Introduction: The aim of this study was to find the best conditions for the extraction of rutin and quercetin from Sophora flower in acacia honey as an extractant, and to formulate a new product - honey with added value. Methods: For determination of the optimal extraction conditions of the above mentioned compounds, 10% m/m of the dried Sophora flower was added to honey and the concentration of rutin and quercetin was monitored, at 4 degrees C and room temperature at 1, 15, 40 and 90 day intervals. The analyses of rutin and quercetin content in honey were performed by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. The content of rutin and quercetin in the flower was also determined to check the quality of the herbal drug used, as well as to evaluate the effectiveness of the extraction. Results: The extraction of rutin and quercetin, in a honey sample using 10% of the Sophora flower, stored at room temperature, for 40 days, was the most intense one (150.24 mg/kg of rutin and 1338.93 mg/kg of quercetin). Sophora flower corresponds to the pharmacopoeia requirements for the quality of an herbal drug. The concentrations of rutin and quercetin in acacia honey were 0.08 and 0.14 mg/kg, respectively. The concentration of rutin in honey from the melliferous pasture of the Sophora flower was 0.40 mg/kg, while quercetin was below the detection limit (0.1 mg/kg). Conclusions: Honey, enriched with rutin and quercetin from Sophora flower, had sensory acceptability and pleasant taste and aroma.

The usage of hawthorn preparations has a long history in the traditional treatment of heart failure, which is considered as the inability of a weakened heart to supply peripheral tissues with the required amount of oxygen and nutrients. The leaf, flower, and fruit of the hawthorn, due to the presence of bioactive compounds such as flavonoids, procyanidins, organic acids, amines, etc., are showing a wide range of the effects on the cardiovascular system. The leaf and flower have a positive monograph in Commission E, as well as the EMA monograph, but only for traditional use. Numerous clinical studies have been conducted to evaluate the beneficial effects of dry hawthorn extracts on heart diseases. There are a large number of phytopreparations in various galenic forms on the market trade. According to the recent guidelines, usage of hawthorn preparations as a monotherapy in heart failure is not recommended. Instead, it should be used as an adjunct therapy.

Lately, in the group of medicinal herbs, a significant place in the prevention and treatment of certain diseases is taken by Aronia, black chokeberry, Aronia melanocarpa (Michx.) Elliot, Rosaceae. Aronia is a rich source of phenolic substances, mainly flavonoids from the anthocyanin subclass which are responsible for its high antioxidant activity. In addition to the above mentioned compounds, in this plant species heavy metals such as lead and cadmium can be found in the extraction. The aim of this paper was to determine the content of Zn, Cd, Cu, and Pb in samples of fresh Aronia berries, in the juice obtained by cold washing of fresh berries, as well as in the plant material that is left behind after squashing. To determine the content of these metals in these samples, the potentiometric stripping analysis (PSA) with oxygen as an oxidant was used. Zinc and copper biometals were detected in the tested samples of Aronia (berry, juice and the rest of the Aronia berry after squeezing). The content of Zn determined ranged within the limits prescribed for this biometal, while in the tested samples the content of Cu was higher than those prescribed as recommended daily doses. In the tested Aronia samples, heavy toxic metals, lead and cadmium were also detected. The contents of these metals were low, and significantly below the content that could have harmful and toxic effects on human health. All results were confirmed by parallel ICP-OES comparative analysis . The low values for the average relative deviation (delta) indicate that there is a good agreement between the results for the content of all detected metals (Zn, Cd, Pb and Cu) obtained using the PSA technique with oxygen as an oxidant, and the ICP-OES technique.

The paper presents the necessary analytical and technological procedures for the formulation of a new product, refreshing non-alcoholic beverage, with extracts of medicinal plants. The possibility of using ethanol-aqueous extracts of Satureja kitaibelii, Origanum vulgare, Nepeta nuda and Hyssopus officinalis for the preparation of the beverage was analysed. The base of refreshing non-alcoholic beverage was an aqueous solution with 7.0% sucrose and 0.2% citric acid. The volatile fractions of the extracts were analysed using GC-FID/GC-MS. The content of total phenols and flavonoids was determined by the spectrometric methods. Antioxidant activity was assessed by FRAP and DPPH test. Antimicrobial activity of the extracts was analysed by broth microdilution test. The extracts of O. vulgare and S. kitaibelii had the highest content of total phenols and flavonoids (129.2 +/- 12.71 and 90.0 +/- 10.56 mg GA/g DE; 110.7 +/- 2.47 and 63.3 +/- 0.58 mg RU/g DE, respectively), enhancing antioxidant activity (EC50= 4.77 +/- 0.49 and EC50=18.85 +/- 0.31 mu g/mL, respectively) and antimicrobial activity. Refreshing non-alcoholic beverage with Oregano extract at a concentration of 1 g/L proved to be the best combination, since it exhibited the highest antioxidant and antimicrobial activity, and sensory was the most acceptable. The formulated product had a pleasant, attractive and harmonious taste and aroma, as well as additional health benefits for consumers.

