Natural Product Communications

The priority ergot alkaloids ergometrine and ergometrinine are highly toxic mycotoxins naturally occurring in different types of grains (i.e. rye, wheat, rice), as well as grain-based foods and, therefore, have gained increasing importance for food safety over the last years. The application of HPLC-MS/MS for the analysis of ergot alkaloids in food presupposes the availability of isotopically labelled internal standards. Thus, a multistep synthesis was developed for ergometrine-(N- ¹³CD₃) and its epimer ergometrinine-(N- ¹³CD₃) with a mass shift of four units compared with the parent compounds. The synthesis is based on the preparation of stable isotope labelled lysergic acid that was coupled with (S)-alaninol. The chemical synthesis of both compounds has been achieved in six steps with an overall yield of 1 % (ergometrine-(N- ¹³CD₃)) and 0.6 % (ergometrinine-(N- ¹³CD₃)), respectively. Structural identification was performed by MS analysis as well as ¹H and ¹³C NMR.

Essential oil of Mentha spicata L. var. viridis ‘Ganga’, an indigenously developed variety, was chemically profiled using various gas chromatographic techniques. Piperitenone oxide was characterized as the most exclusive constituent (69.7%) along with a new C₃–oxygenated p-menthane alcohol, diosphenolene (1.6%). Enantiomeric discrimination revealed (4S)-(-)-limonene, (R)-(-)-linalool and (1S,2S)-(+)-piperitenone oxide as predominant enantiomers. The oil contained mainly C₃–oxygenated p-menthane monoterpenoids, which are distinctive of peppermint, instead of the characteristic C₆–oxygenated class of spearmint. The present findings will aid in understanding the pathway and cause of C₃-oxygenation in a spearmint taxon. The essential oil and pure piperitenone oxide showed growth inhibiting properties and thus, may be utilized in antifungal preparations for disease management of medicinal and aromatic plants.

A phytochemical study of the ethanolic extract of the leaves of Ligularia virgaurea led to the isolation of a new eremophilane-type sesquiterpene lactone, (4S,5R,6S,8S,10R)–6β-angeloyloxy-eremophil-7(11)-en-10βH-8α,12-olide (1), along with a known eremophilane-type sesquiterpene, (4S,5R,6S,10S)–6β-angeloyloxy-10βH-furanoeremophil-9-one (2). Their structures were elucidated by extensive spectroscopic methods, including 1D and 2D nuclear magnetic resonance and high-resolution electrospray ionization mass spectrometry experiments, and the absolute configurations were confirmed by single-crystal X-ray diffraction analysis using the anomalous scattering of Cu Kα radiation.

A new cyclodepsipeptide, isarin (1), was isolated from the insect pathogenic fungus Isaria japonica Yasuda, together with one known compound, beauveriolide I (2). The structure of this new compound was characterized using a combination of spectroscopic and spectrometric analyses.

The solution-phase synthesis of a proline and glycine-rich plant-derived cyclic heptapeptide, gypsophin E (8), is reported via coupling of a tetrapeptide unit Glycyl-L-leucyl-L-valyl-L-proline-OMe with a tripeptide unit Boc-L-isoleucyl-glycyl-L-proline-OH, followed by cyclization of the linear fragment having seven amino acid units. The structure of the newly synthesized cycloheptapeptide was confirmed by means of chemical and spectroscopic methods. The newly synthesized cyclopolypeptide displayed potent antifungal and anthelmintic activities against the pathogenic yeast Candida albicans, the dermatophytes Trichophyton mentagrophytes and Microsporum audouinii at the 6 μg/mL level, and the earthworms Megascoplex konkanensis, Pontoscotex corethruses and Eudrilus eugeniea at a concentration of 2 mg/mL.

Two new hygroline and tropane alkaloids, 4-hydroxybenzenepropanoylhygroline (1) and 3α,4β-dihydroxy-6β-angeloyoxytropane (2) have been isolated from the aerial parts of Schizanthus hookeri and S. tricolor, respectively, two plants indigenous from Chile. Their structures were elucidated by spectroscopic methods and high resolution mass spectrometry. Their antiparasitic activity and cytotoxicity were measured.

