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Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L. a
b
c
c
d
N. Metoui , S. Gargouri , I. Amri , T. Fezzani , B. Jamoussi & L. c
Hamrouni a
Centre technique des agrumes , Tunisie
b
Laboratoire de Protection des Végétaux, Institut National de la Recherche Agronomique de Tunisie , Rue Hédi Karray, 2080 Ariana, Tunisie
Click for updates
c
Laboratoire de Gestion et Valorisation des Ressources Forestières, Institut National de Recherches en Génie Rural , Eaux et Forêts. BP 10, 2080 Ariana, Tunisie d
Institut Supérieur d'Education et de Formation Continue , Tunis, Tunisie Published online: 24 Jul 2015.
To cite this article: N. Metoui, S. Gargouri, I. Amri, T. Fezzani, B. Jamoussi & L. Hamrouni (2015): Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L., Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2015.1007136 To link to this article: http://dx.doi.org/10.1080/14786419.2015.1007136
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [University of Otago] at 05:03 26 July 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2015 http://dx.doi.org/10.1080/14786419.2015.1007136
SHORT COMMUNICATION Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L. N. Metouia, S. Gargourib, I. Amric, T. Fezzanic, B. Jamoussid and L. Hamrounic* a
Centre technique des agrumes, Tunisie; bLaboratoire de Protection des Ve´ge´taux, Institut National de la Recherche Agronomique de Tunisie, Rue He´di Karray, 2080 Ariana, Tunisie; cLaboratoire de Gestion et Valorisation des Ressources Forestie`res, Institut National de Recherches en Ge´nie Rural, Eaux et Foreˆts. BP 10, 2080 Ariana, Tunisie; dInstitut Supe´rieur d’Education et de Formation Continue, Tunis, Tunisie
Downloaded by [University of Otago] at 05:03 26 July 2015
(Received 1 August 2014; final version received 9 January 2015)
Our study is about the essential oil of Citrus aurantium L. in Tunisia and its plant extract. The yield of this essential oil is 0, 56% but the yield of the extract of plant was 17.1% for the aqueous extract ant 18.3% for the ethanolic extract. The analysis of chemical composition by using GC and GC/MS showed the essential oil of C. aurantium L. species to be rich in monoterpenes such as a-terpineol, lianolyl acetate, linalool and limonene. The antifungal activity of this oil showed us an inhibition of the germination of mushrooms, in the same way we could note that the biologic activities are generally assigned to the chemotypes high content in oxygenated monoterpene. Keywords: Citrus aurantium L; essential oil; antifungal activity; herbicides; weed; chemotypes; chemical pesticides; oxygenated monterpene
1. Introduction In Tunisia, Citrus aurantium L., commonly named sour or bitter orange, is grown in warm regions shielded from strong winds, mainly in Cap Bon region which covers an area of 1500 ha (Ammar et al. 2012). Sour orange is among the species most frequently used for both traditional and modern medicinal purposes. The space occupied by the sour orange in Tunisia allows the extraction of large quantities of essential oils from leaves, flowers and fruits. These essential oils contain substances that can be used in food (flavouring agents), in perfumery (odourants), cosmetics (substances treating skin and hair), especially in the food industry and organic agriculture by reducing the use of harmful pesticides and chemical compounds relative to the *Corresponding author. Email: [email protected] This work was presented at the 5th International Congress on Medicinal and Aromatic Plants, CIPAM 2014, 17 – 20 March 2014, Zarzis-Dejerba, Tunisia. q 2015 Taylor & Francis
