Phytochemical Screening, Antioxidant Assay and Cytotoxic Profile for Different Extracts of Chrysopogon zizanioides Roots.

The Chrysopogon zizanioides plant possesses multiple traditional uses, especially in therapeutics, but only a few articles have reported its biological activity. Hence, the present study was planned to explore the phytochemical constituents, cytotoxic potential, radical scavenging activity, and GC/MS (Gas chromatography & Mass spectrometry) analysis of the vetiver root extracts. The roots extracted with different solvents exhibited more significant phytochemical constituents in polar solvents in comparison to non-polar ones, favoring the extraction of a greater number of components in highly polar solvents. All the extracts were tested for their cytotoxicity using SRB (Sulforhodamine B) assay. They confirmed ethanolic extract as a potent extract with GI50 56±0.5 µg/ml in oral cancer (SCC-29B) along with no cytotoxicity in healthy cells (Vero cells), making it a safer therapeutic option in comparison to standard Adriamycin. This extract was also analyzed for its antioxidant potential by DPPH (1,1-Diphenyl-2-picrylhydrazyl) assay with IC50 value 10.73 µg/ml, which was quite comparable to Ascorbic acid having IC50 value 4.61 µg/ml. The quantitative analysis of ethanolic extract exhibited 107 compounds amongst which Khusenic acid, Ascorbic acid, Junipen, gamma-Himachalene, alpha-Guaiene were the majorly occurring compounds that can be explored further for their cytotoxic activity.

Root essential oil of Chrysopogon zizanioides relaxes rat isolated thoracic aorta - an ex vivo approach.

Root of Chrysopogon zizanioides (L.) Roberty has been used in Siddha system of medicine to treat hypertension. The present study was therefore to investigate the vasorelaxation effect of root essential oil of C. zizanioides using rat isolated thoracic aortic rings. Chemical characterization of root essential oil was carried out using Gas Chromatography-Flame Ionization Detector (GC-FID) and Gas Chromatography-Mass Spectrometry (GC-MS). Essential oil nanoemulsion (EONE) was prepared and characterized. Vasorelaxant effect of EONE in endothelium-intact aortic rings precontracted with phenylephrine (PE) (1 µM) or KCl (80 mM) was investigated. Role of Ca2+, nitric oxide and K+ channels in precontracted aortic rings were investigated to elucidate the mechanism of action of the essential oil. Further, the role of muscarinic and prostacyclin receptors in EONE induced relaxation was studied. The EONE significantly induced relaxation (Emax 77.1 ± 4.87%) in PE precontracted aortic rings. The nitric oxide synthase, and cyclooxygenase inhibitors and potassium channel blockers have not significantly inhibited the vasorelaxation induced by EONE. However, EONE induced relaxation in precontracted endothelium-intact aortic rings was significantly inhibited by muscarinic receptor and calcium channel. The root essential oil of C. zizanioides possesses vasorelaxant effect through muscarinic pathway as well as acts as calcium channel blocker.

Metabolic response of vetiver grass (Chrysopogon zizanioides) to acid mine drainage.

Acid mine drainage (AMD) is a sulfuric discharge containing metals and particulates that can spread to nearby water sources, imposing toxicity and physical stress to living things. We have shown that vetiver grass (Chrysopogon zizanioides) is capable of tolerating and treating AMD-impacted water from the abandoned Tab-Simco mining site from southern Illinois, though little is known about its tolerance mechanisms. We conducted metabolomic analyses of vetiver shoots and roots after relatively short- and long-term periods of exposure to Tab-Simco AMD. The metabolic shift of vetiver shoots was dramatic with longer-term AMD exposure, including upregulation of amino acid and glutathione metabolism, cellular respiration and photosynthesis pathways, with downregulation of phosphorylated metabolites. Meanwhile, the roots demonstrated drastic downregulation of phospholipids and phosphorylated metabolites, cellular respiration, glyoxylate metabolism, and amino acid metabolism. Vetiver accumulated ornithine and oxaloacetate in the shoots, which could function for nitrogen storage and various intracellular functions, respectively. Organic acids and glutathione were secreted from the roots for rhizospheric metal-chelation, whereas phosphorylated metabolites were recycled for phosphorus. These findings reveal AMD-induced metabolic shifts in vetiver grass, which are seemingly unique in comparison to independent abiotic stresses reported previously.

