Evaluation of Anticancer and Antioxidant Activity of a Commercially Available CO2 Supercritical Extract of Old Man's Beard (Usnea barbata)

There is a worldwide ongoing investigation for novel natural constituents with cytotoxic and antioxidant properties. The aim of this study was to investigate chemical profile and stated biological activities of the supercritical CO2 extract (SCE) of old man's beard compared to the extracts obtained using the conventional techniques (Soxhlet extracts and macerate). The most abundant compound identified was usnic acid, which content was inversely proportional to the polarity of the solvent used and was the highest in the SCE, which was the sample revealing the highest cytotoxic activity in tested tumor cell lines (B16 mouse melanoma and C6 rat glioma), with lower IC50 values compared to pure usnic acid. Further investigations suggested both SCE and usnic acid to induce apoptosis and/ or autophagy in B16 and C6, indicating higher cytotoxicity of SCE to be related to the higher degree of ROS production. A good correlation of usnic acid content in the extracts and their antioxidant capacity was established, extricating SCE as the most active one. Presented results support further investigations of SCE of old man's beard as a prospective therapeutic agent with potential relevance in the treatment of cancer and/ or in oxidative stress-mediated conditions

Physicochemical characterization of mineral mud from spa Vrujci

Mineralogical and physicochemical analysis of healing mud from spa Vrujci was performed by ICP-OES analysis and FTIR and micro-Raman spectroscopy. The therapeutic effect of mud was improved by plant extracts implementation

Sadržaj ulja divljih plodova različitih biljnih vrsta dobijen konvencionalnom Soxhlet tehnikom ekstrakcije

Oils from wild fruits have gained attention due to the presence of the wide variety of natural bioactive lipid components (essential polyunsaturated fatty acids, phytosterols, lipid-soluble vitamins), which have positive effects on human health. The present study describes the extraction of oils from fruits of different plant species, Sambucus nigra, Prunus spinosa, Sorbus aucuparia and Rosa canina, using the conventional Soxhlet extraction technique with petroleum ether. Before the extraction, mechanical processing of fruit samples (exposure to elevated pressure and temperature) was performed. The yield of oil (%) was expressed as mass of obtained oil per mass of dry plant material (w/w). Moisture content of fruit samples was also determined. Wild fruits showed different moisture contents, ranging from 6.53 to 71.52 g/100 g of fresh weight, in R. canina and S. aucuparia fruits, respectively. S. nigra and P. spinosa fruits revealed similar moisture contents, 68.48 g/100 g and 68.89 g/100 g of fresh weight, respectively. Oil yields of the investigated fruit samples showed statistically significant differences (p lt 0.05) and ranged from 0.38 to 3.22% (w/w) in the R. canina and P. spinosa fruits, respectively. Dry fruits of S. nigra had 2.55% (w/w) of oil, while the oil content in S. aucuparia fruits was 0.68% (w/w). The findings obtained from this study might be useful from the aspect of potential utilization of these wild fruits, in particular fruits of P. spinosa and S. nigra, as raw materials for the extraction of valuable oil for the application in food, cosmetic and pharmaceutical industry.Ulja dobijena iz plodova biljaka sve više dobijaju na značaju, upravo zbog prisustva velikog broja prirodnih, bioaktivnih lipidnih komponenata (esencijalnih, polinezasićenih masnih kiselina, fitosterola, liposolubilnih vitamina), koji ispoljavaju pozitivne efekte na ljudsko zdravlje. U ovom radu opisan je postupak ekstrakcije ulja iz plodova različitih biljnih vrsta (Sambucus nigra, Prunus spinosa, Sorbus aucuparia i Rosa canina), primenom konvencionalne Soxhlet ekstrakcione tehnike sa petrol-etrom. Pre ekstrakcije izvršena je mehanička obrada uzoraka plodova, njihovim izlaganjem povišenom pritisku i temperaturi. Prinos ulja (%) je izražen kao masa dobijenog ulja u odnosu na masu suvog biljnog materijala (w/w). Određen je i sadržaj vlage u uzorcima plodova. Divlji plodovi su se razlikovali po sadržaju vlage, čije su vrednosti bile u granicama od 6,53 g/100 g svežih plodova (R. canina) do 71,52 g/100 g svežih plodova (S. aucuparia). Biljne vrste S. nigra i P. spinosa pokazale su sličan sadržaj vlage u plodovima, i to 68,48 g/100 g i 68,89 g/100 g svežih plodova. Prinosi ulja ispitivanih uzoraka plodova razlikovali su se statistički značajno (p lt 0,05), a vrednosti su bile od 0,38% (kod R. canina plodova) do 3,22% (plodovi P. spinosa). Kod suvih plodova S. nigra dobijeni prinos ulja je iznosio 2,55%, dok je sadržaj ulja u plodovima S. aucuparia iznosio 0,68%. Dobijeni rezultati su korisni zbog mogućnosti upotrebe ispitivanih divljih plodova, naročito plodova P. spinosa i S. nigra (imale su najveći sadržaj ulja), kao polaznih sirovina za ekstrakciju biljnog ulja za primenu u prehrambenoj, kozmetičkoj i farmaceutskoj industriji

