Antioxidant and genotoxic activity of Pistacia terebinthus and Ephedra alata crude extracts 

Antioxidant and genotoxic activity of Pistacia terebinthus and Ephedra alata crude extracts | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Antioxidant and genotoxic activity of Pistacia terebinthus and Ephedra alata crude extracts Mohammed HARIR, Hamdi Bnedif, hauker Reimann, Silva Filomena, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4648793/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In the last decades, cases of poisoning due to medicinal plants have been very common in many countries including Algeria. Algerian herbal medicines have been used in effectively treating several health problems, but their toxicity limits their use. Pistacia terebinthus and Ephedra alata herbal medicine collected from Algeria have been used for a long time to treat several diseases. Thus, this study aimed to investigate the chemical composition and evaluate the genotoxicity and the antioxidant activity of crude extracts of these two herbal medicines ( E. alata and P. terebinthus ) traditionally used in Algeria and North Africa. For this purpose, genotoxicity was assessed using the micronucleus test. Additionally, compounds with antioxidant and genotoxic activities of these two plants were identified by gas chromatography coupled to mass spectrometry (GC-MS). The highest antioxidant capacity of the extracts was found in P. terebinthus ethanol extract owing to its richness of daphnetin, and oleic acid amide. Our results showed that most of these plants exhibited antioxidant activities and genotoxic effects, depending on the level of administered doses. To our knowledge, this is the first report on these effects caused by of both plants. Pistacia terebinthus Ephedra alata Antioxidants Genotoxicity Micronucleus test GC-MS Figures Figure 1 Figure 2 INTRODUCTION Several herbal drugs have yielded important modern therapeutic agents. In Algeria, folk medicinal herbs such as Pistacia terebinthus (family of Anacardiaceae ) are widely distributed. Pistacia terebinthus L. (turpentine tree, terebinth) is a perennial shrub or small tree widely grown in southern and western Turkey, and in western and eastern Algeria on dry rocky slopes and hillsides or in pine forests (Özcan et al., 2009 , Bellifa et al., 2021 ). In fact, P. terebinthus fruits are commonly consumed as a coffee-based beverage, which is prepared with milk, known as ‘menengiç’ coffee in Turkish (Baytop, 1984). P. terebinthus fruits have been the focus of several research activities due to their antimicrobial, antioxidant (Topçu et al., 2007 ), and anti-inflammatory properties (Giner-Larza et al., 2000 ), as well as their high oil content (approximately 40%) ( Matthaus and Ozcan, 2006). These fruits contain high concentrations of unsaturated fatty acids and carotenoids, phenolic compounds and tocopherols, tannins, resinous substances, and dietary fibers (10%) (Özcan, 2004; Matthäus and Özcan, 2006 ). Terebinth has been described as an antiseptic and antihypertensive, and it has also been used to treat gastrointestinal and respiratory disorders (Bozorgi et al., 2013 ). As previously reported, various parts of this species (stems, flowers, and leaves) have antioxidant and other pharmacological properties. Due to their richness in secondary compounds such as flavonoids and polyphenols, radical scavenging activity of Pistacia terebinthus extracts was found to be fairly high (Topçu et al., 2007 ), indicating a possible preventive role in carcinogenesis by reducing oxidative stress ( Kavak et al., 2010 ). Another common folk remedy, Ephedra alata , a medicinal plant of the Ephedracea e family known in Algeria as Alenda, is a perennial, xerophytic, gymnosperm and delicious shrub with erect non-climbing stems and short leaves that are united towards the base and yellowish-green bracts that flowers in July (Bell, A. and Bachman, S., 2011) and has a strong pine-like smell (Blumenthal, M. and King, P, 1995). This shrub grows in temperate, subtropical, and dry environments and can be found in the North African countries of Algeria, Tunisia, Morocco, Egypt, Chad, Mauritania, and Mali, as well as in Asia (China, Palestine, Lebanon, Jordan, Saudi Arabia, Iran, and Iraq) and Europe (Spain) (Ebadi, M.,2007). E. alata is used in traditional medicine for treating respiratory disorders and rheumatism (Aidi et al 2023). The plant is also used in the traditional Chinese Pharmacopeia against cough, hay fever, cold, chills, asthma, allergies, and edema (Al-Snafi,.2017) , while Algerians and Tunisians use it as anti-cancer treatment (Sioud, F et al , 2020) , in addition to its effects in reducing the side effects of chemotherapy (Jaradat, N.A et al ., 2016). The primary active molecules in E. alata are the ephedrine alkaloids which have major side effects such as adverse cardiovascular and cerebrovascular events if used in high doses, which explains why the US Food and Drug Administration (FDA) has banned any drugs that contain ephedrine alkaloids (Ibragic, S. et al. , 2015) . The major compounds detected in E. alata growing in Algeria are isoflavones and flavonol derivatives with hydroxypuerarin isomer as the major molecule. For the evaluation of genotoxic and cytotoxic effects of many herbal medicines, extracts of different plant parts have been used, ranging from leafy vegetables, fruits, and underground storage organs to whole plants. The extracts are prepared mainly in water or organic solvents. Several of these toxicity assays have indicated the involvement of certain factors that are intrinsic components of the extracts, ranging from specific compounds like ascorbic acid to vegetable fibres which could act as nonspecific redox agents, free radical scavengers, or ligands for binding metals or toxic principles. Oxidative stress can cause DNA damage in the cells. If DNA repair is unable to modify or repair the DNA damage, genomic instability may lead to mutation, cancer, aging, and many other diseases. However, cells have special mechanisms to counteract and minimize these damages. Recently, research has shown that dietary patterns and food constituents can revert this toxicity in cells. A considerable body of epidemiological evidence indicates that a diet with high intake of fruits and vegetables is inversely related to the risk of chronic, degenerative diseases such as coronary heart disease and certain types of cancer. Much research effort has focused on the identification of phytochemicals in fruits and vegetables that exert beneficial effects and on the elucidation of the mechanisms by which they inhibit cellular damage and degeneration (Sarkar et al ., 2008). Several experimental systems (e.g., membranes, plants, cell cultures, animal models and human clinical trials) were used to study the bioactivity of these plant-derived compounds. The effects of toxic compounds can for example be observed at the level of chromosomes by quantifying alterations in chromosome structure (chromosomal aberrations) and number (aneuploidy, polyploidy). Assays with several crude vegetable and fruit extracts have shown their clastogenic effect or protective activities against known genotoxic agents such as plants containing aristolochic acid. Additionally, single extract components such as sulfhydryl and flavonoid compounds, gallic acid, ellagic acid, mucic acid, citric acid, reducing sugars, and tannins have been described to have an additive interaction with the major constituent’s chlorophyll and ascorbic acid when modulating the effects of the clastogens (Leme&Marin-Morales, 2009). The increase in chromosomal aberrations may result from interactions of a different variety of chemical agents with DNA. According to ( Ishidate et al . 1988) , agents that induce an increase in the chromosomal aberration’s frequency by direct or indirect mechanism may also be cytotoxic, by causing damage to both DNA and other cellular targets (enzymes, membranes, structural proteins). The present study aimed to establish a phytochemical profile, and evaluate the antioxidant, cytotoxicity, and genotoxicity potential of E. alata and P. terebinthus grown in Algeria. MATERIAL AND MATHODS Plant materials and samples preparation Pistacia terebinthus and Ephedra alata leaves, stems and flowers were collected from Ouled Hamdane of Bordj Ghedir (Bordj Bou Arreridj province, North of Algeria, 1100 m meters in altitude., 35.9125° N, 4.9247° E). The taxonomic identification of plant materials was performed using Flora of Algeria (Quezel and Santa 1962 ), while herbarium specimens were archived in the Herbarium of Tiaret University. The harvested plant material was dried at room temperature (20–25°C) for about fifteen days in the open air until the stabilization of its weight to avoid possible risks of polyphenol oxidation and to preserve as much as possible the integrity of the molecules. Then, the samples were collected in clean bags and stored away from light and moisture in a tightly closed container for further use. Preparation of crude extracts by maceration For the preparation of the crude extracts, the solvent used was ethanol 95% (v/v), which has been chosen to allow the solubilisation of different compounds. This solvent has been used at a solid:liquid ratio of 1:10 g plant material / mL solvent of extraction ( Michel, 2011 ). For this purpose, 5 g of the plant powder from the aerial parts of both plants (leaves, stems, and flowers) were mixed with 50 mL of ethanol in an Erlenmeyer flask. The mixture was kept under magnetic stirring for 24 h at room temperature and the obtained solution was filtered with filter paper (Wattman No. 1 of pore diameter 0.2 µm) under vacuum. The filtrate was recovered and the ethanolic extracts were concentrated under a vacuum at 45°C using a rotary evaporator (Buchi Labortechnik AG, Switzerland) for one hour. Afterwards, the obtained crude extract was lyophilized and stored at 4 C°. Gas chromatography-Mass spectrometry analysis of extracts The phytochemical investigation of 1% (w/w) ethanolic extracts was performed on an Agilent HP 6890 series gas chromatograph system (Agilent Technologies, Spain) connected to an Agilent HP 5973 series mass selective detector. The sample (1 µL) was injected at the injection port (250 ºC) in splitless mode. Chromatographic separation was carried out on a DB-5 column (30 m x 0.25 mm x 0.25 µm) (Agilent Technologies, Spain). The initial column temperature was set at 40°C and raised to 250°C at a rate of 5°C/min and held for 2 minutes resulting in a total chromatographic run time of 45 minutes. Helium (99.999%, Air Liquid, Madrid, Spain) was used as carrier gas at a flow rate of 1.0 mL/min. MS detection was performed in Scan mode at a range of 50 to 650 m/z and the results were compared by using NIST 14 Mass Spec Library search program. Cell culture Hela H2B GFP cells expressing green fluorescent protein fused to histone H2B were provided by Noriaki Shimizu, Graduate School of Integrated Sciences for Life, Hiroshima University, Japan and seeded at a density of 10.000 cells/ml in 75 cm 2 tissue culture flasks, grown in Dulbecco’s modified Eagle’s medium (Gibco BRL, Cergy-Pontoise, France) supplemented with 10% fetal bovine serum (Gibco BRL), 1% L-glutamine and 1% penicillin/streptomycin mixture (Gibco BRL). Cultures were maintained in a humidified atmosphere with 5% CO 2 at 37°C to sub-confluence. Fresh medium was supplied every 48 h. Genotoxicity Evaluation The genotoxicity activity of plant extract solutions was assessed using the micronucleus