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Everlasting flowers (Helichrysum plicatum L.), traditionally used for gastrointestinal disorders such as abdominal pain, jaundice, and hepatic disorders are a rich source of sensitive polyphenolic compounds whose preservation is of great importance. A critical point in the development of formulations with bioactive phenolic compounds, especially liquid extracts is their limited stability at elevated temperatures, pH variations, exposure to oxygen, light, and moisture. In order to overcome the extract limitations, drying method could be the optimal technique for preservation of bioactive compounds, in order to obtain stable powders with appropriate characteristics. One of the most advanced techniques based on the phenomenon of sublimation is a freeze drying, which enabling the long-term preservation of heat-sensitive compounds. In this study, extract obtained by percolation method was lyophilized by freeze drying method (firstly extract was frozen at -80 oC for 1 h, then freeze- dried at -60 oC with pressure of 0.011 mbar for 24 h, and finally main dried at -60 oC with pressure of 0.0012 mbar, for an additional hour in order to remove the capillary water residues). The obtained freeze-dried extract (FHE) was characterized by drying efficiency, the contents of total polyphenols-TPC and flavonoids-TFC. The most dominant polyphenolic compounds were determined and quantified by HPLC method. Differential scanning calorimetry (DSC) and scanning electron microscopy (SEM) analyzes were also performed. The stability of individual compounds, quantified by the HPLC method, was investigated after 30 days, whereby the sample was exposed to a temperature of 40 °C. Drying efficiency was 94.68%, while TPC and TFC were 113.22 mg gallic acid/g and 28.16 mg catechin/g od FHE, respectively. The most dominant individual compounds were naringenin (22.47 mg/g) and kaempferol (21.47 mg/g) of FHE. The DSC method confirmed the FHE stability, and the SEM analysis confirmed that the extract was

Rose hip seed oil is a rich source of polyunsaturated fatty acids, as well as tocopherols, carotenoids, sterols, phospholipids, and phenolic com­pounds. On the other hand, due to the high content of polyunsaturated fatty acids, this oil is prone to oxidation. The aim of this study was to investigate the influence of a natural antioxidant, i.e., pomegranate peel extract, and its com­bination with butylated hydroxytoluene as a commonly used synthetic anti­oxidant, on the stability of rose hip oil. The stability of samples without and with different antioxidants was monitored through analysis of the fatty acid composition and measurement of the quality and stability parameters of the oil (peroxide value, p-anisidine value, thiobarbituric acid reactive substances inhi­bition, total phenolic content and antiradical activity) during a storage period of 12 days at 65 °C. Pomegranate peel extract (0.1 %) inhibited more effectively the second stage of oxidation than butylated hydroxytoluene (0.02 %), while the first stage of oxidation was better prevented by the synthetic antioxidant. Furthermore, the addition of pomegranate peel extract increased the total phen­olic content of the rose hip oil as well as its antiradical activity. Thus, pome­granate peel extract could be used as a potent natural antioxidant for the stabil­ization of beneficial but unstable rose hip oil.

Perennial plant Gentiana lutea L. is used worldwide for the preparation of pharmaceutical and food products. Health benefits of G. lutea roots are associated with the presence of major bitter-tasting secoiridoid gentiopicroside and xanthone isogentisin. The aim was to optimize the heat-assisted extraction of gentipicroside (GP), isogentisin (ISG) and total phenolics (TP) from G. lutea roots and develop models with high accuracy and prediction capacity by response surface methodology (RSM) and artificial neural networks (ANN). Extracts were prepared according to central composite design. Significant independent variables which were previously identified by Plackett-Burman screening design were varied at five levels - temperature (20−80 °C), time (8−180 min), solid-to-solvent ratio (1:10−1:50) and ethanol concentration (10–70 %). Contents of GP, ISG (by HPLC-DAD) and TP (by Folin–Ciocalteu method) were analyzed. The optimal conditions for the extraction were temperature of 65 °C, time of 129.08 min, solid-to-solvent ratio of 1:40, and ethanol concentration 49.33 %. Under these conditions, experimentally obtained results for GP (18.03 mg/g dw), ISG (8.15 mg/g dw) and TP (17.46 mg of gallic acid equivalents/g dw) content were in agreement with the values predicted by RSM and ANN. Comparison of models through the coefficient of determination (R2) and the root mean square error (RMSE) showed that ANN approach was superior to RSM in predicting and modelling GP, ISG and TP content, simultaneously. Effective heat-assisted extraction method for the extraction of GP, ISG and TP from the roots of G. lutea was designed, and models with high accuracy and good prediction capacity were developed.

