Gas Chromatography Coupled To Mass Spectrometry Research Articles

Polar Metabolites Profiling of Wheat Shoots (Triticum aestivum L.) under Repeated Short-Term Soil Drought and Rewatering.

The response of wheat (Triticum aestivum L.) plants to the soil drought at the metabolome level is still not fully explained. In addition, research focuses mainly on single periods of drought, and there is still a lack of data on the response of plants to short-term cyclical periods of drought. The key to this research was to find out whether wheat shoots are able to resume metabolism after the stress subsides and if the reaction to subsequent stress is the same. Gas chromatography coupled with mass spectrometry (GC-MS) is one of the most valuable and fast methods to discover changes in the primary metabolism of plants. The targeted GC-MS analyses of whole shoots of wheat plants exposed (at the juvenile stage of development) to short-term (five days) mild soil drought/rewatering cycles (until the start of shoot wilting) enabled us to identify 32 polar metabolites. The obtained results revealed an accumulation of sugars (sucrose, fructose, glucose, and 1-kestose), proline, and malic acid. During five days of recovery, shoots regained full turgor and continued to grow, and the levels of accumulated metabolites decreased. Similar changes in metabolic profiles were found during the second drought/rewatering cycle. However, the concentrations of glucose, proline, and malic acid were higher after the second drought than after the first one. Additionally, the concentration of total polar metabolites after each plant rewatering was elevated compared to control samples. Although our results confirm the participation of proline in wheat responses to drought, they also highlight the responsiveness of soluble carbohydrate metabolism to stress/recovery.

A Novel Assessment of Metabolic Pathways in Peritoneal Metastases from Low-Grade Appendiceal Mucinous Neoplasms.

There is a paucity of targeted therapies for patients with pseudomyxoma peritonei (PMP) secondary to low-grade appendiceal mucinous neoplasms (LAMNs). Dysregulated metabolism has emerged as a hallmark of cancer, and the relationship of metabolomics and cancer is an area of active scientific exploration. We sought to characterize phenotypic differences found in peritoneal metastases (PM) derived from LAMN versus adenocarcinoma. Tumors were washed with phosphate-buffered saline (PBS), microdissected, then dissociated in ice-cold methanol dried and reconstituted in pyridine. Samples were derivatized in tert-butyldimethylsilyl (TBDMS) and subjected to gas chromatography-coupled mass spectrometry. Metabolites were assessed based on a standard library. RNA sequencing was performed, with pathway and network analyses on differentially expressed genes. Eight peritoneal tumor samples were obtained and analyzed: LAMNs (4), and moderate to poorly differentiated adenocarcinoma (colon [1], appendix [3]). Decreases in pyroglutamate, fumarate, and cysteine in PM from LAMNs were found compared with adenocarcinoma. Analyses showed the differential gene expression was dominated by the prevalence of metabolic pathways, particularly lipid metabolism. The gene retinol saturase (RETSAT), downregulated by LAMN, was involved in the multiple metabolic pathways that involve lipids. Using network mapping, we found IL1B signaling to be a potential top-level modulation candidate. Distinct metabolic signatures may exist for PM from LAMN versus adenocarcinoma. A multitude of genes are differentially regulated, many of which are involved in metabolic pathways. Additional research is needed to identify the significance and applicability of targeting metabolic pathways in the potential development of novel therapeutics for these challenging tumors.

Essential oils of Lauraceae: antimicrobial activity and constituents of Cinnamomum auricolor Kosterm. and Cinnamomum petelotii Kosterm leaves essential oils from Vietnam

