Investigation of essential oils and their potential use in the food industry and pharmacy

Essential oils are complex mixtures of volatile compounds produced by aromatic plants and characterized by distinctive, strong fragrance. Essential oils and their constituents are widely used in a number of food products and beverages, primarily as flavoring agents. Due to the antimicrobial and antioxidant properties of essential oils and their components, they may represent a promising natural alternative to chemical preservatives in the food industry. Similarly, the insecticidal/repellent potential of essential oils may be worth exploring in view of food storage. In addition to antimicrobial and antioxidant activity, essential oils demonstrate various pharmacological activities including anti-inflammatory, expectorant, spasmolytic, carminative, diuretic, rubefacient activity and are hence used in the treatment of respiratory, gastrointestinal, skin disorders as well as rheumatism, neuralgia and many other ailments. The investigation of essential oils is a wide field of research addressing different aspects. In light of these considerations, the essential oils of different plants, mainly from the flora of the Balkan Peninsula, were investigated in order to find new sources of raw material and constituents with potential use in the pharmacy and food industry. The study focused on the content, chemical characterization (e.g. Valeriana sp., Anthemis sp.) and evaluation of their antimicrobial (e.g. Thymus sp., Achillea sp., Armoracia sp.), spasmolytic (Ferula sp., Cymbopogon sp.), antiedematous and antihyperalgesic (Matricaria sp., Laserpitium sp.) properties, as well as their insecticidal properties (Cymbopogon sp.). Since many factors have an influence on the composition of essential oils, chemical analyses provide insight into the variability and possible existence of chemotypes, which is important from the aspect of defining raw material and their quality control. Although of natural origin and usually considered safe, some essential oils contain compounds that bear potential risks (e.g. essential oils of Salvia officinalis, Artemisia absinthium, Ocimum basilicum); therefore, the safety aspect of essential oils should not be neglected.Predavanje po pozivu sa skupa međunarodnog značaja: Unifood conference, Belgrade, September 24-25, 2021UNIFood2021 Conference, 24th-25th September 2021 University of Belgrade, 2nd International UNIfood Conferenc

Safety aspects of herbal products containing compounds with a potential risk

Pojedine biljne droge i preparati biljnih droga sadrže jedinjenja sa potencijalnim rizikom primene, kao što su pirolizidinski alkaloidi (npr. u korenu gaveza, Symphyti radix, listu podbela, Farfarae folium, rizomu lopuha, Petasitidis rhizoma), kumarin (u kori kineskog cimeta, Cassiae cortex, herbi ždraljevine, Meliloti herba), estragol (u plodovima gorkog i slatkog morača, Foeniculi amari/dulcis fructus, plodu anisa, Anisi fructus, herbi bosiljka, Basilici herba), tujon (u listu žalfije, Salviae officinalis folium, herbi belog pelena, Absinthii herba) ili azaron (u rizomu iđirota, Calami rhizoma). Za ove sastojke i droge/preparate droga koji ih sadrže utvrđeni su prihvatljivi, ograničeni dnevni unosi kao i preporuke za dužinu primene. Farmaceut ima ključnu ulogu u savetovanju pacijenata o primeni proizvoda na bazi biljnih droga i preparata biljnih droga, uključujući i one koji sadrže jedinjenja sa potencijalnim rizikom i veoma je važno da bude upoznat sa karakteristikama i pravilnom primenom ovih biljnih proizvoda.Certain herbal drugs and herbal drug preparations contain compounds with a potential risk of use, such as pyrrolyzidine alkaloids (in Comfrey root, Symphyti radix, Coltsfoot leaf, Farfarae folium, Butterbur root, Petasitidis rhizoma), coumarin (in the Cassia bark, Cassiae cortex, Melilot, Meliloti herba), estragole (in Basil, Basilici herba, bitter/sweet fennel, Foeniculi amari/dulcis fructus, Aniseed, Anisi fructus), thujone (in Sage leaf, Salviae officinalis folium, Wormwood, Absinthii herba) or asarone (in Sweet flag rhizome, Calami rhizoma). For these ingredients and herbal drugs/herbal drug preparations which contain them, there are acceptable, limited daily intakes as well as recommendations for duration of use. Since pharmacist plays a key role in advising patients on the use of herbal products, including those containing compounds with a potential risk, it is very important to know the characteristics and proper use of these herbal products

