Aromadendrene Oxide Research Articles

Chemical components and antioxidant potential of essential oil of Rhanterium epapposum growing in sandy habitat in Saudi Arabia: In silico study on cytochrome P450 1A1 and xanthine oxidase

Rhanterium epapposum Oliv. (Family: Asteraceae) is a common plant in Saudi Arabia, possessing a variety of traditional uses. The current targets were studying the comparative chemical profile and antioxidant of the essential oil (EO) of R. epapposum collected from a sandy habitat at Qareenah, Riyadh, Saudi Arabia. The EO profiling was constructed via gas chromatography-mass spectrometry (GC-MS) and GC-flame ionization detection (GC-FID). Forty-eight compounds were found, including monoterpenes (hydrocarbons 3.25% and oxygenated 48.64%), sesquiterpenes (hydrocarbons 12.64 % and oxygenated 32.38%), which are the backbone of the oil along with a minor of other non-terpenoid compounds (1.31%). Linalool (18.02%), α-eudesmol (11.98%), trans-sesquisabinene hydrate (11.63%), aromadendrene oxide-(1) (7.68%), nerol (6.88%), and α-terpineol (5.01%) were identified as the dominant components. The EO was evaluated for its antioxidant activity using in vitro DPPH, ABTS, and FRAP methods. The results demonstrated that R. epapposum EO possesses a dosage-dependent ability for antioxidant scavenging activity as per all three techniques. This EO exhibited IC50 values of 517.59, 350.24, and 713.72 μg/mL for the DPPH, ABTS, and FRAP, respectively, through the comparison with the ascorbic acid with respective values of 36.10, 26.12, and 44.58 μg/mL. The main compounds were subjected to docking analysis on cytochrome P450 1A1 and xanthine oxidase. All evaluated compounds exhibited good binding affinities especially α-eudesmol and linalool for both enzymes, respectively. The current research confirmed the potent free radical scavenging properties of R. epapposum EO. More research on the antioxidant content of this oil and/or its primary constituents in pure form on the two enzymes, cytochrome P450 1A1 and xanthine oxidase, is proposed to develop a better understanding of the oxidative action routes in vivo and in vitro.

Chemical Constituents, Antioxidant, and Antimicrobial Activities of Ethyl Acetate Fractionated Extract from Rhizomes of Zingiber monophyllum Gagnep.: In vitro and in silico Screenings

Objective/Background: Zingiber monophyllum Gagnep., a member of the Zingiberaceae family, is known for its significant biological activities. The current study aimed to determine the volatile components of the ethyl acetate (EtOAc) fractionated extract found in the rhizomes of this species. This is the first report on the chemical composition and bioactivities of Z. monophyllum rhizomes fractionated extract. Methods: The chemical constituents were analyzed and determined using gas chromatography-mass spectrometry (GC-MS). Antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and ferric reducing antioxidant power (FRAP) assays using ascorbic acid as a positive control. Antibacterial and antifungal properties of the EtOAc fractionated extract of Z. monophyllum rhizomes were assessed against Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica, Enterococcus faecalis, Staphylococcus aureus, Bacillus cereus, and Candida albicans. Density functional theory (DFT) and molecular docking were also employed to illustrate antioxidant and antimicrobial activities. Results: Nine components were identified by GC-MS analysis from the EtOAc fractionated extract of Z. monophyllum rhizomes. ( E)-labda-8(17),12-diene-15,16-dial (9), spathulenol (2), and neointermedeol (5) were the major components (21.8%, 16.8%, and 11.9%, respectively). Moderate antioxidant activities of the EtOAc fractionated extract were observed via both the DPPH assay and the FRAP assay using ascorbic acid as the standard compound. The extract demonstrated remarkable antimicrobial activity against all examined microbial strains, except for P. aeruginosa. The DFT study analyzed the antioxidant potential of each component in the fractionated extract. Molecular docking study chose E. faecalis DNA gyrase B, E. coli DNA gyrase B, S. aureus biotin protein ligase, E. faecalis Alanine racemase, and C. albicans N-myristoyltransferase as potential target proteins for antimicrobial activity. Conclusion: In this study, the chemical composition of the EtOAc fractionated extract of Z monophyllum rhizomes was demonstrated through GC-MS analysis for the first time. Nine components, including alloaromadendrene, spathulenol, globulol, τ-cadinol, neointermedeol, aromadendrene oxide-(2), ambrial, (E)-15,16-dinorlabda-8(17),11-dien-13-one, and (E)-lambda-8 (17),12-diene-15,16-dial along with relative content were identified in this fractionated extract. The bioassays revealed that the fractionated extract showed moderate antioxidant activities and significant antimicrobial activities. The antioxidant and antimicrobial potential of each component was also theoretically examined by the DFT study and molecular docking study, respectively.

