Influence of growing conditions on morphological and chemical properties and biological effects of extracts of Arnica montana L.

Arnica montana L. (Asteraceae) je višegodišnja planinska biljka čije se cvetne glavice (Arnicae flos) uglavnom koriste za spoljašnju upotrebu kod modrica i uganuća u fitofarmaceutskim preparatima kao što su masti, kreme i gelovi. Do pre par godina svetsko tržište pomenute sirovine je zadovoljavano isključivo iz spontanog resursa, ali su biljne populacije, zbog prekomernog sakupljanja, znatno degradirane, zbog čega je u većini evropskih zemalja stavljena zabrana za njeno dalje sakupljanje iz prirode. Obzirom da tražnja za ovom sirovinom na tržištu ne opada, kultivacija arnike se nameće kao jedino održivo rešenje zadovoljenja potreba farmaceutske industrije. Budući da A. montana (narodni naziv – arnika) nije autohtona biljna vrsta za područje Srbije, iako se spontano javlja u planinskim oblastima nekih susednih zemalja i Balkanskog poluostrva uopšte, cilj ovih istraživanja je bio da se iznađe tehnološko rešenje njivske proizvodnje na osnovu ispitivanja uticaja vremena zasnivanja, primene đubrenja i tipa sadnica. U istraživanju su pored cvetne glavice, kao glavne droge, posmatrani i podzemni organi arnike: rizom i koren, kao sporedne sirovine. Prva faza eksperimentalnog rada obuhvatila je trogodišnji tro-faktorijalni poljski ogled načina proizvodnje arnike, pri čemu su se kao faktori ispitivali: vreme zasnivanja useva (prolećna i jesenja sadnja), đubrenje (stajnjak i NPK) i način zasnivanja useva (iz semena i klonskom propagacijom – deobom bokora). U drugoj fazi eksperimenta, izvršena je karakterizacija sirovine u smislu njene brze identifikacije, hromatografskim tehnikama identifikovani su prisutni sekundarni metaboliti i određen njihov sadržaj u ispitivanim uzorcima i testirani su biološki efekti odabranih ekstrakata. Ispitivanja su vršena na ogledu postavljenom 2008. godine na lokalitetu Kaluđerskih bara (1008 m n.v.), na planini Tara, u krugu rasadnika Nacionalnog Parka „Tara“. Obzirom na to da arnika prvu vegetaciju provodi u fazi rozete sva merenja morfoloških parametara, prinosa i hemijskih karakterizacija sirovina vršena su tokom 2009. i 2010. godine. Na ogledu su praćeni sledeći parameri: prečnik rozete, visina cvetnog izdanka, broj cvetnih izdanaka, broj cvetnih glavica, prečnik cvetne glavice, broj sekundarnih rozeta. Određeni su sledeći prinosi: prinos cvetne glavice, prinos rizoma, prinos korena i prinos etarskog ulja (rizoma i korena)...Arnica (Arnica montana L., Asteraceae) is a perennial mountain plant whose flower heads (Arnicae flos) are mainly used for topical treatment of bruises and sprains in phytopharmaceutical preparations such as ointments, creams and gels. Until few years ago, the world market demand for this raw material was almost exclusively covered from the spontaneous resources, but plant populations are, due to overcollecting, significantly degraded, what caused prohibition of its collection from the wild in most of European countries. Since the demand for this raw material on the market is constant rise, cultivation arnica is imposed as the only sustainable solution to meet the needs of the pharmaceutical industry. Since the A. montana plant specie is not native to the region of Serbia, although it occurs spontaneously in some mountain areas of neighboring countries and the Balkans in general, the aim of this study was to find a suitable technology solution of field cultivation based on the examination of induced various ecological factor effects. In this research, beside observations connected with flower heads, as the main drug, underground parts of arnica: rhizome and roots, were also studied as side products. The first phase of the experimental work included a three-year three-factorial field experiment in different modes of production, where as the factors examined: time of plantation establishment (spring and autumn planting), fertilization (manure and NPK) and methods of propagation (from seed and clonal propagation - division of the tuft). In the second phase of the experiment, characterization of the raw material was carried out in the terms of its rapid identification, qualitative evaluations based on the content of secondary metabolites and testing of biological effects of extracts obtained. Tests were performed on the field experiment conducted in 2008 at locality of Kaludjerske Bare (1008 m) on mountain Tara, in area of Nursery production department of the National Park "Tara". Since arnica its first vegetation remains in the rosette phenophase, all measurements of morphological parameters, yields and chemical characterizations of raw materials was carried out in 2009 and 2010 year..

Antimicrobial activity of Helichrysum plicatum DC

Dry flower heads of Helichrysum plicatum were characterized by HPLC-DAD and a detailed antimicrobial assay of its ethanol extract was performed. Identification of phenolic compounds indicated the presence of apigenin, naringenin and kaempferol as free aglycones, glycosides of apigenin, naringenin, quercetin and kaempferol as well as chlorogenic acid and chalcone derivate. Antimicrobial activity of the extract was evaluated against various bacteria and fungi as well as yeast Candida albicans using microdilution method. Grampositive bacteria were more sensitive to the tested extract (MIC values were to 0.02 mg/mL) than Gram-negative bacteria (the greatest MIC was 0.055 mg/mL). Regarding pathogenic fungi, our tests demonstrated that fungi were more sensitive to the tested extract than bacteria. The growth of the majority of the tested fungi was inhibited by concentration of 0.005 mg/mL. Moreover, the extract was significantly more active than commercial fungicide, fluconazole. The results of our tests indicate that the extract of H. plicatum has significant antimicrobial activity and may find application in the pharmaceutical and food industry and organic agriculture

Sorbus aucuparia and Sorbus aria as a Source of Antioxidant Phenolics, Tocopherols, and Pigments.