The priority ergot alkaloids ergometrine and ergometrinine are highly toxic mycotoxins naturally occurring in different types of grains (i.e. rye, wheat, rice), as well as grain-based foods and, therefore, have gained increasing importance for food safety over the last years. The application of HPLC-MS/MS for the analysis of ergot alkaloids in food presupposes the availability of isotopically labelled internal standards. Thus, a multistep synthesis was developed for ergometrine-(N-¹³CD₃) and its epimer ergometrinine-(N-¹³CD₃) with a mass shift of four units compared with the parent compounds. The synthesis is based on the preparation of stable isotope labelled lysergic acid that was coupled with (S)-alaninol. The chemical synthesis of both compounds has been achieved in six steps with an overall yield of 1 % (ergometrine-(N-¹³CD₃)) and 0.6 % (ergometrinine-(N-¹³CD₃)), respectively. Structural identification was performed by MS analysis as well as ¹H and ¹³C NMR.

Bursera linanoe cell suspension cultures were initiated from callus grown in Murashige and Skoog medium supplemented with naphthalene acetic acid (3.0 mg L⁻¹) and 6-benzylaminopurine (0.5 mg L⁻¹). In flasks, B. linanoe cell cultures grew over a 9 day period, reaching a maximum biomass of 11.16 g DW L⁻¹. Throughout the growth phase, cell viability was constant at 60 – 70%. In contrast, B. linanoe cells growing in a bioreactor achieved a maximum biomass of 22.26 g DW L⁻¹ (after 7 days), and cell viability was constant at 75 - 85%. Production of linalool and linalyl acetate in the bioreactor (3.02 and 2.40 mg g⁻¹ DW, respectively) was significantly greater than that achieved from cells in flask cultures (1.05 and 0.97 mg g⁻¹ DW, respectively). B. linanoe cell suspension culture has potential as an alternative method for the production of essential oils.

Based on NOE experiments, the structure of naphthodipyranodione from Gentianaceae plants was revised to 1,2-dihydro-4H,6H,8H-naphto[1,2-d:4,5-c'd']dipyrano-4,8-dione. Naphthodipyranodione was assumed to be formed by the degradation of gentiopicroside by enzymatic hydrolysis at low water concentration. The degradation pathway was a unique domino-reaction triggered by enzymatic hydrolysis. Naphthodipyranodione may become an index compound for the drying and/or fermenting procedure of Gentian root.

The compositions of hydrodistillated essential oils of Laser trilobum (L.) Borkh. rhizomes and fruits from Serbia, were investigated using GC and GC/MS. In the dark-blue rhizome oil forty-six compounds (93.1% of the total oil) were identified, with α-pinene (31.5%), γ-terpinene (9.0%), p-cymene (7.9%), β-pinene (6.1%) and 1,4-dimethylazulene (6.0%) as the major components. In the colorless fruits oil, twenty components (96.8% of the total oil) were identified, and the main constituents were limonene (51.6%) and perillaldehyde (26.8%). The antimicrobial activity of the oils was tested using the broth microdilution method against nine bacterial and two fungal strains. The oils revealed significant antimicrobial effect, mainly better than that of thymol, used as a reference compound. The strongest activity was recorded for the rhizome oil against Escherichia coli, Klebsiella pneumoniae and Candida albicans (MICs=25 μg/mL), and the fruit oil against C. albicans ATCC 10259 (MIC=12.5 μg/mL).