Phytochemical profiling and GC-MS analysis of methanolic leaf extract of Persicaria hydropiper (L.) H. Gross: An important ethnomedicinal plant

Medicinal plants are the best source of alternative medicine and are acquiring recognition in the area of pharmacological research for the production of plant-based medicines with improved efficacy. Persicaria hydropiper (L.) H-Gross is known worldwide for its use in traditional medicine systems. It possesses numerous therapeutic properties. The present endeavor was taken up to analyze the phytochemical profile and antioxidant activity by DPPH assay of various extracts of Persicaria hydropiper leaves. The preliminary phytochemical screening of acetone, methanol, and aqueous extract of P. hydropiper leaves showed the presence of important phytochemicals such as phenols, tannins, flavonoids, glycosides, terpenoids, and alkaloids. The quantitative estimation revealed elevated amounts of total phenols, flavonoids, tannins, and total alkaloid content. The antioxidant activity also revealed higher radical scavenging activity by the extracts. The Gas Chromatography-Mass Spectrophotometry assay of methanol extract of P. hydropiper leaves detected and identified altogether 54 phytocompounds including 7 major compounds displaying higher peaks such as Andrographolide, 1H-cyclopropa[a] naphthalene, 1a,2,3,5.6,7a7b-octahydro-1,1,7,7a-tetramethyl-[1aR-(1a?,7?,7a?,7b?)-, Cis-Z alpha bisabolene epoxide, 1,8-cyclopentadecadiyne, Aromadendrene oxide (2), Bicyclo[4.1.0]heptane-3-cyclopropyl 7-hydroxymethyl, (cis) and Alpha-guanine. The concentration of phenols, flavonoids, and tannins revealed in the phytochemical profiling of three extracts might be responsible for the strong antioxidant activity of the extracts and the bioactive compounds identified in the GC-MS investigation exhibit important therapeutic properties required for the treatment of various human diseases. Thus, the medicinal use and properties of Persicaria hydropiper may be attributed to the presence of these phytochemical constituents. Further study of the compounds may provide scope for the production of novel drugs with increased efficacy and potency with the ability to cure harmful human diseases.

Chemical Composition and Antistaphylococcal Activity of Essential Oil of Curcuma mangga Rhizome from Indonesia

This study assessed the antistaphylococcal activity of essential oil (EO) hydrodistilled from the rhizome of Curcuma mangga grown in Indonesia using the broth microdilution volatilization method and standard broth microdilution method modified for evaluation of volatile agents, as well as described its chemical composition using gas chromatography (GC) with mass spectrometry (MS). A fused-silica HP-5MS column and a DB-17MS column were used to separate the components into two columns. The results demonstrated that the EO exhibited antistaphylococcal activity at the minimum inhibitory concentration (MIC) ranging from 128 to 1024 µg/mL. In contrast, the clinical isolate of tetracycline-resistant Staphylococcus aureus was the most sensitive strain (MIC 128 µg/mL). The major constituents of the EO were 15,16-dinorlabda-8(17),11-dien-13-one (24.63/15.78%), followed by ambrial (16.12/10.97%), 13-nor-eremophil-1(10)-en-11-one (7.16/6.21%), 15,16-dinorlabda-8(17),12-dien-14-al (6.61/11.57%), and aromadendrene oxide (5.98/3.77%). These results propose C. mangga rhizome EO as a promising agent for developing natural-based anti-infective preparations.

Antibiofilm Activity of Invasive Plants against Candida albicans: Focus on Baccharis halimifolia Essential Oil and Its Compounds.