In this study, the antimicrobial activity and constituents of essential oils from the leaves of Cinnamomum auricolor Kosterm. and C. petelotii Kosterm collected from Pù Hoạt Natural Reserve (Hạnh Dịch Commune, Quế Phong District), Vietnam were reported. The essential oils obtained by separate hydrodistillation were analyzed by means of gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS). The antimicrobial activities were evaluated by microbroth dilution susceptibility assay. The main constituents of C. auricolor comprised of oxygenated sesquiterpenes mainly caryophyllene oxide (26.0%), spathulenol (11.1%), and (E)-nerolidol (10.7%). However, oxygen-containing monoterpenes were the main compounds identified in S. petelotii and they are linalool (34.3%), 1,8-cineole (20.8%) and terpinene-4-ol (9.9%). The essential oil of C. auricolor displayed prominent antimicrobial activity towards Bacillus cereus ATCC14579 and Staphylococcus aureus ATCC25923 with minimum inhibitory concentration (MIC) value of 16.0 μg/mL. In addition, the essential oil of C. auricolor also exhibited antimicrobial activity against Enterococcus faecalis ATCC29212, and anti-candidal activity towards Candida albicans ATCC10231 with MIC value of 32.0 μg/mL. The leaf essential oil of C. petelotii only exhibited activity towards S. aureus and C. albicans with MIC value of 64.0 μg/mL. The chemical constituents and antimicrobial activity of both essential oils are being reported for the first time.

Bioactive compounds against Moniliophthora roreri (Cif & Par) identified in locally produced liquid amendments (Biols)

The use of liquid organic amendments (biols) is a common practice for farmers due to the multiple benefits in the management and production of their crops, including the control of pests and diseases. The present study analyzes the chemical composition of the pure compound C1 and fractions F2 and F3 of local biols produced in two provinces of Ecuador and their antifungal activity against Moniliophthora roreri (Cif & Par). This work incorporates the use of Gas Chromatography coupled to Mass Spectrometry (GC-MS), Nuclear Magnetic Resonance (NMR) and in vitro inhibition assays for sample analysis. C1 was identified as Mannitol. The percentage of inhibition against M. roreri in F2 and F3 was 44.37% and 8.34%, respectively; and, for C1, 28.63%. The values of the median lethal dose (LD50) obtained corroborate that the F2 fraction was the one with the highest controlling activity against the pathogen. The 1,2-benzenedicarboxylic acid diisooctyl ester compound was the main compound in F2 (30.88%) and the Pentadecanoic acid, 14-methyl-, methyl ester in F3. Finally, all compounds obtained from the biol fractions were identified and it was determined that the fermentative process was suitable for producing bioactive compounds of interest to inhibit the growth of Moniliophthora roreri.

Metabolomic exhibits different profiles and potential biomarkers of Vitis sp co-cultivated with Fusarium oxysporum for short, medium, and long times.

Differential rootstock tolerance to Fusarium sp. supports viticulture worldwide. However, how plants stand against the fungus still needs to be explored. We hypothesize it involves a differential metabolite modulation. Thus, we performed a gas chromatography coupled with mass spectrometry (GC-MS) analysis of Paulsen P1103 and BDMG573 rootstocks, co-cultured with F. oxysporum (FUS) for short, medium, and long time (0, 4, and 8 days after treatment - DAT). In shoots, Principal Component Analysis (PCA) showed a complete overlap between BDMG573 CNT and FUS at 0 DAT, and P1103 treatments showed a slight overlap at both 4 and 8 DAT. In roots, PCA exhibited overlapping between BDMG573 treatments at 0 DAT, while P1103 treatments showed overlapping at 0 and 4 DAT. Further, there is a complete overlapping between BDMG573 and P1103 FUS profiles at 8 DAT. In shoots, 1,3-dihydroxyacetone at 0 and 4 DAT and maltose at 4 and 8 DAT were biomarkers for BDMG573. .For P1103, glyceric acid, proline, and sorbitol stood out at 0, 4, and 8 DAT, respectively. In BDMG573 roots, the biomarkers were β-alanine at 0 DAT, cellobiose and sorbitol at both 4 and 8 DAT. While in P1103 roots, they were galactose at 0 and 4 DAT and 1,3-dihydroxyacetone at 8 DAT. Overall, there is increase in amino acids, glycolysis, and TCA components in tolerant Paulsen P1103 shoots. Thus, it provides a new perspective on the primary metabolism of grapevine rootstocks to F. oxysporum that may contribute to strategies for genotype tolerance and early disease identification. This article is protected by copyright. All rights reserved.

