarly to ketoconazole). CONCLUSIONS: Investigated parsnips represent sources of essential oils and compounds with anticandidal activity. The International Bioscience Conference and the 8th International PSU – UNS Bioscience Conference - IBSC2021, Novi Sad, 25th to 26th November 2021

Kritski origano, Origanum dictamnus L. (Lamiaceae), je endemična biljka grčkog ostrva Krit. Komitet za biljne lekovite proizvode Evropske agencije za lekove (EMA/HMPC) izdao je EU monografiju za tradicionalnu primenu herbe kritskog origana, Origani dictamni herba. Prema ovoj monografiji usitnjena herba se koristi oralno u obliku čaja (infuza) za ublažavanje kašlja povezanog sa prehladom i za simptomatsko ublažavanje blagog gastrointestinalnog diskomfora, kao i dermalno u obliku infuza ili dekokta za tretman manjih inflamacija kože i modrica. U Evropskoj farmakopeji (Ph. Eur.) za sada ne postoje monografije za ovu biljnu drogu i njeno etarsko ulje. Cilj ovog rada bio je ispitivanje hemijskog sastava etarskog ulja izolovanog iz komercijalnog uzorka herbe kritskog origana poreklom iz Grčke, kao i procena njegove antiradikalske aktivnosti. Etarsko ulje je izolovano iz herbe destilacijom vodenom parom u aparaturi po Clevenger-u, prema postupku Ph. Eur. Prinos etarskog ulja iznosio je 1,51% V/m, tj. 1,46% m/m. Hemijski sastav etarskog ulja analiziran je gasnom hromatografijom sa FID i MS detekcijom (GC-FID i GC-MS). Identifikovane su 42 komponente, koje su činile 94,6% sastava etarskog ulja. Procentualno najveći udeo u etarskom ulju činili su oksidovani monoterpeni (58,8%; 9 jedinjenja), a sledili su monoterpenski ugljovodonici (27,4%; 12 jedinjenja), seskviterpenski ugljovodonici (5,6%; 12 jedinjenja) i oksidovani seskviterpeni (2,3%; 6 jedinjenja). Glavna komponenta etarskog ulja bio je monoterpenski fenol karvakrol (51,4%), dok su po količini sledili monoterpenski ugljovodonici p-cimen (14,3%) i γ-terpinen (9,3%). Dobijeni rezultati bili su u skladu sa literaturnim podacima, pri čemu je u slučaju uzorka analiziranog u ovom radu, prinos etarskog ulja bio veći od prethodno publikovanih vrednosti (1, 2). Antiradikalska aktivnost izolovanog etarskog ulja ispitana je spektrofotometrijskim DPPH testom. Stepen antiradikalske aktivnosti je rastao linearno sa povećanjem koncentracije etarskog ulja, pri čemu je koncentracija u kojoj je etarsko ulje inhibiralo 50% DPPH radikala (SC50) iznosila 141,77 μg/mL. Značajan anti-DPPH potencijal etarskog ulja herbe kritskog origana može se barem delom objasniti velikim udelom karvakrola, čiji je antiradikalski potencijal ranije pokazan (3). Rezultati sprovedenih hemijskih ispitivanja mogu biti od značaja za buduće definisanje parametara farmaceutskog kvaliteta droge Origani dictamni herba i njenog etarskog ulja. Cretan dittany, Origanum dictamnus L. (Lamiaceae), is an endemic species from the Greek island of Crete. The Committee on Herbal Medicinal Products of the European Medicines Agency (EMA/HMPC) adopted EU monograph on traditional use of Cretan dittany herb, Origani dictamni herba. According to this monograph, comminuted herb is used orally as herbal tea (infusion) for the relief of cough associated with cold and for mild gastrointestinal disorders, as well as cutaneously as infusion or decoction for the treatment of minor skin inflammations and bruises. European Pharmacopoeia (Ph. Eur.) monographs for this herbal drug and its essential oil do not exist for now. The aim of this work was to investigate chemical composition of the essential oil obtained from a commercial sample of Cretan dittany herb and to evaluate its antiradical activity. The essential oil was isolated from the herb by hydrodistillation in a Clevenger‐type apparatus, according to procedure given in Ph. Eur. The yield was 1.51% V/w, i.e. 1.46% w/w. Chemical composition of the essential oil was analyzed by gas chromatography with FID and MS detection (GC-FID and GC-MS). A total of 42 components were identified, comprising 94.6% of the composition of the essential oil. The dominant were oxygenated monoterpenes (58.8%; 9 compounds), followed by monoterpene hydrocarbons (27.4%; 12 compounds), sesquiterpene hydrocarbons (5.6%; 12 compounds) and oxygenated sesquiterpenes (2.3%; 6 compounds). The main component of the essential oil was monoterpene phenol carvacrol (51.4%), followed by monoterpene hydrocarbons p-cymene (14.3%) and γ-terpinene (9.3%). Obtained results were