Bioguided identification of antimicrobial compounds from Chrysopogon zizaniodes (L.) Roberty root essential oil.

Aim: To determine the group of compounds from Chrysopogon zizaniodes root essential oil that have antimicrobial activity. Materials & methods: Thin-layer chromatography coupled to direct bioautography was used to determinate the fraction(s) having antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREF). Through GC-MS identification, the fractions with the greatest similarity to the active thin-layer chromatography fraction were used to determinate the MIC. Results: The subfraction 8 from column chromatography was responsible for the best MIC for MRSA (62.5 µg/ml) and VREF (125 µg/ml). Five compounds possibly responsible for antimicrobial activity were preliminary identified. Conclusion: We suggest that Cedr-8-en-13-ol, could be the more relevant compound involved in the antimicrobial activity in this study.

Chemical Composition, Antioxidant, and Antimicrobial Activities of Vetiveria zizanioides (L.) Nash Essential Oil Extracted by Carbon Dioxide Expanded Ethanol.

In the present study, the composition of essential oil isolated from the roots of Vetiveria zizanioides (L.) Nash, harvested in China, was studied, along with the bioactivities. A green novel method using an eco-friendly solvent, CO2-pressurized ethanol, or carbon dioxide expanded ethanol (CXE) was employed to isolate the essential oil from the root of Vetiveria zizanioides (L.) Nash with the purpose of replacing the traditional method and supercritical fluid extraction (SFE). After investigating the major operating factors of CXE, the optimal conditions were obtained as follows: 8.4 MPa, 50 °C, 5 mL/min ethanol, and 0.22 mole fraction of CO2, presenting an extraction oil that ranged from 5.12% to 7.42%, higher than that of hydrodistillation (HD) or indirect vapor distillation (IVD). The Gas Chromatography-Mass Spectrometry (GC-MS) analysis showed that three major components, including valerenol (18.48%), valerenal (10.21%), and ß-Cadinene (6.23%), are found in CXE oil, while a total of 23 components were identified, 48 components less than using conventional hydrodistillation. Furthermore, the antimicrobial activities of root oils were evaluated by the microdilution method, which showed that CXE oil exhibited an ability against Gram-positive bacteria, especially Staphylococcus aureus, approximately equivalent to traditional samples. Additionally, the DPPH free radical scavenging assay demonstrated that the antioxidant abilities of root oils were sorted in the descending order: IVD > HD > CXE > SFE. In conclusion, after a comprehensive comparison with the conventional methods, the CXE-related technique might be a promising green manufacturing pattern for the production of quality vetiver oil, due to the modification of ethanol by the variable addition of non-polar compressible CO2, ultimately resulting in a prominent dissolving capability for the extraction of vetiver solutes.

Wound Pathogens: Investigating Antimicrobial Activity of Commercial Essential Oil Combinations against Reference Strains.

This study aimed to determine the antimicrobial activity of 247 essential oil combinations against the reference strains of wound pathogens. Essential oil combinations were investigated for antimicrobial activity against five pathogens. The minimum inhibitory concentration (MIC) assay was used and the fractional inhibitory concentration index (ΣFIC) calculated to determine interactions between selected oils. Twenty-six combinations displayed broad-spectrum antimicrobial activity against all five reference strains and several displayed synergy against more than one pathogen. The combination of Santalum austrocaledonicum (sandalwood) with Commiphora myrrha (myrrh) displayed noteworthy antimicrobial activity against all five reference strains and synergy against four (MIC values 0.03-1.00 mg/ml and ΣFIC values 0.19-1.00 mg/ml) pathogens. No antagonism was observed. Santalum spp. and Vetiveria zizanioides essential oils contributed the most to antimicrobial activity in combination. Essential oil combinations are presented as a viable option in wound therapy.

Decarboxylation Facilitated by Carbocation Formation and Rearrangement during Steam Distillation of Vetiver Oil.

Density functional theory (DFT) calculations are used to probe the validity of mechanistic proposals for the conversion of isozizanoic acid to 12-norisoziza-5-ene, a reaction that occurs during steam distillation of vetiver oil. While this conversion corresponds overall to a simple decarboxylation, a multistep mechanism involving carbocation intermediates is supported by the computational results.

Vetiverianines A, B, and C: Sesquiterpenoids from Vetiveria zizanioides Roots.