In the present study, hydrogels containing the dried extract of tormentil (Potentilla erecta (L.) Raeusch., Rosaceae) were designed and qualitatively evaluated regarding their viscosity, thixotropy, and texture properties. As mucoadhesiveness is a crucial factor determining drug retention within the oral cavity, mucoadhesive properties expressed as the work of adhesion and maximum detachment force under the presence of porcine buccal mucosa and two different models of mucoadhesive layers: mucin and gelatin discs were evaluated. The fingerprints of the analyzed tormentil extract were established by using a LC-ESI-MS method. The dominating compounds of the tested extract are oligomeric proanthocyanidins and agrimoniin. This study reveals that designed hydrogels are promising semi-solid delivery systems for the dry extract of tormentil with beneficial mucoadhesive, thixotropic, and texture characteristics and may be utilized as platforms for tormentil delivery to the oral cavity in periodontal diseases. In vitro evaluation of the efficacy of the tormentil hydrogel against cariogenic Streptococcus mutans biofilms was also performed. The hydrogel significantly reduced artificial biofilm formation. For all Streptococci, complete inhibition was revealed at a final concentration of the extract of 2 mg/mL. These beneficial characteristics, as well as the anti-biofilm activity, enable its use for dental care, but further clinical studies are required.

Rhamnus caroliniana (Rhamnaceae), or Carolina buckthorn, is a plant commonly found in the southeastern United States that was used in Cherokee traditional medicine. To date, there have been no reports on the phytochemical constituents of R. caroliniana. The bark of R. caroliniana was collected and extracted with chloroform. The crude chloroform extract was subjected to preparative column chromatography on silica gel leading to the isolation of two anthraquinones (chrysophanol and physcion), the bianthrone ararobinol, the dihydroxynaphthalene torachrysone, and the fatty alcohol 1-docosanol. The bark essential oil of R. caroliniana was obtained by hydrodistillation and analyzed by gas chromatography – mass spectrometry. The major bark volatiles were the anthrone chrysarobin (24.2%), the piperidine alkaloid piperine (15.4%), and the dibenzoxepin pacharin (7.5%).

Three alkaloids including a new one, N-formyldihydrochelerythrine (1), together with four other known compounds were isolated from the stem bark of Caloncoba glauca. The structure of the new compound was elucidated from spectroscopic and mass spectrometric evidence. This is the first report of alkaloids from the genus Caloncoba. Sesamin (4) [MIC = 256 μg/mL (Candida albicans) and dihydrochelerythrine (2) [MIC = 32 (C. albicans and C. parapsilosis), and 128 μg/mL (C. krusei)] had moderate to weak antifungal activity.

One new flavonol glycoside, patuletin-7-O-[6-O-caffeoyl-2-O-[(S)-3-hydroxyisobutanoyl]glucopyranoside] (astermaritimoside) (1) and four known flavonoids, patuletin-7-O-β-[-6-O-[(S)-3-hydroxyisobutanoyl]glucopyranoside] (2), patuletin-7-O-[6-O-caffeoylglucopyranoside] (3), patuletin-7-O-β-D-glucopyranoside (4) and quercetin-3-O-β-rutinoside (5) have been isolated from Asteriscus maritimus (L.) Less. Chemical structures were elucidated by mass spectrometric, and ¹H NMR, ¹³C NMR, COSY, HMQC and HMBC spectroscopic techniques. The antioxidative effect was evaluated using an electronic spin resonance (ESR) method in order to visualize the inhibition of the DPPH radical. The n-butanol fraction was found to possess a remarkable antioxidant activity that was correlated with the total amount of phenolics. The isolated compounds exhibited good antioxidant activity.

A new marine sterol, 4α-methylergosta-22(E),24(28)-dien-3β-ol (1), was isolated from the octocoral Nephthea columnaris. The structure of 1 was elucidated on the basis of spectroscopic and mass spectrometric methods.

A new 3-methoxybenzensulfonic acid 4-O-β-D-glucopyranoside (1), and ten known compounds (2–11) were isolated from the methanolic extract of the stems of Mallotus microcarpus. The cytotoxicity of the isolated compounds was evaluated by the MTT method. 3-Methoxybenzensulfonic acid 4-O-β-D-glucopyranoside (1) and methyl salicylate 2-rutinoside (5) showed strong cytotoxicity against EGFR-TKI-resistant human lung cancer A549 cells in comparison with camptothecin. Compound 1, leonuriside A (2), 3,4'-dihydroxypropiophenone 3-O-glucoside (6) and (1R,2S)-hovetrichoside A (10) inhibited the growth of human breast cancer MCF-7 cell line with IC₅₀ values in the range of 0.48–1.78 μM. This is the first report on the chemical composition and cytotoxic activity of M. microcarpus.

