Feature-Based Molecular Networking for the Exploration of the Metabolome Diversity of Common Egyptian Centaurea Species in Relation to Their Cytotoxic Activity.

Centaurea is a genus compromising over 250 herbaceous flowering species and is used traditionally to treat several ailments. Among the Egyptian Centaurea species, C. lipii was reported to be cytotoxic against multidrug-resistant cancer cells. In this context, we aimed to explore the metabolome of C. lipii and compare it to other members of the genus in pursuance of identifying its bioactive principles. An LC-MS/MS analysis approach synchronized with feature-based molecular networks was adopted to offer a holistic overview of the metabolome diversity of the Egyptian Centaurea species. The studied plants included C. alexandrina, C. calcitrapa, C. eryngioides, C. glomerata, C. lipii, C. pallescens, C. pumilio, and C. scoparia. Their constitutive metabolome showed diverse chemical classes such as cinnamic acids, sesquiterpene lactones, flavonoids, and lignans. Linking the recorded metabolome to the previously reported cytotoxicity identified sesquiterpene lactones as the major contributors to this activity. To confirm our findings, bioassay-guided fractionation of C. lipii was adopted and led to the isolation of the sesquiterpene lactone cynaropicrin with an IC50 of 1.817 µM against the CCRF-CEM leukemia cell line. The adopted methodology highlighted the uniqueness of the constitutive metabolome of C. lipii and determined the sesquiterpene lactones to be the responsible cytotoxic metabolites.

Chemical composition of the essential oil and botanical study of the flowers of Mentha suaveolens.

CONTEXT: Herbal medicines play a paramount role in the treatment of wide range of diseases, so there is a growing need for their quality control and standardization. Traditionally, histological and morphological inspections have been the usual methods to authenticate herbs intended for medicinal applications. Mentha suaveolens Ehrh. (Lamiaceae) is native to Africa Temperate Asia and Europe and it's cultivated in Egypt. OBJECTIVE: The macro- and micromorphology of the flowers of M. suaveolens Ehrh. cultivated in Egypt were studied to find the diagnostic characters of this species. In addition, the chemical composition of the essential oil of the flowers was also studied to define the chemotype of the plant. MATERIALS AND METHODS: Photographs of macro- and micromorphology were taken using Casio and Leica DFC500 digital cameras, respectively. In addition, the essential oil was prepared by hydrodistillation followed by gas chromatographic/mass spectrometric (GC/MS) analysis for identification of its components. RESULTS: The macro- and micromorphological characteristics of M. suaveolens were determined. The yield of the essential oil obtained by hydrodistillation from M. suaveolens flowers was 1.7% calculated on dry weight basis. GC/MS analysis of the oil resulted in identification of 29 components, which amounted to 99.77% of the total oil composition. The major component was carvone (50.59%) followed by limonene (31.25%). DISCUSSION AND CONCLUSION: The results obtained herein revealed for the macro, micromorphological and chemical composition characteristics of the flowers. The results of GC/MS analysis of the essential oil supported that M. suaveolens cultivated in Egypt could be categorized as carvone-rich chemotype since this compound pertained its high relative percentile.

Chemical composition and biological activities of the essential oil of Mentha suaveolens Ehrh.

Hydrodistilled oils of the fresh aerial parts of Mentha suaveolens Ehrh. cultivated in Egypt were prepared from samples collected along the four seasons. The percentage yields of these essential oils were 0.50%, 0.52%, 0.60%, and 0.47% of the dry weight for winter, spring, summer, and autumn samples. GC/MS analyses of all samples revealed a qualitative and quantitative variability in the oil composition. The total number of compounds identified was 46 among which 15 were common in all samples. The oxygenated compounds constituted about 45%, 46%, 63%, and 44% of the total composition of the oils for winter, spring, summer, and autumn samples, respectively. Carvone was the major constituent in spring, summer, and autumn samples (about 31%, 56%, and 35%, respectively), while limonene (ca. 26%) was the major constituent of the winter sample followed by carvone (ca. 25%). The essential oil of the highest yield (full-flowering summer sample), with the highest oxygenated constituents and carvone contents, was screened for certain biological activities. It exhibited analgesic and acute anti-inflammatory activities (75% and 82% relative to indomethacin). It also showed a potent in vivo antioxidant activity (96% relative to vitamin E). In addition, it exerted moderate cytotoxic, hepatoprotective, and in vitro antioxidant activities. Moreover, the oil had a potent antifungal activity against Candida albicans (MIC = 4 microg/ml), Saccharomyces cerevisiae (MIC = 5.2 microg/ml), and Aspergillus niger (MIC = 6.8 microg/ml).