Sandy everlasting [Helichrysum arenarium (L.) Moench] is herbaceous perennial plant belonging to Asteraceae family and it is native to Europe, Central Asia, and China. It belongs to the section HELICHRYSUM (Asteraceae family, genus Helichrysum) along with H. plicatum DC. Prodr., which very similar phenolic profile and H. italicum (Roth), which is widely used for essential oil extraction. Its flowers have a long tradition in European ethnomedicine as a cholagogue, choleretic, hepatoprotective, and detoxifying herbal drug. The flowers are rich in phenolic compounds including flavonoids, chalcones, phenolic acids, coumarins, and pyrones. Apart frompolyphenols, other compounds such as sterols, lignans, and glycosides of aromatic compounds have been also isolated from H. arenarium. The majority of authors confirm that the most important group of compounds responsible for biological activities is flavonoids. Moreover, significant activities of naringenin, one of the main flavonoids of H. arenarium, were reported. On the other hand, there are no clinical data about testing the extracts or preparations based on H. arenarium. Although H. arenarium is well known in phytotherapy for its potential in the treatment of gallbladder disease and are classified as endangered species in a number of European countries, very few data about its cultivation are available in the literature.

Mešani otpad od lekovitog bilja je kompostiran sa dodatkom novih bakterijskih sojeva koji pripadaju rodovima Streptomyces, Paenibacillus, Bacillus i Hymenobacter. Kompostiranje je praćeno odreĊivanjem hemijskih i bioloških parametara, ukljuĉujući C / N odnos, gubitak organske materije, sadrţaj fosfora i kalijuma, kao i oslobaĊanja CO2 i dehidrogenazne aktivnosti tokom 164 dana. PronaĊeno je da izabrani mezofilni bakterijski sojevi imaju potencijal da znaĉajno redukuju period razgradnje mešanog biljnog otpada od oko 6 meseci do oko 2,5 meseca. Na osnovu merenja indeksa klijanja ĉetiri biljke (Fagopirum esculentum, Thymus vulgaris, Cynara scolimus i Lavandula officinalis) klijavost i rast korena ispitanih biljaka je bio poboljšan inokulisanim kompostom. Indeks klijavosti svih testiranih vrsta na zrelom inokulisanom kompostu je bio u proseku 60% veći u poreĊenju sa kontrolnim (netretiranim) kompostom..., Mixed medicinal plant waste was composted with addition of novel bacterial strains belonging to the genera Streptomyces, Paenybacillus, Bacillus and Hymenobacter. The composting was followed by assessment of chemical and biological parameters including C / N ratio, loss of organic matter, phosphorous and potassium content as well as CO2 generation and dehydrogenase activity during 164 days. The selected mesophilic bacterial starters had a potential to significantly reduce the period of mixed herb waste decomposition, from about 6 months to about 2,5 months. Based on the seed germination index of four plants (Fagopirum esculentum, Thymus vulgaris, Cynara scolimus and Lavandula officinalis) the germination and radial root growth of the investigated plants was improved by the inoculated compost. The germination index of all tested species on the mature inoculated composts was in average 60% higher compared to the control compost. The research indicates that the mesophilic starter addition into the herbs waste can contribute to the speed of waste decomposition and lead to the improvement of biofertilization effect of the obtained compost...