Isolates of medicinal and aromatic herbs are used as additives in non-alcoholic drinks to improve the sensory characteristics and antioxidant potential. The method of drying the plant material, among other factors, has a profound influence on the chemical composition and pharmacological activities of plant extracts. This paper analyzes the effect of the drying technique (I - natural drying, II - in the laboratory oven, III - in low-temperature condensational drier) on the volatile fraction of the wild mint (Mentha longifolia (L.) Hudson) extract, in order to select an extract of the best quality for preparation of non-alcoholic drinks. The volatile profile of the extracts was determined by the GC-FID and GC-MS techniques, the antimicrobial activity by the microdilution technique, the antioxidant activity by the DPPH and FRAP assays, and the sensory acceptance according to the Regulation on quality for refreshing non-alcoholic beverages. The results showed that piperitone was the major component of the volatile fraction extract of the natural drying herb and low-temperature condensational drier herb (53.9% and 61.1%, respectively), while the extract of laboratory oven herb was rich in menthone (35.5%). At the concentrations in the range of 0.8-3.2 mg/mL the extracts better inhibited the Gram (+) bacteria. The beverage to which the extract of naturally dried wild mint was added, showed the antioxidant activity (9.09 +/- 0.17 mu mol Fe2+/mL by FRAP and 14.00 +/- 3.00 mu L/mL by DPPH method) and good sensorial characteristics (concentration of the extract 0.8 g/L).

This study was aimed to compare the efficiency of silymarin extraction from Silybi mariani fructus using different extraction methods. Maceration, percolation, extraction in a water bath, and ultrasonic-assisted extraction have been performed with 60% ethanol. Extracts were analyzed using UV/Vis and HPLC techniques. Our results showed that the highest concentration of silymarin was detected in samples extracted in a water bath with boiling solvent during 30 and 60 minutes (2.33 and 1.95 %, respectively). Since extraction in a water bath is efficient and reduces a lot of time and solvent consumption, this method could present potential replacement for Pharmacopoeia-recommended Soxhlet extraction procedure.

This paper is based on determining the level of the antioxidant activity of beer, to which sensory acceptable amounts of selected extracts of medicinal plants were added, with the aim of obtaining a beer with increased functional and new sensory features. For purposes of this study a commercial lager beer type Pils and extracts of herbal drugs: Melissae folium, Thymi herba, Juniperi fructus, Urticae radix and Lupuli strobuli, were used. Total phenols were analyzed by the method of Folin-Ciocalteu, and the antioxidant activity of samples using FRAP and DPPH test. Sensory evaluation of beer was conducted on 80 subjects, using a nine levels hedonic scale. The results showed that the content of total phenols was the highest in the beer which thyme, juniper and lemon balm were added to (384.22, 365.38 and 363.08 mg GAE/L, respectively), representing the increase of 37.09, 30.36 and 29.55% (respectively) compared to the commercial lager beer. Values of antioxidant activity were correlated with the content of total phenols. The extract of lemon balm blended in the best manner with the baseline, commercial lager beer in terms of sensory acceptability. New beer, enriched with lemon balm, had a pleasant, appealing and harmonious flavor and aroma.

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).

There is a growing research interest on investigation of appropriate alternatives of natural origin to synthetic additives used in food products. The aim of this study was to investigate correlation of chemical composition of the best known biomarker plants from the locality of Vrujci Spa, Serbia with their antioxidant activity in vitro, in terms of their potential usage as feedstock materials in the food industry. Total phenolic (TP), total tannins (TT) and total flavonoid (TF) contents were determined using spectrophotometric methods. Qualitative and quantitative analyses of major phenolics by high-performance liquid chromatography (HPLC) were also used. Major phenolic acids identified in analyzed species were chlorogenic, caffeic and ferulic acid, while predominant flavonoids were flavonols (hyperoside, rutin, isoquercitrin, quercetin) and, in lesser extent flavones (luteolin, apigenin and their 7-O-glucosides). Vitexin 2-O″-rhamnoside was found only in Crataegus species. Antioxidant capacity was estimated by the following methods: 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) test and ferric reducing antioxidant power (FRAP). Majority of investigated plants had high levels of phenolics and exhibited noteworthy antioxidant activity. Moreover, a positive linear correlation was found between TP and TT with antioxidant capacities regarding both methods used. The plants showing the highest antioxidant activities were Veronica officinalis, Mentha pulegium and Fragaria vesca. Hypericum perforatum demonstrated by far the most potent antioxidant activity, as well as TP and TT content. Our results indicate that, in addition to their traditional usage in folk medicine, these plants represent a valuable source of natural antioxidants, and thus may be considered as great potential for the food industry, representing possible alternatives to synthetic additives.