The extracts of five invasive plants were investigated for antifungal and antibiofilm activities against Candida albicans, C. glabrata, C. krusei, and C. parapsilosis. The antifungal activity was evaluated using the microdilution assay and the antibiofilm effect by measurement of the metabolic activity. Ethanol and ethanol-water extracts of Reynoutria japonica leaves inhibited 50 % of planktonic cells at 250 μg mL-1 and 15.6 μg mL-1 , respectively. Ethanol and ethanol-water extracts of Baccharis halimifolia inhibited >75 % of the mature biofilm of C. albicans at 500 μg mL-1 . The essential oil (EO) of B. halimifolia leaves was the most active (50 % inhibition (IC50 ) at 4 and 74 μg mL-1 against the maturation phase and 24 h old-biofilms of C. albicans, respectively). Oxygenated sesquiterpenes were the primary contents in this EO (62.02 %), with β-caryophyllene oxide as the major component (37 %). Aromadendrene oxide-(2), β-caryophyllene oxide, and (±)-β-pinene displayed significant activities against the maturation phase (IC50 =9-310 μ mol l-1 ) and preformed 24 h-biofilm (IC50 =38-630 μ mol l-1 ) of C. albicans with very low cytotoxicity for the first two compounds. C. albicans remained the most susceptible species to this EO and its components. This study highlighted for the first time the antibiofilm potential of B. halimifolia, its EO and some of its components.

SHORT NOTES: A CHEMOTAXONOMIC STUDY OF THE RESINS FROM THREE DIPTEROCARP SPECIES (Shorea macrophylla, Shorea pinanga, and Shorea hopeifolia)

A chemotaxonomic study on the resins of Shorea macrophylla, Shorea pinanga, and Shorea hopeifolia was conducted. The dichloromethane extracts were separated into neutral and acidic fractions and then analyzed using GC-MS. The neutral fraction analysis revealed that spathulenol, caryophyllene oxide, aromadendrene oxide, and isoaromadendrene epoxide were the major constituents in all three species. Furthermore, the main compound in the neutral fraction of Shorea hopeifolia was isocaryophyllene which was undetected in the other two species. The major constituent of the acidic fraction of the three species was hexadecanoic acid, while pentadecanoic acid was the major constituent in the acidic fraction of Shorea pinanga and Shorea hopeifolia. Therefore, the presence of sesquiterpenes as well as fatty acids in S. macrophylla, S. pinanga, and S. hopeifolia was discovered to be a marker for identifying the genus Shorea.

Centaurea triumfetii essential oil chemical composition, comparative analysis, and antimicrobial activity of selected compounds

The essential oils from the Centaurea genus are well known for their pharmacological properties. The most abundant and dominant chemical components in Centaurea essential oils are ß-caryophyllene, hexadecanoic acid, spathulenol, pentacosane, caryophyllene oxide, and phytol. However, whether these dominant components are the key drivers for observed antimicrobial activity remains unclear. Thus, the aim of this study was dual. Here we provide comprehensive, literature-based data to correlate the chemical compounds in Centaurea essential oils with the tested antimicrobial activity. Secondly, we characterized the essential oil of Centaurea triumfettii All. squarrose knapweed using coupled system gas chromatography–mass spectrometry and tested its phytochemicals for antimicrobial activity against E. coli and S. epidermis using disc diffusion assay and monitoring their growth in Muller Hinton broth. The most abundant compounds in C. triumfettii essential oil were hexadecanoic acid (11.1%), spathulenol (10.8%), longifolene (8.8%), germacrene D (8.4%), aromadendrene oxide (6.0%) and linoleic acid (5.3%). Based on our analysis of literature data from other Centaurea essential oils, they were positively correlated with antimicrobial activity. Using an agar disk diffusion method, tested chemical constituents did not show experimental evidence to support this positive correlation to antimicrobial activity when we tested them as pure components. The antibacterial effect of essential oil constituents may be related to a complex synergistic, rather than a single component as suggested by performed network pharmacology analysis, underlying the theoretical interactions between the essential oil phytochemicals listed as potentially responsible for antimicrobial activity and should be confirmed in further in-depth studies. This is the first report on the comparative analysis of Centaurea essential oils with good antimicrobial activity, as well as the first analysis of chemical components of the essential oil from C. triumfettii and the first report of antimicrobial activity of the representative, pure components: aromadendrene, germacrene D, spathulenol, longifolene, and the mixture of selected chemical compounds. This work contributes to the body of knowledge on the genus Centaurea and C. triumfettii species.

