Search

The application of high performance thin layer chromatography (HPTLC)-based non-targeted metabolomics as a holistic approach to compare fingerprints of metabolite changes during Artemisia annua development is described. Images of HPTLC chromatograms obtained after derivatization with anisaldehyde-sulphuric acid reagent were used as a dataset for multivariate analysis. Principal component analysis and orthogonal partial least squares discriminant analysis confirmed the differentiation of samples belonging to vegetative phase, flowering stage, and seed formation stage of the plant development. The obtained results showed that the HPTLC-based metabolomics approach can be a very reliable technique for the investigation of metabolic changes during plant development, complementary to gas chromatography–mass spectrometry and nuclear magnetic resonance-based metabolomics.

Eleven 31-methylenlanostane triterpenoids, i.e., seven 21- and four 26-oic acids, as well as a lupane triterpenoid betulin, isolated from the fruiting bodies of the mushroom Fomitopsis betulina, were tested for in vitro protective effect on chromosome aberrations in peripheral human lymphocytes using cytochalasin-B blocked micronucleus (CBMN) assay. Most of the tested compounds showed a beneficial effect by reducing DNA damage of human lymphocytes more effectively than amifostine, a radioprotective agent, used as a positive control. All the tested compounds decreased MN frequency in the concentration dependent manner, with the concentration of 2.0 μg mL-1 being the most effective – with increase of the concentration the activity slightly decreases. The structure–activity relationship (SAR) studies indicated that the lanostanes containing a conjugated 7,9 (11)-diene system exhibit lower activity than 8-analogues. It was also demonstrated that the DNA protective activities within the 8-lanostane-26-oic acid group are affected by the substitution in position 3 pattern. In the 8 series the oxygenation at C-12 or 16 as well as 21- or 26-oic acid functionality proved beneficial for in vitro protective effect on chromosomal aberrations. Betulin exhibited the lowest protective activity, but it is still comparable to that of amifostine.

Euphorbia seguieriana ssp. seguieriana Necker (ES) and Euphorbia cyparissias (EC) with a habitat in the Deliblato Sands were the subject of this examination. The latexes of these so far insufficiently investigated species of the Euphorbia genus are used in traditional medicine for the treatment of wounds and warts on the skin. To determine their chemical composition, non-targeted screening of the latexes' chloroform extracts was performed using liquid chromatography coupled with quadrupole time-of-flight mass spectrometry employing an electrospray ionization source (LC-ESI QTOF MS). The analysis of the obtained results showed that the latexes of ES and EC represent rich sources of diterpenes, tentatively identified as jatrophanes, ingenanes, tiglianes, myrsinanes, premyrsinanes, and others. Examination of the anticancer activity of the ES and EC latex extracts showed that both extracts significantly inhibited the growth of the non-small cell lung carcinoma NCI-H460 and glioblastoma U87 cell lines as well as of their corresponding multi-drug resistant (MDR) cell lines, NCI-H460/R and U87-TxR. The obtained results also revealed that the ES and EC extracts inhibited the function of P-glycoprotein (P-gp) in MDR cancer cells, whose overexpression is one of the main mechanisms underlying MDR. [ABSTRACT FROM AUTHOR]

The aerial parts of three endemic Centaurea L. species, namely C. tomorosii Micevski, C. soskae Hayek and C. galicicae Micevski, afforded sesquiterpene lactone cnicin (1) and seven flavonoids: apigenin (2), isokaempferide (3), hispidulin (4), eupatorin (5), cirsimaritin (6), santaflavone (7) and salvigenin (8). The structures of the isolated compounds were determined by UV, 1H NMR and 13C NMR spectroscopy and HR-ESI-MS spectrometry. 1H NMR spectroscopy was used as a method for quantitative analysis of sesquiterpene lactone cnicin. [Projekat Ministarstva nauke Republike Srbije, br. 172053 ]

Seven elements, namely, arsenic, lead, cadmium, copper, zinc, iron and manganese were determined in 31 samples of Serbian plum brandies by applying atomic spectrometry techniques. Flame atomic absorption spectrometry was used for quantification of copper, iron, zinc, manganese, lead and cadmium; and hydride generation atomic spectrometry absorption for arsenic quantification. Measured concentrations of heavy metals and arsenic was assessed according to the Serbian regulations, official regulations of some other countries and in respect of microelements content in other similar distilled alcoholic beverages. Amounts of microelements in maximal recommended daily and weekly intake of plum brandy were determined. The influence of production (home made and industrial), type of wooden barrel (oak and mulberry), and duration of ageing process on the content of Zn, Cu, Fe and Mn in plum brandies, as well as coefficient of correlation between Cu content and pH value were also studied. [Projekat Ministarstva nauke Republike Srbije, br. 172053]

From the seeds of white currant (Ribes rubrum, cv. White Champagne), a new sesquiterpenoid glucoside (1) was isolated, along with two known compounds: dihydrophaseic acid 3’-O-β-Dglucopyranoside (2), and 3-carboxymethyl-indole-1-N-β-Dglucopyranoside (3). The structure of the new compound was identified as dihydrophaseic acid 3’-O-β-D-glucopyranosyl-1,6-β-D-glucopyranoside, based on extensive NMR and MS spectral studies.