Three new sesquiterpenoids-vetiverianines A (1), B (2), and C (3)-and a known eudesmane sesquiterpenoid (4) were isolated from the roots of Vetiveria zizanioides. Vetiverianine A (1) has a unique carbon framework comprising a rigid tricyclic ring system. Vetiverianines B (2) and C (3) are new eremophilane sesquiterpenoids. The structures of sesquiterpenoids 1-3, including the absolute configurations, were determined by NMR spectroscopic, X-ray crystallography, and vibrational circular dichroism data analysis. Vetiverianine C (3) exhibited weak cytotoxic activity against HL-60 cells.

Phytostabilization potential of two ecotypes of Vetiveria zizanioides in cadmium-contaminated soils: greenhouse and field experiments.

Soil contamination by cadmium (Cd) poses a serious environmental and public health concern. Phytoremediation, i.e., the use of plants to remove contaminants from soil, has been proposed for treatment of Cd-contaminated ecosystems. In this study, we demonstrated the potential of Vetiveria zizanioides, commonly known as vetiver, to serve as an effective phytoremediation agent. Two ecotypes, i.e., India and Sri Lanka, were grown in greenhouse pots and in the field. Soils were amended with cow manure, pig manure, bat manure, and an organic fertilizer. Among all amendments, pig manure performed best in both greenhouse and field studies in terms of increasing total V. zizanioides biomass production in both ecotypes. In both greenhouse and in the field, tissue of the Sri Lanka ecotype had higher Cd concentrations than did the India ecotype. In the greenhouse, the presence of Cd did not affect total biomass production or root dry weight. The Sri Lanka ecotype had 2.7 times greater adventitious root numbers and 3.6 times greater Cd accumulation in roots than did the India ecotype. In the field study, the Sri Lanka ecotype offers potential as an excluder species, as it accumulated Cd primarily in roots, with translocation factor values <1 and a bioconcentration coefficient for roots >1 for all experiments except for the pig manure amendment. In addition, the highest Cd concentration in the Sri Lanka ecotype root (71.3 mg kg(-1)) was consistent with highest Cd uptake (10.4 mg plant(-1)) in the cow manure treatment. The India ecotype contained lower root Cd concentrations, and Cd accumulation was slightly higher in shoots compared to roots, with translocation factor (TF) values >1. The India ecotype was therefore not considered as an excluder in the Cd-contaminated soil. With the use of excluder species combined with application of organic amendments, soil contamination by Cd may be treated by alternative remediation methods such as phytostabilization.

Determination of Prometryn in Vetiver Grass and Water Using Gas Chromatography-Nitrogen Chemiluminescence Detection.

Nitrogen chemiluminescence detector (NCD) is a nitrogen-specific detector that responds to ammonia, hydrazine, hydrogen cyanide and nitrogen oxide. A method to analyze the herbicide prometryn in plant and water samples was developed using gas chromatograph (GC) coupled with NCD. Extracts from plant (vetiver grass) and water matrices were analyzed for prometryn using an Agilent 7890A GC coupled with an Agilent 255 NCD in a split injection mode with a ratio of 2 : 1. Separation was carried out at 200°C and combustion at 1,018°C with H2 and O2 following optimized method development conditions. The percent recovery of prometryn in the two different matrices tested ranged from 81 to 107%, with relative standard deviations varying from 0.10 to 3.30% for spiked samples. Detection limit of the proposed method was 0.02 µg mL(-1) and the limit of quantification was 0.06 µg mL(-1). The proposed GC-NCD method was successfully applied to determine prometryn extracted from plant and water samples without potential interference of S-triazine, a pesticide from the same group.

A New Neolignan Glycoside from Vetiveria zizanioides Roots.

A new neolignan glycoside (1) and four known aromatic compounds (2-5) were isolated. from the roots of Vetiveria zizanioides. The structure of compound 1 was determined based on spectroscopic analysis and hydrolysis. The structure of known flavonoid glycoside 3 was confirmed by X-ray crystallography. Compound 5 showed weak cytotoxic activity against HL-60 cells with an IC50 value of 13.1 ± 0.04 µM.

Anxiety-like behaviour and c-fos expression in rats that inhaled vetiver essential oil.

