The chemical composition and bioactivities of dry methanol extracts from roots, leaves and fruits of Prangos trifida (Apiaceae), collected in Serbia, were investigated. LC-DAD-QTOF-MS/MS analysis revealed 30 compounds, primarily polyphenols and coumarins. The root and leaf extracts were rich in chlorogenic and/or 3,5-di-O-caffeoylquinic acid (18.20-26.14 mg/g extract), and the fruit extract in oxypeucedanin hydrate and prantschimgin (46.50 and 71.64 mg/g). The leaf extract exhibited the highest total phenolic content (62.86 mg quercetin equivalents/g), total antioxidant activity (FRAP = 0.71 mmol Fe2+/g) and DPPH radical scavenging ability (44.08 mg quercetin equivalents/g). Antimicrobial activity testing (11 bacteria and three yeasts, microdilution method) showed that the most active were the root and leaf extracts against Micrococcus luteus, Staphylococcus aureus, S. epidermidis and Candida albicans (MIC = 0.625-5 mg/mL). The fruit extract showed the strongest cytotoxicity against tested stomach, colon and hypopharynx cancer cell lines (MTT test), with the highest selectivity toward hypopharynx cancer FaDu cells (selectivity index 4.71; determined in relation to non-cancerous VERO cells). No antiviral activity against herpesvirus type 1 was found. The results indicate that P. trifida represents a promising source of polyphenols and coumarins, notably expanding current knowledge on its chemical composition and supporting its potential relevance for pharmaceutical and food industry applications.

Background/Objectives: Lamiaceae species are valuable sources of bioactive natural products, often associated with anti-infective properties. This study investigated chemical composition and bioactivities of dry hydroethanolic extracts and essential oils from Micromeria nervosa (Desf.) Benth. aerial parts from two localities. Methods: Extracts and essential oils were analyzed using LC-DAD-QTOF-MS/MS and GC-FID/MS, respectively. Antimicrobial activity was assessed against 14 strains (microdilution method), and antiviral activity against three viruses by determining cytopathic effects, viral titers (end-point dilution assay) and viral loads (qPCR/RT-qPCR). Cytotoxicity was evaluated on three cancer cell lines (MTT assay) and antioxidant potential using three colorimetric tests. Composition-activity correlation was statistically analyzed; in silico molecular docking/dynamics simulations were performed. Results: Thirty-five compounds were annotated in extracts, including 30 reported for the first time in this species, with rosmarinic acid as the main component. Essential oils contained 31 constituents, dominated by carvacrol. Newly detected phenolics included lithospermic acid and several salvianolic and clinopodic acids. Extracts and oils exhibited notable antibacterial activity, especially against five Gram-positive strains (MIC = 0.313-2.5 mg/mL), and oils showed marked anticandidal effects (MIC = 0.313-0.625 mg/mL) and enhanced cytotoxicity against colon, gastric and hypopharyngeal cancer cells (selectivity indices ≥ 1.66). Extracts displayed potent antiviral activity against human herpesvirus 1 (HHV-1) and adenovirus Ad5, reducing cytopathic effects and viral titers, with qPCR revealing decreased HHV-1 load. In silico analysis suggested HHV-1 glycoprotein D binding. Extracts also showed strong antioxidant potential. Conclusions: These findings demonstrate that M. nervosa is a rich source of compounds with antimicrobial/antiviral, cytotoxic and antioxidant activities, warranting further research.

