Roberta Ascrizzi

Ionic liquids, ultra-sounds and microwaves: an effective combination for a sustainable extraction with higher yields. The cumin essential oil case

This paper deals with the concept of process intensification applied to the extraction of essential oil (EO). Microwave hydrodistillation (MWHD) and simultaneous ultrasound MW-assisted hydrodistillation (US-MWHD) were intensified by coupling them with a green tool: ionic liquids (ILs). The yield and chemical composition of the cumin EO obtained by MWHD and US-MWHD were compared with those from conventional hydrodistillation (HD) using water and three mixtures of water with three different ILs synthesized ad hoc, as maceration and extraction media, and analysed by a multivariate statistical analysis approach. The cumin EO was chemically characterized by GC and GC-MS analysis. The interaction of the ILs and ILs–H2O mixtures with MW was experimentally investigated and discussed, while the ILs dipole moments and optimized geometry in vacuo and in water were calculated at the DFT level. The different approaches were also compared in terms of energy and time savings. All the data clearly showed that the most promising approach was US-MWHD using a 1,3-dimethylimidazolium dimethylphosphate mixture as maceration and extraction medium. A total yield increase was achieved of up to 75% and an energy saving of 46% compared to the classical HD. The proposed technology, using ILs as green solvents, which fits well with the MW and US technology, enabled a continuous-flow and batch extractor to be constructed which would be useful for industrial applications.

Chemical composition and biological studies of the essential oil from aerial parts of beta vulgaris subsp. maritima (L.) arcang. growing in Tunisia

The chemical composition, antioxidant, cytotoxic, anticholinesterase and anti‐tyrosinase activities of the hydrodistilled essential oil of the aerial parts of Beta vulgaris subsp. maritime (L.) Arcang. from Tunisia have been evaluated. The chemical composition of the oil (yield 0.037% [w/w]), determined by GC‐FID and GC/MS is reported for the first time. Twenty five components, accounting for 98.1% of the total oil have been identified. The oil was characterized by a high proportion of oxygenated sesquiterpenes (39.2%), followed by sesquiterpene hydrocarbons (30.3%) and one apocarotenoids (26.3%). The main compounds were γ‐irone (26.3%), α‐cadinol (12.1%), T‐cadinol (10.6%), bicyclogermacrene (10.4%) and δ‐cadinene (6.0%). The isolated oil was tested for its antioxidant activity using the DPPH·, ABTS+·, catalase, and paraoxonase assays and also for its cytotoxic, anticholinesterase, and anti‐tyrosinase activities. The essential oil exhibited high antioxidant activity (IC50 = 0.055 ± 0.006 mg/ml) and important result oncatalase (524.447 ± 2.58 Units/mg protein). Furthermore, it exerted a significant cytotoxic effect against A549 cell line, with IC50 = 42.44 ± 1.40 μg/ml. The results indicate that the essential oil of B. vulgaris subsp. maritima (L.) Arcang. aerial parts may be used in future as an alternative to synthetic antioxidant agents, with potential application in the food and pharmaceutical industries.

Photochemical response of parsley (Petroselinum crispum (Mill.) Fuss) grown under red light: The effect on the essential oil composition and yield

The effect of different wavelengths on plants morphological characters has been widely described, but also the chemical composition of the essential oil is influenced by the lighting conditions in which they are grown. In the present study, the effect of both the enrichment (reverse Emerson effect) and the monochromatic lighting treatments with red light has been evaluated on the essential oil compositions of parsley (Petroselinum crispum (Mill.) Fuss). Multivariate statistical analysis was performed on the results, with both the hierarchical cluster and principal component analyses. Whilst the red-enrichment of the light spectrum did not induce major changes in the essential oil composition, the end of the day monochromatic red (660 nm) treatment caused a chemotype switch in the essential oil and relevant differences in the overall composition, with an increment of the relative abundance of oxygenated compounds, coupled with a relevant decrement in the abundance of phenylpropanoids. The extraction yields remained unchanged in all the three tested conditions of light (control, red-enriched and monochromatic red). Different lighting conditions could be used as a tool to modulate the compounds present in the essential oil, but further studies would be advisable to assess the effects on different species and chemical classes of compounds.