Due to its nutritive and medicinal properties, berries of some Sorbus species are used for the preparation of jams and jelly as well as in traditional medicine. On the other hand, their chemical composition is not much studied especially of those grown in Balkan Peninsula. We have analyzed individual phenolics, tocopherols, carotenoids and chlorophylls using HPLC in berries from Sorbus aucuparia and Sorbus aria collected in different localities in Serbia and Montenegro together with the amounts of total phenolics and proanthocyanidins as well as their radical scavenging activity against DPPH radical. Berries of S. aucuparia were richer source of polyphenolics in comparision with S. aria and, regardless the species and locality, caffeoylquinic acids such as neochlorogenic and chlorogenic acid were the most abundant compounds. Among analyzed tocopherols the most abundant in all samples was α-tocopherol (0.48 - 19.85 μg/g dw) as it was β-carotene among carotenoids (mean concentration of 0.98 μg/g dw in S. aucuparia and 0.40 μg/g dw in S. aria, respectively). Correlation between total phenolics and DPPH radical scavenging activity was noticed. Our study represents comprehensive report on chemical composition of S. aucuparia and S. aria which could contribute to a better understanding of their quality.Chemistry & Biodiversity, 2017, 14, 12, e170032

Crataegus orientalis leaves and berries: Phenolic profiles, antioxidant and anti-inflammatory activity

The present study was designed to define the phenolic content, antioxidant and anti-inflammatory activity of Crateagus orientalis Pall. Ex M. Bieb., traditionally used by local people in southern parts of F.Y.R. Macedonia. The presence and content of 7 phenolics in ethanolic extracts of leaves and berries were studied using HPLC-DAD, where the most dominant compounds were hyperoside, isoquercitrin and chlorogenic acid. The leaf extract was more effective as a DPPH radical scavenger (IC50 = 29.7 μg/g) than the berry extract, as well as in the relative reducing power on Fe3+. Anti-inflammatory potential was studied by means of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibitory activity; both extracts evinced activity. Furthermore, C. orientalis leaf extract showed a concentration dependent inhibition of COX-1 pathway products 12-HHT and TXB2, reaching IC50 values below the lowest applied concentration (68.9% and 55.2% of 12-HHT and TXB2 production inhibition, respectively, at concentration of 0.4 mg/mL). Although inhibitors such as acetylsalicylic acid and quercetin showed higher activity, this study demonstrates that the investigated extracts are potential anti-inflammatory agents.Natural Product Communications (2017), 12(2): 159-16

Optimization of the extraction process of polyphenols from Thymus serpyllum L. herb using maceration, heat- and ultrasound-assisted techniques

Despite the fact that Thymus serpyllum is well-known medicinal plant and its chemical profile and biological activity have been investigated, there is no detailed study regarding the influence of different techniques and conditions on the extraction of polyphenolic compounds from Serpylli herba. The aim of this study was optimization of the extraction parameters that improves the efficiency of polyphenols extraction from T. serpyllum: particle size, solid-to-solvent ratio, solvent type and extraction time, by using maceration, heat- and ultrasound-assisted extraction (HAE and UAE). The extraction efficiency was expressed via total polyphenol content (TPC) and total flavonoid content (TFC). The statistical analysis (one-way ANOVA and full factorial design) has revealed that the optimal conditions for achieving the best polyphenols yield were particle size of 0.3 mm, 1:30 solid-to-solvent ratio and 50% ethanol, as environmentally friendly extraction medium, while extraction time has not shown statistically significant influence on polyphenols concentration, in all procedures. Under these conditions, the measured TPC was 26.6 mg GAE/L in maceration, 29.8 mg GAE/L in HAE and 32.7 mg GAE/L in UAE, which was in agreement with the predicted values, while TFC was 143 mg CE/L, 12.4 mg CE/L and 16.7 mg CE/L for maceration, HAE and UAE, respectively. According to total polyphenols yield, the efficiency of the extraction methods for all variables was ranked by significance in the following order: UAE gt HAE gt maceration, whereas total flavonoids yield was the highest in UAE, although there was no statistically significant difference between maceration and HAE. According to our results, UAE could be selected as the most successful and suitable technique for extraction of bioactive polyphenolic compounds from Serpylli herba. Using LC/MS and HPLC analysis, 9 polyphenolic compounds were identified and quantified: 6,8-Di-C-glucosylapigenin, chlorogenic acid, 6-hydroxyluteolin 7-O-glucoside, caffeic acid, luteolin 7-O-glucuronide, apigenin glucuronide, salvianolic acid g isomer, rosmarinic acid and salvianolic acid I. This study was an initial step in production of polyphenols-rich wild thyme extracts aimed to be used for formulation of foodstuffs and medicines