Compounds of the methanolic leaf extract as chemotaxonomic markers for the Campanula pyramidalis complex (Campanulaceae)

During the past few years, the isophylloid Campanula pyramidalis complex has been the subject of studies aimed at an improved understanding of the relationships within it. The center of distribution of the C. pyramidalis complex is in the Balkan Peninsula with some smaller parts of the area located in the south Apennines. Although 21 taxa of the C. pyramidalis complex were described, only four species are accepted: C. pyramidalis, C. versicolor, C. secundiflora and C. austroadriatica. In the present study, we propose compounds of the methanolic leaf extract as possible chemotaxonomic markers for the C. pyramidalis complex. Eleven flavonoids and two phenolic acids were detected in leaf extract using high-performance liquid chromatography with diode-array detection analysis. The investigated taxa of the C. pyramidal is complex differ in terms of the composition of the methanolic leaf extract. Clustering of investigated taxa is not completely consistent with the previously reported molecular and morphometric dat

Biljni lekoviti proizvodi: Uslovi za bezbednu primenu

As the use of herbal medicinal products is constantly growing, their safety became an important public health issue. In spite of common belief that herbal medicinal products are safe and harmless because they are natural, it is not always so and their usage is not devoid of certain risks. The most common reasons for herbal adverse reactions (ADRs) are: improper use (overdose or prolonged use), interactions with conventional drugs and poor quality of herbal products. In this work were discussed the most important issues on their safe usage. Classification of herbal medicinal products (herbal medicine, traditional herbal medicine) and basic informations on their registration and marketing authorization according to current European and Serbian legislation were also given. Manufacturing of quality herbal medicinal products is highlighted as one of the most important requirements for their safety. Having in mind that most of these products are sold without prescription (OTC) and that they are mostly used in self-medication, the role of health professionals in consulting patients on choosing of herbal medicinal product and its usage was emphasized. The importance of patient's collaboration with health professionals on recognizing and monitoring herbal ADRs and avoiding possible interactions was also pointed out. Examples of the most frequent herbal medicinal products ADRs occurring in course of their recommended and improper use, on their interactions with drugs as well as on their poor quality were given. The importance of involving herbal medicinal products in international pharmacovigilance systems was also explained

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

Heracleum ternatum from Mt. Durmitor: furanocoumarins, polyphenols and antioxidant activity of leaf and flower extracts

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 202

Pimpinella tragium Vill. (Apiaceae) – anatomy and essential oil composition

Pimpinella tragium Vill. is a perennial, herbaceous plant inhabiting dry grasslands, limestone rocky areas and screes of wide sense Mediterranean area, east European lowlands and Caucasus. This study explores the anatomical structure, content and composition of the essential oil of the vegetative organs and fruit of this species. The plant material was collected on two localities: the village Izvor (Bosilegrad, Serbia) and Mt. Galičica (FYR Macedonia). The anatomical studies were conducted on permanent slides obtained by standard method of preparation for viewing under a light microscope. The essential oils obtained by hydrodistillation from roots (collected during flowering and fruiting period), aerial parts with inflorescences, and fruits, were analysed by GC-FID and GC-MS. The anatomical analysis revealed a secondary structure of root and primary structure of stem with closed collateral vascular bundles. Leaves are isobilateral, amphistomatic; petiole is with arched vascular bundles and the fruit (mericarp) is semi-circular and lightly ribbed in cross section. Non-glandular unicellular and bicellular cuneate trichomes are sparsely distributed on the stem, leaves and petiole, but very dense on the fruit. Secretory channels are present in all organs: in the parenchyma of the root and stem cortex, stem pith, in the phloem of root and vascular bundles of stem and petiole, by the leaf vascular bundles and in the fruit pericarp. Essential oil yields from different parts of P. tragium ranged from 0.2-1.1% (v/w). The main compounds in the oils from roots (in both stages, from both localities), as well as from aerial parts and fruits from Mt. Galičica are C-12 norsesquiterpenes (trinorsesquiterpenes): pregeijerene (29.0-56.2%) and gejerene (14.1-22.9%), whereas those from aerial parts and fruits from village Izvor are β-bisabolene (19.1-57.2%) and a phenylpropanoid epoxy-pseudoisoeugenyl-2-metohybutyrate (17.4-22.2%). Trinorsesquiterpenes, which were also found to be characteristic constituents in some other Pimpinella species oils, are the most dominant constituents in all investigated oils, except in the oil of fruit from village Izvor. In addition, phenylpropanoids of pseudoisoeugenol type are present in all oils confi rming them as chemical markers of Pimpinella species analysed so far.7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, Serbi