assay in Hela H2B GFP cells as previously described by Michael Fenech ( 2007 ) with slight modifications. Briefly, test extract solutions were prepared using a stock solution of 200 mg/mL in double distilled water and diluted in a culture medium to give a final concentration of 0.1 to 2 mg/mL. Cells were incubated with extract solutions for 4 h and afterwards subjected to treatment with cytochalasin B (3 µg/mL). Etoposide (14 µg/l) was used as a positive control. After 24 hours of incubation with cytochalasin B, cells were harvested, and slides were prepared by cytospin centrifugation at 100 rpm for 5 min and fixed on a slide. The slides were stored overnight in ice-cold methanol (-20°C) and stained with Gel Green staining solution (Biotium Glowing products for science, 1:100 dilutions in water) for 6 min in the dark. Finally, slides were mounted using diazabicyclo-octane (DABCO) and coverslips. Two replicate experiments were performed independently and the results are shown as the mean of these replicates. Evaluation of micronuclei was performed at 400-fold magnification under a fluorescence microscope using a fluorescein isothiocyanate (FITC) filter (Nikon GmbH, Japan). The number of mononucleated, binucleated, multinucleated, apoptotic, and mitotic cells (was calculated. Also, the number of micronuclei in 1000 binucleated cells was counted. The cytokinesis block proliferation index (CBPI) was determined in 1000 cells that were counted for mononucleated cells, binucleated cells, and multinucleated cells by using the following equation: CBPI= ((No. mononucleate cells) +(2×No.binucleat cells) +(3×No.multinucleate cells))/Total number of cell DPPH radical scavenging assay The method is based on the reduction of alcoholic 1,1-diphenyl-2-picrylhydrazyl (DPPH) solutions in the presence of a hydrogen-donating antioxidant. The effect of each extract on DPPH was measured using a method previously described (Que et al., 2006 ). In brief, 1 mL of various concentrations (0–50 mg/mL) of the ethanolic extract or standard solution was added to 1 mL of freshly prepared DPPH (0.1 mM) methanolic solution. The mixture was mixed vigorously and allowed to stand in the dark for 30 min at room temperature. After this incubation period, the absorbance (A) at 517 nm was recorded using a UV–VIS spectrophotometer (Shimadzu). A calibration curve was prepared with ascorbic acid as a control (1–10 mg/mL methanol the free radical-scavenging activity of each solution was then calculated as percent inhibition according to the following equation: % inhibition = 100 (A (control) - A (sample)) / A (control) The antioxidant activity of the samples was expressed as Efficient Concentration 50 (EC50, in µg/ml) value. EC50 is defined as the concentration of the extract equivalent to the effective antioxidant activity at 50%. Antioxidant activity is also expressed as the Antioxidant Activity Index (AAI), calculated as follows: AAI = final concentration of DPPH in the control sample/IC50Thus, the AAI was determined considering the mass of DPPH and the mass of the ethanolic extract in the reaction. This gives a constant for ethanol extract independent of the concentration of DPPH or the ethanol extract used. The AAI allows the following classification of the antioxidant activity of ethanol extracts: poor (AAI ≤ 0.5), moderate (0.5 < AAI ≤ 1.0), strong (1.0 < AAI < 2.0) or very strong (AAI ≥ 2.0) (Luís et al., 2016 ). Tests were performed in triplicate and DPPH solutions were prepared daily. RESULTS Chemical analysis We have observed that the extraction yield of the different samples was quite similar (nearly 12%). The GC-MS analysis of the compounds with their retention time (RT), molecular formula, and percentage are presented in Tables 1 and 2 . Among the compounds identified in P. terebinthus , the most abundant ones were oleic acid amide (21.46%) followed by daphnetin (13.66%) , a natural coumarin derivative which is a protein kinase inhibitor ( Table 1 ). E. alata extract contains twenty-seven identifiable compounds, with oleic acid amide, ethyl gallate, and pyrogallic acid being the major compounds (Table 2 ) . Table 1 Compounds identified from ethanolic extract of P. terebinthus using GC-MS. N° Compound Molecular formula Retention time (min) % a P. terebinthus 1. Furfural C 5 H 4 O 2 5.981 0.15 2. (E)-Conipheryl alcohol Loliolide C 10 H 12 O 3 28.695 0.17 3. 2,3-Dihydro-3,5-dihydroxy-6-methyl-4H pyran-4-one C 6 H 8 O 4 13.231 0.93 4. 5Hydroxymethylfurfural C 6 H 6 O 3 15.641 0.14 5. Daphnetin C 11 H 16 O 3 29.51 13.66 6. Dihydroxyacetone C 3 H 6 O 3 6.904 1.01 7. Eicosane C 20 H 42 43.59 0.13 8. Furanmethanol C 5 H 6 O 2 6.334 0.16 9. Myristamide C 16 H 32 O 2 32.83 0.49 10. Neophytadiene C 9 H 6 O 30.25 0.35 11. Oleic acid amide C 18 H 35 NO 39.727 21.46 12. Palmitic acid C 20 H 38 32.66 3.11 13. Palmitic acid amide C 16 H 33 NO 36.604 1.20 14. Palmitic acid β-monoglyceride C 19 H 38 O 4 42.001 0.19 15. Palmitic acid, butyl ester C 20 H 40 O 2 36.76 0.22 16. Stearic acid C 18 H 36 O 2 36.333 0.84 17. trans-Phytol C 14 H 29 NO 35.47 1.56 18 α –linolenic acid C 18 H 30 O 2 35.932 2.86 Table 2 Compounds identified from ethanolic extract of E. alata using GC-MS N° Compound Molecular formula Retention time (min) %a E. alata 1. Furfural C 5 H 4 O 2 5.987 0.05 2. (1S) -eno)-(-)-Borneol C 10 H 18 O 13.93 0.24 3. 2,4-Bis(1,1-dimthylethyl) phenol C 14 H 22 O 23 0.75 4. 2-Formyl-1-methylpyrole C 6 H 7 NO 9.531 0.14 5. 2-Methylheptadecane C 18 H 38 31.234 0.05 6. 3,4-Dimethylbenzaldehyde C 9 H 10 O 15.267 0.44 7. 3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one C 6 H 8 O 4 13.19 0.22 8. 4-Methyldecane C 11 H 24 10.047 0.06 9. Benzenemethanol C 7 H 8 O 10.278 0.23 10. Ethyl gallate C 9 H 10 O 5 32.443 14.48 11. Hexadecane C 16 H 34 17.128 0.45 12. n-Cetane C 16 H 34 27.568 0.24 13. n-Heneicosane C 21 H 44 43.576 0.31 14. n-Heptacosane C 27 H 56 28.689 0.13 15. n-Hexacosane C 26 H 54 31.329 0.08 16. n-octacosane C 28 H 58 30.467 0.12 17. n-octacosane C 18 H 38 31.465 0.10 18. o-Cymene C 10 H 14 10.095 0.03 19. Oleic acid amide C 18 H 35 NO 39.713 21.40 20. Palmitic acid C 18 H 36 O 2 33.271 0.09 21. Palmitic acid amide C 16 H 33 NO 36.597 1.11 22. Palmitic acid, butyl ester C 20 H 40 O 2 36.76 0.06 23. p-Methyl cyclohexanone C 7 H 12 O 9.361 0.07 24. Pyrogallic acid C 6 H 6 O 3 19.558 10.47 25. Pyrogallol C 6 H 6 O 3 8.94 0.37 26. Stearic acid C 18 H 36 O 2 36.319 0.78 27. Trans-phytol C 20 H 40 O 35.47 0.77 28 α –linolenic acid C 18 H 30 O 2 35.891 0.59 a The percentage of compound was calculated as the percentage ratio between compound peak area and total peak area. The genotoxicity analysis of the crude extracts from leave and flowers of P. terebinthus on HeLa cells showed that all concentrations tested yielded a micronucleus frequency slightly higher than the control ( Fig. 1) . The increase in micronuclei was not very pronounced and was not comparable to the response to the positive control etoposide, which approximately showed a 10-fold increase in micronuclei. No clear dose-response could be observed. Also, the CBPI did not show any relevant alterations. With regards to the crude extracts of Ephedra alata on HeLa cells, we observed an increase in micronuclei at the lowest concentration, which was not observed at the higher concentrations (Fig. 2 ). As shown in Table 3 , we can rank the samples in order of decreasing reactivity: ascorbic acid > Ephedra alata extract > Pistacia terebinthus extract. Table 3 DPPH radical scavenging activities Table 3 : Antioxidant properties (EC 50 µg/ml) of Ephedra alata and Pistacia terebinthus ethanolic extracts . DPPH free radical scavenging assay Strength of Antioxidant Activity Sample EC 50 (µg/mL) AAI Ephedra alata 0.46 ± 0.968 85.71 ± 3.22 Very strong Pistacia terebinthus 2.90 ± 0.978 13.59 ± 1.66 Very strong Ascorbic acid 0.08 ± 0.839 492.87 ± 09.23 Very strong The EC50 values of the extracts are 0.46 ± 0.96 and 2.90 ± 0.97, with an AAI value of 85.71 ± 3.22 and 13.59 ± 1.66 respectively, which allows the ethanolic extracts of Ephedra alata and Pistacia terebinthus to be classified as having very strong antioxidant activity. DISSCUSSION In the present investigation, more than twenty bioactive chemical constituents were identified in each plant crude extract. Pistacia terebinthus contains daphnetin (DAP) as a major compound (13.6%) which is a natural coumarin derivative isolated from different medicinal herbs. It possesses abundant biological activities, such as antioxidant and anti-inflammatory properties. In addition, the oleic acid amide, present in both crude extracts (21.5%), has been demonstrated to interfere with 5-HT2a, 5HT2c, and GABAa receptors ( Mendelson & Basile,2001) , These two major compounds have all been shown to have hypocholesterolemic and antioxidant activity. Ephedra alata contained twenty-seven compounds with Pyrogallic acid (10.47%), ethyl gallate (14.48%) and oleic acid amide (21.40%) as major compounds. The ethyl ester of gallic acid is an approved food additive (E313) that is added to foods due to its antioxidant activity (Belifa , et al . 2021). Many studies have confirmed the importance of plant antioxidants ( Aissani & Sebai 2022; Aissani et al , 2020 ). In our study, it was important to evaluate Ephedra alata and Pistacia terebinthus ethanolic extracts via the DPPH test to get more informative results on their antioxidant potential. The EC 50 values results confirmed that both E. alata and P. terebinthus ethanolic extracts were active regarding antioxidant capacity. Our finding is in agreement with other studies about other species of Ephedra genus collected in other countries. As reported by Jaradat et al in 2015 , a better antioxidant activity with lower IC 50 of Palestinian E. alata was shown, which can be attributed to their alkaloid content. Our phytochemical results revealed that Algerian E. alata did not contain alkaloids, which can be associated with a safer toxicological profile since Ephedra alkaloids (ephedrine and pseudoephedrine) are the main compounds responsible for its toxicity ( Ibragic, S et al 2015 ). Methanolic extracts of the aerial parts of Palestinian Ephedra alata (Jaradat et al., 2015 ) were also reported to have high antioxidant properties as free radical inhibitors and scavengers, and showed lower EC 50 values (16.03 µg/mL for DPPH). To our best knowledge, this is the first report on the antioxidant activity of the ethanolic extracts of E. alata . However, another work on E. alata from Palestine (Al-rimawi et al., 2017 ) has reported antioxidant effects of the extract, with higher contents in polyphenols, using higher polarity solvents. Thus, the highest activity was reported for the hydroethanolic extract (78 µg/mL for DPPH). This discordance noted in relation to our sample from Algeria is probably due to the effect of the region and climatic conditions affecting the biosynthesis of phenolic compounds, and to the effect of the extraction solvent used. This can generate quantitative differences in the bioactive molecules and therefore influences the antioxidant activity. Parsaeimehr and collaborators in 2010 studied Ephedra wild culture species from Iran ( E. procera Fisch. & Mey., E. pachyclada Boiss. and E. strobilacea Bunge) and found that the methanolic extract of E. strobilacea had higher antioxidant activity (FRAP assay) in comparison to the other species ( Al-Rimawi et al., 2017 ). The same trend was obtained with the hydroalcoholic mixture (ethanol, methanol and distilled water 7:2:1) of E. intermedia Schrenk ex Meyer from