Chemical Composition of Kickxia aegyptiaca Essential Oil and Its Potential Antioxidant and Antimicrobial Activities.

The exploration of new bioactive compounds from natural resources as alternatives to synthetic chemicals has recently attracted the attention of scientists and researchers. To our knowledge, the essential oil (EO) of Kickxia aegyptiaca has not yet been explored. Thus, the present study was designed to explore the EO chemical profile of K. aegyptiaca for the first time, as well as evaluate its antioxidant and antibacterial activities, particularly the extracts of this plant that have been reported to possess various biological activities. The EO was extracted from the aerial parts via hydrodistillation and then characterized by gas chromatography-mass spectrometry (GC-MS). The extracted EO was tested for its antioxidant activity via the reduction in the free radicals, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). In addition, the EO was tested as an antibacterial mediator against eight Gram-negative and Gram-positive bacterial isolates. Forty-three compounds were identified in the EO of K. aegyptiaca, with a predominance of terpenoids (75.46%). Oxygenated compounds were the main class, with oxygenated sesquiterpenes attaining 40.42% of the EO total mass, while the oxygenated monoterpenes comprised 29.82%. The major compounds were cuminic aldehyde (21.99%), caryophyllene oxide (17.34%), hexahydrofarnesyl acetone (11.74%), ar-turmerone (8.51%), aromadendrene oxide (3.74%), and humulene epoxide (2.70%). According to the IC50 data, the K. aegyptiaca EO revealed considerable antioxidant activity, with IC50 values of 30.48 mg L−1 and 35.01 mg L−1 for DPPH and ABTS, respectively. In addition, the EO of K. aegyptiaca showed more substantial antibacterial activity against Gram-positive bacterial isolates compared to Gram-negative. Based on the minimum inhibitory concentration (MIC), the EO showed the highest activity against Escherichia coli and Bacillus cereus, with an MIC value of 0.031 mg mL−1. The present study showed, for the first time, that the EO of K. aegyptiaca has more oxygenated compounds with substantial antioxidant and antibacterial activities. This activity could be attributed to the effect of the main compounds, either singular or synergistic. Thus, further studies are recommended to characterize the major compounds, either alone or in combination as antioxidants or antimicrobial agents, and evaluate their biosafety.

GC-MS analysis and in silico activity prediction of phytocompounds in the roots of Chrysopogon zizanioides (L.) Roberty

Chrysopogon zizanioides (L.) Roberty (Poaceae) commonly known as Ramachamis an aromatic, vigorous growing perennial grass with medicinal properties. The plant is tolerant to extreme soil and climatic conditions and is known for its cooling properties. Roots of the plant are widely used as body scrubber and is suggested for skin diseases in Ayurveda. The present work aims to identify the components in the crude methanolic root extract of C. zizanioides using GC-MS and also to predict the pharmacokinetic behaviour of selected compounds in silico using Swiss ADME online server . 41 compounds were identified of which sesquiterpenes formed the major group. Sesquiterpene Vetivenic acid was the compound with a maximum peak area of 38.9%. Components identified is reported to possess a range of biological activities like anti oxidant, antibacterial, anti cancer, anti inflammatory, anti ulcer, analgesic and insecticidal activities. Compounds with higher peak area like Vetivenic acid, beta vatirenene, beta.-Cedren-9-.alpha.-ol, D Viridiflorol, Gamma muurolene, (Z,E)-alpha-farnesene, Nootkatone, Aromadendrene oxide-(2), 7-Acetyl-2-hydroxy-2methyl-5isopropylbicyclo[4.3.0] nonane, Rosifoliol, 9,10-dehydro isolongifolene, Ylangenol, 4,7,10,13,16,19-Docosahexaenoic acid methyl ester, Carbonic acid, propargyl 2,2,2-tri chloroethyl ester, Oxacyclotetradeca-4,11-diyne, beta eudesmol and longifolene were evaluated in silico. All these compounds proved to obey Lipinski's rule-of-five and were water soluble. Vetivenic acid showed a good bioavailability score of 85% while the others showed 55%. None of the compounds were substrates to P glycoprotein. The values predicted may be used for preliminary evaluation of pharmacological properties of C. zizanioides and also as monographs for the development of potential semisynthetic or synthetic drugs.