Due to its high content of phenolics, black chokeberry has been proposed as a health-promoting material, especially use of chokeberry extract as a food or pharmaceuticals. Chokeberry extract was spray-dried using gum Arabic as a carrier. The optimal conditions for the efficient microencapsulation of chokeberry extract using spray-drying technique have been investigated. The aim of our study was to obtain microparticles with the best potential to improve functionality and stability of extracted chokeberry polyphenols, and to investigate the possibility of microbeads to protect active compounds during simulated digestion process. Morphological characteristics of microbeads were analyzed using FTIR and SEM techniques. Zeta potential, particle size and moisture content were determined. Released total phenolics and total anthocyanins content as well as concentration of individual anthocyanins were quantified before and after digestion process. Microparticles exhibited high encapsulation efficiency up to 87%, and high content of released polyphenols was achieved. After in vitro simulated digestion phenolic compounds decreased by 11-24%, exhibited higher protective effect of gum Arabic. Our results showed that chokeberry microparticles obtained by spray drying method could be useful supplements or functional food. .

Medicinal plant extracts (MPE) have a broad antimicrobial activity, including antiviral effects. In our study, eleven dry extract of ten different medicinal plant prepared with two solvents (5%, dimethyl sulfoxide (DMSO) and 30% ethanol in 5% DMSO) in different concentrations have been examined for anti-noroviral activity. The reduction of norovirus > 1 log10 genome equivalents (GE) has been obtained with 1 mg/mL of Aronia melanocarpa leaf extract (AMLE), dried wine Prokupac (WPE) and Hypericum perforatum extract (HyPE). Some of MPE i.e. HyPE, Aronia melanocarpa fruit extract (AMFE) and Punica granatum (PPE) showed better activity when dissolved in 30% ethanol with 5% DMSO. All results of anti-noroviral activity of tested MPE which achieve lt 0.5 log10 GE are considered as not effective.

In recent years, drug delivery systems are especially interesting because they allow therapeutic efficiency and reduce side effects. Conjugates of hyaluronic acid and chemotherapeutic agents, especially taxol, have been recognized as excellent solution for many therapeutic indications. The aim of conjugation of taxol with hyaluronic acid was the toxicity reduction on healthy cells and increase of selectivity on tumor cells. Hyaluronic acid could be used in various pharmaceutical preparations, also in parenteral, because of its biocompatibility and biodegradability. Hyaluronic acid as a carrier belongs to in situ gelling systems, which is passing a phase transition in response to external conditions such as temperature, different pH value or the presence of ions, enabling prolonged effect of the drug substances. Many tumor cells have a large number of receptors for hyaluronic acid, which has a great affinity for the tumor cells. The incorporation of taxol in gelling system of hyaluronic acid provides sustained drug release, increased anticancer activity, reduced damage of healthy cells and increased selectivity on cancer cells., Preparati sa produženim oslobađanjem su poslednjih godina posebno aktuelni jer omogućavaju veću terapijsku efikasnost, smanjuju pojavu neželjenih efekata i omogućavaju ciljano delovanje. Posebno su poslednjih godina interesantni konjugati hijaluronske kiseline i citostatika, a jedan od njih je i taksol, koji pored mnogobrojnih terapijskih indikacija ispoljava i niz neželjenih efekata. Cilj konjugovanja taksola sa hijaluronskom kiselinom je smanjenje toksičnosti na zdravim i povećanje selektivnosti na tumorskim ćelijama. Hijaluronska kiselina se može primenjivati u različitim farmaceutskim preparatima, pa i u parenteralnim, jer ima niz prednosti kao što su biokompatibilnost i biodegradibilnost. Spada u in situ gelirajuće sisteme koji se podvrgavaju faznom prelazu u odgovoru na spoljne stimulanse kao što su temperatura, pH ili prisustvo jona, čime se omogućava produženo dejstvo lekovite supstance. Mnoge tumorske ćelije pokazuju ekspresiju velikog broja receptora za hijaluronsku kiselinu, pa ona ima veliki afinitet za tumorske ćelije. Inkorporiranjem taksola u gelirajući sistem sa hijaluronskom kiselinom omogućava se postepeno oslobađanje leka, povećanje antikancerskog delovanja, smanjuje se oštećenje zdravih ćelija i povećava selektivnost leka.