Balochistan, Pakistan in DPPH free radical scavenging assay (Gul, Jan, Faridullah, Sherani, & Jahan, 2017 ). The present results show that our ephedra extract has a strong antioxidant potential, which is consistent with previous results on the aerial parts of several Ephedra plants. Furthermore, the aerial parts of E. alata from Tunisia showed high DPPH radical scavenging activity and EC 50 values in methanol extracts ( Dbeibia et al ., 2022 ). It has been reported that the antioxidant properties of plant extracts may be attributed to phenolic compounds and their possible additive, synergistic or antagonistic interactions ( Kurin et al., 2012 ). The demonstrated antioxidant potential may be even more interesting for biotechnological applications and industrial uses. The antioxidant potential of Pistacia extract has been studied by other researchers (Abidi et al 2017 ) and (Rajaei et al. 2010 ), who have reported that the water extract showed a strong DPPH antioxidant activity when compared to THBQ and BHT standards. Kilic et al . (2016 ), have also reported that the DPPH antioxidant activity of pistachio hull extract was 137.16 mg Trolox Equivalent/g extract. Comparing our results with those reported previously on P. atlantica leaf and stem extracts ( Peks el et al. , 2008; Toul et al. , 2017; Amri et al., 2018 ), it can be stated that all extracts were very effective in the DPPH assay. The genotoxicity findings are difficult to interpret. For the P. terebinthus samples, a small increase in micronuclei was observed compared to the control, which might indicate a clastogenic potential of the extracts. However, the increase is fairly small, particularly when compared to the response of the positive control, which yielded a much higher increase in micronuclei. Additionally, no clear dose response could be observed. All in all, genotoxic properties of the extract cannot be clearly concluded from the data. A similar situation can be observed for the E. alata extract samples. There is a clear increase of micronucleus frequency at the lowest concentration, but this increase comes along with a high intra-experimental variability shown by the large standard deviation. It is unclear whether this increase is a real effect or due to an experimental outlier. At all other concentrations, a very small increase in micronuclei can be observed, the relevance of this small increase is unclear. CONCLUSION Our study indicates that both Pistacia terebinthus and Ephedra alata extracts might have a genotoxic effect, arising from the use of herbal medicinal products. Antioxidant activity is also confirmed. This is the first report regarding the genotoxicity activity of both plants. This finding provides new insights into the existing drugs and may help to facilitate the development of antitumor agents. As the global use of herbal medicinal products continues to grow and many more new products are introduced into the market, public health issues and concerns surrounding their safety are also increasingly recognized. Although some herbal medicines have promising potential and are widely used, many of them remain untested and their use is also not monitored. This makes knowledge of their potential adverse effects very limited and identification of the safest and most effective therapies as well as the promotion of their rational use more difficult. In most countries, herbal medicines and related products are introduced into the market without any mandatory safety or toxicological evaluation. Many of these countries also lack effective mechanisms to regulate manufacturing practices and quality standards. These herbal products are continuously made available to consumers without prescription in most cases and the potential hazards of an inferior product are hardly recognized. Declarations DECLARATION OF CONFLICT OF INTEREST The authors declare no conflict of interest. ACKNOWLEDGMENTS The authors are grateful to Mrs. Heike Keim-Heusler for providing technical assistance. FUNDING This work was supported by the German Academic Exchange Service by a Postdoc fellowship. ETHICAL APPROVAL Not applicable References Abidi A, Aissani N, Sebai H, Serairi R, Kourda N, Ben Khamsa S. Protective effect of Pistacia lentiscus oil against bleomycin-induced lung fibrosis and oxidative stress in rat. Nutr Cancer. 2017;69(3):490–7. Aidi Wannes W, Tounsi MS. Phytochemicals, Traditional Uses, Biological Effects, and Possible Molecular Mechanisms of Ephedra alata. Futur Integr Med. 2023;2(4):189–99. Aissani N, Albouchi F, Sebai H. Anticancer effect in human glioblastoma and antioxidant activity of Petroselinum crispum L. methanol extract. Nutr Cancer. 2021;73(11–12):2605–13. Al-Rimawi F, Abu-Lafi S, Abbadi J, Alamarneh AAA, Sawahreh RA, Odeh I. Analysis of phenolic and flavonoids of wild Ephedra alata plant extracts by LC/PDA and LC/MS and their antioxidant activity. African J Tradit Complement Altern Med. 2017;14(2):130–41. Al-Snafi AE. Pharmacology and therapeutic potential of Euphorbia hirta (Syn: Euphorbia pilulifera)-A review. IOSR J Pharm. 2017;7(3):7–20. Amri O, Zekhnini A, Bouhaimi A, Tahrouch S, Hatimi A. Anti-inflammatory activity of methanolic extract from Pistacia atlantica Desf. leaves. Pharmacogn J. 2018;10(1). Askew RR, Blasco-Zumeta J. Parasitic Hymenoptera inhabiting seeds of Ephedra nebrodensis in Spain, with descriptions of a phytophagous pteromalid and four other new species of Chalcidoidea. J Nat Hist. 1997;31(6):965–82. Bellifa N, Benhaddou AI, Ferkous HE, Selka MA, Toumi H, Achouri MY. Antioxidant activity and polyphenol composition of Pistacia terebinthus fruit from Tessala (Western Algeria). North African J Food Nutr Res. 2021;5(11):30–4. Bozorgi M, Memariani Z, Mobli M, Salehi Surmaghi MH, Shams-Ardekani MR, Rahimi R. Five Pistacia species (P. vera, P. atlantica, P. terebinthus, P. khinjuk, and P. lentiscus): a review of their traditional uses, phytochemistry, and pharmacology. Sci World J. 2013;2013. Fenech M. Cytokinesis-block micronucleus cytome assay. Nat Protoc. 2007;2(5):1084–104. Giner-Larza EM, Máñez S, Giner-Pons RM, Recio MC, Rı́os JL. On the anti-inflammatory and anti-phospholipase A2 activity of extracts from lanostane-rich species. J Ethnopharmacol. 2000;73(1–2):61–9. Gul R, Jan SU, Faridullah S, Sherani S, Jahan N. Preliminary phytochemical screening, quantitative analysis of alkaloids, and antioxidant activity of crude plant extracts from Ephedra intermedia indigenous to Balochistan. Sci World J. 2017;2017. Ibragic S, Sofić E. Chemical composition of various Ephedra species. Bosn J basic Med Sci. 2015;15(3):21. Jaradat N, Hussen F, Al Ali A. Preliminary phytochemical screening, quantitative estimation of total flavonoids, total phenols and antioxidant activity of Ephedra alata Decne. J Mater Environ Sci. 2015;6(6):1771–8. Kaya F, Özer A. Characterization of extracted oil from seeds of terebinth (Pistacia terebinthus L.) growing wild in Turkey. Turkish J Sci Technol. 2015;10(1):49–57. Kouider H, Belkacem DB, Lakhdar G. Therapeutic importance of the plant Ephedra alata subsp. Alenda in traditional medicine for the population of the region of Guettara (Djelfa, Algeria). 2020; Li A, Zhao M, Yin F, Zhang M, Liu H, Zhou D, et al. Antioxidant effects of gallic acid alkyl esters of various chain lengths in oyster during frying process. Int J Food Sci Technol. 2021;56(6):2938–45. Luís Â, Neiva DM, Pereira H, Gominho J, Domingues F, Duarte AP. Bioassay-guided fractionation, GC–MS identification and in vitro evaluation of antioxidant and antimicrobial activities of bioactive compounds from Eucalyptus globulus stump wood methanolic extract. Ind Crops Prod. 2016;91:97–103. Matthäus B, Özcan MM. Quantitation of Fatty Acids, Sterols, and Tocopherols in Turpentine (Pistacia terebinthu s Chia) Growing Wild in Turkey. J Agric Food Chem. 2006;54(20):7667–71. Mendelson WB, Basile AS. The hypnotic actions of the fatty acid amide, oleamide. Neuropsychopharmacology. 2001;25(1):S36–9. Michel T, Destandau E, Elfakir C. Evaluation of a simple and promising method for extraction of antioxidants from sea buckthorn (Hippophaë rhamnoides L.) berries: Pressurised solvent-free microwave assisted extraction. Food Chem. 2011;126(3):1380–6. Özcan MM, Tzakou O, Couladis M. Essential oil composition of the turpentine tree (Pistacia terebinthus L.) fruits growing wild in Turkey. Food Chem. 2009;114(1):282–5. Que F, Mao L, Pan X. Antioxidant activities of five Chinese rice wines and the involvement of phenolic compounds. Food Res Int. 2006; 39(5):581–7. Quezel P. et Santa S. Nouvelle flore de l’Algérie et des régions désertiques méditerranéennes. Centre national de la recherche scientifique, Paris, France, 1963, 2, 19-23. Rajaei A, Barzegar M, Mobarez AM, Sahari MA, Esfahani ZH. Antioxidant, anti-microbial and antimutagenicity activities of pistachio (Pistachia vera) green hull extract. Food Chem Toxicol. 2010;48(1):107–12. Topçu G, Ay M, Bilici A, Sarıkürkcü C, Öztürk M, Ulubelen A. A new flavone from antioxidant extracts of Pistacia terebinthus. Food Chem. 2007;103(3):816–22. Valverde JL. The problem of herbal medicines legal status. IOS Press; 1999. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4648793","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":324864500,"identity":"384f79e7-09fc-4c7e-a1b2-50b93385c016","order_by":0,"name":"Mohammed HARIR","email":"data:image/png;base64,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","orcid":"","institution":"Université des Sciences et de la Technologie d'Oran Mohamed Boudiaf","correspondingAuthor":true,"prefix":"","firstName":"Mohammed","middleName":"","lastName":"HARIR","suffix":""},{"id":324864501,"identity":"5e578c4b-53ef-43b7-b1e4-ea2fb4ce77fb","order_by":1,"name":"Hamdi Bnedif","email":"","orcid":"","institution":"Université de M'Sila","correspondingAuthor":false,"prefix":"","firstName":"Hamdi","middleName":"","lastName":"Bnedif","suffix":""},{"id":324864502,"identity":"09ee434a-5cf4-4a30-82ef-fe77e9d2b069","order_by":2,"name":"hauker Reimann","email":"","orcid":"","institution":"University of Würzburg","correspondingAuthor":false,"prefix":"","firstName":"hauker","middleName":"","lastName":"Reimann","suffix":""},{"id":324864503,"identity":"13b1c3ef-c8e5-435b-92a0-d80532e54bdf","order_by":3,"name":"Silva Filomena","email":"","orcid":"","institution":"University of Zaragoza","correspondingAuthor":false,"prefix":"","firstName":"Silva","middleName":"","lastName":"Filomena","suffix":""},{"id":324864504,"identity":"e39e7a00-53c4-4514-9baa-e639282a430a","order_by":4,"name":"Cristina Nerin","email":"","orcid":"","institution":"Centro de Investigación y Tecnología Agroalimentaria de Aragón","correspondingAuthor":false,"prefix":"","firstName":"Cristina","middleName":"","lastName":"Nerin","suffix":""},{"id":324864505,"identity":"48a0f194-4528-43c6-ba72-d5d60f326da3","order_by":5,"name":"Henning Hintzsche","email":"","orcid":"","institution":"University of Bonn","correspondingAuthor":false,"prefix":"","firstName":"Henning","middleName":"","lastName":"Hintzsche","suffix":""}],"badges":[],"createdAt":"2024-06-27 13:14:51","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4648793/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4648793/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":61005318,"identity":"d8b450e2-708e-4607-98b0-d71826600a19","added_by":"auto","created_at":"2024-07-24 13:43:56","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":42091,"visible":true,"origin":"","legend":"\u003cp\u003eGenotoxicity of the crude extracts of \u003cem\u003eP. terebinthus\u003c/em\u003e aereal parts on HeLa cells.