The genus Senecio (family Asteraceae) is one of the largest in the world. It comprises about 1100 species which are the rich source of pyrrolizidine alkaloids. Plants containing pyrrolizidine alkaloids are among the most important sources of human and animal exposure to plant toxins and carcinogens. The pyrrolizidine alkaloids of seven Senecio species (S. erucifolius, S. othonnae, S. wagneri, S. subalpinus, S. carpathicus, S. paludosus and S. rupestris) were studied. Fourteen alkaloids were isolated and their structures determined from spectroscopic data (1H- and 13C-NMR, IR and MS). Five of them were identified in S. erucifolius, four in S. othonnae, two in S. wagneri, four in S. subalpinus, two in S. carpathicus, three in S. paludosus and three in S. rupestris. Seven pyrrolizidine alkaloids were found for the first time in particular species. The results have chemotaxonomic importance. The cytotoxic activity and antimicrobial activity of some alkaloids were also studied.

4-Chlorobenzoic acid (1), quercitrin (2), astilbin (3), along with β-sitosterol, γ-sitosterol, friedelin and β-amyrin were isolated from the aerial parts of Hypericum monogynum. Whereas compound 1 was isolated for the first time from natural sources, flavanonol 3 was not found before in these species.

In this study, volatile compounds were analyzed in five samples of home-made spirit beverage made by the distillation of fermented fruits of cornelian cherry (Cornus mas L.). The major volatile compounds, besides ethanol, identified and quantified were: methanol, acetaldehyde, 1-propanol, ethyl acetate, 2-methyl-1-propanol, 1-butanol, amyl alcohols, 1-hexanol and 2-phenylethanol. The minor volatiles were submitted to liquid-liquid extraction with dichloromethane and analyzed by gas chromatography and gas chromatography/ /mass spectrometry (GC/MS). A total of 84 compounds were identified. The most abundant compounds were straight-chain free fatty acids, ethyl esters of C6-C18 acids, limonene, 2-phenylethanol and 4-ethylphenol. Most of the compounds found in the 'Drenja' spirits investigated in this study are similar to those present in other alcoholic beverages.

The chemical composition of the essential oil of fresh young needles with twigs of Douglas fir (Pseudosuga menziesii Mirb. Franco) obtained by hydrodistillation were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Ten compounds, accounting for 94.26 % of the oil, were identified. The main compounds found were bornyl acetate (34.65 %), camphene (29.82 %), α-pinene (11.65 %) and santene (5.45 %). The antifungal activity of the essential oil was tested against various fungal species. The minimum inhibitory concentration of Douglas fir essential oil ranged from 1.5 to 4 μg mL-1. The fungi most sensitive to the tested oil were Phomopsis helianthi, while Penicillium species, along with Microsporum canis, were the most resistant. Compared to the commercial fungicidal agent bifonazole, the studied essential oil demonstrated higher antifungal activity.

Rutin (1, main constituent) and two flavone C-glycosides, vitexin (2) and vitexin 2''-O-α-rhamnopyranoside (3) were isolated from the aerial parts of Onobrychis montana subsp. scardica. They were identified by 1H-NMR, 13C-NMR and UV-Vis spectroscopy (procedure with shift reagents), and high resolution ESI-MS. A relatively high content of 1 (5.27 mg/g of dry plant material), measured by HPLC, indicated O. montana subsp. scardica as a new natural source of this biologically active compound. The isolated flavonoid compounds might be of value as chemotaxonomic markers.

Resveratrol, which occurs in two isomeric forms, trans and cis, is a phytoalexin with numerous pharmacological activities, such as anti-cancer, antiviral, neuroprotective and anti-aging. Red wine is the main source of the compound and an easy way of including resveratrol in the human diet. In this study, the most popular commercial Serbian wines (red, white and rosé-type) were analyzed for their content of trans- and cis-resveratrol. The analysis was performed by HPLC with a UV detector. Prior to the injection, phenolic compounds were extracted onto a LiChrolut RP18 bonded silica cartridge. The concentration of trans-resveratrol ranged from 0.11 to 1.69 mg L-1 and cis-resveratrol from 0.12 to 1.49 mg L-1.

Essential oils from leaves with young branches of Cotinus coggygria Scop. from two localities in Serbia (Deliblatska peščara and Zemun), obtained by hydrodistillation, were analyzed by GC-MS. Thirty-one component were identified from both oils and among them monoterpenic hydrocarbons were the dominant class (87.4 and 93.1 %). The dominant constituent in both essential oils was limonene (47.0 and 39.2 %). Both oils were also tested for antibacterial and antifungal activities. In comparison to streptomycin, both oils showed slightly higher activity (against most Gram-positive bacteria) in the disc diffusion method and slightly lower activity when the microdilution method was employed. They also exhibited antifungal potential higher than that of the commercial fungicide bifonazole.