Nowdays, the use of products based on medicinal herbs (phytopreparates) are in scientific focus, both in prevention or in treatment of various diseases. Herbal products have been recognized as an excellent source of bioactive compounds which have positive effects on human health. Beside teas, as standard herbal products, phytopreparations are much more represented as final pharmaceutical form on the market today. Phytopreparations must satisfy standard quality, which means that they must be physically, chemically and microbiological stable and have a high degree of purity. In order to determine the stability, storage conditions, shelf-life of the products, stability tests are conducted, which involving tests of environmental factors influence: temperature, relative humidity, light. Stability tests are performed at different stages of development and production. In accordance with the EMEA (European Agency for Drugs and Medical Devices) for a variety of herbal preparations different specific stability tests are conducted., Danas, sve je veći akcenat na upotrebi preparata na bazi lekovitog bilja (fitopreparata), kako u prevenciji tako i u lečenju različitih oboljenja. Biljni preparati su prepoznati kao odličan izvor bioaktivnih komponenti, koji imaju pozitivne efekte na zdravlje ljudi. Pored čajeva, kao standardnih biljnih proizvoda, danas su na tržištu mnogo više zastupljeni fitopreparati kao finalne farmaceutske forme. Fitopreparati moraju biti standardnog kvaliteta, što podrazumeva da moraju biti fizički, hemijski i mikrobilološki stabilni, da su standardizovanog sastava i da su visokog stepena čistoće. U cilju utvrđivanja koliko je jedan proizvod stabilan, definisanja uslova čuvanja, određivanja roka trajanja, sprovode se ispitivanja stabilnosti, koja podrazumevaju ispitivanja uticaja faktora okoline: temperature, relativne vlažnosti vazduha, svetlosti na promenu kvaliteta gotovog proizvoda. Ispitivanja stabilnosti se izvode u različitim fazama razvoja i proizvodnje. U skladu sa zahtevima EMEA (Evropska agencija za lekove i medicinska sredstva) za različite biljne preparate postavljaju se različiti specifični testovi stabilnosti.

Dormant buds are recognized as valuable side product of the blackcurrant cultivation. Four blackcurrant varieties cultivated in Serbia, i.e., Ben Sarek, Ometa, Ben Lomond, and Ben Nevis, were evaluated for the content, chemical composition, and antimicrobial activity of their bud essential oils. The oil yields of buds harvested during two different growth periods ranged from 1.2-2.0%, and the variety Ometa had the highest yield among the tested varieties. GC-FID and GC/MS analysis of the oils allowed the identification of eight main components, i.e., -pinene (1.6-5.4%), sabinene (1.9-38.4%), -car-3-ene (13.0-50.7%), -phellandrene (2.9-18.0%), terpinolene (6.6-11.9%), terpinen-4-ol (0.9-6.6%), -caryophyllene (3.8-10.4%), and -humulene (0.2-4.1%). In addition, the similarity degree of the essential-oil compositions of buds harvested from the upper and lower parts of the shrubs was investigated by hierarchical clustering. All essential oils originating from the same genotype were grouped in the same cluster, indicating the reliability of essential oils as chemotaxonomic markers. For more detailed chemotaxonomic investigations, the three compounds with the greatest variance were chosen, i.e., sabinene, -car-3-ene, and -phellandrene, which proved to be efficient for the variety distinction. Factor analysis showed that the essential-oil composition as chemotaxonomic marker in blackcurrants was more reliable for variety Ben Sarek than for variety Ben Nevis. Moreover, it was demonstrated that the essential oils had very strong inhibitory activity against all tested microorganisms. Fungi were more sensitive than bacteria; indeed their growth was completely inhibited at much lower concentrations. In comparison to commercial antibiotics, significantly lower concentrations of the oils were necessary for the complete inhibition of fungal growth.