Identification of Chemical Constituents of Zingiber zerumbet Rhizome Extract Using GC/MS

Zingiber zerumbet (L.) Roscoe ex Sm. (family-Zingiberaceae) is an important medicinal herb. Though rhizome extract of this species has been used in traditional herbal medicine across the globe for the treatment of numerous diseases like worms, cough, leprosy, asthma and other skin diseases, the composition of the extract is not characterised properly till date. Thus, acetone extract of Zingiber zerumbet rhizome was subjected to Gas Chromatography-Mass Spectrometry (GC/MS) to analyze the phytoconstituents. A total of nine compounds were obtained out of which zerumbone was found to be the major constituent covering 64.58 % of the total area. Beside zerumbone, other important compounds like diacetone alcohol (10.64 %), α-humulene (8.93 %), caryophyllene oxide (5.68 %), humulene epoxide (3.18 %), Eudesm-7(11)-en-4-ol (2.87 %), aromadendrene oxide (1.98 %) were also found in the tested extract with some other compounds in trace amount. Among the compounds, terpenes were found to be the primary constituent of the extract. This finding revealed the presence of the high amount of terpenes in the rhizome of Z. zerumbet for the first time. The study indicated that Z. zerumbet is a good source of sesquiterpenes and oxygenated sesquiterpenes which can be used at industrial level for the production of pharmaceuticals, perfumes and flavouring agents.

Phytochemical, Antioxidant and Antibacterial Potential of Ducrosia anethifolia in Northern Border Region of Saudi Arabia

Ducrosia anethifolia (D. anethifolia) is a drought-tolerant plant widely distributed over Arar valley at the Northern region of Saudi Arabia. The aerial parts of this plant were investigated for its phytochemical constituents, antioxidant and antibacterial potential. GC-MS analysis of the ethyl acetate fraction of methanol extract revealed the presence of some major compounds such as 8-Ethoxypsoralen (6.5%), Prangenin (6.26%), Isoaromadendrene epoxide (7.5%), Aromadendrene oxide (0.96%) and Ferulic acid methyl ester (0.46%). FRAP and DPPH method were used to test the antioxidant capacity of ethyl acetate fraction of D. anethifolia, the results revealed the presence of high reduction capacity (EC50 equals 0.63±0.03g/L), compared with the reducing capacity of the standard ascorbic acid and quercetin which were 0.091±0.002 g/L and 0.026±0.002 g/L, respectively. Moreover, the results of the DPPH test showed that the extract presented a remarkable antioxidant capacity with an IC50 of 0.38±0.02 g/L, This considerable antioxidant capacity is attributed to its richness of some bioactive phytochemical compounds. The antibacterial potential was evaluated by disc-diffusion test, the plant extract was tested on nine different bacterial strains. Results exhibited that, only Gram-positive bacteria recorded good to moderate susceptibility, namely Staphylococcus epidermidis ATCC 49461, Bacillus cereus ATCC 10876, Staphylococcus aureus clinical isolate and Staphylococcus aureus ATCC 25923, which recorded 14.5, 14.0, 9.5- and 7.5-mm zone of inhibition, respectively. In conclusion, the aerial parts of D. anethifolia are rich in some important phytochemical molecules and could be used in the formulation of antioxidant drugs. Whereas, its efficacy against some Gram-positive bacteria only should be studied in-depth. Further studies are also recommended to these phytochemical molecules against various physiological disorders and diseases.

Identification of Anticancer Active Compound from GC-MS Test Results of Zodia Leaves (Evodia suaveolens) Ethanol Extract

Cancer is estimated to be the second leading cause of death globally in 2018. The background of the study was the prevalence of cancer in Indonesia in 2018 reaching 775.120 people. Treatments such as surgery, chemotherapy, radiotherapy, immunotherapy, and hormone therapy have negative side effects. Herbal medicine is an alternative treatment with lower side effects that can reduce the side effects of existing treatments. The aim of this study was to identify the results of GC-MS active compounds from ethanol extract of zodia leaves which have the potential as anticancer. The method of this research was observational research through scientific literature. GC-MS results showed 22 active compounds contained in ethanol extract of zodia leaves. Based on the literature searching, there are 17 active compounds that have potential to be anticancer. Menthofuran; evodone; Bicyclopentene; Delta-Selinene; 3,5,7-Octatrien-2-Ol, 2,6-Dimethyl; Aromadendrene oxide 2; Alpha-bisabolol; 2,4-Dimethyl-2,4-Heptadienal; Phytol; Squalene; Beta-Tocopherol; D-alpha-Tocopherol; Stigmasta-5,23-dien-3.beta.-ol; (23S)-ethylcholest-5-en-3.beta.ol; 24,25-Dihydrolanosterol; Lanosterol; and Obtusifoliol. The compounds are targeted in breast, lung, and ovarian cancer.