Downloaded by [University of Otago] at 05:03 26 July 2015
2
N. Metoui et al.
human health and the environment. However, plants containing high levels of polyphenols have a great importance as natural antioxidants (Hsouna et al. 2013). The DPPH test demonstrated maximum antioxidant activity in the essential oil obtained from old leaves with 94.36%, followed by the flowers 53.98%, young leaves 22% and the peel 19.29% (Sarrou et al. 2013). Essential oil can be extracted from the leaves, which can be used fresh, dried or in the form of extracts. These are composed of some secondary metabolites with fungicidal, weed-killing or antibacterial activities, which could play a significant role in crop protection and ensure sustainable agriculture facing environmental problems. In fact, Neroli essential oil (extracted from the fragrant blossoms of bitter orange tree) exhibited marked antibacterial activity compared with standard antibiotic (Nystatin) (Ammar et al. 2012). Therefore, our work is to study the biological activities of the essential oils extracts from bitter orange to better exploit and enhance the use of this species. The antifungal activity of these essential oils is tested on five fungal species identified microscopically, namely Fusarium oxysporum (causing wilting of plants), Fusarium solani (causing soft dark brown to black cankers on stems, either at the nodes or at the wound), Botrytis cinerea (appearance of brown spots on the leaves, stems and fruits along with a greyish down), Bipolaris sorokinian (elliptical spots, uniformly dark brown on the leaves, can lie down and be delimited by the veins) and Fusarium avenaceum (appearance of a silky white mycelium covering partially the inflorescence).
2. Results and discussion 2.1. Study performance of aqueous and ethanol extracts The extraction yield of essential oil obtained by using hydrodistillation from the fresh leaves of bitter orange harvested in February was calculated and expressed in grams of essential oil/100 gram of plant material. Yields of ethanoic and aqueous extracts studied were, respectively, about 17.1 and 18.3.
2.2. Study of physico-chemical properties of aqueous and ethanol extracts To control the quality of the essential oil studied, we conducted a physico-chemical and organoleptic study, which is an analysis and verification of the quality of oil means. For the refractive index of the bitter orange is about 1.46, it is greater than the index of refraction of water, which is equal to 1.333. This explains the significant ability of the essential oil of bitter orange to reflect light, which makes it possible to be used in the field of cosmetics (Kanko et al. 2004)
2.3. Chemical composition of the essential oil The essential oils obtained were analysed by using gas chromatography with FID and MS detector. The results showed a qualitative and quantitative variability of the chemical composition. Thanks to this method of analysis, it was possible to identify the presence of 21 components representing 99.21% of the total (Table 1). In Tunisia, the chemical composition of essential oils of sour orange was studied by BenDaoued et al. (2010). The results are in agreement with our results; in fact they showed that these oils were rich in linalool, linalyl acetate and a-terpineol. Other compounds, against, with low variability in the percentages of the compounds of the oil that can be attributed to the variability pedoclimatic and the method of extraction and drying.
Natural Product Research
3
Table 1. Chemical composition of the essential oil of bitter orange.
Downloaded by [University of Otago] at 05:03 26 July 2015
No
Retention index (RI)
1 931 2 939 3 991 4 980 5 1018 6 1031 8 1088 9 1103 10 1178 11 1161 12 1260 13 1248 14 1349 15 1353 16 1418 17 1434 18 1452 19 1480 20 1511 21 1581 Total Monoterpenes hydrocarbons Monoterpenes oxygenated Sesquiterpenes hydrocarbons Sesquiterpenes oxygenated
Compounds
%
a-Thujene a-Pinene b-Myrcene b-Pinene a-Terpinene Limonene a-Terpinolene Linalool Terpinen-4-ol a-Terpinol Linalyl acetate Geraniol Terpenyl acetate Geranyl acetone Z-carophyllene trans-a-Bergamotene a-Humulene d-Germacrene D g-Cadinene Carophyllene oxide
Identification methods
0.14 0.69 0.26 0.22 1.58 5.28 0.16 39.30 0.94 10.75 32.10 2.04 4.39 0.32 0.40 0.25 0.11 0.32 0.16 0.10 99.51% 8.33% 1.24% 89.84% 0.1%
MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS,
RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI
2.4. Antifungal activity of essential oil We chose five fungi: F. oxysporum, F. solani, B. cinerea, Bipolaris sorokiniana and F. avenaceum. They were treated with the essential oil of bitter orange and then inhibition of growth of each fungus was determined. Thereafter, it was found that the growth inhibition and fungi are in relation to the dose of the essential oil which can be observed by the decreasing growth with the increasing the dose of the essential oil. Antifungal activity was studied by the method of direct contact of phytopathogenic fungi; the percentages of inhibition are calculated relative to the control (Table 2). Group averages of homogeneous subsets are displayed and used as the sample size of the harmonic means. The application of the essential oil of bitter orange on several fungi results in a significant antifungal activity. But this activity is variable depending on the applied dose of the essential oil and the fungal species.
