966 resultados para capillary gas chromatography-mass spectrometric detection with selective ion monitoring


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- A combined increase in seawater [CO2] and [H+] was recently shown to induce a shift from photosynthetic HCO3- to CO2 uptake in Emiliania huxleyi. This shift occurred within minutes, whereas acclimation to ocean acidification (OA) did not affect the carbon source. - To identify the driver of this shift, we exposed low- and high-light acclimated E. huxleyi to a matrix of two levels of dissolved inorganic carbon (1400, 2800 lmol kg-1) and pH (8.15, 7.85) and directly measured cellular O2, CO2 and HCO3 fluxes under these conditions. - Exposure to increased [CO2] had little effect on the photosynthetic fluxes, whereas increased [H+] led to a significant decline in HCO3- uptake. Low-light acclimated cells overcompensated for the inhibition of HCO3- uptake by increasing CO2 uptake. High-light acclimated cells, relying on higher proportions of HCO3- uptake, could not increase CO2 uptake and photosynthetic O2 evolution consequently became carbon-limited. - These regulations indicate that OA responses in photosynthesis are caused by [H+] rather than by [CO2]. The impaired HCO3- uptake also provides a mechanistic explanation for lowered calcification under OA. Moreover, it explains the OA-dependent decrease in photosynthesis observed in high-light grown phytoplankton.

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Geochemical studies of organic "biomarker" compounds were applied to Eemian sediments cored at Dagebuell (DA-1) on the west coast, and at Krummland (KR-1) in the east of the Baltic Zone of Schleswig-Holstein, Germany. 10 samples from the early stage of the Eemian Transgression to the high Eem at Krummland, and 24 samples from the peak and late phases of the Eemian at Dagebuell provide new insights on the development of the Eemian Sea in the region. C37-C39-ethyl- and methyl-ketones in the Krummland sediments indicated unstable conditions at the onset of the marine trangression, and freshwater influence in keeping with their shallow nearshore environment. In the Dagebuell deposits, patterns typical of marine to brackish conditions were observed, comparable to those found today in the Skagerrak and Belt Sea areas. The sea-surface temperatures estimated from the alkenone unsaturation ratio UK37 at DA-1 corroborate the evidence from "standard" faunal and pollen assemblages, and lithological successions. Here, the temperature maximum attained in pollen assemblage zone PAZ Illc, indicates the early onset of very warm conditions, preceding the highest sea level of the penultimate interglacial by 8,000 years, based on previously published U/Th ages.

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Atmospheric dust samples collected along a transect off the West African coast have been investigated for their lipid content and compound-specific stable carbon isotope compositions. The saturated hydrocarbon fractions of the organic solvent extracts consist mainly of long-chain n-alkanes derived from epicuticular wax coatings of terrestrial plants. Backward trajectories for each sampling day and location were calculated using a global atmospheric circulation model. The main atmospheric transport took place in the low-level trade-wind layer, except in the southern region, where long-range transport in the mid-troposphere occurred. Changes in the chain length distributions of the n-alkane homologous series are probably related to aridity, rather than temperature or vegetation type. The carbon preference of the leaf-wax n-alkanes shows significant variation, attributed to a variable contribution of fossil fuel- or marine-derived lipids. The effect of this nonwax contribution on the d13C values of the two dominant n-alkanes in the aerosols, n-C29 and n-C31 alkane, is, however, insignificant. Their d13C values were translated into a percentage of C4 vs. C3 plant type contribution, using a two-component mixing equation with isotopic end-member values from the literature. The data indicate that only regions with a predominant C4 type vegetation, i.e. the Sahara, the Sahel, and Gabon, supply C4 plant-derived lipids to dust organic matter. The stable carbon isotopic compositions of leaf-wax lipids in aerosols mainly reflect the modern vegetation type along their transport pathway. Wind abrasion of wax particles from leaf surfaces, enhanced by a sandblasting effect, is most probably the dominant process of terrigenous lipid contribution to aerosols.