Flower scent bouquet variation and bee pollinator visits in Stevia rebaudiana Bertoni (Asteraceae), a source of natural sweeteners

Pollinators provide a key service to both natural and agricultural ecosystems. Little is reported on the pollination chemoecology of Stevia rebaudiana (Asteraceae), a hermaphroditic species producing self-incompatible florets in small corymbs. We investigated the chemistry of volatiles potentially involved in its pollination system. The VOCs emitted by the corymbs of 27 F1 open-pollinated genotypes were collected by solid-phase micro-extraction and analyzed by gas chromatography–mass spectrometry (GC–MS), as well as morphometric data of the genotypes were recorded. Finally, we quantified the abundance of pollinators for each genotype. S. rebaudiana flowers were mainly visited by bees (Apidae and Halictidae), followed by hoverflies (Diptera: Syrphidae). GC–MS indicated that S. rebaudiana was characterized by a complex scent profile with large variability among F1 plants. Discriminant analysis showed that limonene, δ-elemene and bicyclogermacrene were the compounds explaining most of the scent bouquet difference between high attractive (>40 pollinators/plant) from low attractive pollinator power (<40 pollinators/plant). Limonene was the most representative VOC among plants that are more attractive to pollinators, while high emissions of δ-elemene and bicyclogermacrene were linked to plants that are less attractive to pollinators. S. rebaudiana morphometric data highlighted that, besides floral VOCs, corymb abundance and size, as well as plant height, may route pollinator visits. Overall, this study adds knowledge on floral phenology and pollinator ecological traits of S. rebaudiana, allowing a deeper understanding of its chemical ecology and pollination.

Patterns in Volatile Emission of Different Aerial Parts of Caper (Capparis spinosa L.)

We analyzed the spontaneous volatile emission of different aerial parts of the caper (Capparis spinosa L.) by HS‐SPME‐GC/MS. We identified 178 different compounds of which, in different proportions based on the sample type, the main ones were (E)‐β‐ocimene, methyl benzoate, linalool, β‐caryophyllene, α‐guaiene, germacrene D, bicyclogermacrene, germacrene B, (E)‐nerolidol, isopropyl tetradecanoate, and hexahydrofarnesyl acetone. The multivariate statistical analyses seem to point out that the parameter leading the emission patterns is the function of the analyzed sample; the flower samples showed differences in the emission profile between their fertile and sterile portions and between the other parts of the plant. The green parts emission profiles group together in a cluster and are different from those of seeds and fruits. We also hydrodistilled fully bloomed caper flowers, whose volatile oil showed significant differences in the composition from those of other parts of the plant reported.

Salvia verticillata: Linking glandular trichomes, volatiles and pollinators

Plants have developed a plethora of signals to interact with other organisms, finally building up a sophisticate language for communication. In this context, we investigated Salvia verticillata L. (Lamiaceae), with the primary goal to link secondary metabolites and actual biotic relationships. We specifically analysed the volatile organic compounds (VOC) spontaneously emitted by leaves and flowers and determined the composition of the essential oils obtained from the aerial parts across 2015 and 2016. We merged information of chemical analyses to a micromorphological investigation on the glandular indumentum and to focal observations on the pollinator assemblage. The VOC profiles were highly variable, with the floral bouquet being the most complex. Flowers and leaves showed 37 and 20 exclusive compounds, dominated by 1,8-cineole (10.4%) and germacrene D (38.4%), respectively. Sesquiterpene hydrocarbons prevailed (83.3% leaves; 73.7% flowers) and 19 common compounds were detected. The oil profiles proved to be consistent across the two years: sesquiterpene hydrocarbons invariably dominated, with germacrene D, bicyclogermacrene and β-caryophyllene as main compounds. The whole plant epidermis is thickly covered by two types of glandular hairs: peltates and small capitates, both responsible for the synthesis of terpenes, finally resulting in the VOC emission and in the essential oil production. S. verticillata attracted mainly bees belonging to two functional groups: medium-sized and large bees, notwithstanding the small size of its flowers. At the site, Apis mellifera and different Bombus species were recorded, mainly interested in feeding on nectar. The literature survey on the isolated volatile compounds confirmed the hypotheses on the seduction strategies towards Apoidea.