Vetiver essential oil (VEO) has been used in aromatherapy for relaxation. This study aimed to investigate the effects of VEO on an anxiety-related behavioural model (the elevated plus-maze, EPM) and immediate-early gene c-fos in amygdala, known to be involved in anxiety. Male Wistar rats were administered diazepam (1 mg/kg i.p.) for 30 min or inhalated with VEO (1%, 2.5% or 5% w/w) for 7 min prior to exposure to the EPM. Then, the effects of 2.5% VEO, the anxiolytic dose, on c-fos expression in amygdala were investigated. The rats given either 2.5% VEO or diazepam exhibited an anxiolytic-like profile in the EPM. VEO and diazepam significantly increased c-fos expression in the lateral division of the central amygdaloid nucleus (CeL). Therefore, the anxiolytic properties of VEO might be associated with altering neuronal activation in CeL. However, future studies are needed to investigate the precise mechanism of action of VEO.

Effect of Vetiveria zizanioides essential oil on melanogenesis in melanoma cells: downregulation of tyrosinase expression and suppression of oxidative stress.

The major objective of this study was to estimate the hypopigmentation function of the essential oil from Vetiveria zizanioides (VZ-EO). Our results indicated that VZ-EO exhibits potent lipid peroxidation inhibitory activity to moderate the bleaching of ß-carotene and to maintain the cellular glutathione (GSH) levels. VZ-EO can markedly decrease melanin production and tyrosinase activity in α-melanin-stimulating-hormone- (α-MSH-) stimulated B16 cells. The effect of VZ-EO on melanogenesis is achieved by the suppression of cellular tyrosinase expression. The results demonstrated that the activity of VZ-EO on melanogenesis might be the result of its potent antioxidative ability, which was reflected in the decreased cellular oxidant and malondialdehyde (MDA) levels and the recovered activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) in α-MSH-stimulated B16 cells. The most abundant compound in VZ-EO is cedr-8-en-13-ol (12.4%), which has a strong capability to inhibit lipid peroxidation. Therefore, VZ-EO has the potential to become an ingredient in future hypopigmentation drugs, foods, and cosmetics.

Qualitative and quantitative analysis of vetiver essential oils by comprehensive two-dimensional gas chromatography and comprehensive two-dimensional gas chromatography/mass spectrometry.

Vetiver essential oils (VEO) are important raw ingredients used in perfume industry, entering the formula of numerous modern fragrances. Vetiver oils are considered to be among the most complex essential oils, resulting most of the time in highly coeluted chromatograms whatever the analytical technique. In this context, conventional gas chromatography has failed to provide a routine tool for the accurate qualitative and quantitative analysis of their constituents. Applying comprehensive two-dimensional gas chromatography techniques (GC×GC-FID/MS) afforded the mean to separate efficiently vetiver oil constituents in order to identify them in a more reliable way. Moreover, this is the first time that a complete true quantitation of each constituent is carried out on such complex oils by means of internal calibration. Finally, we have studied the influence of the injection mode on the determined chemical composition, and showed that several alcohols underwent dehydration under defined chromatographic conditions (splitless mode) usually recommended for quantitation purposes.

Effect of calcium on growth performance and essential oil of vetiver grass (Chrysopogon zizanioides) grown on lead contaminated soils.

The aim of this study was to investigate effect of calcium on growth, survival, essential oil yield and chemical compositions of vetiver grass grown on lead contaminated soils. Calcium inform of CaCO3 (0, 2000, 4000, 6000 mg Ca kg(-1)) was added to river sand soils containing 4000 mg Pb kg(-1) dry soil. Results showed that, in the absence of calcium treatment, no plants survived after 2 weeks of cultivation, while the rest grew well to the end of the experimental period (42 weeks). Calcium treatments generally resulted in a slight decrease in biomass. Interestingly, an increase in calcium over 2000 mg kg(-1) did not result in a decrease in accumulation of lead in vetiver roots and shoots. The levels of lead in roots and shoots under calcium treatments were around 2000 and 90 mg kg(-1) dry weight, respectively. The addition of CaCO3 did not improve vetiver essential oil yield and chemical composition compared to the control. A level of applied CaCO3 about half of the lead concentration in soils was sufficient to improve vetiver growth and survival, and accumulate high concentrations of lead in the roots. This finding can be applied for re-vegetation of lead contaminated soils using vetiver.

Comprehensive two-dimensional gas chromatography-mass spectrometry analysis of volatile constituents in Thai vetiver root oils obtained by using different extraction methods.