in accordance with the literature data; it should be noted that in the case of the sample analyzed in this work, the essential oil yield was higher than previously published values (1, 2). The antiradical activity of the essential oil was investigated by spectrophotometric DPPH test. The percentage of antiradical activity increased linearly with the increase of essential oil concentration. The concentration at which the essential oil inhibited 50% DPPH radicals (SC50) was 141.77 μg/mL. Significant anti-DPPH potential of the essential oil of Cretan dittany herb can be at least partly explained by the presence of high quantity of carvacrol, which antiradical potential was demonstrated previously (3). The results of performed chemical investigations may be important for defining pharmaceutical quality parameters of the herbal drug Origani dictamni herba and its essential oil in the future. Drugi naučni simpozijum Saveza farmaceutskih udruženja Srbije sa međunarodnim učešćem, 28. 10. 2021. Beograd

Velika mečja šapa, Heracleum sibiricum L. (Apiaceae), autohtona je u severnoj, istočnoj i centralnoj Evropi, kao i na Balkanskom poluostrvu i u Sibiru (1). Infuz pripremljen od nadzemnih delova ove biljke se u narodnoj medicini na Balkanskom poluostrvu koristi kao aperitiv i za tretman dijareje i drugih gastrointestinalnih tegoba (2). Prethodna ispitivanja H. sibiricum nisu uključivala analizu polifenola, dok su furanokumarini do sada analizirani u korenu, plodovima i listovima (1, 3). Cilj ovog rada bio je ispitivanje hemijskog sastava i antioksidantne aktivnosti metanolnih i dihlormetanskih ekstrakata listova i cvasti ove biljke. Osušen biljni materijal (sakupljen na Staroj Planini u Srbiji u julu 2014. god.) ekstrahovan je sukcesivno dihlormetanom i metanolom (u oba slučaja bimaceracijom 72+48 h, sa odnosom droga rastvarač 1:10 m/V). Rastvarači su uklonjeni pod sniženim pritiskom. Hemijski sastav dobijenih suvih ekstrakata analiziran je LC-MS metodama. Dodatno, u slučaju metanolnih ekstrakata spektrofotometrijski je utvrđen sadržaj ukupnih polifenola pomoću Folin‐Ciocalteu reagensa. Antioksidantna aktivnost metanolnih ekstrakata ispitana je spektrofotometrijskim FRAP i DPPH testovima, a dihlormetanskih ekstrakata DPPH testom. LC-MS analiza pokazala je da su suvi metanolni ekstrakti listova i cvasti bili vrlo slični. U njima su određene značajne količine heterozida kvercetina i kemferola (154,30 i 144,20 mg/g suvog ekstrakta). Najzastupljeniji su bili vincetoksikozid A (kvercetin 7-O-ramnozil 3-O-glukozid; 26,83 i 29,61 mg/g), kemferitrin (kemferol 3,7-di-O-ramnozid; 33,62 i 34,68 mg/g) i jedan kvercetin dezoksiheksozil dezoksiheksozid (69,06 i 79,91 mg/g). U ekstraktu listova detektovano je još šest, a u ekstraktu cvasti još pet heterozida ovih flavonola. Takođe, u oba ekstrakta identifikovana je hlorogenska kiselina (19,31 i 4,12 mg/g). Spektrofotometrijski utvrđen sadržaj ukupnih polifenola iznosio je 126,08 i 125,33 mg ekvivalenata galne kiseline/g. U suvom dihlormetanskom ekstraktu listova LC-MS metodom detektovane su male količine furanokumarina izopimpinelina i bergaptena (ukupno 0,22 mg/g suvog ekstrakta), čije je prisustvo i ranije utvrđeno (3). U ekstraktu cvasti, pored dva navedena, identifikovano je još šest furanokumarina i svi su takođe bili prisutni u malim količinama (ukupno 20,07 mg/g); najzastupljeniji je bio bergapten (4,84 mg/g), a po količini ga je sledio biakangelikol (3,29 mg/g). Ukupna antioksidantna aktivnost suvih metanolnih ekstrakata listova i cvasti (FRAP vrednosti 1,08 i 0,94 mmol Fe2+/g), kao i antiradikalska aktivnost (DPPH SC50 vrednosti 64,10 i 64,77 μg/mL) bile su u korelaciji sa sadržajem ukupnih polifenola. Anti-DPPH aktivnost suvih dihlormetanskih ekstrakata bila je očekivano niža (SC50 vrednosti 636,19 i 790,20 μg/mL). Može se zaključiti da listovi i cvasti H. sibiricum predstavljaju nove izvore jedinjenja sa poznatom antioksidantnom aktivnošću. Siberian cow parsnip, Heracleum sibiricum L. (Apiaceae), is autochthonous to northern, eastern and central Europe, Balkan Peninsula and Siberia (1). Infusion prepared from aerial parts of this plant is used in folk medicine on Balkan Peninsula as aperitif and to treat diarrhea and other gastrointestinal complaints (2). Previous studies of H. sibiricum have not included polyphenol analysis, while furanocoumarins were analyzed in roots, fruits and leaves (1, 3). In this