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The Mediterranean species Cynara cardunculus L. is recognized in the traditional medicine, for their hepatoprotective and choleretic effects. Biomass of C. cardunculus L. var. altilis (DC), or cultivated cardoon, may be explored not only for the production of energy and pulp fibers, but also for the extraction of bioactive compounds. The chemical characterization of extractable components, namely terpenic and phenolic compounds, may valorize the cultivated cardoon plantation, due to their antioxidant, antitumoral and antimicrobial activities. In this study, the chemical composition of lipophilic and phenolic fractions of C. cardunculus L. var. altilis (DC), cultivated in the south of Portugal (Baixo Alentejo region) was characterized in detail, intending the integral valorization of its biomass. The biological activity of cultivated cardoon extracts was evaluated in terms of antioxidant, human tumor cell antiproliferative and antibacterial effects. Gas chromatography-mass spectrometry (GC-MS) was used for the chemical analysis of lipophilic compounds. Sixty-five lipophilic compounds were identified, from which 1 sesquiterpene lactone and 4 pentacyclic triterpenes were described, for the first time, as cultivated cardoon components, such as: deacylcynaropicrin, acetates of β- and α-amyrin, lupenyl acetate and ψ-taraxasteryl acetate. Sesquiterpene lactones were the major family of lipophilic components of leaves (≈94.5 g/kg), mostly represented by cynaropicrin (≈87.4 g/kg). Pentacyclic triterpenes were also detected, in considerably high contents, in the remaining parts of cultivated cardoon, especially in the florets (≈27.5 g/kg). Taraxasteryl acetate was the main pentacyclic triterpene (≈8.9 g/kg in florets). High pressure liquid chromatography-mass spectrometry (HPLC-MS) was utilized for the chemical analysis of phenolic compounds. Among the identified 28 phenolic compounds, eriodictyol hexoside was reported for the first time as C. cardunculus L. component, and 6 as cultivated cardoon components, namely 1,4-di-O-caffeoylquinic acid, naringenin 7-O-glucoside, naringenin rutinoside, naringenin, luteolin acetylhexoside and apigenin acetylhexoside. The highest content of the identified phenolic compounds was observed in the florets (≈12.6 g/kg). Stalks outer part contained the highest hydroxycinnamic acids abundance (≈10.3 g/kg), and florets presented the highest flavonoids content (≈10.3 g/kg). The antioxidant activity of phenolic fraction was examined through 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Stalks outer part, and receptacles and bracts extracts demonstrated the highest antioxidant effect on DPPH (IC50 of 34.35 μg/mL and 35.25 μg/mL, respectively). (cont.) abstract (cont.) The DPPH scavenging effect was linearly correlated with the total contents of hydroxycinnamic acids (r = -0.990). The in vitro antiproliferative activity of cultivated cardoon lipophilic and phenolic extracts was evaluated on a human tumor cells line of triple-negative breast cancer (MDA-MB-231), one of the most refractory human cancers to conventional therapeutics. After 48 h of exposition, leaves lipophilic extract showed higher inhibitory effect (IC50 = 10.39 μg/mL) than florets lipophilic extract (IC50 = 315.22 μg/mL), upon MDA-MB-231 cellular viability. Pure compound of cynaropicrin, representative of the main compound identified in leaves lipophilic extract, also prevented the cell proliferation of MDA-MB-231 (IC50 = 17.86 μM). MDA-MB-231 cells were much more resistant to the 48 h- treatment with phenolic extracts of stalks outer part (IC50 = 3341.20 μg/mL) and florets (IC50 > 4500 μg/mL), and also with the pure compound of 1,5-di-O-caffeoylquinic acid (IC50 = 1741.69 μM). MDA-MB-231 cells were exposed, for 48 h, to the respective IC50 concentrations of leaves lipophilic extract and pure compound of cynaropicrin, in order to understand their ability in modelling cellular responses, and consequently important potentially signaling pathways for the cellular viability decrease. Leaves lipophilic extract increased the caspase-3 enzymatic activity, contrarily to pure compound of cynaropicrin. Additionally, leaves lipophilic extract and pure compound of cynaropicrin caused G2 cell cycle arrest, possibly by upregulating the p21Waf1/Cip1 and the accumulation of phospho-Tyr15-CDK1 and cyclin B1. The inhibitory effects of leaves lipophilic extract and cynaropicrin pure compound, against the MDA-MB-231 cell proliferation, may also be related to the downregulation of phospho-Ser473-Akt. The antibacterial activity of cultivated cardoon lipophilic and phenolic extracts was assessed, for the first time, on two multidrug-resistant bacteria, such as the Gram-negative Pseudomonas aeruginosa PAO1 and the Gram-positive methicillin-resistant Staphylococcus aureus (MRSA), two of the main bacteria responsible for health care-associated infections. Accordingly, the minimum inhibitory concentrations (MIC) were determined. Lipophilic and phenolic extracts of florets did not have antibacterial activity on P. aeruginosa PAO1 and MRSA (MIC > 2048 μg/mL). Leaves lipophilic extract did not prevent the P. aeruginosa PAO1 growth, but pure compound of cynaropicrin was slightly active (MIC = 2048 μg/mL). Leaves lipophilic extract and pure compound of cynaropicrin blocked MRSA growth (MIC of 1024 and 256 μg/mL, respectively). The scientific knowledge revealed in this thesis, either by the chemical viewpoint, or by the biological viewpoint, contributes for the valorization of C. cardunculus L. var. altilis (DC) biomass. Cultivated cardoon has potential to be exploited as source of bioactive compounds, in conciliation with other valorization pathways, and Portuguese traditional cheeses manufacturing.