The chemical composition of the essential oil (EO), microwave extract (ME) and hydrolate extract (HE) from the same batch of leaves of Fortunella japonica, was investigated by combination of chromatographic (GC, CC) and spectroscopic techniques (GC-MS, ¹³C NMR). F. japonica essential oil and extracts are complex mixtures of 28–60 compounds being mainly oxygenated sesquiterpenes. The EO composition was dominated by germacrene D (14.9%), β-elemol (9.1%), cis-guai-6-en-10p-ol (6.3%), β-eudesmol (5.5%), and δ-elemene (5.2%). Limonene was the unique monoterpene identified at appreciable amount (7.1%). The extract obtained by microwave assisted hydrodistillation contained as main components: β-elemol (12.4%), germacrene D (9.9%), cis-guai-6-en-10β-ol (9.0%), β-eudesmol (8.2%), germacra-1(10),5-dien-4α-ol (7.1%) and α-eudesmol (6.4%). Finally, the highest content of oxygenated sesquiterpenes (near 92%) was found in the hydrolate extract displaying cryptomeridiol (23.3%, but totally absent in the EO and ME), β-eudesmol (20.6%) and α-eudesmol (10.7%). Combined analysis by chromatographic and spectroscopic techniques appeared useful for identification of various sesquiterpenols bearing a tertiary alcohol function.

Eugenol (EU) – PEG adduct was synthesized to improve the chemical and physical properties of eugenol. The phenolic group was covalently bound to the carboxyl group of PEG and the release kinetics were studied in vitro in buffer solution at pH 7.4, in simulated gastric fluid and in mouse plasma. Studies in vitro on the release of the parent drug from the prodrug in various media indicate that the adduct may be sufficiently stable to pass intact into the gastrointestinal tract and release EU into the circulation. The antioxidant activity of PEG-EU adduct was also evaluated. Scavenging activity was absent in the original PEG-EU adduct but gradually increased on the basis of drug delivery.

Screening and chromatography of extracts from a terrestrial and a marine-derived streptomycete yielded two new nitrogenous benzene derivatives, namely (S)-N-[3-hydroxy-1-(4-hydroxyphenyl)-propyl]-acetamide (1a), and (R)-2-(1-methyl-2-oxopropylamino)-benzoic acid (2). Additionally, eight known compounds were isolated, 2-acetamidophenol, phenazine-1-carboxylic acid, phenazine-1-carboxylic acid methyl ester, perlolyrin, tyrosol, uracil, and anthranilic acid. The structures of the new compounds were deduced from high resolution mass, 1D and 2D NMR spectra and by comparison with related compounds from the literature. The absolute configuration of 1a and 2 was determined by comparison of experimental and calculated CD and ORD data.

Phytochemical analysis of the mature fruits of Furcraea tuberosa (Agavaceae) resulted in the isolation of a new bisdesmosidic spirostanol saponin (1), along with eight known steroidal glycosides (2–9), one known phenolic carboxylic acid ester (10) and three known flavonol glycosides (11–13). The structures of these compounds were assigned using a combination of 1D and 2D NMR techniques including 1H, ¹³C, COSY, TOCSY, HSQC and HMBC NMR, and confirmed by mass spectrometry. Thus the new saponin was elucidated as (25R)–6α-(β-D-glucopyranosyloxy)-5α-spirostane-3β-O-[(6-O-hexadecanoyl)-β-D-glucopyranoside]. The literature survey revealed that most of the steroidal saponins isolated have shown potent cytotoxic effects against various human cancer cell lines and the results are herein reviewed.