The objective of this study was to investigate chemical composition, antimicrobial and cytotoxic activities of dried methanol and dichloromethane extracts of the flowering tops (herb) of yarrow, Achillea millefolium L. (Asteraceae) collected from natural habitat in Serbia. This plant, predominantly distributed in the northern hemisphere, is widely used in folk medicine, but also as an important crop for pharmaceutical and cosmetic industries, primarily due to its appetite-stimulating, choleretic, gastroprotective, spasmolytic, analgesic, antiinflammatory, astringent, antioxidant and/or antimicrobial activities. GC-FID and GC-MS analysis of unsaponifiable fraction of dichloromethane extract showed that the most abundant compounds were alpha-amyrin, (1-sitosterol, one taraxasterol isomer and (1-amyrin. LC-MS analysis of methanol extract revealed 28 phenolic compounds, predominantly caffeoylquinic acids, as well as apigenin and luteolin type flavonoids. Further, microdilution assay showed that both extracts possess moderate antimicrobial activity against Listeria monocytogenes (both MICs 1.25 mg mL-1) and Candida albicans (MICs 2.5 and 0.3125 mg mL-1, respectively). Investigation of the antibiofilm effects showed that methanol extract significantly inhibited biofilm formation of L. monocytogenes, but did not disrupt L. monocytogenes mature biofilm. Additionally, the reduction in L. monocytogenes motility was induced by methanol extract in dose dependent manner. Furthermore, MTT assay showed that cancer A549 and HCT 116 cell lines were more sensitive to the action of dichloromethane extract (both IC50 values 0.7 mg mL-1). Both extracts exhibited greater toxicity to cancer than to normal MRC-5 cells, as well as selective antimicrobial effect against L. monocytogenes and C. albicans. The results of bioactivity testing of chemically characterized A. millefolium extracts represent good basis for further investigations of this crop with the aim of widening its industrial application and justifying cultivation of the investigated yarrow population.

Plants of the genus Prangos are intensively investigated as potential new sources of bioactive isolated products. In this work, the chemical composition of volatile constituents (essential oils and headspace volatiles) and dichloromethane extracts, as well as antimicrobial and antibiofilm activities of essential oils and MFDEs (methanol fractions of dichloromethane extracts) of Prangos trifida from Serbia, were investigated. Volatiles of roots, leaves, stems and fruits, and fatty acids and phytosterols in dichloromethane extracts of roots and fruits were analyzed by GC-FID-MS, whereas coumarins in MFDEs by LC–MS and some isolated coumarins by 1H-NMR. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations/minimum fungicidal concentrations (MBCs/MFCs) of essential oils and MFDEs were determined against 13 microorganisms. Antibiofilm activity was assessed against four microorganisms. Additionally, congo red and ergosterol binding assays were conducted to elucidate selected mechanisms of antibiofilm action in the case of Candida albicans. Total of 52 volatile constituents, 16 fatty acids, eight phytosterols and 10 coumarins were identified. Essential oils demonstrated significant activity, surpassing that of commercial food preservatives, against six tested molds from the Aspergillus, Penicillium and Trichoderma genera, as well as against bacteria Staphylococcus aureus and Bacillus cereus. Most of the oils strongly inhibited the formation of biofilms by S. aureus, Listeria monocytogenes and Escherichia coli. MFDEs exhibited noteworthy effects against B. cereus and the tested Aspergillus species, particularly A. niger, and significantly inhibited C. albicans biofilm formation. This inhibition was linked to a marked reduction in exopolysaccharide production, while antifungal mechanisms associated with ergosterol remained unaffected.

The aim of this work was to investigate the composition of essential oils isolated from fruits of Peucedanum longifolium, and Rhizomatophora aegopodioides (a species which was previously placed in the genus Peucedanum), as well as to compare the obtained results to those available for other previously investigated related species (including taxa which are also, according to some authors, excluded from the genus Peucedanum). Essential oils were obtained via hydrodistillation in a Clevenger-type apparatus and their composition was analyzed using GC-FID and GC-MS. To compare these data to those of previously investigated taxa, a chemometric approach was applied; the data were analyzed using multivariate statistical methods: non-metric multidimensional scaling (nMDS) and hierarchical cluster analysis. The most abundant in P. longifolium essential oil were monoterpenes (79.7%), mostly α-phellandrene (26.2%), β-phellandrene + limonene (21.0%) and myrcene (9.5%), followed by sesquiterpenes (18.3%), mostly germacrene B (9.5%). On the other hand, dominant in R. aegopodioides essential oil were non-terpenic aliphatic hydrocarbons (46.1%), mainly n-undecane (16.5%) and n-nonane (11.3%). In addition, this essential oil also contained a notable quantity of sesquiterpenes (25.1%), with (E)-sesquilavandulol being the most abundant (10.0%). The results of multivariate statistics revealed a clear separation of the essential oil composition of R. aegopodioides and P. longifolium, as well as of P. longifolium and P. officinale. The clustering of the samples of most of the taxa that do not belong to the Peucedanum in the narrow sense (sensu stricto) was also observed, which is in accordance with their recent inclusion in separate genera.