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Type I Neurofibromatosis (NF1) is an autosomal-dominant inheritable disorder, with an incidence of 1:3,000, and a prevalence of 1:4,000 to 5,000. Pathogenesis is based on mutations of the NF1 gene, a tumor suppressor gene encoding a cytoplasmic protein named neurofibromin that controls cellular proliferation. Patients affected by NF1 typically present with cutaneous neurofibromas, cafè au lait spots and eye involvement, but they can also be affected by various visceral tumors, such as neurofibromas (nodular or plexiform type), gastrointestinal stromal tumors or endocrine tumors, such as pheochromocytomas. Visceral neurofibromas are often asymptomatic but when growing in size they may present with pain, palpable abdominal mass, symptoms secondary to bowel obstruction or main vessels compression, and even gastrointestinal bleeding when mucosa or submucosa are involved. In these cases surgery becomes mandatory in order to remove all neoplastic tissue. The Authors describe a case of a young man affected by NF1 with associated retrocaval abdominal mass with compression and displacement of the inferior vena cava, thus requiring a complex surgical procedure.

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We studied the effect of Silicon (Si) on Casparian band (CB) development, chemical composition of the exodermal CB and Si deposition across the root in the Si accumulators rice and maize and the Si non-accumulator onion. Plants were cultivated in nutrient solution with and without Si supply. The CB development was determined in stained root cross-sections. The outer part of the roots containing the exodermis was isolated after enzymatic treatment. The exodermal suberin was transesterified with MeOH/BF3 and the chemical composition was measured using gas chromatography-mass spectroscopy (GC-MS) and flame ionization detector (GC-FID). Laser ablation-inductively coupled plasma-mass spectroscopy (LA-ICP-MS) was used to determine the Si deposition across root cross sections. Si promoted CB formation in the roots of Si-accumulator and Si non-accumulator species. The exodermal suberin was decreased in rice and maize due to decreased amounts of aromatic suberin fractions. Si did not affect the concentration of lignin and lignin-like polymers in the outer part of rice, maize and onion roots. The highest Si depositions were found in the tissues containing CB. These data along with literature were used to suggest a mechanism how Si promotes the CB development by forming complexes with phenols.