Climatological effects on Chemical Composition, Biological Activity of Essential oil from Eucalyptus globulus Leaves and Seeds

In the last decade, there has been interest worldwide concern about the antibacterial and antioxidant properties of plants. This work aims to study the antibacterial, antioxidant activities and chemical composition of Eucalyptus globulus essential oil leaves and seeds. Essential oil was extracted from Eucalyptus globulus seeds and leaves by hydrodistillation giving yields of 1.8% and 0.8% respectively, Chemical composition analysis by gas chromatography coupled with mass spectrometry (GC-MS) revealed the presence of a total of 33 compounds in seeds and 28 compounds in leaves. Antibacterial activity showed that essential oils was active against all tested bacterial strains. Antioxidant propriety measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) showed that essential oils obtained from leaves have higher antioxidant activity when compared to essential oils of seeds. However, the total antioxidant capacity revealed that seeds (22,68 mg EAA/g MS) have high activity with respect to leaves (5,44 mg EAA/g MS). These results confirmed the richness of the two essential oils in phenolic compounds responsible for their biological activity. A comparative bioclimatic study was carried out in order to investigate the effect of climate on chemical composition of Eucalyptus globulus leaves and seeds in Morocco. Factorial component analysis between chemical composition of essential oils from different countries and climatic parameters revealed that 1.8 cineole is strongly correlated with minimal (m) and maximal temperature values (M) and Emberger quotient (Q). Whereas, essential oils extracted from seeds is not affected by these parameters.&nbsp

Aegle marvels (L.) Correa Leaf Essential Oil and Its Phytoconstituents as an Anticancer and Anti-Streptococcus mutans Agent.

Aegle mamelons (A. marmelos) or Indian Bael leaves possess anti-cancerous and antibacterial properties and are used in the traditional medicine system for the treatment of oral infections. In the present study, the essential oil of the leaves of A. marmelos was explored for its anticancer, antioxidant, and anti-cariogenic properties. The hydro-distilled oil of A. marmelos leaves was analyzed using gas chromatography coupled with mass spectrometry (GC-MS). Monoterpene limonene (63.71%) was found to have the highest percentage after trans-2-Hydroxy-1,8-cineole and p-Menth-2,8-dien-1-ol. The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay was used to investigate the anticancer activity of the extracted oil against human oral epidermal carcinoma (KB), and the results showed significantly higher (**** p < 0.0001) anticancer activity (45.89%) in the doxorubicin (47.87%) when compared to the normal control. The antioxidant activity of the essential oil was evaluated using methods of DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)). The results showed a significant (*** p < 0.001) percentage of inhibition of DPPH-induced free radical (70.02 ± 1.6%) and ABTS-induced free radical (70.7 ± 1.32%) at 100 µg/mL with IC50, 72.51 and 67.33 µg/mL, respectively, comparatively lower than standard compound ascorbic acid. The results of the molecular docking study of the significant compound limonene with the receptors tyrosinase and tyrosine kinase 2 supported the in vitro antioxidant potential. The anti-cariogenic activity was evaluated against Streptococcus mutans (S. mutans). Results showed a significant minimum inhibitor concentration of 0.25 mg/mL and the killing time was achieved at 3 to 6 h. The molecular-docking study showed that limonene inhibits the surface receptors of the S. mutans c-terminal domain and CviR protein. The study found that A. marmelos leaves have potential anti-carcinoma, antioxidant, and anti-cariogenic effects on human oral epidermal health, making them a valuable natural therapeutic agent for managing oral cancer and infections.