Table 2. Effect of essential oils of sour orange on some phytopathogenic fungi.
Fungi F. oxysporum B. sorokiniana F. avenaceum B. cinerea F. solani
2 mL/mL
4 mL/mL
Control Growth (mm)
Growth
Inhibition (%)
Growth
Inhibition (%)
7.4 ^ 0.56 7.15 ^ 0.21 8.15 ^ 0.21 6.5 ^ 0.14 9.25 ^ 0.35
2.9 ^ 0.28 2.25 ^ 0.21 3.8 ^ 0.28 4 ^ 0.28 3.1 ^ 0.14
60.54 ^ 6.83a 68.47 ^ 3.90a 53.31 ^ 4.68a 38.49 ^ 3.01b 66.49 ^ 0.24a
1.2 ^ 0.14 1.3 ^ 0.28 1.25 ^ 0.21 1.55 ^ 0.07 1.65 ^ 0.07
83 ^ 3.15a 81.75 ^ 4.97a 84 ^ 2.2a 76.13 ^ 1a 82 ^ 1.44a
4
N. Metoui et al.
3. Conclusion In this work, we studied the chemical composition and antifungal activity of the essential oil of bitter orange. The qualitative and quantitative analyses of oils identified 21 constituents with aterpineol (10.75%), lianalyl acetate (32.1%), linalool (39.3%) and limonene (5.28%) as major compounds. The study of the antifungal activity of essential oils of bitter orange showed inhibition of the tested pathogenic fungi. Thus, this oil showed significant biological activities. Therefore, these results can be recovered and could be used in the fight against weeds and fungal diseases by farmers and serve as an alternative to chemical pesticides.
Downloaded by [University of Otago] at 05:03 26 July 2015
Supplementary material Experimental details relating to this paper are available online http://dx.doi.org/10.1080/1478 6419.2015.1007136. Acknowledgements We are grateful to the Technical Center of Citrus (CTA) represented by Mr Mohamed Cherif, its director, and his team who gave us the plant material required for the completion of this work.
References Ammar AH, Bouajila J, Lebrihi A, Mathieu F, Romdhane M, Zagrouba F. 2012. Chemical composition and in vitro antimicrobial and antioxidant activities of Citrus aurantium L. Flowers essential oil (Neroli oil). Pak J Biol Sci. 15:1034–1040. doi:10.3923/pjbs.2012.1034.1040. BenDaoued H, Romdane M, Souchard J. P, Cazaux S, Bouajila J. 2010. Chemical composition and anticancer and antioxidant activities of Schinus Molle L. and Schinus terebinthifolius Raddi berries essential oils. J Food Sci. 75:466–472. Hsouna BA, Hamdi NB, Halima N, Abdelkafi S. 2013. Characterization of essential oil from Citrus aurantium L. flowers: antimicrobial and antioxidant activities. J Oleo Sci. 62:763–772. doi:10.5650/jos.62.763. Kanko C, Sawaliho BELH, Kone S, N’Guessan YT. 2004. E´tude des proprie´te´s physico-chimiques des huiles essentielles de Lippia multiflora, Cymbopogon citratus, Cymbopogon nardus, Cymbopogon giganteus. C R Chim. 7:1039–1042. Sarrou E, Chatzopoulou P, Dimassi-Theriou K, Therios I. 2013. Volatile constituents and antioxidant activity of peel, flowers and leaf oils of Citrus aurantium L. growing in Greece. Molecules. 18:10639–10647. doi:10.3390/ molecules180910639.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L. a
b
c
c
d
N. Metoui , S. Gargouri , I. Amri , T. Fezzani , B. Jamoussi & L. c
Hamrouni a
Centre technique des agrumes , Tunisie
b
Laboratoire de Protection des Végétaux, Institut National de la Recherche Agronomique de Tunisie , Rue Hédi Karray, 2080 Ariana, Tunisie
Click for updates
c
Laboratoire de Gestion et Valorisation des Ressources Forestières, Institut National de Recherches en Génie Rural , Eaux et Forêts. BP 10, 2080 Ariana, Tunisie d
Institut Supérieur d'Education et de Formation Continue , Tunis, Tunisie Published online: 24 Jul 2015.