Senna occidentalis (L.) Link and Senna hirsuta (L.) H. S. Irwin & Barneby: constituents of fruit essential oils and antimicrobial activity

Abstract Senna occidentalis and S. hirsuta are mostly gathered from the wild for medicinal use and have a disagreeable odour when crushed. The volatile oils isolated from fresh fruits of S. occidentalis and S. hirsuta were subjected to gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and antimicrobial assays. GC and GC-MS analyses permitted the identification of 58 constituents. S. occidentalis oil was dominated by cyperene (10.8%), β-caryophyllene (10.4%), limonene (8.0%) and caryophyllene oxide (6.8%). The main components of S. hirsuta fruit oil were benzyl benzoate (24.7%), τ-cadinol (18.9%), 2,5-dimethoxy-p-cymene (14.6%) and β-caryophyllene (5.1%). S. occidentalis fruit oil exhibited better antimicrobial activity (MIC 78–312 μg/mL) against Escherichia coli, Staphylococcus aureus, Bacillus subtilis and Aspergillus niger compared with S. hirsuta oil. The compositions and the activities of the fruit essential oils of S. occidentalis and S. hirsuta are reported for the first time.

VOCs as fingerprints for the chemical profiling of hashish samples analyzed by HS-SPME/GC–MS and multivariate statistical tools

PurposeThe statistical evaluation of the chemical profile of seized hashish samples is a valuable tool to aid the estimation of the route through which the material has reached the dealers’ market.MethodsIn this study, the complete volatile organic compound (VOC) emission profiles of 48 seized hashish samples have been analyzed by means of headspace solid-phase microextraction/gas chromatography–mass spectrometry and evaluated with chemometric tools; multivariate statistical analyses, both hierarchical cluster analysis and principal component analysis (PCA) methods have been performed on the results to assess the existence of possible patterns throughout the samples.ResultsThe total VOC emission profiles sharply distributed the samples in clusters based on their batches of origin; this trend was also clearly shown in the PCA plot, in which samples coming from the same seizure were grouped together. The Δ9-tetrahydrocannabinol (THC) content analysis did not show a relevant trend in terms of lot of origin of the samples.ConclusionsThe evaluation of the VOCs released into the headspace traced a much more complete chemical profiling of the samples, as compared to the analysis of cannabinoids only, or the THC titration. The multivariate statistical analyses were very useful to estimate the origin of the seized material.

Chemical Composition and Activity of Essential Oils of Carissa macrocarpa (Eckl.) A.DC. Cultivated in Tunisia and Its Anatomical Features

This is the first study investigating the chemical composition of essential oils (EOs) isolated from different tissues of Carissa macrocarpa (Eckl.) A.DC., their antimicrobial activity and the anatomical characters of the aerial organs and the fruits. The main EO components were pentadecanal and tetradecan‐1‐ol (31.9 and 16.5% in fresh leaf EO, respectively), (E)‐nerolidol and caryophyllene oxide (27.3 and 15.0% in fruit EO, respectively), linalool and hexahydrofarnesyl acetone (30.9 and 24.9% in stem EO, respectively), benzyl benzoate (24.3% in flower EO). The fruit EO was more active against Candida albicans (MIC = 0.46 mg/mL) compared to the reference antibiotic (17.66 mg/mL). Furthermore, at this concentration it inhibited all the Gram‐positive bacteria. Concerning the anatomical features, it is noteworthy to mention the presence of a large cluster of calcium oxalate crystals inside some parenchymatous cells. Large ducts corresponding to non articulated laticifers were identified in the cortex of leaf, stem and fruit pericarp. The laticifers categories and their distribution are taxonomically important to discriminate this species from others acclimated in different countries. Considering the obtained results, EOs of C. macrocarpa can be a good source of antimicrobial compounds, contributing to solve the problem of microbial resistance to antibiotics.