Composition and antimicrobial activity of root, leaf, stem, flower and fruit essential oils from cultivated Pastinaca sativa subsp. sativa, and its two wild-growing relatives P. sativa subsp. urens and P. hirsuta (Apiaceae) were investigated. Twenty-nine hydrodistilled essential oils of plants from different localities and/or years were analysed by GC-FID and GC/MS. Dominant in root oils was myristicin (P. sativa) or apiole (P. hirsuta), in leaf and stem oils myristicin (cultivated plants) or gamma-palmitolactone (wild-growing plants) and in flower and fruit oils aliphatic esters. Multivariate statistics (PCA, nMDS, UPGMA clustering) generally revealed separation of oils of investigated Pastinaca taxa and demonstrated their chemosystematic significance. One oil per each organ of all three plants (fifteen in total) was tested using microdilution method for activity against Candida tropicalis, C. parapsilosis, C. krusei, C. glabrata, C. albicans, Staphylococcus aureus, Bacillus cereus, Listeria monocytogenes, Escherichia coli, Salmonella Typhimurium and Enterobacter cloacae; MIC = 0.25-8 mg/mL, MBC(MFC) = 0.5-16 mg/mL.

The composition and anticholinesterase activity of the dried MeOH extracts of Hieracium scheppigianum and H. naegelianum underground parts (rhizomes and roots), as well as the anticholinesterase activity of the dried, previously chemically characterised MeOH extracts of the flowering aerial parts of these two and 26 other Hieracium species in the strict sense (s. str.), were investigated. Furthermore, the anticholinesterase activity of 12 selected secondary metabolites of these extracts was evaluated. Using semi-preparative LC-MS, five caffeoylquinic acids and the sesquiterpene lactone crepiside E were isolated from H. scheppigianum underground parts extract. All these compounds were also identified in the underground parts extract of H. naegelianum. Quantitative LC-MS analysis showed that the analysed underground parts extracts were rich in both caffeoylquinic acids (139.77 and 156.62 mg/g of extract, respectively) and crepiside E (126.88 and 116.58 mg/g). In the Ellman method, the tested extracts showed an interesting anti-AChE and/or anti-BChE activity (IC50 =0.56-1.58 mg/mL), which can be explained, at least partially, by the presence of some of their constituents. Among the metabolites tested, the best activity was revealed for the flavonoids apigenin, luteolin and diosmetin, and the sesquiterpene lactone 8-epiixerisamine A (IC50 =68.09-299.37 μM).

This paper represents the fourth part of the inventory of the flora of Serbia (Niketić et al. 2018, 2020, 2021), which contains nomenclatural, taxonomic and floristic notes related to taxa from the Magnoliopsida group. At the same time, this contribution is the basis for publication of subsequent volumes of the An annotated checklist of vascular flora of Serbia in order to supplement the data on vascular plants in our country.