\u003c/p\u003e","description":"","filename":"Fig1300.png","url":"https://assets-eu.researchsquare.com/files/rs-4648793/v1/cd65f6903796ceb4b86b43ee.png"},{"id":61005317,"identity":"5e5d634a-7ddd-4ed2-aead-5ebf9e26fe4c","added_by":"auto","created_at":"2024-07-24 13:43:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":39711,"visible":true,"origin":"","legend":"\u003cp\u003eCompounds identified from ethanolic extract of\u003cem\u003e E. alata \u003c/em\u003eusing GC-MS\u003c/p\u003e","description":"","filename":"Fig2300.png","url":"https://assets-eu.researchsquare.com/files/rs-4648793/v1/e29db93b60c8a34f65c77416.png"},{"id":61640343,"identity":"b7e4705c-99f6-4232-9c21-8fb19aaab47c","added_by":"auto","created_at":"2024-08-02 09:49:55","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1048185,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4648793/v1/dab318d1-2429-4569-a89a-86612fd473b6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Antioxidant and genotoxic activity of Pistacia terebinthus and Ephedra alata crude extracts ","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eSeveral herbal drugs have yielded important modern therapeutic agents. In Algeria, folk medicinal herbs such as \u003cem\u003ePistacia terebinthus\u003c/em\u003e (family of \u003cem\u003eAnacardiaceae\u003c/em\u003e) are widely distributed. \u003cem\u003ePistacia terebinthus\u003c/em\u003e L. (turpentine tree, terebinth) is a perennial shrub or small tree widely grown in southern and western Turkey, and in western and eastern Algeria on dry rocky slopes and hillsides or in pine forests (\u0026Ouml;zcan et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2009\u003c/span\u003e, Bellifa et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In fact, \u003cem\u003eP. terebinthus\u003c/em\u003e fruits are commonly consumed as a coffee-based beverage, which is prepared with milk, known as \u0026lsquo;menengi\u0026ccedil;\u0026rsquo; coffee in Turkish \u003cb\u003e(Baytop, 1984).\u003c/b\u003e\u003cem\u003eP. terebinthus\u003c/em\u003e fruits have been the focus of several research activities due to their antimicrobial, antioxidant (Top\u0026ccedil;u et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), and anti-inflammatory properties (Giner-Larza et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2000\u003c/span\u003e), as well as their high oil content (approximately 40%) (\u003cb\u003eMatthaus and Ozcan, 2006).\u003c/b\u003e These fruits contain high concentrations of unsaturated fatty acids and carotenoids, phenolic compounds and tocopherols, tannins, resinous substances, and dietary fibers (10%) \u003cb\u003e(\u0026Ouml;zcan, 2004;\u003c/b\u003e Matth\u0026auml;us and \u0026Ouml;zcan, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2006\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e Terebinth has been described as an antiseptic and antihypertensive, and it has also been used to treat gastrointestinal and respiratory disorders (Bozorgi et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). As previously reported, various parts of this species (stems, flowers, and leaves) have antioxidant and other pharmacological properties. Due to their richness in secondary compounds such as flavonoids and polyphenols, radical scavenging activity of \u003cem\u003ePistacia terebinthus\u003c/em\u003e extracts was found to be fairly high (Top\u0026ccedil;u et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), indicating a possible preventive role in carcinogenesis by reducing oxidative stress (\u003cb\u003eKavak et al., 2010\u003c/b\u003e). Another common folk remedy, \u003cem\u003eEphedra alata\u003c/em\u003e, a medicinal plant of the \u003cem\u003eEphedracea\u003c/em\u003ee family known in Algeria as Alenda, is a perennial, xerophytic, gymnosperm and delicious shrub with erect non-climbing stems and short leaves that are united towards the base and yellowish-green bracts that flowers in July \u003cb\u003e(Bell, A. and Bachman, S., 2011) and\u003c/b\u003e has a strong pine-like smell \u003cb\u003e(Blumenthal, M. and King, P, 1995).\u003c/b\u003e This shrub grows in temperate, subtropical, and dry environments and can be found in the North African countries of Algeria, Tunisia, Morocco, Egypt, Chad, Mauritania, and Mali, as well as in Asia (China, Palestine, Lebanon, Jordan, Saudi Arabia, Iran, and Iraq) and Europe (Spain) \u003cb\u003e(Ebadi, M.,2007).\u003c/b\u003e\u003cem\u003eE. alata\u003c/em\u003e is used in traditional medicine for treating respiratory disorders and rheumatism \u003cb\u003e(Aidi et al 2023).\u003c/b\u003e The plant is also used in the traditional Chinese Pharmacopeia against cough, hay fever, cold, chills, asthma, allergies, and edema \u003cb\u003e(Al-Snafi,.2017)\u003c/b\u003e, while Algerians and Tunisians use it as anti-cancer treatment \u003cb\u003e(Sioud, F\u003c/b\u003e\u003cb\u003eet al\u003c/b\u003e, \u003cb\u003e2020)\u003c/b\u003e, in addition to its effects in reducing the side effects of chemotherapy \u003cb\u003e(Jaradat, N.A\u003c/b\u003e\u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2016).\u003c/b\u003e The primary active molecules in \u003cem\u003eE. alata\u003c/em\u003e are the ephedrine alkaloids which have major side effects such as adverse cardiovascular and cerebrovascular events if used in high doses, which explains why the US Food and Drug Administration (FDA) has banned any drugs that contain ephedrine alkaloids \u003cb\u003e(Ibragic, S.\u003c/b\u003e\u003cb\u003eet al.\u003c/b\u003e, \u003cb\u003e2015)\u003c/b\u003e. The major compounds detected in \u003cem\u003eE. alata\u003c/em\u003e growing in Algeria are isoflavones and flavonol derivatives with hydroxypuerarin isomer as the major molecule.\u003c/p\u003e \u003cp\u003eFor the evaluation of genotoxic and cytotoxic effects of many herbal medicines, extracts of different plant parts have been used, ranging from leafy vegetables, fruits, and underground storage organs to whole plants. The extracts are prepared mainly in water or organic solvents. Several of these toxicity assays have indicated the involvement of certain factors that are intrinsic components of the extracts, ranging from specific compounds like ascorbic acid to vegetable fibres which could act as nonspecific redox agents, free radical scavengers, or ligands for binding metals or toxic principles.\u003c/p\u003e \u003cp\u003eOxidative stress can cause DNA damage in the cells. If DNA repair is unable to modify or repair the DNA damage, genomic instability may lead to mutation, cancer, aging, and many other diseases. However, cells have special mechanisms to counteract and minimize these damages. Recently, research has shown that dietary patterns and food constituents can revert this toxicity in cells. A considerable body of epidemiological evidence indicates that a diet with high intake of fruits and vegetables is inversely related to the risk of chronic, degenerative diseases such as coronary heart disease and certain types of cancer. Much research effort has focused on the identification of phytochemicals in fruits and vegetables that exert beneficial effects and on the elucidation of the mechanisms by which they inhibit cellular damage and degeneration \u003cb\u003e(Sarkar\u003c/b\u003e\u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2008).\u003c/b\u003e Several experimental systems (e.g., membranes, plants, cell cultures, animal models and human clinical trials) were used to study the bioactivity of these plant-derived compounds. The effects of toxic compounds can for example be observed at the level of chromosomes by quantifying alterations in chromosome structure (chromosomal aberrations) and number (aneuploidy, polyploidy). Assays with several crude vegetable and fruit extracts have shown their clastogenic effect or protective activities against known genotoxic agents such as plants containing aristolochic acid. Additionally, single extract components such as sulfhydryl and flavonoid compounds, gallic acid, ellagic acid, mucic acid, citric acid, reducing sugars, and tannins have been described to have an additive interaction with the major constituent\u0026rsquo;s chlorophyll and ascorbic acid when modulating the effects of the clastogens \u003cb\u003e(Leme\u0026amp;Marin-Morales, 2009).\u003c/b\u003e The increase in chromosomal aberrations may result from interactions of a different variety of chemical agents with DNA. According to (\u003cb\u003eIshidate\u003c/b\u003e\u003cb\u003eet al\u003c/b\u003e. \u003cb\u003e1988)\u003c/b\u003e, agents that induce an increase in the chromosomal aberration\u0026rsquo;s frequency by direct or indirect mechanism may also be cytotoxic, by causing damage to both DNA and other cellular targets (enzymes, membranes, structural proteins).\u003c/p\u003e \u003cp\u003eThe present study aimed to establish a phytochemical profile, and evaluate the antioxidant, cytotoxicity, and genotoxicity potential of \u003cem\u003eE. alata\u003c/em\u003e and \u003cem\u003eP. terebinthus\u003c/em\u003e grown in Algeria.\u003c/p\u003e"},{"header":"MATERIAL AND MATHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePlant materials and samples preparation\u003c/h2\u003e \u003cp\u003e \u003cem\u003ePistacia terebinthus\u003c/em\u003e and \u003cem\u003eEphedra alata\u003c/em\u003e leaves, stems and flowers were collected from Ouled Hamdane of Bordj Ghedir (Bordj Bou Arreridj province, North of Algeria, 1100 m meters in altitude., 35.9125\u0026deg; N, 4.9247\u0026deg; E). The taxonomic identification of plant materials was performed using Flora of Algeria \u003cb\u003e(Quezel and Santa 1962\u003c/b\u003e), while herbarium specimens were archived in the Herbarium of Tiaret University. The harvested plant material was dried at room temperature (20\u0026ndash;25\u0026deg;C) for about fifteen days in the open air until the stabilization of its weight to avoid possible risks of polyphenol oxidation and to preserve as much as possible the integrity of the molecules. Then, the samples were collected in clean bags and stored away from light and moisture in a tightly closed container for further use.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003ePreparation of crude extracts by maceration\u003c/h2\u003e \u003cp\u003eFor the preparation of the crude extracts, the solvent used was ethanol 95% (v/v), which has been chosen to allow the solubilisation of different compounds. This solvent has been used at a solid:liquid ratio of 1:10 g plant material / mL solvent of extraction (\u003cb\u003eMichel, 2011\u003c/b\u003e). For this purpose, 5 g of the plant powder from the aerial parts of both plants (leaves, stems, and flowers) were mixed with 50 mL of ethanol in an Erlenmeyer flask. The mixture was kept under magnetic stirring for 24 h at room temperature and the obtained solution was filtered with filter paper (Wattman No. 1 of pore diameter 0.2 \u0026micro;m) under vacuum. The filtrate was recovered and the ethanolic extracts were concentrated under a vacuum at 45\u0026deg;C using a rotary evaporator (Buchi Labortechnik AG, Switzerland) for one hour. Afterwards, the obtained crude extract was lyophilized and stored at 4 C\u0026deg;.