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A multi-residue gas chromatography-mass spectrometry method was developed in order to evaluate the presence of 39 pesticides of different chemical families (organophosphorus, triazines, imidazole, organochlorine), as well as some of their transformation products, in surface water samples from Ria de Aveiro. Ria de Aveiro is an estuarine coastal lagoon, located in the northern west region of Portugal, which receives inputs from agriculture, urban and industrial activities. The analytical method was developed and validated according international guidelines and showed good linearity, with correlation coefficients higher than 0.9949 for all compounds, adequate precision and accuracy, and high sensitivity. Pesticides were chosen from the priority pollutants list of the Directive 2008/105/EC of the European Parliament and of the Council (on environmental quality standards in the field of water policy), or were selected due their common use in agricultural practices. Some of these 39 pesticides are, or are suspected to be, endocrine disruptor compounds (EDCs), being capable of altering the endocrine system of wildlife and humans, causing form malfunction and ultimately health problems. Even those pesticides which are not EDCs, are known to be awfully toxic and have a recognised impact in human health. The aquatic environment is particularly susceptible to pollution due to intentional and accidental release of chemicals to water [3]. Pesticide contamination of surface water is a national issue as it is often used as drinking water. This concern is especially important in rural agricultural areas where population uses small private water supplies, regularly without any laboratory surveillance. The study was performed in seven sampling points and the results showed a considerable concern pesticide contamination of all samples.

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Dendritic cells are antigen presenting cells that provide a vital link between the innate and adaptive immune system, providing the initial detection of pathogenic invaders. Research into this family of cells has revealed that they perform information fusion which directs immune responses. We have derived a Dendritic Cell Algorithm based on the functionality of these cells, by modelling the biological signals and differentiation pathways to build a control mechanism for an artificial immune system. We present algorithmic details in addition to experimental results, when the algorithm was applied to anomaly detection for the detection of port scans. The results show the Dendritic Cell Algorithm is successful at detecting port scans.

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In the present study, natural occurrence of fungi and aflatoxin B1 (AFB1) in pellet feed and feed ingredients used for rainbow trout was investigated with emphasis to Aspergillus section Flavi members and medicinal plants inhibitory to Aspergillus growth and/or AF production. The feed samples were cultured on the standard isolation media including dichloran rosebengal chloramphenicol agar (DRCA) and Aspergillus flavus/parasiticus agar (AFPA) for 2 weeks at 28 °C. Identification of fungal isolates was implemented based on the macro- and microscopic morphological criteria. AFs were detected using high performance liquid chromatography (HPLC). Based on the results obtained, a total of 109 fungal isolates were identified of which Aspergillus was the prominent genus (57.0%), followed by Penicillium (12.84%), Absidia (11.01%) and Pseudallscheria (10.10%). The most frequent Aspergillus species was A. flavus (60.66%) isolated from all the feed ingredients as well as pellet feed. Among 37 A. flavus isolates, 19 (51.35%) were able to produce AFB1 on yeast extract-sucrose (YES) broth in the range of 10.2 to 612.8 [tg/g fungal dry weight. HPLC analyses of trout feed showed that pellet feed and all feed ingredients tested except gluten were contaminated with different levels of AFB1 in the range of 1.83 to 67.35 lig/kg. In order to finding natural inhibitors of fungal growth and/or AF production, essential oils (EOs) and extracts of 49 medicinal plants were studied against an aflatoxin-producing A. parasiticus using a microbioassay technique. The EOs was analyzed by gas chromatography/mass spectrometry (GC/MS). Based on the results obtained, Achillea millefolium sub sp. elborsensis, Ferula gummosa, Mentha spicata, Azadirachta indica, Conium maculatum and Artemisia dracunculus remarkably inhibited A. parasiticus growth without affecting AF production by the fungus. Besides of Thymus vulgaris and Citrus aurantifolia, the EO of Foeniculum vulgare significantly inhibited both fungal growth (-70.0%) and AFs B1 and G1 (-99.0%) production. The EO of Carum carvi and ethyl acetate extract of Platycladus orientalis suppressed AFs B1 and G1 by more than 90.0%, without any obvious effect on fungal growth. The IC50 values of bioactive plants for AFs B1 and G1 were determined in the ranges of 90.6 to 576.2 and 2.8 to 61.9 µg/ml, respectively. Overall, results of the present study indicate the importance of AF contamination of trout feed as a risk factor for fish farming and thus, an urgent necessity for constant monitoring of trout feed for any unacceptable levels of AF contamination. Likewise, antifungal activities of bioactive plants introduced here would be an important contribution to explain the use of these plants as effective antimicrobial candidates to protect feeds from toxigenic fungus growth and subsequent AF contamination.