Aim of this work was the chemical profiling of six Stachys taxa from Balkan Peninsula: S. recta subsp. nitens, S. atherocalyx, S. beckeana, S. zepcensis (all four belonging to S. recta group), S. alpina subsp. dinarica and S. plumosa. Dry ethanol extracts of aerial flowering plant parts were qualitatively and quantitatively characterized for their phenolic composition using the HPLC-DAD-ESI-MS technique. In total, 44 phenolic compounds were detected, belonging to three main classes of constituents: phenolic acids, phenylethanol glycosides (PhGs) and flavonoids. Chlorogenic acid was the main phenolic acid in all samples. Regarding PhGs, in all investigated extracts acteoside and lavandulifolioside were present. In S. recta group, extracts of S. atherocalyx, S. recta subsp. nitens and S. beckeana showed similar pattern considering phenylethanol glycosides, while S. zepcensis contained only the two aforementioned compounds. PhGs profile of S. plumosa extract comprised the same compounds from S. recta group, but it was also characterized by the presence of few high-molecular compounds. PhGs composition of S. alpina subsp. dinarica extract was somewhat similar with those of S. recta group and also with S. plumosa, but still specific by the presence of echinacoside and leucosceptoside A. The main flavonoids in extracts of Stachys recta group as well as in S. alpina subsp. dinarica extract were 8-hydroxyflavone glycosides (7-O-allosylglucosides of isoscutellarein and hypolaetin and their 4′-methyl derivatives). In addition, in the extract of S. alpina subsp. dinarica two luteolin-7-O-hexosides were also present. On the other hand, S. plumosa extract differed considerably containing mainly derivatives of chrysoeriol and apigenin, while 8-hydroxyflavones (isoscutellarein derivatives) were detected in traces. The results confirmed the importance of flavone 7-O-allosylglucosides at the sectional level of Stachys genus.

Fast inverted, oil-in-water (o/w) emulsions, also known as SWitch-Oil-Phase (SWOP) emulsions, express the performances of both o/w and water-in-oil (w/o) emulsions during application to the skin, favoring their use as cosmetic carriers in sunscreen products. The objective of this study was to investigate the antioxidant potential (by 2 different methods) and the ultraviolet (UV) absorption ability of SWOP emulsion (S) with incorporated plant-based antioxidants dihydroquercetin (DHQ) and β-carotene (βC), using quercetin (Q) in a reference emulsion, in addition to the evaluation of their physicochemical properties and stability. A new biochemical extracellular model for in vitro assessment of antioxidative properties for the SWOP emulsions (S, SQ, SDHQ, and SDHQβC) was developed and compared with the results of 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay. The analyses were performed at 20 °C and 37 °C, and oxidative stress parameters were monitored and statistically analyzed. The sun protection factor (SPF) of the samples was determined in vitro. Q and DHQ incorporated into the SWOP emulsion exhibited a strong DPPH radical scavenging ability. Neither incorporated nor pure βC showed DPPH radical scavenging ability at the tested concentrations. Contrary to that, in the bioenvironment conditions, SDHQβC showed minor antioxidative effects increase and also a significant decrease in exogenous pro-oxidative effects, caused by pro-oxidant, when compared to SDHQ. The obtained SPFs of SDHQβC, SDHQ, and SQ were 5.19, 4.65, and 3.35, respectively. The physicochemical stability of the emulsions was satisfactory during 1 month storage. The presented results demonstrated that the SWOP emulsion is a suitable carrier for antioxidants with a photoprotective ability. The novel biochemical approach could be used in addition to DPPH assay with several advantages, relevant for the testing of antioxidant activity of potential active ingredients in cosmetic products.