Comparative chemical analysis of essential oils from different organs of three Pastinaca taxa

INTRODUCTION: Pastinaca sativa subsp. sativa L., Apiaceae (parsnip) is cultivated mainly in the temperate regions of the world because of its edible root. The roots of the best quality are obtained from the plants from the first year, in which this biennial plant usually forms only leaf rosette. In the second year, flowering stems emerge (the plant is cultivated for two years in order to obtain fruits for reproduction). Wild-growing P. sativa subsp. urens (Req. ex Godr.) Čelak. is widely distributed in Europe and P. hirsuta Pančić is endemic in the central part of the Balkan Peninsula (east Serbia, North Macedonia and south and west Bulgaria). OBJECTIVES: To investigate and compare the composition of the essential oils obtained from roots, leaves, stems, flowers and fruits of cultivated P. sativa subsp. sativa (from the first and/or the second year) and wild-growing P. sativa subsp. urens and P. hirsuta from Serbia. METHOD / DESIGN: Essential oils were isolated from dried and comminuted plant material by hydrodistillation using Clevenger-type apparatus for 2.5 h. The composition of essential oils was determined by GC-FID and GC-MS and analyzed using multivariate statistical methods: principal component analysis (PCA), non-metric multidimensional scaling (nMDS) and unweighted pair-group arithmetic averages clustering (UPGMA). RESULTS: Twenty-nine parsnip essential oils were investigated: 11 P. sativa subsp. sativa oils (from four localities; the oils of roots and leaves from both the first and the second year, and of the other organs from the second year), 10 P. sativa subsp. urens oils (from two localities) and eight P. hirsuta oils (from one locality, collected in two different years). Pastinaca sativa subsp. sativa roots from the first year provided the highest oil yields (0.51-0.77%, w/w). The roots of this taxon from the second year (0.02%), and the roots of other investigated Pastinaca taxa (0.10-0.14%) had notably lower oil yields. Regarding other plant organs, high oil yields were generally obtained for fruits (1.40-3.90%) and flowers (0.43-0.93%), and low oil yields for leaves (0.07-0.16%) and stems (0.03-0.13%). In general, 13 to 53 compounds were identified in the essential oils; identified components accounted for 89.4-98.6% of the oils. Phenylpropanoid myristicin was the most abundant in the root essential oils of both investigated P. sativa subspecies (39.7-82.5%). It is interesting to note that the oil of P. sativa subsp. sativa roots from the first year also contained high amounts of terpinolene (14.8-28.7%), which significantly decreased in the oil of this taxon from the second year (1.2%). In P. hirsuta root oils another phenylpropanoid apiole (30.9 and 25.8%) was dominant and the quantities of myristicin were somewhat lower (11.6 and 20.3%). Cultivated P. sativa subsp. sativa leaf essential oils were dominated by myristicin (42.8 and 41.4%) and in the leaf oils of wild-growing parsnips (P. sativa subsp. urens and P. hirsuta), γ-palmitolactone was the most abundant (22.6-47.5%). Additionally, the leaf oils of both investigated P. sativa subspecies contained significant amounts of sesquiterpenes (36.0-46.2%), e.g. (E)-β-farnesene (13.8-22.4%). Compared with the leaf oils, corresponding stem oils were qualitatively similar. However, in the stem oils, the contents of sesquiterpenes were lower [e.g. (E)-β-farnesene 4.9-14.4% in P. sativa oils] and the contents of myristicin (64.9 and 63.3% in P. sativa subsp. sativa oils) and γ-palmitolactone (50.6-60.4% in wild-growing parsnips oils) were higher. The flower and fruit essential oils were dominated by aliphatic esters. The most abundant in P. hirsuta oils were hexyl butanoate (61.9% in the flower oil; 22.0 and 58.4% in the fruit oils) and hexyl hexanoate (17.0% in the flower oil; 59.8 and 29.1% in the fruit oils), and in the oils of both P. sativa subspecies the dominant was octyl butanoate (26.1-31.4% in the flower oils; 53.6-79.0% in the fruit oils). In PCA and nMDS analyses of the oils (except fruit oils), the separation of all three investigated Pastinaca taxa was noticed. The same relations were observed in UPGMA analyses of the leaf, stem and flower oils. In the case of statistical analysis of the fruit oils and UPGMA analysis of the root oils, the samples of two subspecies of P. sativa were grouped together. CONCLUSIONS: Wild-growing parsnips are equally interesting sources of essential oils as cultivated parsnip. Locality and year of collection did not significantly influence relations among taxa observed in multivariate statistical analysis.The International Bioscience Conference and the 8th International PSU – UNS Bioscience Conference - IBSC2021, Novi Sad, 25th to 26th November 202