Supercritical and high pressure subcritical fluid extraction from Lemon balm (Melissa officinalis L., Lamiaceae)

Analysis of yield and composition profile of Melissa officinalis (Lemon balm) extract obtained by two-step CO2 extraction at high pressure was analyzed. The first or essential oil fraction, collected at 10 MPa and 40 degrees C, contained mostly the high volatile and aromatic compounds. The second fraction was consenquently extracted at 30 MPa using different temperature: 25 degrees C (higher pressure than critical pressure but at temperature below the critical; HPCP-CO2), 40 degrees C or 100 degrees C (supercritical CO2; SC-CO2). Obtained yield of essential oil fraction was 0.45% at 10 MPa, while depending on applied extraction temperature (HPCP-CO2 or SC-CO2), the yields of the second fractions collected at 30 MPa varied from 0.44% to 0.94%. Within the study, the content of active substances in obtained extracts was analyzed, too. For comparison with CO2 extracts (HPCP or SC) the yield and characterization of essential oil obtained by hydrodistillation was accomplished, as well. A significant difference was noted not only between the essential oil isolated by hydrodistillation and SC-CO2 extraction (40 degrees C, 10 MPa) but also between total extracts obtained by HPCP-CO2 and SC-CO2 extraction. The pseudo kinetic model was applied to define SC-CO2 extraction of Lemon balm to describe the relation between the yield of extraction and CO2 consumption. Antioxidant activity of obtained extracts was determined by free radical scavenging capacity (RSC) and significant correlation of antioxidant capacity and composition profile between obtained extracts was observed

Optimizacija ekstrakcije antioksidantnih komponenata iz herbe hajdučke trave

Recently, research efforts have been directed toward medicinal plants and their extracts, as important sources of natural antioxidant. Lots of biologically active compounds are responsible for the antioxidant effects of yarrow - Achillea millefilium L. extracts. The aim of our study was to determine which of the process parameters of pressure enhanced solvent extraction of Millefolii herba is significant for its efficiency and weather there are interactions between the examined parameters. Compression time, decompression time and the number of cycles were identified as independent variables, while the content of total flavonoids, tannins and total polyphenols were selected as dependent variables. For obtaining the extract of M. herba, rich in antioxidative ingredients, compression time should be set on its higher level (2.0 min), decompression time on its lower level (1.30 min) and the number of cycles on its higher level (99).Oksidativna degradacija sastojaka hrane dovodi do promene njenog mirisa i ukusa, čime se narušava njen nutritivni i senzorni kvalitet. Iz navedenog razloga je neophodno dodavati antioksidante tokom tehnološkog procesa izrade namirnica. Lekovito bilje i njihovi izolati predmet su istraživanja, pored ostalog i kao izvor antioksidantnih materija i alternativa sintetičkim antioksidansima za primenu u industriji hrane. Istraživanja velikog broja biljaka ukazala su da skoro sve poseduju određena antioksidativna svojstava. Hajdučka trava - Achillea millefilium L. (Asteraceae) i njeni ekstrakti, zahvaljujući prisustvu, pre svega, polifenola, flavonoida i tanina, ispoljavaju antioksidativni efekat. Sadržaj ovih materija u izolatima zavisi od njihove koncentracije u polaznom biljnom materijalu ali i od načina/uslova ekstrakcije. Cilj našeg ispitivanja bio je da utvrdimo optimalne vrednosti procesnih parametara za izvođenje pritiskom ubrzane ekstrakcije iz herbe hajdučke trave (Millefolii herba), radi dobijanja ekstrakata bogatih antioksidativnim materijama, kao i da utvrdimo zavisnost koja postoji između procesnih parametara. Ekstrakcija je obavljena u Timatic mikro ekstraktoru. Takozvano vreme kompresije, vreme dekompresije i broj ciklusa kruženja ekstrakcionog sredstva predstavljaju nezavisno promenljive veličine, dok su sadržaj ukupnih flavonoida, tanina, ukupnih polifenola kao i indeks refrakcije, suvi ostatak, relativna gustina, sadržaj etanola i pH vrednost zavisno promenljive veličine. Vreme kompresije bilo je 1 odnosno 2 minuta, vrednost vremena dekompresije 1,3 i 2 min, dok su ekstrakcije obavljene uz minimalno 20 odnosno maksimalno 99 ciklusa. Kao ekstragens korišćen je 70% (V/V) etanol. Droga: ekstrakt odnos iznosio je 1:3. Radni pritisak kretao se u opsegu od 1 do 99 bar. Primenjen je 23 faktorski dizajn, odnosno urađeno je osam ekstrakcija herbe hajdučke trave. Statistička analiza eksperimentalnih rezultata izvršena je upotrebom softvera Design Expert 7.0. U cilju potpune optimizacije primenjena je multivarijantna metodologija upotrebom Derringer funkcije poželjnih odgovora. Proces optimizacije postupka ekstrakcije pokazao je da je za izradu ekstrakata herbe hajdučke trave bogatih sadržajem antioksidativnih materija (flavonoidi, tanini i polifenoli), potrebno podesiti vreme kompresije na najviši nivo (2,0 min), vreme dekompresije na najniži (1,30 min) i vršiti ekstrakciju uz maksimalni broj ciklusa (99)