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A avaliação do aporte de matéria orgânica no ambiente aquático por atividades antrópicas pode ser realizada através da identificação e quantificação de marcadores moleculares. Diversos estudos apontam à aplicação dos marcadores moleculares com esta finalidade, no entanto, poucos avaliam a variação das concentrações desses compostos ao longo do tempo, registrada nas camadas sedimentares. O presente trabalho realiza um estudo a partir de três classes de marcadores moleculares presentes em perfis sedimentares da região do Complexo Estuarino de Paranaguá (CEP) no Paraná (PR), que nos últimos anos vêm sofrendo com o crescente desenvolvimento de atividades antrópicas. Como objetivo, tem-se identificar as principais fontes de matéria orgânica e estudar o histórico destes aportes em colunas sedimentares do CEP, relacionando as taxas de sedimentação com a deposição de origem natural e antrópica. A legislação vigente para o monitoramento ambiental, no que diz respeito à contaminação por esgoto fecal, sugere a avaliação por indicadores microbiológicos, porém, indicadores químicos como os esteróides fecais são uma alternativa bastante promissora, pois estes são persistentes, sendo menos sensíveis a variações ambientais. Outros dois marcadores moleculares de aportes antrópicos ao ambiente que foram determinados neste estudo são os alquilbenzenos lineares (LABs), presente em detergentes, que indicam aportes antrópicos oriundos de esgoto doméstico e a determinação de cafeína, tendo em vista que os esteróides fecais podem ser originários de fezes de animais de sangue quente, podendo indicar outras fontes. Para o presente trabalho foram coletados 12 testemunhos de até 1 m de profundidade em maio de 2006, totalizando 12 pontos de coleta e um montante de 121 amostras. As análises foram realizadas por cromatografia em fase gasosa com detecção por espectrometria de massas (CG-EM). Os esteróides encontrados em maior concentração foram o β- sitosterol (71,4 µg g-1), estigmasterol (8,7 µg g-1), colestanol (3,6 µg g-1) e o estigmastanol (2,8 µg g-1), todos oriundos de fonte natural, indicando que a maior contribuição para o CEP é por aporte biogênico. O coprostanol, que é um esterol fecal, foi encontrado entre as concentrações de 0,001 e 4,10 µg g-1, outros dois esteróides de origem fecal também foram detectados, coprostanona e epicoprostanol, onde as maiores concentrações foram 3,6 e 0,2 µg g-1, respectivamente, sendo encontrados em regiões próximas a centros urbanos, indicando origem antrópica. As maiores concentrações para o ∑LABs também foram encontradas em regiões próximas às cidades de Antonina e Paranaguá, sendo a maior encontrada no testemunho #3 Gererês (208 ng g-1). Para o último marcador molecular analisado, a cafeína, foi encontrada a maior concentração de 18,41 ng g-1, sendo este ponto localizado longe dos centros urbanos, porém este contaminante é bastante solúvel em água podendo ser transportado na coluna d’água e percorrer grandes distâncias. Através dos compostos analisados, pode-se perceber que a intervenção antropogênica foi mais marcante nos testemunhos coletados no eixo leste-oeste do CEP, ficando registrado nas camadas sedimentares.

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The exopolysaccharides are extracellular compounds produced by some species of fungi and bacteria. It is suggested that these molecules, even when in the form of complex polysaccharide-peptide, are the main bioactive molecules of many fungus. Some of the biological activities displayed by these compounds can be accentuated and others may arise when you add chemically polar or nonpolar groups to polysaccharides. The fruiting body of Pleurotus sajor-caju produces a heteropolysaccharide with antineoplastic and antimicrobial activity, but other biological activities of this polymer have not been evaluated. In this work the exopolysaccharide of Pleurotus sajor-caju was sulfated chemically and structurally characterized. We also evaluated the antiproliferative, antioxidant and anticoagulant activities from native exopolysaccharide (PN) and its sulfated derivated (PS). Polyacrylamide gel electrophoresis, infrared spectroscopy and nuclear magnetic resonance (¹³C) proved successful in sulfation of PN to obtain PS. Analysis by gas chromatography-mass spectroscopy showed that PN and PS are composed of mannose, galactose, 3-O-methyl-galactose and glucose in proportion percentage of 44,9:16,3:19,8:19 and 49, 7:14,4:17,7:18,2, respectively. The percentage of sulfate found in PS was 22.5%. Antioxidants assays revealed that the sulfation procedure affects differently the activities of exopolysaccharides, while the total antioxidant capacity, the scavenging activity of superoxide radical and ferric chelating were not affected by sulfation, on the other hand the chemical modification of PN enhanced the scavenging activity of hydroxyl radical and reducing power. PS also showed anticoagulant activity in a dose-dependent manner and clotting time was 3.0 times higher than the baseline value in APTT at 2 mg/mL. The exopolysaccharide not presented antiproliferative activity against HeLa tumor cells, but PS affects the cellular proliferation in a time-dependent manner. After 72 h, the inhibition rate of PS (2.0 mg/mL) on HeLa cells was about 60%. The results showed that PN sulfation increase some of their activities.