Essential oil of Cymbopogon citratus cultivated in Egypt: seasonal variation in chemical composition and anticholinesterase activity.

The yield of essential oil obtained by hydrodistillation of Cymbopogon citratus fresh leaves ranged from 0.15% to 0.46% w/w; being the highest in spring and the lowest in winter. The oil sample obtained in winter exhibited a moderate acetylcholinesterase inhibitory activity (IC50 2.86 ± 0.17 mg/mL), compared to physostigmine (IC50 0.012 mg/mL), while other samples were relatively weak (IC50 values of 2.86-5.40 mg/mL). In all samples, oxygenated monoterpenes were predominating (73.22-89.32%). GC-MS identified a total of 61, 25, 50 and 63 components in oil samples obtained in spring, summer, autumn and winter, respectively. Citral content was the highest in autumn and summer samples (82.02% and 80.01% citral; respectively) and the lowest in winter sample (60.01%). Citral, isolated from the oil demonstrated a relatively potent anticholinesterase activity (IC50 0.21 ± 0.01 mg/mL).

Comparative Study on the Essential Oils from Five Wild Egyptian Centaurea Species: Effective Extraction Techniques, Antimicrobial Activity and In-Silico Analyses.

The genus Centaurea is recognized in folk medicine for anti-inflammatory, anti-itch, antitussive, purgative, astringent, and tonic activities. To study the chemical determinant for antimicrobial activity essential oils (EOs), five Centaurea species were analyzed including: C. scoparia, C. calcitrapa, C. glomerata, C. lipii and C. alexandrina. Conventional hydro-distillation (HD) and microwave-assisted extraction (MAE), as new green technologies, were compared for the extraction of essential oils. GC/MS analysis identified 120 EOs including mostly terpenoid except from C. lipii and C. alexandrina in which nonterpenoids were the major constituents. Major terpenoids included spathulenol, caryophyllene oxide and alloaromadendrene oxide-2. To probe antibacterial activity, potential EO inhibitors of a bacterial type II DNA topoisomerase, DNA gyrase B were screened via an in silico molecular docking approach. Spathulenol and alloaromadendrene oxide-2 possessed the best binding affinity in the ATP- binding pocket of Gyrase B enzyme. Principal component analysis and agglomerative hierarchical clustering were used for sample classification and revealed that sesquiterpenes contributed the most for accessions classification. In vitro antimicrobial activity against Staphylococcus aureus, Escherichia coli and Aspergillus niger for all EOs were also evaluated. EOs from C. lipii, C. glomerata and C. calcitrapa exhibited significant MIC against S. aureus with an MIC value of 31.25 µg/mL.

UPLC-Orbitrap HRMS metabolic profiling of Cymbopogon citratus cultivated in Egypt; neuroprotective effect against AlCl3-induced neurotoxicity in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Cymbopogon citratus (lemongrass) is commonly used in teas, soups and treat inflammatory-based ailments, vascular and nervous disorders. AIM OF THE STUDY: The study aimed to evaluate the neuroprotective effect of Cymbopogon citratus leaves through scientific protocol. The effect of aqueous (AE) and ethanolic (EE) extracts was evaluated against AlCl3-induced Alzheimer's disease (AD) in rats. Metabolic profiling of the plant, isolation of bioactive compounds and standardization of the active fraction were investigated. MATERIALS AND METHODS: AE of Cymbopogon citratus leaves was prepared as per traditional method (infusion), EE was prepared by repeated maceration in 90% ethanol, bioactive fraction (BAEE) was obtained from EE and the active compounds thereof were obtained by column chromatography. Metabolic profiling of Cymbopogon citratus was performed by UPLC-Orbitrap HRMS and HPLC was used for standardization. AlCl3-induced Alzheimer's rats were used to assess neuroprotective effect of the extracts. Neuroprotective mechanism(s) of Cymbopogon citratus extracts was clarified through histopathological examination of brain tissues, estimation of AD biochemical markers, oxidative stress and neuroinflammation in brain homogenates. In addition, antioxidant (using DPPH assay) and anticholinesterase (using modified Ellman's method) activities were investigated. RESULTS: AlCl3-treated rats (17 mg/kg/day) showed histopathological alteration in brain tissues together with elevated levels of Aß, tau proteins, MDA, NF-kB and IL-6. However, treatment with AE and EE of Cymbopogon citratus leaves prevented the pathological changes and maintained the levels of oxidative stress and inflammatory markers. In addition, BAEE significantly inhibited acetylcholinesterase enzyme (2.11 ±â€¯0.11 mg/ml) and exhibited a strong antioxidant activity (24.99 ±â€¯0.00 µg/ml). UPLC-MS of Cymbopogon citratus leaves showed peaks for twenty-eight compounds, twenty-one of them were identified. Three flavonoids; isoorientin, isoschaftoside and luteolin-7-O-neohesperidoside were isolated from BAEE as major constituents. The powdered leaves of Cymbopogon citratus was found to contain remarkable amounts of caffeic acid (3.49 mg/g dry wt.) and isoorientin (7.37 mg/g dry wt.) as determined by HPLC. CONCLUSION: Cymbopogon citratus ethanolic extract attenuates AlCl3-induced neurotoxicity in rats through inhibition of oxidative stress and inflammatory markers. This effect could possibly attributed, in part to its high content of phenolic acids and flavonoids. Accordingly, we recommend Cymbopogon citratus leaves for protection against AD.