1H-NMR spectroscopy was applied for the quantitative analysis of cnicin, a bioactive germacranolide type sesquiterpene lactone, in the aerial parts of seven wild-growing Centaurea species collected in Serbia and Montenegro. The analysis was performed by comparison of the integral of the one-proton signal of cnicin (H-13, δ 5.75) with that of the two-proton singlet ( δ 6.98) of 2,6-bis(1,1-dimethylethyl)-4-methylphenol (BHT), used as the internal standard. Cnicin, within the concentration range 1.06-6.12 mg/g, calculated per weight of the fresh plant material was detected in six species, the exception being C. salonitana. This method allows the rapid and simple quantification of cnicin without any pre-purification step.

The investigation of roots of Anthemis cotula (Asteraceae) from east Serbia revealed, in addition to polyacetylenes previously isolated from the same species, three prenylated 4-hydroxyacetophenones, detected for the first time in any Anthemis species. It is possible that they act as phytoalexins in the plant. From the aerial parts, six linear sesquiterpene lactones (four known and two new ones), as well as two known flavones, apigenin and hispidulin, were isolated.

Two flavonoid glycosides 1 and 2 having the luteolin structure were isolated from flowers of the endemic plant species Cephalaria pastricensis. They were identified by 1H and 13C NMR, as well as UV/Vis spectroscopy. The structures of 1 and 2 were also confirmed by the spectral data of aglycones and TLC of the sugars obtained after acid hydrolysis. Flavones 1 and 2 showed significant antiradical activity in the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay.

Five tetrahydrofuran sesquiterpenes, so-called davanones, and coumarin umbelliferone isolated from Artemisia lobelii All. var. canescens (DC) Briqu, were tested for antifungal activity. All the compounds inhibited the growth of the applied fungi. The overall activity of one of them 2-hydroxy-2,6,10-trimethyl-7,10-epoxydodeca- 3,11-dien-5-on, was comparable to that of the antibiotic bifonazole.

Five lignans with a 2,6-diaryl-3,7-dioxabicycloŠ3.3.0Ćoctane skeleton epieudesmin, kobusin, pinoresinol, fargesin and sesartemin, were isolated from the aerial parts and roots of Achillea lingulata. Their structures were identified by comparison of their 1H-NMR and MS data to those in the literature. Fargesin and pinoresinol have not been isolated previously from any species of the genus Achillea.

The conformations of the C-6 lactonized germacranolides 1–3 were calculated by applying the PM3 semi-empirical method. The low-temperature 1H-NMR spectra of parthenolide (1) were also measured. The relations between the calculated and the experimentally determined geometries were established using a modified Karplus equation. The applied quantum-chemical calculations proved to be an efficient and easy-to-use tool for the elucidation and prediction of the properties of germacranolide-type sesquiterpene lactones.

1. Introduction 2. Results 3. Asteraceae 3.1. Genus Artemisia L. 3.1.1. Artemisia annua L. 3.1.2. Artemisia vulgaris L. 3.1.3. Artemisia absinthium L. (warmwood) 3.1.4. Artemisia scoparia W. et K. 3.1.5. Artemisia camprestris L. 3.2. Genus Ambrosia L. 3.2.1. Ambrosia artemisiifolia L. (the common rag weed) 3.3. Genus Tanacetum L. ( syn. Chrysanthemum L.) 3.3.1. Tanacetum parthenium L. (feverfew) 3.3.2. Tanacetum serotinum L. 3.3.3. Tanacetum vulgare L. (tansy) 3.3.4. Tanacetum macrophyllum Willd. 3.3.5. Tanacetum corymbosum L. 3.4. Genus Telekia Baumg. 3.4.1. Telekia speciosa (Schreb.) Baumg. 3.5. Genus Inula L. 3.5.1. Inula helenium L. 3.5.2. Inula spiraeifolia L. 3.6. Genus Eupatorium L. 3.6.1. Eupatorium cannabinum L. 3.7. Genus Achillea L. 3.7.1. Achillea abrotanoides Vis. 3.7.2. Achillea millefolium subsp. pannonica 3.7.3. Achillea crithmifolia W. et K. 3.7.4. Achillea clypeolata Sibth. et Sm. 3.7.5. Achillea serbica Nyman 3.7.6. Achillea depressa Janka 3.8. Genus Anthemis L. 3.8.1. Anthemis carpatica Willd. 3.8.2. Anthemis cretica L. subsp. cretica 3.9. Genus Centaurea L. 3.9.1. Centaurea derventana Vis. et Pan~. 3.9.2. Centaurea kosaninii Hayek 3.9.3. Centaurea solstitialis L. 4. Apiaceae 4.1. Genus Laserpitium L. 4.1.1. Laserpitium siler L. 4.1.2. Laserpitium marginatum L. 4.1.3. Laserpitium latifolium L. 4.1.4. Laserpitium alpinum W. K. 4.2. Genus Angelica L. 4.2.1. Angelica silvestris L. 4.3. Genus Peucedanum L. 4.3.1. Peucedanum austriacum (Jacq.) Koch