To cite this article: N. Metoui, S. Gargouri, I. Amri, T. Fezzani, B. Jamoussi & L. Hamrouni (2015): Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L., Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2015.1007136 To link to this article: http://dx.doi.org/10.1080/14786419.2015.1007136
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [University of Otago] at 05:03 26 July 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2015 http://dx.doi.org/10.1080/14786419.2015.1007136
SHORT COMMUNICATION Activity antifungal of the essential oils; aqueous and ethanol extracts from Citrus aurantium L. N. Metouia, S. Gargourib, I. Amric, T. Fezzanic, B. Jamoussid and L. Hamrounic* a
Centre technique des agrumes, Tunisie; bLaboratoire de Protection des Ve´ge´taux, Institut National de la Recherche Agronomique de Tunisie, Rue He´di Karray, 2080 Ariana, Tunisie; cLaboratoire de Gestion et Valorisation des Ressources Forestie`res, Institut National de Recherches en Ge´nie Rural, Eaux et Foreˆts. BP 10, 2080 Ariana, Tunisie; dInstitut Supe´rieur d’Education et de Formation Continue, Tunis, Tunisie
Downloaded by [University of Otago] at 05:03 26 July 2015
(Received 1 August 2014; final version received 9 January 2015)
Our study is about the essential oil of Citrus aurantium L. in Tunisia and its plant extract. The yield of this essential oil is 0, 56% but the yield of the extract of plant was 17.1% for the aqueous extract ant 18.3% for the ethanolic extract. The analysis of chemical composition by using GC and GC/MS showed the essential oil of C. aurantium L. species to be rich in monoterpenes such as a-terpineol, lianolyl acetate, linalool and limonene. The antifungal activity of this oil showed us an inhibition of the germination of mushrooms, in the same way we could note that the biologic activities are generally assigned to the chemotypes high content in oxygenated monoterpene. Keywords: Citrus aurantium L; essential oil; antifungal activity; herbicides; weed; chemotypes; chemical pesticides; oxygenated monterpene
1. Introduction In Tunisia, Citrus aurantium L., commonly named sour or bitter orange, is grown in warm regions shielded from strong winds, mainly in Cap Bon region which covers an area of 1500 ha (Ammar et al. 2012). Sour orange is among the species most frequently used for both traditional and modern medicinal purposes. The space occupied by the sour orange in Tunisia allows the extraction of large quantities of essential oils from leaves, flowers and fruits. These essential oils contain substances that can be used in food (flavouring agents), in perfumery (odourants), cosmetics (substances treating skin and hair), especially in the food industry and organic agriculture by reducing the use of harmful pesticides and chemical compounds relative to the *Corresponding author. Email: [email protected] This work was presented at the 5th International Congress on Medicinal and Aromatic Plants, CIPAM 2014, 17 – 20 March 2014, Zarzis-Dejerba, Tunisia. q 2015 Taylor & Francis