Synthesis of chalcones and their antimicrobial and drug potentiating activities

The increased resistance of microorganisms to antimicrobial drugs makes it necessary to search for new active compounds, such as chalcones. Their simple chemical structure makes them molecules easy to synthesize. Therefore, the aim of this study was to evaluate the antimicrobial and potentiating activity of antibiotics and antifungals by synthetic chalcones against strains of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Candida tropicalis. The synthesis of chalcones was carried out by Claisen-Schimidt aldol condensation. Nuclear Magnetic Resonance (NMR) and Gas Chromatography Coupled to Mass Spectrometry (GC/MS) were also performed. Microbiological tests were performed by the broth microdilution method, using gentamicin, norfloxacin and penicillin as standard drugs for the antibacterial assay, and fluconazole for the antifungal assay. Three chalcones were obtained (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one (DB-Acetone), (1E,3E,6E,8E)-1,9-diphenylnone-1,3,6,8-tetraen-5-one (DB-CNM), (1E,4E)-1,5-bis (4-methoxyphenyl) penta-1,4-dien-3-one (DB-Anisal). The compound DB-Acetone was able to inhibit P. aeruginosa ATCC 9027 at a concentration of 1.4 × 102 μM (32 μg/mL), while DB-CNM and DB-Anisal inhibited the growth of S. aureus ATCC 25923 at 17.88 × 102 μM and 2.71 × 101 μM (512 μg/mL and 8 μg/mL) respectively. In the combined activity, DB-Anisal was able to potentiate the effect of the three antibacterial drugs tested against E. coli 06, norfloxacin (128 for 4 μg/mL ±1) against P. aeruginosa 24 and penicillin (1,024 for 16 μg/mL ±1) against S. aureus 10. In antifungal assays, chalcones were not able to inhibit the growth of fungal strains tested. However, both showed potentiating activity with fluconazole, ranging from 8.17 x 10−1 μM (0.4909 μg/mL) to 2.35 μM (13.96 μg/mL). It is concluded that synthetic chalcones have antimicrobial potential, demonstrating good intrinsic activity against fungi and bacteria, in addition to potentiating the antibiotics and antifungal tested. Further studies are needed addressing the mechanisms of action responsible for the results found in this work.

Herbivore induced fruit volatiles guide the solitary tephritid endoparasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae) in selecting an oviposition substrate.

Oviposition in a generalist parasitoid is a complex process that involves interactions among its host, host plant, and spatiotemporal environment. Our study was aimed at exploring the ability of a generalist parasitic wasp, Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), in perceiving odor cues from its host insect [Oriental fruit fly Bactrocera dorsalis (Hendel, Diptera: Tephritidae)] in infested mango fruits. A series of experiments involving behavioral and electrophysiological studies suggest that D. longicaudata is able to distinguish host infested fruit volatiles from uninfested. Additionally, we found that D. longicaudata can distinguish between high and low-level infestations using specific cues emanating from fruits. Coupled gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry (GC-MS) analysis of infested mango fruit head space volatiles identified compounds such as ethyl butanoate, ethyl 3-hydroxybutyrate, β-myrcene, trans-β-ocimene, allo-ocimene, ethyl octanoate, ethyl-α-toulate, β-elemene, β-caryophyllene, humulene, caryophyllene oxide, ethyl dodecanoate, ethyl hexadecanoate, 2-furancarboxylic acid, 3-hydroxy-2-methyl-4-pyrone, and phenethyl alcohol emanated from three different host treatments with different levels of larval infestation which elicited antennal responses in D. longicaudata. We suggest that these cues might play a significant role in attracting D. longicaudata in the field.

Fingerprinting methods for origin and variety assessment of rice: development, validation and data fusion experiments

Several analytical techniques, i.e. Near Infrared (NIR) and Mid-Infrared (MIR) spectroscopy, Hyper Spectral Imaging (HSI), Gas Chromatography coupled to Mass Spectrometry (GC-MS) and Proton-transfer Reaction Time-of-Flight Mass spectrometry (PTR-TOF-MS), combined with chemometrics, are examined to evaluate their potential for solving food authenticity questions on the case of rice. In total, 237 rice samples were analyzed in this study to examine origin and variety assessment and sourced from producing countries (Italy, Spain, Vietnam, Pakistan and Thailand). The Gaussian Process Latent Variable Model (GP-LVM) was applied as technique to obtain a meaningful compressed representation of the data in a two-dimensional space followed by a classification with a nearest neighbour algorithm. What concerns the origin assessment, GC-MS results score good across all countries, resulting in the most accurate method, with prediction rates ranging from 86% to 94%. Data fusion experiments of combination of GC-MS with NIR, or GC-MS with HSI resulted in prediction rates on origin assessment of more than 90%. Variety assessment was performed with these analytical techniques. Using single techniques, HSI achieved prediction values above 90% for all classes (96%–99%). For data fusion experiments of variety assessment combining GC-MS and NIR all prediction values were equal or higher than 92%; for GC-MS combined with HSI, prediction values were all 98%. The presented methodology can be successfully used for the authentication of rice.