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eGas chromatography-Mass spectrometry analysis of extracts\u003c/h2\u003e \u003cp\u003eThe phytochemical investigation of 1% (w/w) ethanolic extracts was performed on an Agilent HP 6890 series gas chromatograph system (Agilent Technologies, Spain) connected to an Agilent HP 5973 series mass selective detector. The sample (1 \u0026micro;L) was injected at the injection port (250 \u0026ordm;C) in splitless mode. Chromatographic separation was carried out on a DB-5 column (30 m x 0.25 mm x 0.25 \u0026micro;m) (Agilent Technologies, Spain). The initial column temperature was set at 40\u0026deg;C and raised to 250\u0026deg;C at a rate of 5\u0026deg;C/min and held for 2 minutes resulting in a total chromatographic run time of 45 minutes. Helium (99.999%, Air Liquid, Madrid, Spain) was used as carrier gas at a flow rate of 1.0 mL/min. MS detection was performed in Scan mode at a range of 50 to 650 m/z and the results were compared by using NIST 14 Mass Spec Library search program.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eCell culture\u003c/h2\u003e \u003cp\u003eHela H2B GFP cells expressing green fluorescent protein fused to histone H2B were provided by Noriaki Shimizu, Graduate School of Integrated Sciences for Life, Hiroshima University, Japan and seeded at a density of 10.000 cells/ml in 75 cm\u003csup\u003e2\u003c/sup\u003e tissue culture flasks, grown in Dulbecco\u0026rsquo;s modified Eagle\u0026rsquo;s medium (Gibco BRL, Cergy-Pontoise, France) supplemented with 10% fetal bovine serum (Gibco BRL), 1% L-glutamine and 1% penicillin/streptomycin mixture (Gibco BRL). Cultures were maintained in a humidified atmosphere with 5% CO\u003csub\u003e2\u003c/sub\u003e at 37\u0026deg;C to sub-confluence. Fresh medium was supplied every 48 h.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eGenotoxicity Evaluation\u003c/h2\u003e \u003cp\u003eThe genotoxicity activity of plant extract solutions was assessed using the micronucleus assay in Hela H2B GFP cells as previously described by \u003cb\u003eMichael\u003c/b\u003e Fenech (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) with slight modifications. Briefly, test extract solutions were prepared using a stock solution of 200 mg/mL in double distilled water and diluted in a culture medium to give a final concentration of 0.1 to 2 mg/mL. Cells were incubated with extract solutions for 4 h and afterwards subjected to treatment with cytochalasin B (3 \u0026micro;g/mL). Etoposide (14 \u0026micro;g/l) was used as a positive control. After 24 hours of incubation with cytochalasin B, cells were harvested, and slides were prepared by cytospin centrifugation at 100 rpm for 5 min and fixed on a slide. The slides were stored overnight in ice-cold methanol (-20\u0026deg;C) and stained with Gel Green staining solution (Biotium Glowing products for science, 1:100 dilutions in water) for 6 min in the dark. Finally, slides were mounted using diazabicyclo-octane (DABCO) and coverslips. Two replicate experiments were performed independently and the results are shown as the mean of these replicates.\u003c/p\u003e \u003cp\u003eEvaluation of micronuclei was performed at 400-fold magnification under a fluorescence microscope using a fluorescein isothiocyanate (FITC) filter (Nikon GmbH, Japan). The number of mononucleated, binucleated, multinucleated, apoptotic, and mitotic cells (was calculated. Also, the number of micronuclei in 1000 binucleated cells was counted. The cytokinesis block proliferation index (CBPI) was determined in 1000 cells that were counted for mononucleated cells, binucleated cells, and multinucleated cells by using the following equation:\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eCBPI= ((No. mononucleate cells) +(2\u0026times;No.binucleat cells) +(3\u0026times;No.multinucleate cells))/Total number of cell\u003c/h2\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003eDPPH radical scavenging assay\u003c/h2\u003e \u003cp\u003eThe method is based on the reduction of alcoholic 1,1-diphenyl-2-picrylhydrazyl (DPPH) solutions in the presence of a hydrogen-donating antioxidant. The effect of each extract on DPPH was measured using a method previously described (Que et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). In brief, 1 mL of various concentrations (0\u0026ndash;50 mg/mL) of the ethanolic extract or standard solution was added to 1 mL of freshly prepared DPPH (0.1 mM) methanolic solution. The mixture was mixed vigorously and allowed to stand in the dark for 30 min at room temperature. After this incubation period, the absorbance (A) at 517 nm was recorded using a UV\u0026ndash;VIS spectrophotometer (Shimadzu). A calibration curve was prepared with ascorbic acid as a control (1\u0026ndash;10 mg/mL methanol the free radical-scavenging activity of each solution was then calculated as percent inhibition according to the following equation:\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e% inhibition\u0026thinsp;=\u0026thinsp;100 (A (control) - A (sample)) / A (control)\u003c/h2\u003e \u003cp\u003eThe antioxidant activity of the samples was expressed as Efficient Concentration 50 (EC50, in \u0026micro;g/ml) value. EC50 is defined as the concentration of the extract equivalent to the effective antioxidant activity at 50%.\u003c/p\u003e \u003cp\u003eAntioxidant activity is also expressed as the Antioxidant Activity Index (AAI), calculated as follows:\u003c/p\u003e \u003cp\u003eAAI\u0026thinsp;=\u0026thinsp;final concentration of DPPH in the control sample/IC50Thus, the AAI was determined considering the mass of DPPH and the mass of the ethanolic extract in the reaction. This gives a constant for ethanol extract independent of the concentration of DPPH or the ethanol extract used. The AAI allows the following classification of the antioxidant activity of ethanol extracts: poor (AAI\u0026thinsp;\u0026le;\u0026thinsp;0.5), moderate (0.5\u0026thinsp;\u0026lt;\u0026thinsp;AAI\u0026thinsp;\u0026le;\u0026thinsp;1.0), strong (1.0\u0026thinsp;\u0026lt;\u0026thinsp;AAI\u0026thinsp;\u0026lt;\u0026thinsp;2.0) or very strong (AAI\u0026thinsp;\u0026ge;\u0026thinsp;2.0) (Lu\u0026iacute;s et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Tests were performed in triplicate and DPPH solutions were prepared daily.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eChemical analysis\u003c/h2\u003e \u003cp\u003eWe have observed that the extraction yield of the different samples was quite similar (nearly 12%). The GC-MS analysis of the compounds with their retention time (RT), molecular formula, and percentage are presented in Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. Among the compounds identified in \u003cem\u003eP. terebinthus\u003c/em\u003e, the most abundant ones were oleic acid amide \u003cb\u003e(21.46%)\u003c/b\u003e followed by daphnetin \u003cb\u003e(13.66%)\u003c/b\u003e, a natural coumarin derivative which is a protein kinase inhibitor \u003cb\u003e(\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e).\u003c/b\u003e \u003cem\u003eE. alata\u003c/em\u003e extract contains twenty-seven identifiable compounds, with oleic acid amide, ethyl gallate, and pyrogallic acid being the major compounds (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e\u003ctable id=\"Tab1\" border=\"1\" style=\"margin-right: calc(10%); width: 90%;\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCompounds identified from ethanolic extract of \u003cem\u003eP. terebinthus\u003c/em\u003e using GC-MS.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cem\u003eN\u0026deg;\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eCompound\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003eMolecular\u003c/p\u003e\n \u003cp\u003eformula\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" rowspan=\"2\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003eRetention\u003c/p\u003e\n \u003cp\u003etime (min)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e%\u003c/em\u003e\u003csup\u003e\u003cem\u003ea\u003c/em\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003eP. terebinthus\u003c/em\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e1.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eFurfural\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e5\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e4\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e5.981\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.15\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e2.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003e(E)-Conipheryl alcohol Loliolide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e10\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e12\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e28.695\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.17\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e3.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003e2,3-Dihydro-3,5-dihydroxy-6-methyl-4H pyran-4-one\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e8\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e4\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e13.231\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.93\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e4.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003e5Hydroxymethylfurfural\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e15.641\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.14\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e5.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eDaphnetin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e11\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e29.51\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e13.66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e6.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eDihydroxyacetone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e6.904\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e1.01\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e7.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eEicosane\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e20\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e42\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e43.59\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.13\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e8.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eFuranmethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e5\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e6.334\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.16\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e9.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eMyristamide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e32\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e32.83\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.49\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e10.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eNeophytadiene\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e9\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e30.25\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.35\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e11.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eOleic acid amide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e35\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e39.727\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e21.46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e12.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003ePalmitic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e20\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e38\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e32.66\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e3.11\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e13.