Distribution of Balkan endemic Euphorbia serpentini

This paper is a survey of distribution of the Balkan endemic obligate serpetinophyte Euphorbia serpentini in Bosnia and Herzegovina and Serbia based on a revision of herbarium specimens from SARA, BEO, BEOU and private collections of the authors, literature data and on field observations. In addition a short morphological description and photographs of the species are presented

Goniolimon dalmaticum Rchb. f. and G. tataricum (L.) Boiss. (Plumbaginaceae) in the Croatian flora and their distribution in the Balkan Peninsula

Goniolimon tataricum (L.) Boiss. is recognized as a new species for the Croatian flora. Besides G. dalmaticum Rchb. f., G. tataricum represents the second species of this genus in Croatia. To confirm the identity of G. tataricum as a new species, morphometric study and multivariate statistical analysis were performed on the data from four Croatian populations of G. dalmaticum and G. tataricum. Based on results of morphometric analysis, an identification key for these two species is provided. Data about distribution in the Balkan Peninsula, habitat and ecology of the newly-recorded species are also presented. Typification of G. dalmaticum was performed for the first time

Fatty acids, phytosterols, furanocoumarins and polyphenols of Prangos trifida s.l. (Apiaceae) roots and fruits

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 202

Composition of Essential Oils from Fruits of Peucedanum longifolium and Rhizomatophora aegopodioides (Apiaceae) with Regard to Other Related Taxa—A Chemometric Approach

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

Sekundarni metaboliti i antiholinesterazna aktivnost herbe Chamaecytisus heuffelii subsp. jankae

Chamaecytisus heuffelii subsp. jankae (Velen.) Niketić (Fabaceae) is a dwarfish shrub, which range includes eastern part of the Balkan Peninsula (Albania, Bulgaria, Serbia) (1). The aim of the investigation was to chemically characterize aerial flowering parts of this plant: the dried methanol extract (TM), its alkaloid (AF) and non-alkaloid (NAF) fractions, and to assess their ability and the ability of detected flavonoids to inhibit enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), in vitro in the Ellman spectrophotometric test. Plant material was collected in eastern Serbia (Jelašnička Klisura gorge), dried, powdered and extracted with methanol, after dichloromethane pre-extraction. Using solvents of different polarities and pH value, AF (yield 14.35%) and NAF (yield 57.78%) were obtained from TM. By LC-MS analysis, flavonoids luteolin-8-C- glucosylpentoside (159.81 mg/g), genistin (22.38 mg/g), vitexin (6.32 mg/g), rutin (4.25 mg/g), and genistein (0.86 mg/g) were detected and quantified in TM, while luteolin-8-C- glucosylpentoside (212.70 mg/g), rutin (1.38 mg/g), and genistin (1.03 mg/g) in NAF. Using GC-FID-MS analysis, quinolizidine alkaloids sparteine, 17-oksosparteine, and lupanine were identified in AF. TM and NAF exhibited significant and comparable anti-cholinesterase activity (IC50AChE 0.54 and 0.72 mg/mL; IC50BuChE 0.45 and 0.68 mg/mL). All detected flavonoids evinced the ability to inhibit enzymes; the most active were genistein and vitexin (IC50AChE 54.75 and 88.35 μg/mL; IC 50BuChE 27.83 and 49.77 μg/mL). Considering the quinolizidine alkaloids toxicity and AF poor anti-cholinesterase activity, it might be concluded, regarding safety profile and medical potential, that the fraction of the herb methanol extract, flavonoid-rich and alkaloid-free (NAF), is good candidate for further research.Chamaecytisus heuffelii subsp. jankae (Velen.) Niketić (Fabaceae) je polužbun, koji raste na području istočnog dela Balkanskog poluostrva (Albanija, Bugarska, Srbija) (1). Cilj istraživanja bio je da se izvrši hemijska karakterizacija nadzemnih delova u cvetu ove biljke: suvog metanolnog ekstrakta (TM), njegove alkaloidne (AF) i frakcije bez alkaloida (NAF), i da se in vitro, u Ellmanov-om spektrofotomerijskom testu, ispita njihova sposobnost, kao i sposobnost detektovanih flavonoida da inhibiraju enzime acetilholinesterazu (AChE) i butirilholinesterazu (BChE). Biljni materijal je sakupljen u istočnoj Srbiji (Jelašnička klisura), osušen, samleven i ekstrahovan metanolom, nakon pre-ekstrakcije dihlormetanom. Primenom rastvarača različite polarnosti uz promenu pH vrednosti, iz TM su dobijene AF (prinos 14,35%) i NAF (prinos 57,78%). LC-MS analizom, u TM detektovani su i kvanitikovani flavonoidi luteolin-8-C-glukozilpentozid (159,81 mg/g), genistin (22,38 mg/g), viteksin (6,32 mg/g) i rutin (4,25 mg/g) i genistein (0,86 mg/g), a u NAF luteolin-8-C- glukozilpentozid (212,70 mg/g), rutin (1,38 mg/g) i genistin (1,03 mg/g). GC-FID-MS analizom, u AF identifikovani su hinolizidinski alkaloidi spartein, 17-oksospartein i lupanin. TM i NAF ispoljili su značajnu i uporedivu antiholinesteraznu aktivnost (IC50AChE 0,54 i 0,72 mg/mL; IC 50BChE 0,45 i 0,68 mg/mL). Svi detektovani flavonoidi su pokazali sposobnost da inhibiraju enzime; najaktivniji su bili genistein i viteksin (IC 50AChE 54,75 i 88,35 μg/mL; IC 50BuChE 27,83 i 49,77 μg/mL). Uzimajući u obzir toksič nost hinolizidinskih alkaloida i slabu antiholinesteraznu aktivnost AF, mož e se zaključ iti da je frakcija metanolnog ekstrakata herbe bogata flavonoidima i oslobođena alkaloida (NAF), u pogledu bezbednosnog profila i lekovitog potencijala dobar kandidat za dalja ispitivanja.VIII Kongres farmaceuta Srbije sa međunarodnim učešćem, 12-15.10.2022. Beogra