Chronic alcohol administration alters metabolomic profile of murine bone marrow.

People with hazardous alcohol use are more susceptible to viral, bacterial, and fungal infections due to the effect of alcohol on immune system cell function. Metabolized ethanol reduces NAD+ to NADH, affecting critical metabolic pathways. Here, our aim was to investigate whether alcohol is metabolized by bone marrow cells and if it impacts the metabolic pathways of leukocyte progenitor cells. This is said to lead to a qualitative and quantitative alteration of key metabolites which may be related to the immune response. We addressed this aim by using C57BL/6 mice under chronic ethanol administration and evaluating the metabolomic profile of bone marrow total cells by gas chromatography-coupled mass spectrometry (GC-MS). We identified 19 metabolites. Our data demonstrated that chronic ethanol administration alters the metabolomic profile in the bone marrow, resulting in a statistically diminished abundance of five metabolites in ethanol-treated animals: uracil, succinate, proline, nicotinamide, and tyrosine. Our results demonstrate for the first time in the literature the effects of alcohol consumption on the metabolome content of hematopoietic tissue and open a wide range of further studies to investigate mechanisms by which alcohol compromises the cellular function of the immune system.

Anaerobic biodegradation of anthracene by oral Firmicutes isolates from smokers and its potential pathway

Exposure to polycyclic aromatic hydrocarbons (PAHs) from tobacco smoke has been linked to many negative health effects. Studies on the biodegradation of PAHs by human microbiota and detailed pathways for their anaerobic biodegradation are scarce despite their importance in getting rid of these toxic compounds. In a previous study for our group, we determined the ability of oral bacterial isolates in the anaerobic biodegradation of anthracene as a model of PAHs. Three isolates with the highest anthracene degradation ability were selected for the present study which include Limosilactobacillus fermentum, Veillonella parvula, and Streptococcus anginosus. In this study, we aimed at exploring and elucidating the anthracene anaerobic biodegradation pathways in selected Firmicutes oral isolates. Metabolites throughout the pathway were detected by gas chromatography coupled with mass spectroscopy (GC-MS) using anthracene as sole source of carbon. After incubation for 3 days, anthracene was undetected in the supernatant of L. fermentum and V. parvula, while a residual of 3% of anthracene was detected in presence of S. anginosus. Results revealed that anaerobic biodegradation by L. fermentum and V. parvula started with hydroxylation and dehydrogenation producing 9,10- anthraquinone and ended up with simpler structures such as catechol, while S. anginosus hydroxylation for anthracene resulted in the production of 1,2-anthracenediol and ended up with catechol and phthalic acid. The biodegradation of anthracene by oral bacteria could convert it to other toxic metabolites such as anthraquinone and catechol which were reported to have potential carcinogenic effects. Moreover, fatty acids detected as biodegradation metabolites could be one of the causes of smokers’ heart-related diseases. Thus, this study explored oral metabolites resulting from smoking under anaerobic conditions towards elucidating the role of oral microbiota in health and disease states.

Quantification of 7 cannabinoids in cannabis oil using GC-MS: Method development, validation and application to therapeutic preparations in Friuli Venezia Giulia region, Italy

The use of therapeutic cannabis preparations in Friuli Venezia Giulia is increasingly expanding. Even if cannabis oil finds its applications in several disorders affecting adults and children, it is not yet a standardized product and, to ensure the quality of the preparation, a quantitative analysis must be carried out before dispensing it to patients. Gas chromatography coupled to mass spectrometry (GC-MS) is a frequently used technique for quantification of cannabinoids, the active compounds of C. sativa. In this context, we developed a GC-MS method for the simultaneous quantification of 7 cannabinoids (CBD, CBDA, CBG, CBN, THCA, THCV and Δ9-THC) that is not time and sample consuming: 10 μL of cannabis oil were used for the sample preparation, that consists in derivatization of analytes through silylation. Calibration curves were built from 0.2 to 2 μg/mL. The percentage of accuracy and precision did not exceed the values recommended by validation guidelines. The limit of detection was 0.01 μg/mL; whereas the lower limit of quantification was 0.2 μg/mL. There was no carry over. The proposed GC-MS method showed good sensitivity, specificity, linearity, accuracy, precision and applicability to therapeutic preparations.