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003ePalmitic acid amide\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e33\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e36.604\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e1.20\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e14.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003ePalmitic acid \u0026beta;-monoglyceride\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e19\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e38\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e4\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e42.001\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.19\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e15.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003ePalmitic acid, butyl ester\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e20\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e40\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e36.76\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.22\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e16.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003eStearic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e36\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e36.333\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e0.84\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e17.\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003etrans-Phytol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e14\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e29\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e35.47\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e1.56\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" style=\"width: 3.7643%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 56.7921%;\"\u003e\n \u003cp\u003e\u0026alpha; \u0026ndash;linolenic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e30\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 12.1113%;\"\u003e\n \u003cp\u003e\u003cem\u003e35.932\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" style=\"width: 15.2209%;\"\u003e\n \u003cp\u003e\u003cem\u003e2.86\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCompounds identified from ethanolic extract of \u003cem\u003eE. alata\u003c/em\u003e using GC-MS\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003eN\u0026deg;\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCompound\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMolecular\u003c/p\u003e \u003cp\u003eformula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eRetention\u003c/p\u003e \u003cp\u003etime (min)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e%a\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eE. alata\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFurfural\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eC\u003csub\u003e5\u003c/sub\u003eH\u003csub\u003e4\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e5.987\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.05\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e2.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e(1S) -eno)-(-)-Borneol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e10\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e13.93\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.24\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e3.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2,4-Bis(1,1-dimthylethyl) phenol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e14\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e22\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e23\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.75\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e4.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2-Formyl-1-methylpyrole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e7\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e9.531\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.14\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e5.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2-Methylheptadecane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e38\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e31.234\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.05\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e6.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,4-Dimethylbenzaldehyde\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e9\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e10\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e15.267\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.44\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e7.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3,5-Dihydroxy-6-methyl-2,3-dihydro-4H-pyran-4-one\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e8\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e4\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e13.19\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.22\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e8.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4-Methyldecane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e11\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e24\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e10.047\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.06\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e9.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBenzenemethanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e7\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e8\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e10.278\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.23\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e10.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEthyl gallate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e9\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e10\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e5\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e32.443\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e14.48\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e11.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHexadecane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e34\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e17.128\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.45\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e12.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-Cetane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e34\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e27.568\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.24\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e13.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-Heneicosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e21\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e44\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e43.576\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.31\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e14.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-Heptacosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e27\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e56\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e28.689\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.13\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e15.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-Hexacosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e26\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e54\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e31.329\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.08\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e16.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-octacosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e28\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e58\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e30.467\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.12\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e17.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003en-octacosane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e38\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e31.465\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.10\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e18.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eo-Cymene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e10\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e14\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e10.095\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.03\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e19.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOleic acid amide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e35\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e39.713\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e21.40\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e20.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePalmitic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e36\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e33.271\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.09\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e21.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePalmitic acid amide\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e16\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e33\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eNO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e36.597\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e1.11\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e22.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePalmitic acid, butyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e20\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e40\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e36.76\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.06\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e23.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ep-Methyl cyclohexanone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e7\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e12\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e9.361\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.07\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e24.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePyrogallic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e19.558\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e10.47\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e25.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePyrogallol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e6\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e3\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e8.94\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.37\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e26.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStearic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e36\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e36.319\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.78\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e27.