Metabolites profiling of Chrysanthemum pacificum Nakai parts using UPLC-PDA-MS coupled to chemometrics.

Methanol-soluble constituents from the flowers, non-flowering aerial parts and roots of Chrysanthemum pacificum Nakai were analysed via high resolution UPLC-PDA-qTOF-MS followed by chemometrics. Forty-seven chromatographic peaks belonging to various metabolite classes were detected. Most metabolite classes showed qualitative and quantitative differences across parts, with luteolin conjugates being mostly enriched in flowers whereas non-flowering aerial parts contained mostly quercetin and methoxylated flavone conjugates. Root sample ranked the lowest for all flavones and dicaffeoylquinic acids. In contrast, 1,5-di-caffeoylquinic acid levels were found at high levels in flowers and aerial parts reaching 3145 and 1390 µg/g, respectively, suggesting that C. pacificum could serve as a natural resource of this well-recognised anti-hepatotoxic phenolic. Principal component analysis was further used for organs classification in an untargeted manner. This study provides the first map of secondary metabolites distribution in C. pacificum Nakai organs.

Comparative DNA profiling, phytochemical investigation, and biological evaluation of two Ficus species growing in Egypt.

AIM AND BACKGROUND: A comparison between two Ficus species, cultivated in Egypt, was carried out in this study. Their DNA analysis revealed that they are not closely related. MATERIALS AND METHODS: The pharmacopoeial constants of the leaves showed higher total ash and acid insoluble ash in F. lyrata than in F. platypoda. The other parameters were close in both species. Preliminary phytochemical screening revealed the presence of carbohydrate and/or glycosides, tannins, flavonoids, sterols, and triterpenes in their leaves and was detected in traces in their stems. RESULTS: Saponification of n-hexane extract of the leaves yielded 46% and 74.8% for the unsaponifiable matters and 20% and 15% for the fatty acids for F. platypoda and F. lyrata, respectively. n-Docosane (21.69%) and n-heptacosane (33.77%) were the major hydrocarbons in F. platypoda and F. lyrata, respectively. b-Sitosterol was the main sterol, palmitic (22.07%) and carboceric (35.72%) acids were the major identified saturated fatty acids in both species, while linoleic acid was the main unsaturated fatty acid (18.66% and 16.7%) in both species, respectively. The acute toxicity study revealed that the two species were safe up to 2 g/kg. The antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and pyrogallol as the standard was more significant for F. platypoda (232.6 µg/ml) than for F. lyrata, (790.9 µg/ml). The oral antihyperglycemic activity in diabetic rats using alloxan revealed that the 80% ethanolic extract of the leaves of F. platypoda was more active than that of the leaves of F. lyrata in decreasing the blood glucose level at 200 mg/kg/day (107.9 ± 5.817, 127.2 ± 4.359) and 400 mg/kg/day (64.11 ± 4.358, 127.7 ± 6.889), respectively, when compared with the diabetic control gliclazide (172.3 ± 2.089). CONCLUSION: The results of this study provide evidence that the two Ficus species have antioxidant and antihyperglycemic activity, in the order F. platypoda and then F. lyrata.