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Este trabalho descreve o isolamento e purificação do ácido α-eleosteárico (α-ESA) a partir do óleo de tungue e sua caracterização por espectroscopia de infravermelho com transformada de Fourier (FTIR), cromatografia gasosa acoplada com espectrometria de massas (GC-MS) e espectroscopia de ressonância magnética nuclear (RMN) de 1H e 13C. O α-ESA apresenta atividades biológicas (antitumorais, anti-inflamatórias e antioxidantes), tornando-se importante compreender sua interação com membranas lipídicas. Assim, este trabalho também descreve resultados referentes ao efeito da incorporação de α-ESA na dinâmica molecular de lipossomos compostos por fosfatidilcolina. O sistema lipossomal puro e contendo α-ESA foi caracterizado através do uso de espectroscopia de UV-visível, FTIR, RMN e calorimetria de varredura diferencial (DSC). Como resultados da purificação do α-ESA, obtivemos uma pureza de 95,9% utilizando acetona como solvente de recristalização em detrimento dos 92,2% em solução etanólica. Na incorporação em lipossomos, observou-se uma maior interação do α-ESA com a parte polar, de interface e os primeiros metilenos da região apolar da fosfatidilcolina. Além disso, α-ESA apresentou um efeito de redução da fluidez de lipossomos. Os resultados contribuem para a geração de conhecimento para o desenvolvimento de novos sistemas farmacológicos.

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Phytochemical analyses as well as antimicrobial and antioxidant activities of the extracts of C. sumatrensis aerial parts were investigated in this study. METHODS: The aerial parts of C. sumatrensis were air dried, weighed and exhaustively extracted with hexane, ethyl acetate and methanol successively. The crude extracts were screened for metabolites. These extracts of the plant were evaluated for antimicrobial and antioxidant activities using agar diffusion and DPPH method respectively. The extracts were also analysed using Gas chromatography – Mass spectrometry, and the chromatogram coupled with mass spectra of the compounds were matched with a standard library. RESULTS: Preliminary phytochemical investigation of crude n-hexane, ethyl acetate and methanol extracts of the aerial parts of Conyza sumatrensis revealed the presence of anthraquinones, flavonoids, terpenoids, phenolics, tannin, glycosides and carbohydrate. All the crude extracts gave a clear zone of inhibition against the growth of the test bacteria ( Staphylococcus aureus , Escherichia coli , Bacillus subtilis , Pseudomona aeruginosa, Salmonella typhi , Klebsiellae pneumonae ) at moderate to high concentrations, as well as test fungi ( Candida albicans , Aspergillus niger , penicillium notatum and Rhizopus stolonifer ) at high concentration. Methanolic extract exhibited significant radical scavenging property with IC50 of 17.08 μg/mL while n-hexane and ethyl acetate extracts showed no significant antioxidant activity. GC-MS of N-hexane extract showed a total number of eleven chemical constituents with α-Farnesene and spathulenol being the most abundance compounds constituting 20.27 and 22.28% of the extract respectively. Ethyl acetate extract revealed thirteen compounds with two most abundant compounds, cis-β-farnesene (16.64 %) and cis-pinane (21.09 %). While methanolic extract affords seventeen compounds with Ephytol being the most abundant compound (19.36 %).