\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTrans-phytol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e20\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e40\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e35.47\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.77\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e28\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eα \u0026ndash;linolenic acid\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003e18\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eH\u003c/em\u003e\u003csub\u003e\u003cem\u003e30\u003c/em\u003e\u003c/sub\u003e\u003cem\u003eO\u003c/em\u003e\u003csub\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sub\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003e35.891\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003e0.59\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003csup\u003ea\u003c/sup\u003e The percentage of compound was calculated as the percentage ratio between compound peak area and total peak area.\u003c/p\u003e \u003cp\u003eThe genotoxicity analysis of the crude extracts from leave and flowers of \u003cem\u003eP. terebinthus\u003c/em\u003e on HeLa cells showed that all concentrations tested yielded a micronucleus frequency slightly higher than the control (\u003cb\u003eFig.\u0026nbsp;1)\u003c/b\u003e. The increase in micronuclei was not very pronounced and was not comparable to the response to the positive control etoposide, which approximately showed a 10-fold increase in micronuclei. No clear dose-response could be observed. Also, the CBPI did not show any relevant alterations. With regards to the crude extracts of \u003cem\u003eEphedra alata\u003c/em\u003e on HeLa cells, we observed an increase in micronuclei at the lowest concentration, which was not observed at the higher concentrations (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAs shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, we can rank the samples in order of decreasing reactivity: ascorbic acid\u0026thinsp;\u0026gt;\u0026thinsp;\u003cem\u003eEphedra alata\u003c/em\u003e extract\u0026thinsp;\u0026gt;\u0026thinsp;\u003cem\u003ePistacia terebinthus\u003c/em\u003e extract.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDPPH radical scavenging activities\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e: Antioxidant properties (EC\u003csub\u003e50\u003c/sub\u003e\u0026micro;g/ml) of \u003cem\u003eEphedra alata and Pistacia terebinthus ethanolic extracts\u003c/em\u003e.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eDPPH free radical scavenging assay\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eStrength of\u003c/p\u003e \u003cp\u003eAntioxidant Activity\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSample\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEC\u003csub\u003e50\u003c/sub\u003e (\u0026micro;g/mL)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAAI\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEphedra alata\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.968\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e85.71\u0026thinsp;\u0026plusmn;\u0026thinsp;3.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVery strong\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePistacia terebinthus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e2.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.978\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e13.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.66\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVery strong\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAscorbic acid\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.839\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e492.87\u0026thinsp;\u0026plusmn;\u0026thinsp;09.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eVery strong\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe EC50 values of the extracts are 0.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.96 and 2.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.97, with an AAI value of 85.71\u0026thinsp;\u0026plusmn;\u0026thinsp;3.22 and 13.59\u0026thinsp;\u0026plusmn;\u0026thinsp;1.66 respectively, which allows the ethanolic extracts of \u003cem\u003eEphedra alata\u003c/em\u003e and \u003cem\u003ePistacia terebinthus\u003c/em\u003e to be classified as having very strong antioxidant activity.\u003c/p\u003e \u003c/div\u003e"},{"header":"DISSCUSSION","content":"\u003cp\u003eIn the present investigation, more than twenty bioactive chemical constituents were identified in each plant crude extract. \u003cem\u003ePistacia terebinthus\u003c/em\u003e contains daphnetin (DAP) as a major compound (13.6%) which is a natural coumarin derivative isolated from different medicinal herbs. It possesses abundant biological activities, such as antioxidant and anti-inflammatory properties. In addition, the oleic acid amide, present in both crude extracts (21.5%), has been demonstrated to interfere with 5-HT2a, 5HT2c, and GABAa receptors (\u003cb\u003eMendelson \u0026amp; Basile,2001)\u003c/b\u003e, These two major compounds have all been shown to have hypocholesterolemic and antioxidant activity. \u003cem\u003eEphedra alata\u003c/em\u003e contained twenty-seven compounds with Pyrogallic acid (10.47%), ethyl gallate (14.48%) and oleic acid amide (21.40%) as major compounds. The ethyl ester of gallic acid is an approved food additive (E313) that is added to foods due to its antioxidant activity \u003cb\u003e(Belifa\u003c/b\u003e, \u003cb\u003eet al\u003c/b\u003e. \u003cb\u003e2021).\u003c/b\u003e\u003c/p\u003e \u003cp\u003eMany studies have confirmed the importance of plant antioxidants (\u003cb\u003eAissani \u0026amp; Sebai 2022; Aissani\u003c/b\u003e\u003cb\u003eet al\u003c/b\u003e, \u003cb\u003e2020\u003c/b\u003e). In our study, it was important to evaluate \u003cem\u003eEphedra alata\u003c/em\u003e and \u003cem\u003ePistacia terebinthus\u003c/em\u003e ethanolic extracts via the DPPH test to get more informative results on their antioxidant potential. The EC\u003csub\u003e50\u003c/sub\u003e values results confirmed that both \u003cem\u003eE. alata\u003c/em\u003e and \u003cem\u003eP. terebinthus\u003c/em\u003e ethanolic extracts were active regarding antioxidant capacity.\u003c/p\u003e \u003cp\u003eOur finding is in agreement with other studies about other species of \u003cem\u003eEphedra\u003c/em\u003e genus collected in other countries. As reported by \u003cb\u003eJaradat\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e \u003cb\u003ein 2015\u003c/b\u003e, a better antioxidant activity with lower IC\u003csub\u003e50\u003c/sub\u003e of Palestinian \u003cem\u003eE. alata\u003c/em\u003e was shown, which can be attributed to their alkaloid content. Our phytochemical results revealed that Algerian \u003cem\u003eE. alata\u003c/em\u003e did not contain alkaloids, which can be associated with a safer toxicological profile since \u003cem\u003eEphedra\u003c/em\u003e alkaloids (ephedrine and pseudoephedrine) are the main compounds responsible for its toxicity (\u003cb\u003eIbragic, S\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e \u003cb\u003e2015\u003c/b\u003e).\u003c/p\u003e \u003cp\u003eMethanolic extracts of the aerial parts of Palestinian \u003cem\u003eEphedra alata\u003c/em\u003e (Jaradat et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) were also reported to have high antioxidant properties as free radical inhibitors and scavengers, and showed lower EC\u003csub\u003e50\u003c/sub\u003e values (16.03 \u0026micro;g/mL for DPPH). To our best knowledge, this is the first report on the antioxidant activity of the ethanolic extracts of \u003cem\u003eE. alata\u003c/em\u003e. However, another work on \u003cem\u003eE. alata\u003c/em\u003e from Palestine (Al-rimawi et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) has reported antioxidant effects of the extract, with higher contents in polyphenols, using higher polarity solvents. Thus, the highest activity was reported for the hydroethanolic extract (78 \u0026micro;g/mL for DPPH). This discordance noted in relation to our sample from Algeria is probably due to the effect of the region and climatic conditions affecting the biosynthesis of phenolic compounds, and to the effect of the extraction solvent used. This can generate quantitative differences in the bioactive molecules and therefore influences the antioxidant activity. Parsaeimehr and collaborators \u003cb\u003ein 2010\u003c/b\u003e studied \u003cem\u003eEphedra\u003c/em\u003e wild culture species from Iran (\u003cem\u003eE. procera\u003c/em\u003e Fisch. \u0026amp; Mey., E. \u003cem\u003epachyclada\u003c/em\u003e Boiss. and \u003cem\u003eE. strobilacea\u003c/em\u003e Bunge) and found that the methanolic extract of \u003cem\u003eE. strobilacea\u003c/em\u003e had higher antioxidant activity (FRAP assay) in comparison to the other species ( Al-Rimawi et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The same trend was obtained with the hydroalcoholic mixture (ethanol, methanol and distilled water 7:2:1) of \u003cem\u003eE. intermedia\u003c/em\u003e Schrenk ex Meyer from Balochistan, Pakistan in DPPH free radical scavenging assay (Gul, Jan, Faridullah, Sherani, \u0026amp; Jahan, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe present results show that our ephedra extract has a strong antioxidant potential, which is consistent with previous results on the aerial parts of several Ephedra plants. Furthermore, the aerial parts of E. alata from Tunisia showed high DPPH radical scavenging activity and EC\u003csub\u003e50\u003c/sub\u003e values in methanol extracts (\u003cb\u003eDbeibia\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e., \u003cb\u003e2022\u003c/b\u003e). It has been reported that the antioxidant properties of plant extracts may be attributed to phenolic compounds and their possible additive, synergistic or antagonistic interactions (\u003cem\u003eKurin et al., 2012\u003c/em\u003e). The demonstrated antioxidant potential may be even more interesting for biotechnological applications and industrial uses.\u003c/p\u003e \u003cp\u003eThe antioxidant potential of \u003cem\u003ePistacia\u003c/em\u003e extract has been studied by other researchers (Abidi et al \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and (Rajaei et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), who have reported that the water extract showed a strong DPPH antioxidant activity when compared to THBQ and BHT standards. \u003cb\u003eKilic\u003c/b\u003e \u003cb\u003eet al\u003c/b\u003e. \u003cb\u003e(2016\u003c/b\u003e), have also reported that the DPPH antioxidant activity of pistachio hull extract was 137.16 mg Trolox Equivalent/g extract. Comparing our results with those reported previously on \u003cem\u003eP. atlantica\u003c/em\u003e leaf and stem extracts (\u003cb\u003ePeks el\u003c/b\u003e \u003cb\u003eet al.\u003c/b\u003e, \u003cb\u003e2008; Toul\u003c/b\u003e \u003cb\u003eet al.\u003c/b\u003e, \u003cb\u003e2017;\u003c/b\u003e Amri et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), it can be stated that all extracts were very effective in the DPPH assay.\u003c/p\u003e \u003cp\u003eThe genotoxicity findings are difficult to interpret. For the \u003cem\u003eP. terebinthus\u003c/em\u003e samples, a small increase in micronuclei was observed compared to the control, which might indicate a clastogenic potential of the extracts. However, the increase is fairly small, particularly when compared to the response of the positive control, which yielded a much higher increase in micronuclei. Additionally, no clear dose response could be observed. All in all, genotoxic properties of the extract cannot be clearly concluded from the data.