To investigate the preventive effects of tea on hyperglycemia and vascular complications of diabetes, we report the extraction and composition as well as the vasculoprotective effects of black tea extract (BTE), green tea extract (GTE), and dark tea extract (DTE). High Performance Liquid Chromatography (HPLC) and colorimetric methods were conducted to analyze for tea catechins, caffeine, polyphenols, amino acids and polysaccharides of BTE, GTE and DTE. The inhibitory effects of α-glucosidase, aldose reductase (AR), advanced glycation end-products (AGEs) and glucose uptake promotion effect in BTE, GTE and DTE were explored in vitro. Contents of six major catechin forms and total catechin as well as polyphenols are higher in GTE and DTE than BTE. BTE, GTE, and DTE showed the inhibitory effects of α-glucosidase, AR, and AGEs, but only DTE exhibited the glucose uptake promotion effect in HepG2 cells. The results suggest that regular consumption of tea can help prevent the progression of hyperglycemia and the vascular complications of diabetes.

Garcinia dulcis (Roxb.) Kurz is a tropical fruit tree native to Southeast Asia where it has a long history of use as a traditional medicine for the treatment of ailments such as lymphatitis, parotitis, struma, scurvy, cough, and sore throat. Despite its medicinal values, this plant is not well known and rarely found nowadays. Research on the phytochemical constituents and biological activities of G. dulcis have demonstrated that various parts of the plant contain an abundance of bioactive compounds mainly xanthones and flavonoids, with significant pharmacological properties such as anti-atherosclerosis, anti-bacterial, anti-cancer, anti-hypertension, and anti-malarial. In the present review, current knowledge of the phytochemistry of G. dulcis and biological activities of its active constituents based on the available literature are summarized in order to explore application potentials and prospective research works on this plant.

Extracts and infusions of wild artichoke (Cynara cardunculus L.) and globe artichoke (C. scolymus L.) (heads, bracts and stems) were examined for their total phenolic content (TPC) and their antioxidant activity after performing Classical Extraction (CE) and Ultrasound-Assisted Extraction (UAE). UAE proved to be more effective, since extracts exhibited higher antioxidant activity and TPC values than CE extracts and infusions. Moreover C. cardunculus heads extract using UAE, displayed the maximum TPC values (1.57 mg gallic acid equivalents (GAE) g⁻¹ fresh weight (fw)), the highest DPPH^• scavenging activity (IC₅₀; 0.91mg mL⁻¹) and the highest ABTS^•+ radical scavenging capacity (2.08 mg Trolox Equivalents (TE) g⁻¹ fw). Moreover, the effect of different concentrations of C. cardunculus head extracts (showing the highest TPC and antioxidant activity) on Aspergillus parasiticus growth was estimated in AFPA medium. The maximum inhibition was found to be ~42.1% in comparison with the control.

The content of two groups of compounds with biological activity (non-hallucinogenic indole compounds and free phenolic acids) were analyzed in extracts of fruiting bodies of four species of Phellinus: P. igniarius, P. pini, P. pomaceus and P. robustus. The presence of indole compounds in methanolic extracts was analyzed by high-performance liquid chromatography and thin-layer chromatography coupled with densitometric detection. Three metabolites (serotonin, tryptamine, and L-tryptophan) were identified. The contents of individual indole compounds ranged from 1.70 (tryptamine in P. robustus) to 8.32 mg x 100 g¹ dry weight (L-tryptophan in P. robustus). Four free phenolic acids were detected in methanolic extracts by the HPLC method. The total content ranged from 9.9 mg x 100 g¹ DW (P. igniarius) to 32.5 mg x 100 g¹ DW (P. robustus).

Indonesia is one of the richest countries with respect to plants resources. People from various ethnic, language, and religious groups have used the plants as alternative medicines, health foods and beverages for hundreds of years. To establish modern application for these understudied plant resources, ethnopharmacological data from more than 40 leguminous plants in Indonesia, spanning the western to the eastern parts of the Indonesian archipelago, were reviewed. In particular, bioactive secondary metabolites, including flavonoids, were described in detail to promote research into these plants as functional foods, nutraceuticals, and medicines.