The way of drying the fresh herbal material influences the chemical content and the biological activities of their essential oils. The influence of the different drying methods of the herb Mentha longifolia (L.) Hudson on the antioxidant and antimicrobial activity of the extracted essential oils has been analyzed in this study. Drying has been carried out in three ways: in the natural way, in the laboratory oven (45 degrees C) and in the absorptional low-temperature condensational drier (35 degrees C). The antioxidant activity of the essential oil has been estimated by FRAP and DPPH assays, while the antimicrobial activity has been estimated by the diffusible and micro-delusional method, testing on the nine types of bacteria and two types of fungi. The essential oil obtained from the herb dried in the natural way has shown the highest antioxidant activity and the lowest from the herb dried in the laboratory oven. Bacillus subtilis, Micrococcus luteus and Enterococcus faecalis have shown the highest sensitivity on the three samples. The oil obtained from the herb dried in the absorptional low-temperature drier has shown the strongest antimicrobial effect.

SUMMARY Medicinal herbs and their mixtures, which are widely used for prevention and treatment of some disease, can also present health risks due to the presence of toxic metals such as Pb and Cd. The application of different agrotechnical practices during plant growing season, as well as the process of circulation of substances in nature, may be the cause of plant contamination. The aim of this study was to determine the content of lead, cadmium and copper from soil which are bioavailable for absorption by plant, as well as the total content of these metals in some medicinal herbs that were cultivated in two different locations. The presence of these metals in the samples was detected using highly sensitive microanalytical technique potentiometric stripping analysis (PSA). The mean value of lead total content in the analyzed samples of medicinal herbs Chamimillae flos, Urticae folium, Menthae folium, Altheae radix and Basilici herba, which were grown at the sites near the industrial zone, was about 1.55 μg/g, 1.82 μg/g, 1.90 μg/g, 1.99 μg/g and 2.74 μg/g, respectively. Contrary to this, the total content of this toxic metal in the analyzed plant samples grown on rural areas was detected only on some sites. Based on the results of this study, it can be concluded that medicinal herbs contained a certain amount of lead and that its content varied depending on the location at which the plant species were grown as well as on plant affinity to certain metal. Cadmium and copper were not detected in the tested plant material.

SUMMARY In this study we investigated the effects of four emollients (cetearyl octanoate, decyl oleate, C12-15 alkyl benzoate and olive oil) incorporated at a concentration of 10% on the structural properties of the cosmetic oil/water (o/w) creams (creams K1-K4, respectively) as well as the in vivo effects of those creams on pH and hydration level of the healthy volunteer’s skin after a single application. Creams’ structure was investigated throughout polarization microscopy. Anisotropic structures were found, i.e. stabilization of creams’ structure was performed by forming lamellar gel-crystalline and liquid-crystalline phase, different for different creams, which was probably caused by the presence of different emollients. The effects of creams (K1-K4) on skin hydration level and pH (30, 90, 150 and 300min after single application) were examined using Multi Probe Adapter (MPA ® 9, Courage& Khazaka Electronic, Germany), with appropriate sonde, Corneometer ® CM 825 for measuring capacitance as a measure of skin moisture level, while the skin pH was measured using Skin-pH-Meter, PH 900 sonde.The results indicate that the presence of the different emollients in o/w creams (10%) led to the creams'different effects on the skin hydration level. Cream with decyl oleate leads to a good initial hydration effect and probably rapid release of large amounts of water from the cream. Creams containing both C12-C15 alkyl benzoate or olive oil had a weaker initial effect on the skin moisture, showing good occlusive properties. A cream containing 10% cetearyl octanoate showed the lowest skin hydration potential.