The essential oil of Lippia alba Mill (Lamiales:Verbenaceae) as mosquitocidal and repellent agent against Culex quinquefasciatus Say (Diptera: Culicidae) and Aedes aegypti Linn (Diptera: Culicidae)

BackgroundBeing a low-risk insecticide, plant essential oils emerge as competent mosquitocidal and repellent candidates. However, essential oil may act differently in different mosquito species and different developmental stages of same mosquito species. In the current investigation, we evaluated the ovicidal, larvicidal, adulticidal and repellent activities of essential oil extracted from the leaves of Lippia alba against two medically important mosquito species, Aedes aegypti and Culex quinquefasciatus. The efficacy of the oil was assessed under laboratory conditions against different developmental stages of the selected species.ResultsFrom the findings, it can be inferred that L. alba oil is more effective as a repellent against both the targeted mosquito species. Results also demonstrated that ovicidal, larvicidal and repellent properties of the essential oil were higher against Culex quinquefasciatus than Aedes aegypti. GC-MS analysis of the oil showed the presence of aromadendrene oxide, caryophyllene oxide, etc. as major constituent compounds.ConclusionThe outcomes of this study clearly indicated that the essential oil of L. alba has the potentiality to act more effectively as a repellent, followed by a larvicidal agent against mosquito and at the same time the results revealed differential vulnerability of different mosquito species and their life stages against a particular plant essential oil.

Antibacterial and antiproliferative activities of the fresh leaf essential oil of Psidium guajava L. (Myrtaceae).

This study evaluated the antibacterial and antiproliferative activities of the essential oil of Psidium guajava leaves (PG-EO), traditionally used in folk medicine. The essential oil was obtained from fresh leaves by hydrodistillation, using a modified Clevenger apparatus. The major PG-EO chemical constituents were identified by GC-MS and GC-FID as being β-caryophyllene (16.1%), α-humulene (11.9%), aromadendrene oxide (14.7%), δ-selinene (13.6%), and selin-11-en-4α-ol (12.5%). The antibacterial activity of the essential oil of P. guajava leaves was determined in terms of its minimum inhibitory concentrations (MIC) using the broth microdilution method in 96-well microplates. PG-EO had moderate activity against Streptococcus mutans (MIC = 200 µg/mL), S. mitis (MIC = 200 µg/mL), S. sanguinis (MIC = 400 µg/mL), S. sobrinus (MIC = 100 µg/mL), and S. salivarius (MIC = 200 µg/mL). The antiproliferative activity was evaluated against different tumor cell lines: breast adenocarcinoma (MCF-7), human cervical adenocarcinoma (HeLa), and human gliobastoma (M059J). A normal human cell line (GM07492A, lung fibroblasts) was included. The antiproliferative activity was evaluated using the XTT assay and the results were expressed as IC50. The essential oil showed significantly lower IC50 values against MCF-7 and M059J lines than that obtained for the normal line, showing selectivity. Our results suggest that the essential oil of Psidium guajava L. has promising biological activities and can be considered a new source of bioactive compounds.

UFLC-Q-TOF-MS fingerprints of rhizome of Curcuma phaeocaulis and its vinegar processed products and inhibitory effect on thrombosis

Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.