The composition of four samples of essential oil isolated from the leaves of individual Monodora crispata trees growing wild in an Ivorian forest (Adiopodoumé) were investigated by a combination of chromatographic (GC(RI)) and spectrometric (GC-MS, ¹³C NMR) techniques. In total, fifty-seven compounds accounting for 92.3–98.7% of the whole composition were identified. These oils were characterized by the preeminence of sesquiterpene hydrocarbons and the content of the main components varied substantially from sample to sample. Two samples were largely dominated by germacrene D (67.3% and 76.3%, respectively), a third sample was represented by germacrene D (28.3%), germacrene C (14.3%), δ-elemene (12.1%) and β-elemene (9.3%) and the fourth sample was characterized by 3-dimethylallylindole (15.1%), germacrene D (12.0%) and cyperene (7.8%). ¹³C NMR spectroscopy was particularly efficient for the identification of heat-sensitive compounds.

A new monoterpenoid indole alkaloid, ochroborbone (1), along with five known alkaloids (2–6), were isolated from the stems and leaves of Ochrosia borbonica. Among them, ochroborbone (1) is a rare C17-nor monoterpenoid indole alkaloid, and the known compounds (2-6) were isolated from Ochrosia for the first time. These structures were established on the basis of extensive spectroscopic methods. All isolated compounds were evaluated for their cytotoxicities against five human cancer cell lines: HL-60, SMMC-7721, A-549, MCF-7 and SW480 in vitro. Compounds 1 and 2 exhibited inhibitory effects with IC₅₀ values comparable with those of cisplatin.

A new stigmasterone 7α,15α-dihydroxystigmast-4-en-3-one (1), along with fifteen known compounds was isolated from the EtOAc and n-BuOH fractions of the EtOH extract of Agathis macrophylla. The structure of these compounds 1-16 were established mainly by spectroscopic analysis and by comparison of the spectroscopic data with those reported for structurally related compounds. The isolated compounds 1-8 were screened for cytotoxicity in A549 tumor cell line using RSB assay. Compounds 1 and 2 displayed moderate cytotoxicity against A549 tumor cell line with IC₅₀ value of 36.5 and 16.0 μmol/L, respectively.

Ganoderma applanatum is a fungus used for the prevention and treatment of a variety of disorders in China. In the present study, four new compounds, named shushe acids A-D (1-4), were isolated from the fruiting bodies of this species. Their structures were identified on the basis of spectroscopic methods. Compounds 1-4 are all natural product hybrids composed of derivatives of gallic acid, glycerol and succinic acid. None of the four compounds showed activity against the MCF-7 cell line.

Bacopa monnieri (L.) Pennell (water hyssop) from the Scrophulariaceae family represents one of the best known plants of the Ayurveda system, with reported procognitive, antioxidant, anti-inflammatory, neuroprotective, and anticonvulsant activity. It is considered to be a central nervous system modulating agent. However, recent studies have indicated its potential use in cancer prevention and treatment. Here we report the findings from a study of the effects of B. monnieri extracts derived from artificial digestive juices on physiological traits of prostate cancer cells (DU 145), such as viability and migratory activity. The safety profile and chemopreventive potential of B. monnieri extracts were investigated in Vibrio harveyi mutagenicity assays. Additionally, in this study for the first time the content of phenolic compounds and bacosides released from B. monnieri biomass to artificial digestive juices was determined. The investigated extracts were cytotoxic to DU 145; however, in non-cytotoxic concentrations, they significantly reduced cancer cell motility, thereby demonstrated anti-invasive activity. All extracts under study exhibited very strong antimutagenic activity against nitroquinoline-N-oxide. Bacoside A and phenolic acids were determined, but qualitative and quantitative differences between extracts were found.