Downloaded by [University of Otago] at 05:03 26 July 2015
2
N. Metoui et al.
human health and the environment. However, plants containing high levels of polyphenols have a great importance as natural antioxidants (Hsouna et al. 2013). The DPPH test demonstrated maximum antioxidant activity in the essential oil obtained from old leaves with 94.36%, followed by the flowers 53.98%, young leaves 22% and the peel 19.29% (Sarrou et al. 2013). Essential oil can be extracted from the leaves, which can be used fresh, dried or in the form of extracts. These are composed of some secondary metabolites with fungicidal, weed-killing or antibacterial activities, which could play a significant role in crop protection and ensure sustainable agriculture facing environmental problems. In fact, Neroli essential oil (extracted from the fragrant blossoms of bitter orange tree) exhibited marked antibacterial activity compared with standard antibiotic (Nystatin) (Ammar et al. 2012). Therefore, our work is to study the biological activities of the essential oils extracts from bitter orange to better exploit and enhance the use of this species. The antifungal activity of these essential oils is tested on five fungal species identified microscopically, namely Fusarium oxysporum (causing wilting of plants), Fusarium solani (causing soft dark brown to black cankers on stems, either at the nodes or at the wound), Botrytis cinerea (appearance of brown spots on the leaves, stems and fruits along with a greyish down), Bipolaris sorokinian (elliptical spots, uniformly dark brown on the leaves, can lie down and be delimited by the veins) and Fusarium avenaceum (appearance of a silky white mycelium covering partially the inflorescence).
2. Results and discussion 2.1. Study performance of aqueous and ethanol extracts The extraction yield of essential oil obtained by using hydrodistillation from the fresh leaves of bitter orange harvested in February was calculated and expressed in grams of essential oil/100 gram of plant material. Yields of ethanoic and aqueous extracts studied were, respectively, about 17.1 and 18.3.
2.2. Study of physico-chemical properties of aqueous and ethanol extracts To control the quality of the essential oil studied, we conducted a physico-chemical and organoleptic study, which is an analysis and verification of the quality of oil means. For the refractive index of the bitter orange is about 1.46, it is greater than the index of refraction of water, which is equal to 1.333. This explains the significant ability of the essential oil of bitter orange to reflect light, which makes it possible to be used in the field of cosmetics (Kanko et al. 2004)
2.3. Chemical composition of the essential oil The essential oils obtained were analysed by using gas chromatography with FID and MS detector. The results showed a qualitative and quantitative variability of the chemical composition. Thanks to this method of analysis, it was possible to identify the presence of 21 components representing 99.21% of the total (Table 1). In Tunisia, the chemical composition of essential oils of sour orange was studied by BenDaoued et al. (2010). The results are in agreement with our results; in fact they showed that these oils were rich in linalool, linalyl acetate and a-terpineol. Other compounds, against, with low variability in the percentages of the compounds of the oil that can be attributed to the variability pedoclimatic and the method of extraction and drying.
Natural Product Research
3
Table 1. Chemical composition of the essential oil of bitter orange.
Downloaded by [University of Otago] at 05:03 26 July 2015
No
Retention index (RI)
1 931 2 939 3 991 4 980 5 1018 6 1031 8 1088 9 1103 10 1178 11 1161 12 1260 13 1248 14 1349 15 1353 16 1418 17 1434 18 1452 19 1480 20 1511 21 1581 Total Monoterpenes hydrocarbons Monoterpenes oxygenated Sesquiterpenes hydrocarbons Sesquiterpenes oxygenated
Compounds
%
a-Thujene a-Pinene b-Myrcene b-Pinene a-Terpinene Limonene a-Terpinolene Linalool Terpinen-4-ol a-Terpinol Linalyl acetate Geraniol Terpenyl acetate Geranyl acetone Z-carophyllene trans-a-Bergamotene a-Humulene d-Germacrene D g-Cadinene Carophyllene oxide
Identification methods
0.14 0.69 0.26 0.22 1.58 5.28 0.16 39.30 0.94 10.75 32.10 2.04 4.39 0.32 0.40 0.25 0.11 0.32 0.16 0.10 99.51% 8.33% 1.24% 89.84% 0.1%
MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS, MS,
RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI RI
2.4. Antifungal activity of essential oil We chose five fungi: F. oxysporum, F. solani, B. cinerea, Bipolaris sorokiniana and F. avenaceum. They were treated with the essential oil of bitter orange and then inhibition of growth of each fungus was determined. Thereafter, it was found that the growth inhibition and fungi are in relation to the dose of the essential oil which can be observed by the decreasing growth with the increasing the dose of the essential oil. Antifungal activity was studied by the method of direct contact of phytopathogenic fungi; the percentages of inhibition are calculated relative to the control (Table 2). Group averages of homogeneous subsets are displayed and used as the sample size of the harmonic means. The application of the essential oil of bitter orange on several fungi results in a significant antifungal activity. But this activity is variable depending on the applied dose of the essential oil and the fungal species.
