Metabolic profiling of Vitex agnus castus leaves, fruits and sprouts: analysis by LC/ESI/(QqQ)MS and (HR) LC/ESI/(Orbitrap)/MS n.

Food supplements based on Vitex agnus castus L. (Verbenaceae) fruits, also known as chasteberry, are routinely used by women against somatic and psychic premenstrual symptoms such as depression, sadness or irritability. With the aim of highlighting the differences in the chemical profiles of cultivated fruits and different parts of wild plants (fruits, leaves and sprouts) of V. agnus castus, a method concerning with the quali-quantitative study of the derived hydroalcoholic extracts was carried out by using high-performance liquid chromatography coupled to electrospray negative ionization Orbitrap multicollisional high resolution mass spectrometry (LC/ESI/(Orbitrap)MS(n)) and high-performance liquid chromatography coupled to electrospray negative ionization triple quadrupole tandem mass spectrometry (LC/ESI/(QqQ)MS) in multiple reaction monitoring (MRM) mode.

Mosquitocidal essential oils: are they safe against non-target aquatic organisms?

In latest years, the importance of the Melaleuca alternifolia essential oil (EO) has been greatly empathised due to its anti-microbial and anti-inflammatory effects, as well as to its toxic properties towards many arthropods of great medical and veterinary importance. In this research, the EO extracted from aerial parts of M. alternifolia was evaluated for its toxicity against larvae of the most invasive mosquito worldwide, Aedes albopictus (Diptera: Culicidae), and towards adults of the water flea, Daphnia magna (Cladocera: Crustacea), a non-target aquatic organism that share the same ecological niche of A. albopictus. The chemical composition of M. alternifolia EO was investigated by GC-MS analysis. Tea tree EO was mainly composed by oxygenated monoterpenes, with 1,8-cineole as the major constituent. M. alternifolia EO exerted toxic activity against A. albopictus larvae, with a LC50 = 267.130 ppm. However, this EO had a remarkable acute toxicity also towards adults of the non-target arthropod D. magna, with a LC50 = 80.636 ppm. This research provide useful information for the development of newer and safer mosquito control tools, highlighting that the non-target effects against aquatic organisms that share the same ecological niche of A. albopictus larvae are crucial in the development of ecofriendly mosquito control strategies. Further research is needed to investigate the chronic and/or reproductive toxicity of M. alternifolia EO both towards target and non-target aquatic arthropods.

Determination of six steviol glycosides of Stevia rebaudiana (Bertoni) from different geographical origin by LC-ESI-MS/MS.

Liquid chromatography electro-spray tandem mass spectrometry (LC-ESI/MS/MS) was applied to the determination of sweet glycosides (steviol glycosides), and toxic aglycon steviol in 24 samples of Stevia rebaudiana (Bertoni) aerial parts, which had been experimentally cultivated in Italy, although derived from seeds of different geographical origin. On the basis of the specific fragmentation of these compounds, an LC-MS/MS method was developed with the aim of quantifying analytes in plant material. Although toxic steviol was not detectable in all the samples, the samples with the highest levels of steviol glycosides were identified. Analysis of the different samples revealed that they were good quality samples, quality being directly linked to the presence of sweet glycosides in the plants cultivated in Italy, although there were differences in the content of these compounds according to the origin of the seeds, and in particular, a major concentration of compounds with major sweetness activity and minor toxicity was found in the population coming from Brazil (for example: sample 10, stevioside content 15.74±2.0% p/p and rebaudioside A content 3.09±0.39% p/p of dried plant). Finally, based on this metabolomic targeted approach, the results obtained for the samples were treated by Principal Component Analysis, identifying specific genotypic differences based on the geographic origin of the seeds.

Abelmoschus esculentus (L.) Moench. and Abelmoschus moschatus Medik: seeds production and analysis of the volatile compounds.