Chemosystematic evaluation of the composition of leaf and flower essential oils of eight Heracleum L. taxa from Southeastern Europe

The genus Heracleum L. (Apiaceae) is represented by a dozen of native sub(species) in Southeastern Europe. In this study, the composition of the essential oils obtained from the leaves and the flowers of eight taxa of this genus, collected in Serbia, Montenegro, Macedonia and Slovenia, was statistically analyzed to evaluate its chemosystematic significance. Investigated taxa included H. orphanidis Boiss. and the representatives of H. sphondylium group: H. sphondylium L., H. sibiricum L., H. montanum Schleich. ex Gaudin, H. ternatum Velen., H. pyrenaicum subsp. pollinianum (Bertol.) F. Pedrotti & Pignatti, H. pyrenaicum subsp. orsinii (Guss.) F. Pedrotti & Pignatti and H. verticillatum Pančić. Essential oils were hydrodistilled using Clevenger-type apparatus and analyzed by GC-FID and GC-MS. Chemosystematic significance of their components was evaluated using multivariate statistics: principal component analysis (PCA), non-metric multidimensional scaling (nMDS) and unweighted pair-group arithmetic averages clustering (UPGMA). The analyses included our previously published data on the oils of eight samples of the leaves and three of the flowers, as well as additionally analyzed oils of eight samples of the leaves and five of the flowers. Leaf and flower oils of investigated members of H. sphondylium group were dominated by various sesquiterpenes [(E)-caryophyllene, (E)-nerolidol, (E)-β-farnesene, α-trans-bergamotene, germacrene D, β-bisabolene and/or β-sesquiphellandrene], phenylpropanoids [apiole, methyl eugenol, elemicin and/or (Z)-isoelemicin], and/or monoterpene limonene. On the other hand, leaf and flower oils of H. orphanidis were rich in aliphatic esters, mostly octyl acetate. Separate statistical analyses of the compositions of the leaf oils and the flower oils demonstrated segregation of H. orphanidis from investigated representatives of H. sphondylium group, and grouping of the subspecies of H. pyrenaicum within this group. Morphologically related species H. sibiricum and H. ternatum were closely located in PCA and nMDS, and in UPGMA even shared the same cluster. PCA showed that some of both aforementioned dominant constituents and those present in lower amounts influenced the separation of investigated taxa. It can be concluded that applied multivariate statistical methods demonstrated the grouping of investigated Heracleum taxa according to their current systematics, and justify further similar study on the essential oils of more species of this genus.7th Balkan Botanical Congress, 10-14th September 2018, Novi Sad, SerbiaSaopštenje sa međunarodnog skupa štampano u izvodu