Table 2. Effect of essential oils of sour orange on some phytopathogenic fungi.
Fungi F. oxysporum B. sorokiniana F. avenaceum B. cinerea F. solani
2 mL/mL
4 mL/mL
Control Growth (mm)
Growth
Inhibition (%)
Growth
Inhibition (%)
7.4 ^ 0.56 7.15 ^ 0.21 8.15 ^ 0.21 6.5 ^ 0.14 9.25 ^ 0.35
2.9 ^ 0.28 2.25 ^ 0.21 3.8 ^ 0.28 4 ^ 0.28 3.1 ^ 0.14
60.54 ^ 6.83a 68.47 ^ 3.90a 53.31 ^ 4.68a 38.49 ^ 3.01b 66.49 ^ 0.24a
1.2 ^ 0.14 1.3 ^ 0.28 1.25 ^ 0.21 1.55 ^ 0.07 1.65 ^ 0.07
83 ^ 3.15a 81.75 ^ 4.97a 84 ^ 2.2a 76.13 ^ 1a 82 ^ 1.44a
4
N. Metoui et al.
3. Conclusion In this work, we studied the chemical composition and antifungal activity of the essential oil of bitter orange. The qualitative and quantitative analyses of oils identified 21 constituents with aterpineol (10.75%), lianalyl acetate (32.1%), linalool (39.3%) and limonene (5.28%) as major compounds. The study of the antifungal activity of essential oils of bitter orange showed inhibition of the tested pathogenic fungi. Thus, this oil showed significant biological activities. Therefore, these results can be recovered and could be used in the fight against weeds and fungal diseases by farmers and serve as an alternative to chemical pesticides.
Downloaded by [University of Otago] at 05:03 26 July 2015
Supplementary material Experimental details relating to this paper are available online http://dx.doi.org/10.1080/1478 6419.2015.1007136. Acknowledgements We are grateful to the Technical Center of Citrus (CTA) represented by Mr Mohamed Cherif, its director, and his team who gave us the plant material required for the completion of this work.
References Ammar AH, Bouajila J, Lebrihi A, Mathieu F, Romdhane M, Zagrouba F. 2012. Chemical composition and in vitro antimicrobial and antioxidant activities of Citrus aurantium L. Flowers essential oil (Neroli oil). Pak J Biol Sci. 15:1034–1040. doi:10.3923/pjbs.2012.1034.1040. BenDaoued H, Romdane M, Souchard J. P, Cazaux S, Bouajila J. 2010. Chemical composition and anticancer and antioxidant activities of Schinus Molle L. and Schinus terebinthifolius Raddi berries essential oils. J Food Sci. 75:466–472. Hsouna BA, Hamdi NB, Halima N, Abdelkafi S. 2013. Characterization of essential oil from Citrus aurantium L. flowers: antimicrobial and antioxidant activities. J Oleo Sci. 62:763–772. doi:10.5650/jos.62.763. Kanko C, Sawaliho BELH, Kone S, N’Guessan YT. 2004. E´tude des proprie´te´s physico-chimiques des huiles essentielles de Lippia multiflora, Cymbopogon citratus, Cymbopogon nardus, Cymbopogon giganteus. C R Chim. 7:1039–1042. Sarrou E, Chatzopoulou P, Dimassi-Theriou K, Therios I. 2013. Volatile constituents and antioxidant activity of peel, flowers and leaf oils of Citrus aurantium L. growing in Greece. Molecules. 18:10639–10647. doi:10.3390/ molecules180910639.