Nine accessions of Abelmoschus esculentus (L.) Moench. and three of Abelmoschus moschatus Medik were both grown in Central Italy for the evaluation of the production of seeds. Furthermore, the volatiles emitted by the mature seeds were sampled by mean of SPME. Seventy compounds were detected in the headspace of the seeds of A. esculentus. The principal constituents common to all the nine accessions were isopentyl 2-methyl butanoate (24.5-59.1%) and heptanoic acid 2-methylbutyl ester (6.6-13.5%). In the headspace around the seeds of A. moschatus 93 components were detected. Among the main volatiles shared by the three accessions, n-tridecane (1.5-26.9%), isopentyl 2-methyl butanoate (0.2-14.3%) and decanal (1.6-5.7%) should be mentioned. Many differences were present in the volatiles emitted by the various accessions and between the two Abelmoschus species.

Larvicidal and repellent activity of Hyptis suaveolens (Lamiaceae) essential oil against the mosquito Aedes albopictus Skuse (Diptera: Culicidae).

Lamiaceae have traditionally been used in developing countries for their insecticidal and repellent properties against several insect species. In our research, the essential oil (EO) extracted from fresh leaves of Hyptis suaveolens (Lamiaceae), and its main constituents were evaluated for larvicidal and repellent activity against the Asian tiger mosquito, Aedes albopictus Skuse (Diptera: Culicidae), currently the most invasive mosquito worldwide. H. suaveolens EO had insecticidal activity against A. albopictus larvae and mortality was dosage dependent. At the highest dosages of 450 and 400 ppm, there were no significant differences on larval mortality, as mortality ranged between 98.33% and 93.33%, respectively. At dosages ranging from 250 to 350 ppm, mortality rates were lower and not significantly different from each other. Terpinolene was found to be the most effective pure compound. Efficacy protection from H. suaveolens EO, at dosages ranging from 0.03748 to 0.7496 µg cm(-2) of skin, was evaluated during 150 min of observation. Results indicated that this EO had a significant repellent activity (RD(50) = 0.00035 µg cm(-2); RD(90) = 0.00048 µg cm(-2)), with differences in repellency rates, as a function of both concentration and observation time. Protection time ranged from 16 to 135 min. These results clearly evidenced that the larvicidal and repellent activity of H. suaveolens EO could be used for the development of new and safer products against A. albopictus.

Profiling of phenolics from Tephrosia cinerea.

An HPLC-ESIMS(n) method, based on high-performance liquid chromatography coupled to electrospray positive ionization multistage ion trap mass spectrometry, has been used to rapidly identify and guide the isolation of phenolic constituents from the methanol extract of the aerial parts of T. cinerea. On the basis of the results of the online screening by HPLC-ESIMS (n), 17 phenolic constituents, including two new compounds, namely demethylapollinin 7- O- ß-D-glucopyranoside (1) and cineroside A (17), were isolated, and their structures were unambiguously assigned by the extensive use of 1D- and 2D-NMR experiments.

Metabolite fingerprinting of Camptotheca acuminata and the HPLC-ESI-MS/MS analysis of camptothecin and related alkaloids.

The major phytochemical constituents, namely, alkaloids, flavonoids and ellagic acid derivatives, of leaves of Camptotheca acuminata were identified using high performance liquid chromatography (HPLC) coupled with electrospray mass spectrometry (ESI-MS) in extracts of plants cultivated in Italy and collected at different growth stages. Alkaloids related to camptothecin were identified and quantified by HPLC coupled with ESI-tandem mass spectrometry (MS/MS) employing, respectively, an ion trap and a triple quadrupole mass analyser. The fragmentation patterns of alkaloids related to camptothecin were analysed and a specific Multiple Reaction Monitoring HPLC-MS/MS method was developed for the quantitative determination of these constituents. The described method provides high sensitivity and specificity for the characterisation and quantitative determination of the alkaloids in C. acuminata.

Composition and content of seed flavonoids in forage and grain legume crops.