The chemical constituents and anti-inflammatory activity of the essential oil from the leaf of Jatropha curcas L (Euphorbiaceae) collected from Nigeria are reported. The analysis of the chemical constituents of the essential oil was achieved by using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). All constituents of the essential oil, namely neophytadiene (35.8%), phytol (23.1%), trans-pinane (12.7%), 6,10,14-trimethyl-2-pentadecanone (12.3%) and citronellyl propanoate (11.2%), were present in significant amounts. The anti-inflammatory activity of the leaf oil was determined on Wistar rats using egg-albumin as phlogistic agent; significant inhibition (P< 0.05) was shown at a dose of 2%, v/v. Percentage inhibition of the anti-inflammation increased steadily to 76.6% in the 4^th hour.

This study evaluated the in vitro antimicrobial effect of 3β-acetoxy-norlup-20-one (1) and 3-chloro-4α,14α-dimethyl-5α-cholest-8-ene (2), triterpene derivatives from Euphorbia officinarum latex against fungal and bacterial phytopathogens. Results showed that although mycelial growth of several strains of Verticillium dahlia, and Fusarium oxysporum fsp. melonis and Penicillium expansum was affected only moderately, the two compounds were able to reduce highly conidia formation and germination, suggesting that they act as fungistatic compounds. Their antibacterial activity was tested against Pseudomonas syringae pv. syringae (Pss), P. syringae pv. tabacci (Pst), Erwinia amylovora (Ea) and Agrobacterium tumefaciens (At) using disc diffusion method. Results showed that compound 2 was more effective in inhibiting the growth of Pss, Pst and Ea than compound 1.

Continuous investigations of the roots of Aconitum nagarum var. lasiandrum led to the isolation of two new C₁₉-diterpenoid alkaloids, lasiandrine (1) and lasiandroline (2). Their structures were elucidated on the basis of extensive interpretation of spectroscopic and mass spectrometric data.

Papaver degenii (Urum. & Jav.) Kuzmanov is a Bulgarian endemic and glacial relict growing in the alpine zone of Pirin Mountains and it is included in the Red Data Book of Bulgaria. Numerous plants were in vitro multiplied in order to study the alkaloid composition of the species and to test the potential for green biotechnology establishment. Percentages of the crude alkaloid mixture in dried in vitro shoots and ex vitro adapted plants to room phytotron were similar to that in seeds (0.49%) and 5 to 6 times higher than in the aerial parts of wild-growing plants. Amurensine was the only alkaloid detected in the seeds. Four isoquinoline alkaloids were isolated from the regenerated plants, belonging to two alkaloid types: isopavines (amurensine and O-methylthalisopavine) and protopines (allocryptopine and protopine), amurensine being the main alkaloid: 63.4% in in vitro shoots, up to 88.1% in the aerial parts of ex vitro adapted plants. Allocryptopine was in higher amount (30%) in the roots of the ex vitro adapted plants.

The aim of the present study was the chemical analysis of the essential oils from Juniperus phoenicea and J. drupacea female cones and evaluation of their biological effects. Fresh samples, collected in Greece, were subjected separately to hydrodistillation and the oils obtained analyzed by GC-FID and GC-MS. The oils were assessed using the CAM (chorioallantoic membrane) assay to evaluate their anti-angiogenic potential and the lack of irritant effects in topical application. GC analysis showed that mainly quantitative differences among the samples were observed: limonene was the most abundant compound in J. drupacea (27.0%) compared with J. phoenicea oil (1.6%); the content of α-pinene was high in both essential oils (J. phoenicea 22.1%, J. drupacea 26.1%) followed by germacrene D (J. phoenicea 7.4%, J. drupacea 7.1%, respectively). Nevertheless, qualitative differences were also detected as the diterpene 4-epi-abietal was present in a considerable amount (13.2%) in J. phoenicea essential oil, but was not detected in J. drupacea oil. In the CAM assay, only J. phoenicea essential oil evidenced a rather weak anti-angiogenic activity compared with the standard retinoic acid, but no irritant effect was observed for either essential oil suggesting their safety for topical application.