Fast inverted, oil-in-water (o/w) emulsions, also known as SWitch-Oil-Phase (SWOP) emulsions, express the performances of both o/w and water-in-oil (w/o) emulsions during application to the skin, favoring their use as cosmetic carriers in sunscreen products. The objective of this study was to investigate the antioxidant potential (by 2 different methods) and the ultraviolet (UV) absorption ability of SWOP emulsion (S) with incorporated plant-based antioxidants dihydroquercetin (DHQ) and β-carotene (βC), using quercetin (Q) in a reference emulsion, in addition to the evaluation of their physicochemical properties and stability. A new biochemical extracellular model for in vitro assessment of antioxidative properties for the SWOP emulsions (S, S Q , S DHQ , and S DHQβC ) was developed and compared with the results of 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay. The analyses were performed at 20 °C and 37 °C, and oxidative stress parameters were monitored and statistically analyzed. The sun protection factor (SPF) of the samples was determined in vitro . Q and DHQ incorporated into the SWOP emulsion exhibited a strong DPPH radical scavenging ability. Neither incorporated nor pure βC showed DPPH radical scavenging ability at the tested concentrations. Contrary to that, in the bioenvironment conditions, S DHQβC showed minor antioxidative effects increase and also a significant decrease in exogenous pro-oxidative effects, caused by pro-oxidant, when compared to S DHQ . The obtained SPFs of S DHQβC , S DHQ , and S Q were 5.19, 4.65, and 3.35, respectively. The physicochemical stability of the emulsions was satisfactory during 1 month storage. The presented results demonstrated that the SWOP emulsion is a suitable carrier for antioxidants with a photoprotective ability. The novel biochemical approach could be used in addition to DPPH assay with several advantages, relevant for the testing of antioxidant activity of potential active ingredients in cosmetic products. Keywords SWOP emulsion , plant-based antioxidants , flavonoids , antioxidant activity , bioenvironment , photoprotection , statistics

This work is focused on nine Heracleum taxa (Apiaceae): H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii and H. verticillatum, belonging to the H. sphondylium group, and H. orphanidis, all from sect. Heracleum, as well as H. austriacum subsp. siifolium from sect. Wendia. The aim was to isolate the headspace (HS) fractions from 17 fruit samples (collected from all nine Heracleum taxa) and 13 root samples (collected from all eight taxa belonging to sect. Heracleum), to investigate their composition and chemosystematic significance, as well as to compare these results with those previously obtained for the essential oils isolated by hydrodistillation from the same fruit and root samples. The HS fractions were isolated using an automatic static HS sampler and analysed by GC-FID and GC-MS. The fruit HS fractions were dominated by octyl acetate and/or ?-pinene (the taxa from the sect. Heracleum), or n-octanol (H. austriacum), whereas the root HS fractions mainly contained ?-pinene and/or (Z)-?-ocimene (the taxa from the H. sphondylium group), or n-nonane (H. orphanidis). The chemosystematic significance was evaluated using multivariate statistical methods: principal component analysis (PCA), non-metric multidimensional scaling (nMDS) and agglomerative hierarchical clustering based on the unweighted pair-group arithmetic average (UPGMA) algorithm. As in the case of previously investigated essential oils, the statistical analysis of the fruit and root HS fractions generally resulted in the grouping of the investigated H. sphondylium group representatives, and within this group, the grouping of morphologically related H. sphondylium and H. montanum and the isolated position of H. verticillatum. The statistical analysis of the root HS fractions resulted in better separation of the taxa (even compared to previous analysis of the essential oils), i.e. isolated positions of H. ternatum and H. pyrenaicum were observed.

Dry MeOH extract of Ferula heuffelii (Apiaceae) underground parts was tested for spasmolytic, gastroprotective and antioxidant activities. HPLC analysis revealed that chlorogenic acid (CGA; 34.6 mg/g) was its main constituent. Extract in vitro exhibited notable total antioxidant activity (FRAP value=1.0 μmol Fe2+ /mg), and scavenging of DPPH (SC50 =62.5 μg/ml) and • OH radicals (49.5 % at 20 μg/ml in 2-deoxyribose assay). In vitro on isolated rat ileum, extract exhibited significant spasmolytic activity, i. e., it showed 124.6 % of maximal atropine effect on spontaneous contractions (at 100 μg/ml), and reduced spasmogenic effect of KCl (80 mm) to 44.4 % (at 60 μg/ml) and of highest applied concentration of ACh to 26.3 % (at 120 μg/ml). In parallel experiments, spasmolytic effect of CGA was also demonstrated. In acute EtOH-induced gastric ulceration model in rats, extract (100 mg/kg p.o.) showed significant gastroprotective effect (gastric damage score 0.50), similar to ranitidine (20 mg/kg p.o.). Obtained results showed that tested F. heuffelii polar extract represents new herbal preparation with potential use against some gastrointestinal complaints.