Distribution of the genus Galanthus L. (Amaryllidaceae) in Serbia

The distribution of species from the genus Galanthus in Serbia is presented, and notes on their taxonomy and ecology are given in the paper. To date, only two species from the genus are known to be present, namely G. nivalis and G. elwesii. Further research on delimitation of the taxa occurring in Serbia is needed

Chemical composition of essential oils from fruits of Peucedanum longifolium and P. aegopodioides (Apiaceae)

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 202

Preliminary analysis of specialised metabolites of Cytisus jankae flowering aerial part

Cytisus jankae Velen. (Fabaceae) is a dwarfish shrub (10–25 cm height) with 3-foliolate leaves and capitate inflorescence with tubular calyces. The whole plant is densely covered with silky hairs. Its range includes central and east part of the Bal- kan Peninsula and Romania. It inhabits dry rocky places on limestone or ultramafic soils. The objective of this study was to establish the phytochemical profile of flowering aerial parts of this plant. The plant material was collected in eastern Ser- bia (Jelašnička Klisura gorge). In the air-dried herb, by the direct gravimetric method, the total alkaloids content (0.32%) was determined, and using colorimetric assays the total poly- phenols (43.8%), tannins (1.13%), and the total flavonoids (1.31%) were quantified. Additionally, the powdered dried plant material was successively extracted with dichlorometh- ane (maceration) and methanol (bimaceration) at room tem- perature. After the solvents were evaporated under reduced pressure, dried extracts were subjected to the further analy- sis. The total alkaloids content in both the dichloromethane (7.01%) and methanol (3.47%) extracts, and the contents of total polyphenols (13.75%), tannins (2.45%) and flavonoids (4.65%) in the methanol extract, were determined by afore- mentioned tests. Applying the LC-MS method, in the metha- nol extract flavonoids quercetin 3-O-rutinoside, luteolin, api- genin and genistein were identified. GC-MS analysis resulted in the identification of quinolizidine alkaloids sparteine, 17-oxosparteine and lupanine (2-oxosparteine) in both the methanol and dichloromethane extracts, as well as of one tri- terpene, β-amyrin, in the dichloromethane extract. This is the first report on specialised metabolites from flowering aerial parts of C. jankae.7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, Serbi

DPPH radical scavenging potential of the root essential oils of five Heracleum L. taxa