U ovoj doktorskoj disertaciji izvršena je karakterizacija suvih ekstrakata cvasti smilja, Helichrysum plicatum DC. i ispitana njihova antioksidativna, citotoksična, spazmolitička i antimikrobna aktivnost. Posebna pažnja posvećena je pronalaženju korelacija između hemijskog sastava i farmakoloških aktivnosti suvih ekstrakata u cilju utvrđivanja mogućih aktivnih supstanci ili aktivnih markera. U okviru preformulacionih ispitivanja, biljna droga cvasti smilja - Helichrysi flos (biološki izvor Helichrysum plicatum DC.) je precizno definisana, identifikovana i ispitana. Suvi ekstrakti dobijeni su ekstrakcijom usitnjene biljne droge, primenom metode trostruke perkolacije, prečišćavanjem dobijenog tečnog ekstrakta metodom reekstrakcije i uparavanjem pod vakuumom. U toku procesa ekstrakcije varirana su tri faktora: stepen usitnjenosti biljne droge, vrsta ekstragensa (40%, 50%, 60% etanol) i vrsta rastvarača za re-ekstrakciju (5:5, 9:1, 100:0 etilacetat:etanol; v:v), na tri nivoa. Razvijena je HPTLC fingerprintna metoda za brzu i pouzdanu identifikaciju glikozida i aglikona flavonoida, prisutnih u biljnoj drogi i ekstraktima. Njihova identifikacija je bazirana na različito obojenim fluorescentnim mrljama uz primenu standarda. Tentativna LC-UV-MS analiza ekstrakata dobijenih re-ekstrakcijom je pokazala prisustvo 18 jedinjenja. Identifikovane su fenolne kiseline i flavonoidi. Flavonoidi su bili zastupljeni sa 4 grupe jedinjenja: flavanoni (naringenin i njegovi derivati), flavoni (apigenin, derivati apigenina i luteolina), flavonoli (kvercetin i kempferol i njihovi derivati) i halkoni (izosalipurpozid). Tentativna LC-UV-MS analiza biljne droge i ekstrakta cvasti smilja pokazala je da ima razlike u njihovim hromatogramima, zato što su pojedine komponente u biljnoj drogi zastupljene u nižim koncentracijama i detektuju se tek nakon procesa prečišćavanja ekstrakta. Takođe, identifikovana je komponenta cinarin, koja do sada nije identifikovana ni u jednoj Helichrysum vrsti. Dobijeni, In this doctoral dissertation, the characterization of dry extracts of everlasting inflorescences, Helichrysum plicatum DC., and assay of their antioxidant, cytotoxic, spasmolytic and antimicrobial activity were carried out. Special attention was paid to investigation of correlation between chemical composition and pharmacological effects of dry extracts to determine potentially active substances or active markers. In the preformulation study, the herbal drug Helichrysi flos (Helichrysum plicatum DC.) was precisely defined, identified and tested. The dry extracts were prepared from powdered herbal drug by triple percolation method and purification of obtained liquid extracts by means of re-extraction method and vacuum evaporation. The following factors varied in the extraction procedures at three levels: degree of fineness of herbal drug, type of extragens (40%, 50%, 60% ethanol/water ratio) and re-extraction solvent (5:5, 9:1, 100:0 ethilacetate/ethanol ratio; v/v). The HPTLC fingerprint method was developed for rapid and reliable identification of glycosides and aglicone flavonoids present in the herbal drugs and extracts H.plicatum. This identification was based on the color fluorescence, and application of the standards. The tentative LC-UV-MS analysis of purified extracts revealed the presence of 18 compounds. Phenolic acids and flavonoids were also identified. As for flavonoids, there were four groups of compounds present: flavanones (naringenin and derivates), flavones (apigenin and derivates, luteolin), flavonols (quercetin and derivates; kaempeferol and derivates) and halcones (isosalipurpozide). The tentative LC-UV-MS analysis of drug and extract of everlasting inflorescences showed the difference between chromatograms of herbal drugs and extracts, because some compounds in herbal drug were present in small amounts and their presence was detected after extract purification process. A cynarin component was also identified that has not been identified in any