Preparation, physicochemical characterization and computational studies of Plectranthus ornatus codd essential oil/β-cyclodextrin inclusion complex

This study aims to prepare and chemically characterize inclusion complexes (IC) between β-cyclodextrin (β-CD) and Plectranthus ornatus essential oil (OEPO). Samples were evaluated by gas chromatography coupled with mass spectrometry (GC-MS) and Fourier transform infrared spectroscopy (FTIR). Moreover, a theoretical study concerning inclusion complexes of β-CD with the four main compounds of OEPO through Molecular Dynamics (MD) simulations, Density Functional Theory (DFT) and Parametric Method-6 (PM6) calculations was performed. The GC-MS analysis of OEPO resulted in the identification of 36 volatile compounds, with the four main components of this essential oil being: β-caryophyllene (21.52%), terpinen-4-ol (6.89%), phytol (4.00%) and caryophyllene oxide (3.32%). The GC-MS results indicated the presence of OEPO volatile compounds in the ICs prepared by the water/ethanol kneading (KWE) and water kneading (KW) method. More specific, β-CD inclusion complexes with β-butoxyethanol (1.15%), 1,3,8-ρ-menthatriene (0.45%), β-caryophyllene (0.20%), phytol (0.41%) and δ-cadinol (0.20%) prepared by the KWE method were detected, while the KW inclusion method resulted in the encapsulation of β-butoxyethanol (3.98%) only. The higher inclusion content of OEPO molecules in KWE was also confirmed by FTIR, with the KWE complex showing more OEPO absorption bands than KW. The PM6 study showed that hydrogen bonds are important, but hydrophobic interactions are the main driving force for triggering the complex formation of the main OEPO volatile compounds with β-CD. The intermolecular host-guest interactions and the investigation for the most suitable inclusion mode were examined by conducting several Molecular dynamics simulations. Moreover, PM6 and DFT approaches revealed the most stable cyclodextrin inclusion complex structures. These findings are in accordance with the experimental data from GC-MS analysis. The effect of solvent on the interaction energy was also studied through simulations and the results showed that the most stable structures are characterized by the formation of hydrogen bonds between guests and cyclodextrin hosts. The developing of such preparation methods that results in ICs like OEPO/β-CD may be useful in the optimization of existing formulations and therefore, be adopted by the pharmaceutical industry

The antimicrobial activity and essential oil constituents of the leaves and trunks of Aquilaria banaensis P.H.Hô (Thymelaeceae) from Vietnam

The aim of the present paper was to report the chemical constituents and antimicrobial activity of essential hydrodistilled from the leaves and trunk of Aquilaria banaensis P.H.Hô (Thymelaeceae) from Vietnam. The essential oils were analysed comprehensively for their constituents by using Gas chromatography coupled with Mass spectrometry (GC/MS). The antimicrobial activity was determined by agar well diffusion and broth microdilution methods. The leaf essential oil comprised mainly of sesquiterpenes while fatty acids constitutes the bulk of the trunk essential oil. The main constituents of the leaf essential oil were β-caryophyllene (17.11%), α-selinene (10.99%), α-humulene (8.98%), β-selinene (8.01%), β-guaiol (6.69%) and β-elemene (5.65%). However, hexadecanoic acid (48.46%), oleic acid (19.80%) and tetradecanoic acid (5.32%) were the major compounds identified in the trunk essential oil. The trunk essential oil displayed antimicrobial activity against Staphylococcus aureus, with the minimum inhibitory concentration (MIC) value of about 256.0 µg/mL.

Conventional vs. Microwave-Assisted Hydrodistillation: Influence on the Chemistry of Sea Fennel Essential Oil and Its By-Products.