Flavonoid content and composition in 124 accessions of forage and grain legumes was assessed by means of HPLC-DAD and HPLC-ESI-MS analyses of seed extracts. Excluding soybean, only 7 out of 77 grain legume accessions exhibited a total flavonoid content higher than 0.1 mg/g fresh seed weight (mean content 0.33 +/- 0.31 mg/ g fresh seed weight); while in 38 out of 47 forage legume, the mean flavonoid content was 1.49 +/- 0.89 mg/g fresh seed weight. On the basis of their higher seed flavonoid content and agronomic relevance, two grain and five forage legume accessions were selected for further HPLC-ESI-MS analyses aimed to identify bioactive compounds. Chemical analysis of the selected accessions permitted the identification of several flavonoids with pharmaceutical properties such as vitexin in Vigna angularis and Trigonella foenum-graecum, quercetin in all Trifolium spp and Medicago sativa, taxifolin in Trifolium pratense and Trifolium alexandrinum, myricetin in Trifolium repens, kaempferol in Phaseolus vulgaris and T. repens and luteolin in M. sativa and T. alexandrinum. Since much of literature information is regarding flavonoid content and composition of the green part of legume species, the obtained results contributed to fill the gap in knowledge about flavonoid content at the seed level.

Essential oils from Mediterranean lamiaceae as weed germination inhibitors.

The essential oils obtained from rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.), and savory (Satureja montana L.) and the four monoterpenes that are their major constituents have been analyzed by GC and GC-MS and tested for their allelopathic properties on the seeds of three different annual weeds (Chenopodium album, Portulaca oleracea, and Echinochloa crus-galli) and three crops (Raphanus sativus, Capsicum annuum, and Lactuca sativa), with the aim to evaluate in vitro their potential as germination inhibitors. The essential oil composition varied with the species, thymol being the main constituent (44%) of thyme and carvacrol (57%) that of savory oil. Differences in essential oil composition were observed within two different rosemary ecotypes, type A, with alpha-pinene (37%) and 1,8-cineole (23%), and type B, characterized by a 2-fold content of 1,8-cineole (47%). This latest essential oil inhibited completely the germination of weeds while concurrently displaying little effect on pepper. The other two oils showed less selective action. S. montana essential oil, with 57% carvacrol, is the most active compound, completely inhibiting germination both of crops and weeds. Borneol, one of the main constituents of the oil of rosemary type B, showed an activity comparable to that of the whole oil. Crop and weed seeds treated with 1,8-cineole showed germination values that were not significantly different from controls, even if a slowing of the germination process expressed in terms of a significant increase in mean germination time was observed. Monoterpene compounds also present in the essential oils mainly represented the volatile fraction released from the crops and their residues into the soil.

Main agronomic-productive characteristics of two ecotypes of Rosmarinus officinalis L. and chemical composition of their essential oils.

The productive potential of two different ecotypes of Rosmarinus officinalis (Cevoli and Lunigiana) cultivated in the littoral area near Pisa (northern Tuscany, Italy) and the differences in the yield and composition of the essential oils of leaves, flowers, and stems obtained from different positions of the plants were used to characterize the two ecotypes. The Cevoli ecotype plant produced the highest yield of dry matter (221 g plant-1) in comparison to the Lunigiana ecotype (72 g plant-1). There were significant differences in dry matter production of different organs of both ecotypes. The essential oil contents of Cevoli and Lunigiana ecotypes were similar. In contrast, the oil contents of the different plant parts showed marked differences. The apical part of the plant and the leaves gave the highest essential oil yields. The major difference between the oils of the two ecotypes consisted in the 1,8-cineole contents (6.6 and 37.9% in Cevoli and Lunigiana, respectively). The Cevoli ecotype was determined to be the most suitable for essential oil extraction because it was characterized by a preponderance of flowers and leaves in the apical portion. The Cevoli ecotype could be classifited as an alpha-pinene chemotype, whereas Lunigiana is a 1,8-cineole chemotype.