INTRODUCTION: Vetiver root oil is known as one of the finest fixatives used in perfumery. This highly complex oil contains more than 200 components, which are mainly sesquiterpene hydrocarbons and their oxygenated derivatives. Since conventional GC-MS has limitation in terms of separation efficiency, the comprehensive two-dimensional GC-MS (GC x GC-MS) was proposed in this study as an alternative technique for the analysis of vetiver oil constituents. OBJECTIVE: To evaluate efficiency of the hyphenated GC x GC-MS technique in terms of separation power and sensitivity prior to identification and quantitation of the volatile constituents in a variety of vetiver root oil samples. METHODOLOGY. Dried roots of Vetiveria zizanioides were subjected to extraction using various conditions of four different methods; simultaneous steam distillation, supercritical fluid, microwave-assisted, and Soxhlet extraction. Volatile components in all vetiver root oil samples were separated and identified by GC-MS and GC x GC-MS. The relative contents of volatile constituents in each vetiver oil sample were calculated using the peak volume normalization method. RESULTS: Different techniques of extraction had diverse effects on yield, physical and chemical properties of the vetiver root oils obtained. Overall, 64 volatile constituents were identified by GC-MS. Among the 245 well-resolved individual components obtained by GC x GC-MS, the additional identification of 43 more volatiles was achieved. CONCLUSION: In comparison with GC-MS, GC x GC-MS showed greater ability to differentiate the quality of essential oils obtained from diverse extraction conditions in terms of their volatile compositions and contents.

Antioxidant potential of the root of Vetiveria zizanioides (L.) Nash.

Vetiveria zizanioides, an aromatic plant commonly known as vetiver has been used for various ailments. The essential oil of vetiver root has been shown to possess antioxidant activity. However, antioxidant potential of spent root extract has not been reported. Hence, in the present study, ferric reducing, free radical scavenging and antioxidant activity of two genotypes namely KS1 and gulabi of V. zizanioides L. Nash root were investigated using in vitro assays - the ferric reducing antioxidant power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), total phenolic content (TPC), total antioxidant capacity (TAC) and reducing power (RP). KS1 genotype showed higher FRAP values, DPPH inhibition, TPC and RP potential compared to gulabi and the antioxidant activity increased with the concentration of the extract (10-1000 microg/mL). A significant protective effect of cv KS1 (100 microg/mL) extract was also observed in reduced glutathione (GSH) and malondialdehyde (MDA) concentrations of erythrocytes subjected to oxidative stress by tert-butyl hydroperoxide (t-BHP) and hydrogen peroxide (H202). The cv KS1 showed better antioxidant activity, compared to cv gulabi indicating the possibility of exploring the presence of different phytoconstituents in the two varieties.

Release properties of chemical and enzymatic crosslinked gelatin-gum Arabic microparticles containing a fluorescent probe plus vetiver essential oil.

Oil-containing gelatin-gum Arabic microparticles were prepared by complex coacervation followed by crosslinking with glutaraldehyde or transglutaminase. A fluorescent mixture, khusimyl dansylate (KD) as the fluorescent compound mixed to the vetiver essential oil, was used as oil model. The effect of the type of crosslinking of the coacervated gelatin-gum Arabic membrane, the physical state of microparticles, wet or freeze-dried and the type of release media, aqueous with surfactants, Sodium Dodecyl Sulphate (sds) or Tween 80 (tw) and anhydrous ethanol as organic media on the release rate of the KD from the microparticles, was experimentally investigated. It was shown that the oil was dispersed uniformly throughout the microparticles and the chemical crosslinked microparticles were more resistant to swelling, presenting smaller sizes after hydration. Also the crosslinking effect, transglutaminase or glutaraldehyde, could be confirmed by the integrity of the crosslinked gelatin-gum Arabic microparticles after incubation in the aqueous sds media, compared to complete dissolution of the uncrosslinked microparticles in this media. The cumulative fluorescent KD release from the gelatin-gum Arabic microparticles decreased in the following order of dissolution media: anhydrous ethanol>tw>sds and the wet microparticles have shown a faster KD release than freeze-dried ones. A mathematical model was used to estimate the diffusion coefficient (D). The chemically crosslinked gelatin-gum Arabic microparticles ensured a pronounced retard effect in the KD diffusion, presenting a D varying from 0.02 to 0.6 x 10(-11)cm(2)/s, mainly in an aqueous media, against D varying from 1.05 to 13.9 x 10(-11)cm(2)/s from the enzymatic crosslinked microparticles.