In the Balkan Peninsula, Heracleum L. taxa (Apiaceae) were traditionally used for the treatment of various digestive and respiratory diseases, epilepsy, hypertension and sexual weakness. The purpose of this work was to investigate 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability of the root essential oils of some of the Balkan Heracleum taxa, i.e. H. sibiricum L., H. ternatum Velen., H. verticillatum Pančić, H. pyrenaicum subsp. pollinianum (Bertol.) F. Pedrotti & Pignatti and H. pyrenaicum subsp. orsinii (Guss.) F. Pedrotti & Pignatti. GC-FID and GC-MS analysis of these oils, obtained by hydrodistillation using Clevenger-type apparatus, revealed the domination of monoterpenes, mostly β-pinene (26.2-47.3%). Additionally, H. sibiricum root oil was rich in phenylpropanoids, mainly elemicin (25.6%) and methyl eugenol (22.3%). In colorimetric DPPH assay, the strongest activity was exhibited by H. sibiricum oil (SC50=5.19 μL/mL), followed by H. pyrenaicum subsp. orsinii, H. ternatum, H. pyrenaicum subsp. pollinianum and H. verticillatum oils (SC50=7.85-12.33 μL/mL). In TLC-DPPH (dot-blot) test, three the most active root oils, i.e. those of H. sibiricum, H. ternatum and H. pyrenaicum subsp. orsinii revealed yellow anti-DPPH zones (Rf=0.30-0.42), which were then eluted and analyzed by GC-FID and GC-MS. It was shown that elemicin and methyl eugenol, dominant in H. sibiricum oil, were also the most abundant in its anti-DPPH zone (64.5 and 19.5%). β-Pinene and other monoterpene hydrocarbons were not detected in this, and also in the active zones of H. ternatum and H. pyrenaicum subsp. orsinii oils (both contained two closely located anti-DPPH zones, which were eluted together). Anti-DPPH zones of H. ternatum oil were dominated by trans-sabinol (21.8%), spathulenol (21.7%) and (E)-sesquilavandulol (13.1%), and those of H. pyrenaicum subsp. orsinii oil by (E)-sesquilavandulol (14.5%) and intermedeol (13.6%). These oxygenated terpenes were detected in H. ternatum and H. pyrenaicum subsp. orsinii oils only in small quantities. Another minor constituent of H. pyrenaicum subsp. orsinii root oil, (Z)-falcarinol, was among the dominant ones in its anti-DPPH zones (11.6%).7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, SerbiaSaopštenje sa međunarodnog skupa štampano u izvod

Procena bezbednosnog profila etarskih ulja osam taksona roda Heracleum (Apiaceae) u odnosu na utvrđeni sadržaj furanokumarina

For essential oils of roots, leaves, flowers and fruits of eight Heracleum taxa (H.sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii, H. verticillatum and H. orphanidis), we previously demonstrated antimicrobial, cytotoxic (selective to cancer cells) and/or antioxidant activities. In this work, for these essential oils maximum daily intake related to total furanocoumarins (FCs) content was estimated, according to Committee on Herbal Medicinal Products of European Medicines Agency (EMA/HMPC) recommendations. FCs were quantified using gas chromatography, and their sum equivalent to xanthotoxin (8-methoxypsoralen, 8-MOP) was calculated. It was shown that daily intake, not contributing significantly to overall risk (equivalent to intake of 1.5 mg FCs) for root essential oils was in the range of 1.94-5.23 mL, for fruit oils of 5.23-15.68 mL and for leaf or flower oils of 2.90-15.68 mL. Daily intake, not posing any unacceptable risk (equivalent to intake of 15 μg FCs) for root oils was in the range of 0.02-0.05 mL, for fruit oils of 0.05-0.16 mL and for leaf or flower oils of 0.03-0.16 mL. This work demonstrates the application of current EMA/HMPC recommendations, in order to establish safety profile of herbal preparations containing FCs.Za etarska ulja korena, listova, cvasti i plodova osam taksona roda Heracleum (H. sphondylium, H. sibiricum, H. montanum, H. ternatum, H. pyrenaicum subsp. pollinianum, H. pyrenaicum subsp. orsinii, H. verticillatum i H. orphanidis) prethodno su pokazane antimikrobna, citotoksična (selektivna prema tumorskim ćelijama) i/ili antioksidantna aktivnost. Cilj ovog rada bio je da se ustanovi maksimalni dozvoljeni dnevni unos etarskih ulja u odnosu na utvrđeni sadržaj ukupnih furanokumarina, u skladu sa preporukama Komiteta za biljne lekovite proizvode Evropske agencije za lekove (EMA/HMPC). Furanokumarini su kvantifikovani gasnom hromatografijom i njihov ukupni sadržaj izražen je kao ksantotoksin (8- metoksipsoralen, 8-MOP). Dnevni unos ispitivanih etarskih ulja koji ne doprinosi značajno ukupnom riziku (ekvivalentan unosu 1,5 mg furanokumarina izraženih kao 8-MOP), kreće se u opsegu 1,94-5,23 mL za etarska ulja korena, 5,23-15,68 mL za ulja plodova i 2,90-15,68 mL za ulja listova ili cvasti. Dnevni unos etarskih ulja koji ne predstavlja nikakav rizik (ekvivalentan unosu 15 μg furanokumarina izraženih kao 8-MOP), kreće se u opsegu 0,02-0,05 mL za etarska ulja korena, 0,05-0,16 mL za ulja plodova i 0,03-0,16 mL za ulja listova ili cvasti. Ovaj rad demonstrira primenu aktuelnih EMA/HMPC preporuka u cilju utvrđivanja bezbednosnog profila biljnih preparata u kojima su detektovani furanokumarini