\u003c/p\u003e \u003cp\u003eA similar situation can be observed for the \u003cem\u003eE. alata\u003c/em\u003e extract samples. There is a clear increase of micronucleus frequency at the lowest concentration, but this increase comes along with a high intra-experimental variability shown by the large standard deviation. It is unclear whether this increase is a real effect or due to an experimental outlier. At all other concentrations, a very small increase in micronuclei can be observed, the relevance of this small increase is unclear.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eOur study indicates that both \u003cem\u003ePistacia terebinthus and Ephedra alata\u003c/em\u003e extracts might have a genotoxic effect, arising from the use of herbal medicinal products. Antioxidant activity is also confirmed. This is the first report regarding the genotoxicity activity of both plants. This finding provides new insights into the existing drugs and may help to facilitate the development of antitumor agents. As the global use of herbal medicinal products continues to grow and many more new products are introduced into the market, public health issues and concerns surrounding their safety are also increasingly recognized. Although some herbal medicines have promising potential and are widely used, many of them remain untested and their use is also not monitored. This makes knowledge of their potential adverse effects very limited and identification of the safest and most effective therapies as well as the promotion of their rational use more difficult. In most countries, herbal medicines and related products are introduced into the market without any mandatory safety or toxicological evaluation. Many of these countries also lack effective mechanisms to regulate manufacturing practices and quality standards. These herbal products are continuously made available to consumers without prescription in most cases and the potential hazards of an inferior product are hardly recognized.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eDECLARATION OF CONFLICT OF INTEREST\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eACKNOWLEDGMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to\u0026nbsp;Mrs. Heike Keim-Heusler\u0026nbsp;for providing technical assistance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFUNDING\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by\u0026nbsp;the German Academic Exchange Service\u0026nbsp;by a Postdoc fellowship.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eETHICAL APPROVAL\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNot applicable\u003c/em\u003e\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAbidi A, Aissani N, Sebai H, Serairi R, Kourda N, Ben Khamsa S. Protective effect of Pistacia lentiscus oil against bleomycin-induced lung fibrosis and oxidative stress in rat. Nutr Cancer. 2017;69(3):490\u0026ndash;7. \u003c/li\u003e\n\u003cli\u003eAidi Wannes W, Tounsi MS. Phytochemicals, Traditional Uses, Biological Effects, and Possible Molecular Mechanisms of Ephedra alata. Futur Integr Med. 2023;2(4):189\u0026ndash;99. \u003c/li\u003e\n\u003cli\u003eAissani N, Albouchi F, Sebai H. Anticancer effect in human glioblastoma and antioxidant activity of Petroselinum crispum L. methanol extract. Nutr Cancer. 2021;73(11\u0026ndash;12):2605\u0026ndash;13. \u003c/li\u003e\n\u003cli\u003eAl-Rimawi F, Abu-Lafi S, Abbadi J, Alamarneh AAA, Sawahreh RA, Odeh I. Analysis of phenolic and flavonoids of wild Ephedra alata plant extracts by LC/PDA and LC/MS and their antioxidant activity. African J Tradit Complement Altern Med. 2017;14(2):130\u0026ndash;41. \u003c/li\u003e\n\u003cli\u003eAl-Snafi AE. Pharmacology and therapeutic potential of Euphorbia hirta (Syn: Euphorbia pilulifera)-A review. IOSR J Pharm. 2017;7(3):7\u0026ndash;20. \u003c/li\u003e\n\u003cli\u003eAmri O, Zekhnini A, Bouhaimi A, Tahrouch S, Hatimi A. Anti-inflammatory activity of methanolic extract from Pistacia atlantica Desf. leaves. Pharmacogn J. 2018;10(1). \u003c/li\u003e\n\u003cli\u003eAskew RR, Blasco-Zumeta J. Parasitic Hymenoptera inhabiting seeds of Ephedra nebrodensis in Spain, with descriptions of a phytophagous pteromalid and four other new species of Chalcidoidea. J Nat Hist. 1997;31(6):965\u0026ndash;82. \u003c/li\u003e\n\u003cli\u003eBellifa N, Benhaddou AI, Ferkous HE, Selka MA, Toumi H, Achouri MY. Antioxidant activity and polyphenol composition of Pistacia terebinthus fruit from Tessala (Western Algeria). North African J Food Nutr Res. 2021;5(11):30\u0026ndash;4. \u003c/li\u003e\n\u003cli\u003eBozorgi M, Memariani Z, Mobli M, Salehi Surmaghi MH, Shams-Ardekani MR, Rahimi R. Five Pistacia species (P. vera, P. atlantica, P. terebinthus, P. khinjuk, and P. lentiscus): a review of their traditional uses, phytochemistry, and pharmacology. Sci World J. 2013;2013. \u003c/li\u003e\n\u003cli\u003eFenech M. Cytokinesis-block micronucleus cytome assay. Nat Protoc. 2007;2(5):1084\u0026ndash;104. \u003c/li\u003e\n\u003cli\u003eGiner-Larza EM, M\u0026aacute;\u0026ntilde;ez S, Giner-Pons RM, Recio MC, Rı́os JL. On the anti-inflammatory and anti-phospholipase A2 activity of extracts from lanostane-rich species. J Ethnopharmacol. 2000;73(1\u0026ndash;2):61\u0026ndash;9. \u003c/li\u003e\n\u003cli\u003eGul R, Jan SU, Faridullah S, Sherani S, Jahan N. Preliminary phytochemical screening, quantitative analysis of alkaloids, and antioxidant activity of crude plant extracts from Ephedra intermedia indigenous to Balochistan. Sci World J. 2017;2017. \u003c/li\u003e\n\u003cli\u003eIbragic S, Sofić E. Chemical composition of various Ephedra species. Bosn J basic Med Sci. 2015;15(3):21. \u003c/li\u003e\n\u003cli\u003eJaradat N, Hussen F, Al Ali A. Preliminary phytochemical screening, quantitative estimation of total flavonoids, total phenols and antioxidant activity of Ephedra alata Decne. J Mater Environ Sci. 2015;6(6):1771\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eKaya F, \u0026Ouml;zer A. Characterization of extracted oil from seeds of terebinth (Pistacia terebinthus L.) growing wild in Turkey. Turkish J Sci Technol. 2015;10(1):49\u0026ndash;57. \u003c/li\u003e\n\u003cli\u003eKouider H, Belkacem DB, Lakhdar G. Therapeutic importance of the plant Ephedra alata subsp. Alenda in traditional medicine for the population of the region of Guettara (Djelfa, Algeria). 2020; \u003c/li\u003e\n\u003cli\u003eLi A, Zhao M, Yin F, Zhang M, Liu H, Zhou D, et al. Antioxidant effects of gallic acid alkyl esters of various chain lengths in oyster during frying process. Int J Food Sci Technol. 2021;56(6):2938\u0026ndash;45. \u003c/li\u003e\n\u003cli\u003eLu\u0026iacute;s \u0026Acirc;, Neiva DM, Pereira H, Gominho J, Domingues F, Duarte AP. Bioassay-guided fractionation, GC\u0026ndash;MS identification and in vitro evaluation of antioxidant and antimicrobial activities of bioactive compounds from Eucalyptus globulus stump wood methanolic extract. Ind Crops Prod. 2016;91:97\u0026ndash;103. \u003c/li\u003e\n\u003cli\u003eMatth\u0026auml;us B, \u0026Ouml;zcan MM. Quantitation of Fatty Acids, Sterols, and Tocopherols in Turpentine (Pistacia terebinthu s Chia) Growing Wild in Turkey. J Agric Food Chem. 2006;54(20):7667\u0026ndash;71. \u003c/li\u003e\n\u003cli\u003eMendelson WB, Basile AS. The hypnotic actions of the fatty acid amide, oleamide. Neuropsychopharmacology. 2001;25(1):S36\u0026ndash;9. \u003c/li\u003e\n\u003cli\u003eMichel T, Destandau E, Elfakir C. Evaluation of a simple and promising method for extraction of antioxidants from sea buckthorn (Hippopha\u0026euml; rhamnoides L.) berries: Pressurised solvent-free microwave assisted extraction. Food Chem. 2011;126(3):1380\u0026ndash;6. \u003c/li\u003e\n\u003cli\u003e\u0026Ouml;zcan MM, Tzakou O, Couladis M. Essential oil composition of the turpentine tree (Pistacia terebinthus L.) fruits growing wild in Turkey. Food Chem. 2009;114(1):282\u0026ndash;5. \u003c/li\u003e\n\u003cli\u003eQue F, Mao L, Pan X. Antioxidant activities of five Chinese rice wines and the involvement of phenolic compounds. Food Res Int. 2006; 39(5):581\u0026ndash;7. \u003c/li\u003e\n\u003cli\u003eQuezel P. et Santa S. Nouvelle flore de l\u0026rsquo;Alg\u0026eacute;rie et des r\u0026eacute;gions d\u0026eacute;sertiques m\u0026eacute;diterran\u0026eacute;ennes. Centre national de la recherche scientifique, Paris, France, 1963, 2, 19-23.\u003c/li\u003e\n\u003cli\u003eRajaei A, Barzegar M, Mobarez AM, Sahari MA, Esfahani ZH. Antioxidant, anti-microbial and antimutagenicity activities of pistachio (Pistachia vera) green hull extract. Food Chem Toxicol. 2010;48(1):107\u0026ndash;12. \u003c/li\u003e\n\u003cli\u003eTop\u0026ccedil;u G, Ay M, Bilici A, Sarık\u0026uuml;rkc\u0026uuml; C, \u0026Ouml;zt\u0026uuml;rk M, Ulubelen A. A new flavone from antioxidant extracts of Pistacia terebinthus. Food Chem. 2007;103(3):816\u0026ndash;22. \u003c/li\u003e\n\u003cli\u003eValverde JL. The problem of herbal medicines legal status. IOS Press; 1999. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Pistacia terebinthus, Ephedra alata, Antioxidants, Genotoxicity, Micronucleus test, GC-MS","lastPublishedDoi":"10.21203/rs.3.rs-4648793/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4648793/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn the last decades, cases of poisoning due to medicinal plants have been very common in many countries including Algeria. Algerian herbal medicines have been used in effectively treating several health problems, but their toxicity limits their use. \u003cem\u003ePistacia terebinthus\u003c/em\u003e and \u003cem\u003eEphedra alata\u003c/em\u003e herbal medicine collected from Algeria have been used for a long time to treat several diseases. Thus, this study aimed to investigate the chemical composition and evaluate the genotoxicity and the antioxidant activity of crude extracts of these two herbal medicines (\u003cem\u003eE. alata and P. terebinthus\u003c/em\u003e) traditionally used in Algeria and North Africa. For this purpose, genotoxicity was assessed using the micronucleus test. Additionally, compounds with antioxidant and genotoxic activities of these two plants were identified by gas chromatography coupled to mass spectrometry (GC-MS). The highest antioxidant capacity of the extracts was found in \u003cem\u003eP. terebinthus\u003c/em\u003e ethanol extract owing to its richness of daphnetin, and oleic acid amide. Our results showed that most of these plants exhibited antioxidant activities and genotoxic effects, depending on the level of administered doses. To our knowledge, this is the first report on these effects caused by of both plants.\u003c/p\u003e","manuscriptTitle":"Antioxidant and genotoxic activity of Pistacia terebinthus and Ephedra alata crude extracts ","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-24 13:43:52","doi":"10.21203/rs.3.rs-4648793/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b7e8520c-5109-43d2-bf2b-52ad00ce4033","owner":[],"postedDate":"July 24th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-08-02T09:41:50+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-24 13:43:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4648793","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4648793","identity":"rs-4648793","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}