Synergistic interaction of β-caryophyllene with aromadendrene oxide 2 and phytol induces apoptosis on skin epidermoid cancer cells

BackgroundPamburus missionis (Wight) Swingle (Rutaceae) is traditionally used in the treatment of swellings, chronic rheumatism, paralysis and puerperal diseases. In a previous study the authors demonstrated apoptotic activity of Pamburus missionis essential oil (EO) on A431 and HaCaT cells. The major components of EO were β-caryophyllene (25.40%), 4(14),11- eudesmadiene (7.17%), aromadendrene oxide 2 (14.01%) (AO-(2) and phytol (6.88%). Purpose of studyTo investigate the role as well as the interactions among EO components inducing apoptosis in A431 and HaCaT cells. MethodsIsobolographic analysis and combination index methods were used to detect the type of interactions among the essential oil (EO) components. Cell viability was used to detect cytotoxic activity. Mechanism of cell death was studied using Annexin V-FITC/PI binding assay, cell cycle analysis, measurement of MMP and ROS generation by flow cytometry. Expression of apoptosis associated proteins was investigated by western blot. ResultsCombination of P. missionis EO components: β-caryophyllene/ aromadendrene oxide 2 (β-C/AO-(2)), β-caryophyllene/phytol (β-C/P) and aromadendrene oxide 2 /phytol (AO-(2)/P) inhibited growth and colony formation ability of skin epidermoid A431 and precancerous HaCaT cells. Synergistic interaction was observed between β-C/AO-(2) and β-C/P combination while AO-(2)/P exhibited an additive effect. Combination of components induced chromatin condensation, phosphatidylserine externalisation, increase in sub-G1 DNA content, cell cycle arrest at G0/G1 phase and intracellular ROS accumulation. Inhibition of intracellular ROS by N-acetyl cysteine treatment blocked apoptosis induced by the combinations. The combinations induced apoptosis in A431 and HaCaT cells mediated by loss of mitochondrial membrane potential (ΔΨm), increase in Bax/Bcl-2 ratio, release of cytosolic cytochrome c and activation of caspases (cleaved form of caspase-3, caspase-8, caspase-9) and by PARP cleavage. ConclusionThe present study demonstrates interactions among β-C, AO-(2) and P in the induction of apoptosis on A431 and HaCaT cells. These data suggest the combination of β-caryophyllene with aromadendrene oxide 2 and phytol could be potential therapeutics for the treatment of skin epidermoid cancer and precancerous cells.

Aromadendrene oxide 2, induces apoptosis in skin epidermoid cancer cells through ROS mediated mitochondrial pathway

AimsAromadendrene oxide 2 (AO-(2)) is an oxygenated sesquiterpene naturally found as a chemical component of essential oils. In the present study anticancer activity of AO-(2) has been investigated on A431 human epidermoid cancer and precancerous HaCaT cells. Material and methodsCell viability was used to detect cytotoxic activity. Mechanism of cell death induced by AO-(2) treatments was studied using Annexin V-FITC/PI binding, cell cycle analysis, measurement of MMP and ROS generation by flow cytometry. Expression of apoptosis related proteins was investigated by western blot. Key findingsAO-(2) inhibited the growth and colony formation ability of A431 and HaCaT cells in concentration dependent manner. It induced cell cycle arrest at G0/G1 phase and apoptosis through intracellular ROS accumulation. Inhibition of intracellular ROS by ascorbic acid and N-acetyl cysteine treatment completely blocked apoptotic effect. N-acetyl cysteine treatment significantly reversed G0/G1 arrest induced by AO-(2). AO-(2) treatment caused loss of mitochondrial membrane potential (ΔΨm), increase in Bax/Bcl-2 ratios, cytochrome c release, activation of caspases (cleaved caspase-3 and caspase-9) and PARP cleavage. AO-(2) also significantly inhibited the growth of multicellular tumor spheroids of A431 and HaCaT cells. SignificanceThe results of the present study reveals that AO-(2) a chemical component of essential oils induces apoptosis in A431 and HaCaT cells.

Comparative GC-MS analysis of all <em>Curcuma</em> species grown in Sri Lanka by multivariate test