work, chemical composition and antioxidant activity of methanol and dichloromethane extracts obtained from leaves and flowers of this plant were investigated. Dried plant material, collected on Mts Stara Planina (Serbia) in July 2014, was extracted successively with dichloromethane and methanol (in both cases by bimaceration 72+48 h; drug/solvent=1:10 w/V). Solvents were removed under reduced pressure. The composition of obtained dry extracts was analyzed by LC-MS methods. Additionally, for methanol extracts, total phenolics contents were determined spectrophotometrically using Folin-Ciocalteu reagent. Antioxidant activity of methanol extracts was tested by spectrophotometric FRAP and DPPH assays, and of dichloromethane extracts by DPPH assay. LC-MS analysis showed that investigated leaf and flower dry methanol extracts were very similar. In both extracts, significant amounts of quercetin and kaempferol glycosides (154.30 and 144.20 mg/g of dry extract) were determined. Dominant were vincetoxicoside A (quercetin 7-O-rhamnosyl 3-O-glucoside; 26.83 and 29.61 mg/g), kaempferitrin (kaempferol 3,7-di-O-rhamnoside; 33.62 and 34.68 mg/g) and one quercetin deoxyhexosyl deoxyhexoside (69.06 and 79.91 mg/g). Six more glycosides of these flavonols were detected in the leaf extract and five more in the flower extract. Also, chlorogenic acid (19.31 and 4.12 mg/g) was identified in both extracts. Spectrophotometrically determined total phenolics contents were 126.08 and 125.33 mg gallic acid equivalents/g. In the leaf dry dichloromethane extract, by LC-MS method, small amounts of furanocoumarins isopimpinellin and bergapten were detected (totally 0.22 mg/g of dry extract). Their presence was also previously shown (3). In the flower extract, besides these two, six more furanocoumarins were identified; all present in small amounts (totally 20.07 mg/g). The most abundant was bergapten (4.84 mg/g), followed by byakangelicol (3.29 mg/g). Total antioxidant (FRAP values 1.08 and 0.94 mmol Fe2+/g) and antiradical activities (DPPH SC50 values 64.10 and 64.77 μg/mL) of the leaf and flower dry methanol extracts were correlated with their total phenolics contents. Anti-DPPH activities of the dry dichloromethane extracts were expectedly lower (SC50 values 636.19 and 790.20 μg/mL). Heracleum sibiricum leaves and flowers represent new sources of compounds with known antioxidant activity. Drugi naučni simpozijum Saveza farmaceutskih udruženja Srbije sa međunarodnim učešćem, 28. 10. 2021. Beograd

According to current EU monograph of the Committee on Herbal Medicinal Products of European Medicines Agency (EMA/HMPC) for ginkgo leaf, Ginkgonis folium (Ginkgo biloba, Ginkgoaceae), herbal medicines prepared from refined and quantified dry extract of ginkgo leaf are used for the improvement of (age-associated) cognitive impairment and of quality of life in mild dementia. It was demonstrated in clinical trials that long-term use of the extract is effective mainly in patients more than 50 years old. The precise mode of action is not known. According to Ph. Eur., the extract should contain 22.0-27.0% of flavonoids, 2.8-3.4% of ginkgolides A, B and C, and 2.6-3.2% of bilobalide, which are active constituents, and maximum 5 ppm of ginkgolic acids, which are potential allergens. These herbal medicines are characterized by corresponding posology, duration of use, contraindications, special warnings, and precautions for use, interactions and possible undesirable reactions. Prema aktuelnoj monografiji EU Komiteta za biljne lekovite proizvode Evropske agencije za lekove (EMA/HMPC) za list ginka, Ginkgonis folium (Ginkgo biloba, Ginkgoaceae), biljni lekovi na bazi suvog prečišćenog kvantifikovanog ekstrakta lista ginka koriste se za ublažavanje (sa starošću povezanog) kognitivnog oštećenja i za poboljšanje kvaliteta života kod blage demencije. U kliničkim studijama je pokazana efikasnost hronične primene ekstrakta prevashodno kod pacijenata starosti preko 50 godina. Precizan mehanizam delovanja nije poznat. Prema Ph. Eur., ekstrakt treba da sadrži 22,0-27,0% flavonoida, 2,8-3,4% ginkolida A, B i C i 2,6-3,2% bilobalida, koji spadaju u aktivne sastojke, kao i najviše 5 ppm ginkolnih kiselina koje su potencijalni alergeni. Ove biljne lekove karakterišu odgovarajući režim doziranja i dužina primene, kontraindikacije, posebna upozorenja i mere opreza, interakcije i moguće neželjene reakcije.