Usnea barbata CO2-supercritical extract in alkyl polyglucoside-based emulsion system: contribution of Confocal Raman imaging to the formulation development of a natural product

Topical treatment of skin infections is often limited by drawbacks related to both antimicrobial agents and their vehicles. In addition, considering the growing promotion of natural therapeutic products, our objective was to develop and evaluate naturally-based emulsion system, as prospective topical formulation for skin infections-treatment. Therefore, alkyl polyglucoside surfactants were used for stabilization of a vehicle serving as potential carrier for supercritical CO2-extract of Usnea barbata, lichen with well-documented antimicrobial activity, incorporated using two protocols and three concentrations. Comprehensive physicochemical characterization suggested possible involvement of extract's particles in stabilization of the investigated system. Raman spectral imaging served as the key method in disclosing extract's particles potential to participate in the microstructure of the tested emulsion system via three mechanisms: (1) particle-particle aggregation, (2) adsorption at the oil-water interface and (3) hydrophobic particle-surfactant interactions. Stated extract-vehicle interaction proved to be correlated to the preparation procedure and extract concentration on one hand and to affect the physicochemical and biopharmaceutical features of investigated system, on the other hand. Thereafter, formulation with the best preliminary stability and liberation profile was selected for further efficiency and in vivo skin irritation potential evaluation, implying pertinent in vitro antimicrobial activity against G+ bacteria and overall satisfying preliminary safety profile

Pasji trn - potencijalni sastojak funkcionalne hrane

Since nowadays the link between nutrition and health has been established, the industry of food and pharmaceuticals developed a new category- functional food. Medicinal plants became significant for production of functional food, because the biological activities. Sea buckthorn - Hippophae rhamnoides L., is a shrub with fruits that are false, soft and flashy berries. Fresh ripe fruits are commonly used as a rich source of vitamin C and B and fatty oil from fruits and seeds, which is a significant source of carotenoids, vitamins A, D, E and essential fatty acids (EFA). The aim of this paper is to estimate commercially available sample of sea buckthorn as potential constituent of functional food by quantification of EFA and total tocopherols content. The dominant components are oleic and linoleic acid (22,9% and 35,4%), while total tocopherols were 88,6mg/100g. So, tested oil sample could be used in functional food. .Zbog potvrđene povezanosti ishrane i zdravlja, prehrambena i farmaceutska industrija su razvile novu kategoriju proizvoda - funkcionalnu hranu. Lekovite biljke, kao nosioci bioloških aktivnosti, postale su značajne u proizvodnji funkcionalne hrane. Pasji trn - Hippophae rhamnoides L. (Eleagnaceae) je listopadni žbun sa plodovima u obliku mesnate koštunice. Koristi se svež zreo plod bogat vitaminima B i C i masno ulje iz plodova i semena koje sadrži visok procenat karotenoida, vitamina A, D i E i esencijalnih masnih kiselina. Cilj rada je procena kvaliteta komercijalno dostupnog uzorka masnog ulja pasjeg trna kao potencijalne sirovine za funkcionalnu hranu. Određen je sadržaj esencijalnih masnih kiselina (EMK) i tokoferola. Rezultati su pokazali da su dominantne komponente oleinska i linolna kiselina zastupljene u količini od 22,9%, odnosno 35,4%, dok je količina ukupnih tokoferola 88,6 mg/100g. Zbog visokog sadržaja EMK i značajnog tokoferola, ovo ulje se može koristiti kao dodatak u proizvodnji funkcionalne hrane.