This study examines the essential oil and the ethanol extract of Artemisia alba Turra herb, in terms of chemical composition, antioxidant and antimicrobial activities in order to select the active pharmacological isolate for making phytopreparation from the group of bitter aromatics and/or bitter liqueurs. Chemical composition of the essential oils and volatile fractions of the extract was determined by GC-FID/GC-MS and non-volatile fractions of the extract by HPLC. The content of total phenolics and flavonoids in the extracts was determined by Folin-Ciocalteu reagent and aluminum chloride, respectively. The antioxidant activity of isolates was tested by FRAP and DPPH tests and the antimicrobial by microdilution method. Major constituents in the essential oil were camphor, artemisia ketone and 1,8-cineole (23.7%, 15.2% and 14.1%, respectively), and in the volatile fraction of the extract, scopoletin (14.0%) and corymbolone (10.3%). The main components in non-volatile fraction of the extract were kaempferol 3-O-(6 "-O-malonylglucoside)- 7-O-rhamnoside, chlorogenic acid and rutin (16.1%, 11.4% and 9.5% respectively). The content of total phenolics in the extract, expressed as gallic acid was 77.18 mg/g and flavonoids, expressed as rutin 53.80 mg/g of dry extract. Extract exhibited stronger antioxidant activity than essential oil and better antimicrobial activity on a larger number of tested microorganisms.

Introduction. Rational phytotherapy is a modern concept of using plant-originated drugs which has emerged from the need to improve phytotherapy in order to make the use of herbal remedies more efficient and safer. The aim of this study was to give the health-care workers more information on the manufacturing process of high quality phytopreparation following principles of Good Manufacturing Practice and Good Laboratory Practice on the example of herbal sedative, Odoval S? capsules. Material and Methods. This study was designed to reflect the production process of a high-quality and safe herbal remedy, starting from defining the formulation and the production procedure to the quality control of raw materials, characterization of the final product, and testing stability of active ingredients in the capsules. Results. Formulation of the phytopreparation, validation of the production process, quality control and stability testing, all together have resulted in the production of capsules with defined valeric acid content (1 mg valeric acid per capsule). Discussion. The preparation is recommended to relieve the symptoms caused by chronic stress (anxiety, irritability, fatigue, lack of concentration, heart palpitations) and for mild insomnia. Conclusion. This paper presents the complete cycle of the production of a phytopreparation on the example of a new herbal sedative - Odoval S? capsules.

The purpose of this study was to examine the chemical composition of the es­sential oil of Thymus serpyllum L. collected on the slopes of Kopaonik and to compare it with the results of the analysis of essential oils of the species which originate from other parts of Europe. The isolation of the essential oil was carried out through the process of hydrodistillation in the Clevenger type apparatus while the chromatographic analysis of the essential oil was carried out by using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). 26 constituents representing 98.4 % of the detected components were identified out of 27 registered constituents. Sesquiterpenes are a dominant class of the compounds in tested essential oils with trans-caryophyllene as the principal component. The content of identified monoterpenes was 37, 9 % (α-pinene as the main component), sesquiterpenes 60,5 % (trans-caryophyllene, γ-muurolene and α-humulene as the major constituents). Apart from dominant constituents, trans-caryophyllene, γ-muurolene and α-humulene, camphor (3.6 %), limonene (2.7 %) and β-bisabolol (2.6 %) were also found in larger quantities.

In this study, we have examined the yield of extracted substances obtained by means of extraction using 70 % ethanol (v/v), the content of total phenols and flavonoids, as well as the antioxidant activity of the extracts obtained from the samples of the herbs dried by means of different techniques. Wild mint Mentha longifolia (L.) Hudson was dried naturally in a laboratory oven at a temperature of 45 °C and in an absorptive low temperature condensation oven at 35°C. The highest yield of extracts was obtained from the naturally dried herbs and the lowest from the herbs dried in the low temperature condensation drying oven. The content of total phenols and flavonoids was determined by spectrophotometric methods with an FC reagent and by the complexation reaction with aluminium-chloride, respectively. The extract of the naturally dried herbs had the highest overall content of phenols (113.8±2.0 mg of gallic acid/g of the dry extract) and flavonoids (106.7±0.3 mg of rutin/g of the dry extract). The highest antioxidant activity determined by the FRAP and DPPH assay was determined in the extracts obtained from naturally dried herbs (2.76±0.15 mmol Fe2+/mg of the dry extract and EC50=0.022±0.001 mg/ml), while the lowest was obtained from the extracts of herbs dried in the laboratory oven (1.13±0.11 mmol Fe2+/mg of the dry extract and EC50=0.033±0.001 mg/ml). The HPLC-DAD analysis result show that the greatest content of phenolic compounds show extract obtained from naturally dried plant material. The dominant phenolic component in the all extracts is Kaempferol 3-O-glucoside. The content of all phenolic compound strongly depend on the drying conditions.