Essential oils obtained by hydrodistillation of different parts of Iris persica L. were investigated for the first time by GC-FID and GC-MS; moreover, their antifungal activities were determined. 34, 32, 27, and 17 compounds were identified in the oils from air-dried flowers, leaves, rhizomes and fresh bulbs, respectively, representing ≥ 98% each oil. The major constituents of the flower essential oil were phenylethanol (24.8%) and furfural (13.8%), which, as the main component, constituted also 39.0% and 22.2% of the leaf and rhizome volatile fractions, respectively. Phenylacetaldehyde (37.1%) was the main constituent of the bulb volatile fraction. In in vitro tests, moderate antifungal activity was detected for the oils against strains of the human pathogenic fungal species Candida albicans, Microsporum canis, and Trichophyton mentagrophytes, the plant-fungal pathogen Pyricularia oryzae, and the fungal food contaminant Aspergillus carbonarius,. The highest activity was exhibited by the essential oils from leaves and flowers, suggesting that they could be considered natural antimicrobial agents.

In this study we isolated two polyphenolic acids of m/z 639, called catharinol A and catharinol B, from Plantago catharinea L. (Plantaginaceae) leaves. Although presenting very similar structures, catharinol A showed higher antioxidant activity when compared with gallic acid and quercetin standards. These compounds are position isomers and present in their chemical structure the rare sugar D-allose. Molecules with similar constitution are known to have important biological activities such as antitumor and immunosuppressive. These compounds were isolated by high-performance liquid chromatography (HPLC) and characterized by mass spectrometry (FIA-ESI-IT-MS/MS) and nuclear magnetic resonance (NMR). This work is the first study on the chemical composition of P. catharinea and encourages the production of Plantago species as a good source of bioactive molecules.

Fourteen neo-clerodane diterpenoids isolated from Scutellaria altissima (Lamiaceae) were tested for insect antifeedant activity against Leptinotarsa decemlineata Say. Potato leaf disks treated with small amounts of the compounds (concentration 1000, 100, 10 ppm) resulted in good to very good antifeedant activity. Clerodin (1), scutecyprin (11) and 11-epi-scutecolumnin C (12) showed strong feeding inhibition at 1000 ppm and exhibited significant antifeedant activity at a concentration of 100 ppm. Activity was established by calculating the feeding ratio (FR) between the consumed areas of treated disks (CTD) and control disks (CCD). For comparison, FR₅₀ values were determined as the FR at a CCD of 50%. Structural features of the compounds associated with the changes in activity and structure-antifeedant activity relationships are discussed. For the first time the anti-feedant activity has been evaluated of neo-clerodane diterpenoids with an unusual R-configuration of the carbon atom C-11.

A high performance thin layer chromatographic (HPTLC) method was developed and validated for simultaneous determination of rhein and aloe-emodin, major anthraquinone constituents, in S. alata leaves. The separation was performed on a silica gel 60 F₂₅₄ HPTLC plate using ethyl acetate: methanol: water (100:17:10, v/v/v) as the development system. Validation of the analytical method for rhein and aloe-emodin promoted acceptable parameters. Good linearity in the range of 40–480 ng/band was obtained while intra-day and inter-day precisions were shown to be precise with relative standard deviations of less than 5%. The average percentage recoveries of rhein and aloe-emodin were 98.8% and 98.9%, suggesting acceptable accuracy. The content of rhein and aloe-emodin in S. alata leaves collected from 5 different provinces in Thailand analyzed by the validated HPTLC method were in the ranges of 0.098 ± 0.017 −0.30 ± 0.02%, w/w, and 0.081 ± 0.0006 - 0.34 ± 0.0009 %, w/w, respectively. Five commercial products of S. alata tea available in the market were purchased and analyzed for rhein and aloe-emodin contents. The contents of rhein and aloe-emodin in the tea samples were in the ranges of 0.085 ± 0.004 - 0.23 ± 0.04%, w/w, and 0.096 ± 0.006 - 0.30 ± 0.01 %, w/w, respectively. The HPTLC method is rapid, reliable, sensitive and economical for routine analysis of rhein and aloe-emodin contents in S. alata leaf raw materials and its commercial products.