KINETIČKO-SPEKTROFOTOMETRIJSKI PRISTUP HIDROLITIČKOJ DEGRADACIJI AMPICILINA PRIMENJEN ZA ODREĐIVANJE HISTIDINA

The objective of this research was to develop a kinetic-spectrophotometric method for the determination of microquantities of L-histidine in pure form and dietary supplements. The method was based on the kinetics of ampicillin degradation by Ni(II) ion as a catalyst in the presence of L-histidine in a strongly alkaline medium. The rate of this reaction was monitored spectrophotometrically by measuring the increase in absorbance at 265 nm as a function of time. The same approach was used for the investigation of the reaction rate in the absence of histidine. A differential variant of the tangent method was used to process the kinetic data. Beer’s law was obeyed in the interval of histidine concentration from 1.24 µg/ml to 11.63 µg/mlwith the relative standard deviation ranging from 8.1% to 0.7%. The detection limit of 0.46 µg/ml was estimated based on the 3S0 criterion. The interference effects of some metal ions, anions, and other molecules on the reaction rate were studied to assess method selectivity. Herein described method was applied for the quantification of histidine in dietary supplements. The point hypothesis test confirmed that there was no significant difference between the proposed and the reference method.Cilj ovog istraživanja je bio razvoj kinetičko-spektrofotometrijske metode za određivanje mikrokoličina L-histidina u čistom obliku i dijetetskim suplementima. Metoda je zasnovana na reakciji degradacije ampicilina koja je katalizovana Ni(II) jonom u prisustvu L-histidina u jako baznoj sredini. Brzina reakcije praćena je spektrofotometrijski merenjem povećanja apsorbance na 265 nm sa vremenom. Isti pristup je korišćen za praćenje brzine reakcije u odsustvu histidina. Diferencijalna varijanta tangensne metode je upotrebljena za obradu kinetičkih podataka. Metoda je zadovoljavala Beer-ov zakon u intervalu koncentracija od 1,24 µg/ml do 11,63 µg/ml sa relativnom standardnom devijacijom od 8,1% do 0,7%. Detekcioni limit od 0,46 µgcm-3 je procenjen korišćenjem 3S0 kriterijuma. Selektivnost metode je ispitana na osnovu uticaja pojedinih metalnih jona, anjona i organskih molekula na brzinu reakcije. Opisana metoda je primenjena za kvantitativno određivanje histidina u dijetetskim suplementima. Primenom t-testa potvrđeno je da nema značajne razlike između rezultata predložene i referentne metode

TYROSINASE INHIBITORY AND ANTIOXIDANT ACTIVITY OF WILD ROSA CANINA L. AND SORBUS AUCUPARIA L. FRUIT EXTRACTS

In the present work, fruits from two plant species, Rosa canina L. and Sorbus aucuparia L., popular in traditional folk medicine in Serbia, were studied. The aim was to examine and compare the efficiency of the ultrasonic extraction with different solvents regarding physicochemical properties, polyphenolic profile of extracts, as well as their tyrosinase inhibitory and antioxidant activity. The polyphenols evaluation indicated that water was the best solvent for a thorough extraction of bioactive compounds from the R. canina fruits, while propylene glycol-water (45:55, v/v) was the most efficient regarding S. aucuparia fruits, followed by ethanol-water (7:3, v/v). Only flavonoids were more abundant in S. aucuparia fruit extracts. R. canina water extracts showed a higher antioxidant activity, using several in vitro tests with different working principles. However, S. aucuparia ultrasonic extracts with propylene glycol-water (45:55, v/v) demonstrated a higher potential concerning tyrosinase inhibitory and chelating activity. Therefore, these ultrasonic extracts, being great sources of natural anti-tyrosinase inhibitors and antioxidants, can be considered as promising candidates suitable for pharmaceutical application, as great sources of natural anti-tyrosinase inhibitors and antioxidants