Sideritis scardica Griseb., Lamiaceae (ironwort, mountain tea), an endemic plant of the Balkan Peninsula, has been used in traditional medicine in the treatment of antimicrobial infections, gastrointestinal complaints, inflammation and rheumatic disorders. This study reports a comparison between conventional (hydrodistillation HD and solvent extraction SE) and alternative (supercritical carbon dioxide SC CO2) extraction methods regarding the qualitative and quantitative composition of the obtained extracts as analyzed by GC and GC-MS techniques and their anitimicrobial activity. Different types of extracts were tested, the essential oil EO obtained by HD, EO-CO2 and AO-CO2 obtained by SC CO2 at different preasures 10 and 30 MPa, at 40 °C, respectively, and the fractions A, B, C and D obtained by successive solvent extraction (SE) A: ethanol, B: diethyl ether, C: ethyl acetate and D: n-butanol). While EO was characterized by the presence of the high percentage of oxygenated monoterpenes and sesquiterpenes (30.01 and 25.54%, respectively), the rest of the investigated samples were the most abundant in fatty acids and their esters and diterpenes (from 16.72 to 71.07% for fatty acids and their esters, and from 23.30 to 72.76%, for diterpenes). Microbial susceptibility tests revealed the strong to moderate activity of all investigated extracts against the tested microorganisms (MIC from 40 to 2,560 μg/mL). Although differences in the chemical compositions determined by GC and GC-MS analysis were established, the displayed antimicrobial activity was similar for the all investigated extracts.

The present study describes investigation on chemical profile and pharmacological properties of hydromethanolic extracts of Carlina acaulis and C. acanthifolia roots and herbs. In order to investigate the antioxidant properties several methods were applied: FRAP (Ferric Reducing Antioxidant Power) assay, TBA assay (Fe2+/ascorbic induced lipid peroxidation) and scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and OH radicals. In assessing anti-inflammatory activity the carrageenan-induced rat paw oedema test was used, with indomethacin as a reference drug. Anti-ulcer activity of the extracts was investigated using ethanol-induced stress ulcer in rats and ranitidine as a reference drug. Antimicrobial activity was tested on five bacterial strains and one strain of fungi using the agar diffusion method. Investigated extracts exhibited considerable dose-dependent antioxidant, anti-inflammatory and anti-ulcer activities. Extracts of herbs were more active than the corresponding root extracts. All extracts exhibited moderate activity against all tested microorganisms. Herb extracts of C. acaulis and C. acanthifolia, as well as their root extracts exhibited similar bioactivity, which could be explained with their similar chemical composition revealed by HPLC analysis. The pharmacological activities of investigated Carlina extracts demonstrated here will serve as the basis for their further investigations in order to estimate their impact on human health and nutrition.

According to Erg. B. 6, C. acaulis , is the biological source of Carlinae radix . The root is traditionally used in the treatment ofcatarrh of upper respiratory tract, as diuretic, stomachic, diaphoretic and tonic. Externally it was used for the treatment of different skin conditions. Though not described in any modern pharmacopoeia, C. radix is present in the modern market of medicinal plants. Drug on domestic and international markets only partly come from the species C. acaulis , and in major part from the species of C. acanthifolia . Recent studies have shown that because of the very similar chemical compositions of C. acaulis and C. acanthifolia root essential oils and their respective methanol root and herb extracts, they exhibit similar pharmacological activities (antimicrobial, antioxidant, anti-inflammatory and gastroprotective).The taxonomic classification of the genus is very interesting. The Serbian Flora lists three Carlina species: C. acaulis L., C. utzka Hacq. ( C. acanthifolia All.)and C. vulgaris L. The first two species are biological sources of the drug Carlinae radix, while C.vulgaris has no importance in the traditional medicine. Genus Carlina in the European Flora, is divided into two subgenus ( Lyrolepis and Carlina ), which include 13 species with many subspecies.According Meusel and Kastner, genus includes five subgenus ( Carlowizia , Lyrolepis , Mitina , Heracantha and Carlina ) with 28 species. In the Serbian Flora infraspecific classification of C. acaulis is not shown. According to the European flora the species includes two subspecies: ssp. acaulis i ssp. simplex . According to Meusel-Kastner the species C. acaulis is divided into: ssp. acaulis (var. acaulis , var. alpina ), ssp. caulescens . Our flora does not indicate infraspecific classification of C. utzka Hacq. (syn. C. acanthifolia All.). According to the Flora of Europe, C. acanthifolia ,isdivided into: ssp. acanthifolia ( C. utzka ) and ssp. c ynara , and Meusel and Kastner: ssp. acanthifolia , ssp. utzka and ssp. c ynara . According the literature mentioned above the classification of C.vulgaris is diferent. Based on the morpho-anatomical characteristics of the genus Carlina represented in our flora ( C. acaulis , C. utzka , C. vulgaris ), they can be easily and reliably differentiated.