Antimicrobial and cytotoxic activities were tested for dried MeOH extracts of Hieracium calophyllum (CAL), H. coloriscapum (COL), H. pseudoschenkii (PSE), H. valdepilosum (VAL) and H. glabratum (GLA) herbs (flowering aerial parts), their 2 sesquiterpene lactones (SLs) 8-epiixerisamine A and crepiside E, and dried CH2 Cl2 extract of H. scheppigianum (SCH) herb. In microdilution test, extracts showed activity on all tested microorganisms (8 bacteria, 10 fungi). The best effect was exhibited by SCH and CAL on Salmonella Typhimurium (MIC=1.7-2.5 mg/mL MBC=3.4-5.0 mg/mL), and SCH and VAL on Candida albicans (MIC=2.5 mg/mL MFC=5.0 mg/mL). SLs showed notable effect on all tested fungi Aspergillus ochraceus, Penicillium funiculosum, C. albicans and C. krusei (MIC=0.15-0.4 mg/mL MFC=0.3-0.8 mg/mL). In MTT test, extracts inhibited growth of all tested cancer cells (HeLa, LS174 and A549), with the best effect on HeLa (IC50 =148.1 μg/mL for SCH, and 152.3-303.2 μg/mL for MeOH extracts); both SLs were active against HeLa cells (IC50 =46.2 μg/mL for crepiside E and 103.8 μg/mL for 8-epiixerisamine A). Extracts and SLs showed good safety profile on normal MRC-5 cells.

Dry MeOH extracts of the twig barks of Pyrus communis subsp. pyraster, P. spinosa and their hybrid P.×jordanovii nothosubsp. velenovskyi, collected in wild in Serbia, were analyzed. By LC/MS, the contents of arbutin (99.9-131.0 mg/g), chlorogenic acid (2.2-6.3 mg/g), catechin (1.0-5.3 mg/g) and total dimeric and trimeric procyanidins (42.2-61.3 mg/g), including procyanidin B2 (8.9-17.2 mg/g), were determined. Colorimetrically, high contents of total phenolics (436.2-533.4 mg GAE/g) and tannins (339.4-425.7 mg GAE/g), as well as strong total antioxidant activities (FRAP values 4.5-5.9 mmol Fe2+ /g), and DPPH (SC50 =6.6-7.1 μg/ml) and hydroxyl radical (SC50 =447.1-727.7 μg/ml) scavenging abilities were revealed. In vitro, all extracts exhibited notable inhibition of α-amylase (IC50 =310.8-617.7 μg/ml) and particularly strong inhibition of α-glucosidase (IC50 =2.1-3.7 μg/ml). Molecular docking predicted that among identified compounds procyanidin B2 is the best inhibitor of these carbohydrate-digesting enzymes. Obtained results showed that the barks of investigated Pyrus hybrid and its parent taxa have similar composition and bioactivity.

Aromatic plants and essential oils have many applications in medicine, pharmaceuticals, cosmetics, and the food industry. The essential oil of the flowering aerial parts of Achillea grandifolia, obtained by hydrodistillation, was analyzed for its constituents and investigated for antimicrobial and radical scavenging activity. The essential oil was characterized by a high amount of oxygenated monoterpenes (72.7%) with 1,8-cineole (29.2%) and camphor (23.4%) being the most abundant. Sesquiterpenes were present in smaller quantities (4.8%). Antimicrobial activity was tested against eight ATCC bacterial strains and two ATCC strains of Candida albicans. The essential oil exhibited highly pronounced antimicrobial activity against Micrococcus luteus with a MIC value of 3.50 ?g/mL, as well as significant antimicrobial activity (<100 ?g/mL) against Staphylococcus aureus, S. epidermidis and Bacillus subtilis. Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa were resistant. Achillea grandifolia essential oil exhibited concentration-dependent antiradical activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical with an SC50 value of 5.4 mg/mL. The TLC-DPPH assay revealed two main light yellow spots indicating components with anti-DPPH activity, which after isolation were identified as 1,8-cineole and camphor.