Curcuma is clinically valuable genus in Traditional Medicine. People use various plants under the same vernacular name may lead to adulteration or substitution. Whole plants of Curcuma species were collected in 2016 in the flowering season. Voucher specimens of the plants were authenticated from the National Herbarium, Peradeniya. Essential oils were extracted from Clevenger’s apparatus and analyzed separately by GC-MS. The analyses were carried out with RTX WAX capillary column. Sampling and experiments were done according to WHO guidelines. One hundred sixty four phytochemicals were analyzed by simple correspondence and by cluster variable method. By cluster varibale as per phytochemicals present, mainly two groups were identified. C. albiflora and C. oligantha were identified as one group and the rest of the three plants were kept in the other group. A total of 64 constituents of essential oil obtained from whole plant of C. albiflora were identified by GC-MS, where α-pinene (10.87 %), caryophyllene oxide (8.85 %), alcanfor (5.12 %), aromadendrene oxide-(1) (4.81 %), n-hexadecanoic acid (4.74 %), α-famesene (3.93 %), camphene (3.52 %), and isoborneol (3.4 %) were detected as major compounds. The essential oil of C. aromatica possesses 7–methanoazulene (13.75 %) and curcumene (25.71%). Caryophyllene (15.07%), phytol (13.38%), humulene (8.24%), elemene (6.11%), caryophyllene oxide (5.82%) were found in C. oligantha. This preliminary study has identified chemical markers present in all Curcuma species grown in Sri Lanka. Keywords: Curcuma species, essential oils, GC-MS, multivariate.

Induction of apoptosis by essential oil from P. missionis in skin epidermoid cancer cells

BackgroundThe genus Pamburus (Rutaceae) comprises the only species, Pamburus missionis (Wight) Swingle. Pamburus missionis is traditionally used in the treatment of swellings, chronic rheumatism, paralysis and puerperal diseases. PurposeThe present study investigates the cancer chemotherapeutic potential of essential oil (EO) from P. missionis. MethodsEO was isolated by steam distillation and chemical composition was determined by GC-MS. Cell viability was used to detect cytotoxic activity. Mechanism of cell death was studied using Annexin V-FITC/PI binding, cell cycle analysis, measurement of MMP and ROS generation by flow cytometry. Expression of apoptosis related proteins was investigated by western blot. ResultsGC-MS analysis of the essential oil revealed the presence of 51 components. The major components were β-Caryophyllene, 4(14),11-Eudesmadiene, Aromadendrene oxide-(2) and Phytol. EO inhibited the growth and colony formation ability of A431 and HaCaT cells. EO treatment induced nuclear condensation and loss of membrane integrity, DNA fragmentation, increase in sub-G1 DNA content and increase in intracellular ROS level. Inhibition of intracellular ROS by ascorbic acid and N-acetyl cysteine treatment blocked EO induced apoptosis, revealing that apoptotic activity was by ROS accumulation. EO induced apoptosis was found to be due to the loss of mitochondrial membrane potential (ΔΨm), increase in Bax/Bcl-2 ratio, release of cytochrome c and activation of caspases (cleaved form of caspase-3, caspase-8, caspase-9) and by PARP cleavage. ConclusionThe present study revealed cancer chemotherapeutic potential of EO from P. missionis. EO induces cell death through intrinsic (mitochondrial) and extrinsic apoptotic pathway in A431 and HaCaT cells. These results suggest that EO could be used as a potential therapeutic agent for the treatment of skin epidermoid cancer.

Antioxidative Activities of Essential Oils and Ethanol Extrations from Ornamental Zingiberaceae Species

The essential oils from five different Zingiberaceae species [Costus speciosus (J. Konig.) Smith, Hedychium coronarium J. Koenig, Hedychium gardnerianum Griff., Zingiber zerumbet (Linn.) Roscoe ex Smith, Hedychium chrysoleucum Hook.] were extracted through hydro-distillation process and their chemical compounds were analyzed by GC-MS. In addition, the antioxidant activities of essential oils and ethanol extractions were described. The yields of essential oils ranged from 0.20 % in C. speciosus to 1.52 % in Z. zerumbet. A total of 48 components were identified, of which majority were ⟨-santalo (0.22-41.83 %), aromadendrene oxide I (39.63-55.40 %) and zerumbone (1.67-84.07 %). The essential oils showed an effective DPPH• scavenging activities with IC50 values ranging between 5.28 and 13.26 μg/mL. The result showed that the ethanol extractions had different DPPH• scavenging (IC50, 0.94-9.92 μg/mL) and most of them were lower than ascorbic acid (IC50, 8.37 μg/mL) and trolox (IC50, 10.19 μg/mL). The results indicate that the essential oils and ethanol extractions from the five ornamental Zingiberaceae species could serve as an important bio-resource of antioxidant for using in the functional food, cosmetic, and pharmaceutical industries.