The main objectives of this study were to investigate the effects of the applied essential oil (EO) isolation method, conventional hydro-distillation (HD), and microwave-assisted hydro-distillation (MHD) on the chemical profile of sea fennel (Crithmum maritimum L.) essential oil and to investigate the main constituents present in the liquid by-products of EOs isolation (hydrolate and residual wastewater). Headspace-solid phase microextraction (HS-SPME) was used to isolate hydrolate components, while gas chromatography coupled with mass spectrometry (GC-MS) was used to detect and analyse the chemical constituents of the essential oils and hydrolates. The phenolic composition of the wastewater extracts was analysed by high performance liquid chromatography (HPLC). The EO obtained by MHD had a higher yield of limonene and sabinene. The chemical composition of the hydrolates differed from the EO compositions. The content of terpinen-4-ol in the MHD hydrolate was higher, while several compounds were detected in relatively high proportions only in the HD hydrolate. MHD also resulted in a higher phenolic content of the wastewater, where an increase in the concentration of chlorogenic acid was also observed. It can be concluded that the isolation method had a great influence on the profile of sea fennel EOs, especially on their corresponding hydrolates and residual wastewater extracts. Due to their valuable chemical composition, these by-products can be a cost-effective source of bioactive compounds that have great potential for use in various industries.

The phenological stage of hemp inflorescences affects essential oil yield and its chemical composition

The inflorescences of Cannabis sativa L. represent a high-value by-product of industrial hemp cultivation, and their volatile fraction has recently attracted attention due to the applications of hemp-derived Essential Oil (EO) in several fields. Following a previous evaluation of the EOs profile in diverse hemp genotypes, the current study investigated how the phenological stage of inflorescences affects the EO chemical composition and hydrodistillation yield. Two hemp varieties with different flower behaviour, Codimono (monoecious) and Fibrante (dioecious), were cultivated in open field, and the flowering tops were collected at five developmental stages. The air-dried samples were hydrodistilled with a Clevenger apparatus, and the obtained EOs were analysed by Gas Chromatography coupled with Mass Spectrometry (GC-MS). All the samples showed terpenes and phytocannabinoids as major chemical classes. Among the former class, sesquiterpene hydrocarbons were the most abundant components in the EOs at all phenological stages, ranging from 34.1% to 67.6% and 33.8–73.8% in Codimono and Fibrante, respectively, while oxygenated sesquiterpenes were highest at the final stage (27.7% in Codimono and 24.0% in Fibrante). Phytocannabinoids, instead, were the second major chemical class, accounting for 29.3% in Codimono and 30.5% Fibrante. The EO yield increased with phenological development, reaching the highest rate at seed harvest time in both genotypes (0.018–0.265% w/w in Codimono and 0.026–0.265% w/w in Fibrante).

Effect of a polyherbal formulation on L-thyroxine induced hyperthyroidism in a rat model: In vitro and in vivo analysis and identification of bioactive phytochemicals

An excess of thyroid hormones in the blood characterizes hyperthyroidism. Long-term use of prescription medications to treat hyperthyroidism has substantial adverse effects and when discontinued, the symptoms frequently recur. Several plant species have been utilized to cure hyperthyroidism. In the present work, we investigated the impact of polyherbal extract (POH) of four medicinal plants to treat hyperthyroidism. Biochemical analysis revealed the presence of a high concentration of phytochemicals in the POHs. The in vitro antioxidant study revealed their antioxidant and free radical scavenging capacity. The gas chromatography coupled mass spectrometry analysis of the POHs showed the presence of 13 bioactive phytochemical compounds. The effect of various concentrations of POHs on L-thyroxine-induced hyperthyroidism in Wistar albino rats was evaluated for 18 days. The TSH, T3 and T4 levels increased significantly and reduced the increase of liver enzymes caused by hyperthyroidism in POH-treated rats. The data showed that POH therapy could restore thyroid function to normal. The injection of POH increased the size comprising vacuolated cells, columnar follicular cells and highly coloured nuclei with increasing POH content and the number of normal thyroid follicles rose. The findings indicate that polyherbal formulations of these medicinal plants include credible antithyroid compounds that may offer a protective and an effective alternative treatment to synthetic thyroid medications.