Present study investigated triterpene profile, antihyperalgesic and antiedematous activities of Hieracium scheppigianum flowering aerial parts dichloromethane extract (SCH), and antihyperalgesic and antiedematous activities of previously chemically characterised polyphenol-rich H. glabratum and H. calophyllum flowering aerial parts methanol extracts (GLA and CAL, respectively). alpha- and beta-Amyrin and their acetates, and lupeol acetate were identified and quantified in SCH by GC-FID and GC-MS. In carrageenan-induced localised inflammation model in rats, SCH and GLA (50-200 mg/kg, p.o.) produced significant and dose-dependent antihyperalgesic effect of 26.9%-56.2% (ED50=163.0 +/- 26.5 mg/kg) and 25.3%-51.6% (ED50=211.6 +/- 70.6 mg/kg), respectively, and CAL (200 mg/kg, p.o.) exhibited effect of 38.1%. Extracts did not significantly reduce paw edema. SCH and GLA, which demonstrated higher (over 50%) antihyperalgesic efficacy, were tested in a rotarod test (200 mg/kg, p.o.) and no alteration of motor coordination was observed. Also, acute administration of SCH and GLA in mice (2000 mg/kg, p.o.) caused neither mortality nor toxicity.

The composition of leaf and flower essential oils of eight Heracleum taxa (populations collected in Serbia, Montenegro, North Macedonia and Slovenia) was statistically analyzed to evaluate its chemosystematic significance. Investigated taxa included H. orphanidis and the representatives of H. sphondylium group: H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii and H. verticillatum. Hydrodistilled essential oils were analyzed by GC–FID and GC–MS. Chemosystematic significance was evaluated using multivariate statistics: principal component analysis (PCA), non-metric multidimensional scaling (NMDS) and unweighted pair-group arithmetic averages clustering (UPGMA). Statistical analyses included our previously published data on the composition of essential oils of eight leaf and three flower samples, as well as the data on the composition of the oils of additional eight leaf and five flower samples obtained in the current work. Leaf and flower essential oils of H. sphondylium group members were dominated by various sesquiterpenes, phenylpropanoids and/or monoterpenes. Heracleum orphanidis leaf and flower essential oils were rich in aliphatic esters, mostly octyl acetate. Statistical analysis of the composition of leaf essential oils, as well as of flower oils, demonstrated the grouping of investigated populations of these Heracleum taxa according to their systematics, i.e., separation of H. orphanidis from the representatives of H. sphondylium group, and grouping of H. pyrenaicum subspecies within this group. Morphologically related H. sibiricum and H. ternatum were closely located in PCA and NMDS and in UPGMA even shared the same cluster.

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.

According to legislation of EU, as well as of our country, herbal medicinal products (HMPs) and traditional herbal medicinal products (THMPs) are categories of medicinal products. As active ingredients, they contain herbal drugs (herbal substances) and/or herbal drug preparations (herbal preparations). HMPs are authorized based on proved quality, clinical efficacy and safety. THMPs are registered based on proved quality and sufficient data on traditional use (acceptable safety level and plausible efficacy). The Committee on Herbal Medicinal Products (HMPC) of the European Medicines Agency (EMA) publishes EU herbal monographs, which comprise preclinical and clinical data on HMPs with well-established use (WEU), as well as data on traditional use (TU) for THMPs. HMPC also develops drafts of EU list entries for THMPs, which are adopted and their final versions are published by the European Commission. By the end of July 2019, HMPC issued 160 final monographs, which are available on the EMA website (157 monographs for 156 herbal drugs and herbal drug preparations, two monographs for two herbal drug combinations, and monograph for herbal tea combinations); EU list entries on 13 herbal drugs and preparations were finalized. This paper provides an overview of the therapeutic areas and indications approved in these final EU monographs. In addition, overview of HMPs and THMPs, which are authorized/registered in the Republic of Serbia is given.