IntroductionHieracium s. str. represents one of the largest and most complex genera of flowering plants. As molecular genetics seems unlikely to disentangle intricate relationships within this reticulate species complex, analysis of flavonoids and phenolic acids, known as good chemosystematic markers, promise to be more reliable. Data about pharmacological activity of Hieracium species are scarce. ObjectiveEvaluation of the chemosystematic significance of flavonoids and phenolic acids of methanol extracts of aerial flowering parts of 28 Hieracium species from the Balkans. Additionally, investigation of antioxidant potentials of the extracts. MethodsComparative qualitative and quantitative analysis of flavonoids and phenolic acids was performed by LC-MS. Multivariate statistical data analysis included non-metric multidimensional scaling (nMDS), unweighted pair-group arithmetic averages (UPGMA) and principal component analysis (PCA). Antioxidant activity was evaluated using three colorimetric tests. ResultsDominant phenolics in almost all species were luteolin type flavonoids, followed by phenolic acids. Although the investigated Hieracium species share many compounds, the current classification of the genus was supported by nMDS and UPGMA analyses with a good resolution to the group level. Hieracium naegelianum was clearly separated from the other investigated species. Spatial and ecological distances of the samples were likely to influence unexpected differentiation of some groups within H. sect. Pannosa. The vast majority of dominant compounds significantly contributed to differences between taxa. The antioxidant potential of the extracts was satisfactory and in accordance with their phenolics composition. ConclusionsComparative LC-MS analysis demonstrated that flavonoids and phenolic acids are good indicators of chemosystematic relationships within Hieracium, particularly between non-hybrid species and groups from the same location. Copyright

Although some widespread, native cow parsnips (Heracleum L. spp., Apiaceae) had broad medicinal and culinary applications throughout history, the knowledge about their volatile constituents is insufficient. This work investigates the composition and bioactivities of H. sphondylium L. (HSPH), H. sibiricum L. (HSIB) and H. montanum Schleich. ex Gaudin (HMON) essential oils. The composition was tested by GC and GC-MS. (Z)-β-Ocimene was the most abundant in HSPH (28.9%) and HMON (20.4%) root oils, while in HSIB root oil, β-pinene (26.2%), methyl eugenol (22.3%) and elemicin (25.6%) prevailed. Leaf and flower oils were dominated by various sesquiterpenes (germacrene D, β-sesquiphellandrene, (E)-β-farnesene and/or (E)-caryophyllene) and/or phenylpropanoids (apiole, methyl eugenol, elemicin and/or (Z)-isoelemicin). Octyl acetate (57.5–67.1%) was the main constituent of all fruit oils. The antimicrobial activity was screened by a microdilution method against eight bacteria and eight fungi. The strongest antimicrobial effect, in several cases better than the activity of antibiotics, was shown by HSPH (MICs = 0.12–3.30 mg mL−1) and HMON (MICs = 0.10–1.30 mg mL−1) flower oils against bacteria, and HSIB fruit oil against fungi (MICs = 0.15–0.40 mg mL−1). The MTT test revealed that the oils were not or weakly cytotoxic against human malignant HeLa, LS174 and/or A549 cells (except HSPH root oil; IC50 = 5.72–24.31 μg mL−1) and that tested oils were not toxic against human normal MRC-5 cells (at 200.00 μg mL−1). Significant activity observed against microorganisms that are the common cause of foodborne diseases, food contamination and/or hospital-acquired infections justifies certain traditional uses of the investigated plants and represents a good basis for further research of these Heracleum oils.