Non-host plant odors influence the tritrophic interaction between tomato, its foliar herbivore Tuta absoluta and mirid predator Nesidiocoris tenuis.

The tomato leafminer, Tuta absoluta is a destructive invasive pest of cultivated tomato and other Solanaceae plants, with yield losses of 80-100%. Mirid predators are key natural enemies of T. absoluta, but they also feed on host plants in the absence of their prey. Management of T. absoluta is a challenge due to its high biotic potential, resistance to many insecticides and the absence of sufficiently adapted auxiliary fauna in its new dispersion zones. Olfaction plays an important role in the tritrophic interaction between tomato, its herbivore pest T. absoluta and its mirid predators, which can be influenced by non-host plant odors. However, how non-host odours shape this interaction is poorly understood. Previously, we had demonstrated belowground crop protection properties of certain Asteraceae plants against the root-knot nematode Meloidogyne incognita, pest of tomato and other Solanaceae plants. Additionally, Asteraceae plants impact negatively on feeding behavior of above-ground pests of Solanaceae plants, including the greenhouse whitefly (Trialeurodes vaporariorum) and green peach aphid (Myzus persicae). Here, we tested the hypothesis that foliar volatiles from some of these non-host Asteraceae plants can influence the tomato-T. absoluta-mirid predator tritrophic interaction. In olfactometer assays, T. absoluta females were attracted to volatiles of the Solanaceae host plants tomato and giant nightshade but avoided volatiles of the Asteraceae plants, blackjack and marigold, and the positive control, wild tomato, when tested alone or in combination with the host plants. Coupled gas chromatography-mass spectrometry analysis showed that host and non-host plants varied in their emission of volatiles, mainly monoterpenes and sesquiterpenes. Random forest analysis combined with behavioral assays identified monoterpenes as the host plant attractive blend to T. absoluta and its mirid predator, with sesquiterpenes identified as the non-host plant repellent blend against T. absoluta. Contrastingly, the mirid predator was indifferent to the non-host plant repellent sesquiterpenes. Our findings indicate that terpenes influence the tomato-T. absoluta-mirid predator tritrophic interaction. Further, our results emphasize the importance of studying crop protection from a holistic approach to identify companion crops that serve multi-functional roles.

Chemical Characterization of Crude Phenolic Leaf Extract of Acalypha wilkesiana

Acalypha wilkesiana is a widely eaten vegetable, reportedly used traditionally in the management of hypertension, diabetes and as an antimicrobial agent, probably because of its rich phenolic composition. The phytochemical and phenolic profiles of the crude leaf extract of the plant were assessed using standard methods, and gas chromatography coupled to mass spectrometry (GC-MS) and flame ionization detector (GC-FID). Qualitative phytochemical screening showed the presence of alkaloids, phenols, flavonoids, cardiac glycosides, tannin, steroids and saponins. GC-FID analysis revealed the presence of dihydrocytisine, ammodendrine, spartein, hydroxylupanine, ribalinidine, anthocyanin, flavone, flavonones, aphylidine, proanthocyanidin, isolupanine, narigenin, sapogenin, cardiac glycoside, tannin, and cyanogenic glycoside in varying proportions. GC-MS analysis identified 14 constituents with major ones being 2,6-Octadienal, 3,7-dimethyl-, (Z) Citral (14.88 %), citral 2,6-Octadienal, 3,7-dimethyl-, (E) (16.9 %), cis-3-Hexenyl cis-3-hexenoate (10.59 %), trans-2,7-Dimethyl-4,6-octadien-2-ol (20.56 %) and 3,8,11-Trioxatetracyclo[4.4.1.0(2,4).0(7,9)] undecane (11.69 %). This study has shown that crude phenolic leaf extract of A. wilkesiana contains pharmacologically active compounds which support its traditional use in the treatment of oxidative stress related diseases and hence a potential candidate for drug discovery. Keywords: Acalypha wilkesiana, phytochemicals, gas chromatography-mass spectrometry, Secondary metabolites.