Brocchia cinerea essential oil from Brezina (Algerian Sahara): Chemical characterization and antibacterial activity

Z Boukhobza1, A Cheriti1*, N Boulenouar1,2, H Djeradi1, M Kacimi El Hassani3, MB Lahreche3 and Ka Sekkoum1

1Phytochemistry and Organic Synthesis Laboratory, Faculty of Medicine, UTMB, 08000, Algeria
2Department of Biology, University Center Nour E., El Bayadh, 32000, Algeria
3Organic Chemistry & Natural Substances Laboratory, Faculty of Sciences, Z. Achour University, Djelfa, 17000, Algeria

Corresponding author: A Cheriti, Phytochemistry and Organic Synthesis Laboratory, Faculty of Medicine, UTMB, 08000, Algeria, E-mail:

Citation: A Cheriti, Z Boukhobza, et al (2020) Brocchia cinerea essential oil from Brezina (Algerian Sahara): Chemical characterization and antibacterial activity, J Chem Sci Chem Engg 1(1): 19-24.

Received Date: : April 30, 2020; Accepted Date: May 05, 2020; Published Date: May 11, 2020


The essential oil of wild-growing medicinal specie Brocchia cinerea (Asteraceae) from Brezina (Algerian Sahara) was studied for chemical composition by GC/MS analysis and antibacterial activity. Twenty-nine identified components constituted 94.07% of the oil. The most important major components in the oil were trans-Thujone (36.11 %), Camphor (12.08 %), Santolina triene (11.25 %), 1,8-Cineol (6.98 %) and Cis-Verbenyl acetate (6.33 %), together reaching 72.75% of the total oil which is characterized by the presence of high content of oxygenated monoterpenes. The antibacterial activity of the essential oil was tested using the disc diffusion assay. The results showed that the essential oil of Brocchia cinerea had antibacterial activity against all tested Gram-positive bacteria (Enterococcus faecalis and Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli, Klebsiella pneumonia, and Pseudomonas aereginosa).

Keywords: Essential Oil; Brocchia Cinerea; GC/MS; Oxygenated Monoterpenes; Antibacterial activity; Sahara;


Since ancient times, volatile essential oils and plant extracts are used for preserving human health in most ancient civilizations. Thus, the World Health Organization (WHO) has recognized the potential utility of traditional remedies and strives to preserve primary health care involving medicinal plants [1]. The importance of natural products to human health was emphasized by the award of the 2015 Nobel Prize in Physiology or Medicine to Professor Satoshi Omura and Dr. William Campbell for the discovery of avermectin from a Streptomyces sp. and its development as a treatment for river blind¬ness and other parasitic diseases, and to Professor Tu Youyou of Beijing for her discoveries leading to the development of the antimalarial drug artemisinin from Artemisia annua [2], a substance which, together with chloroquine, constitutes a candidate drug for the treatment of the current SARSCov-2 (COVID 19) pandemic [3].

Due to its geographic position, with a varied landscape and diversified climate favorable for rich vegetation, Algeria has a varied flora of about 4450 taxa of which 3950 indigenous with 6.5% endemic [4]. In particular, the Sahara part constitutes an important reservoir of many plants that have not been investigated until today. Furthermore, Ozenda noted that Asteraceae, Fabaceae, and Poaceae are everywhere dominant families in the flora of Sahara. This Asteraceae family represent 13,8%, 11,2%, and 7,9% of the total flora respectively in Sptentrional Sahara, Central Sahara, and Meridional Sahara[5].

Among this flora, Brocchia cinerea species from the Asteraceae family have been widely used in the Algerian Sahara ethnopharmacopea for the treatment of various diseases as a medicinal plant [6]. Brocchia cinerea is traditionally used as an infusion and decoction to treat digestive troubles, rheumatoid arthritis, fever, inflammation, headaches, migraines, colic, diarrhea, cough, cooling broncho-pulmonary, urinary and pulmonary infections [7, 8].

Some interesting biological studies have been described in the literature that has examined the antibacterial, antiprotozoal, analgesic, and antipyretic activities of essential oils and some extracts from B. cinerea [9-13].

However, as far as we know, tracing the current literature, nothing was found concerning the chemical composition and biological activity of B. cinerea essential oil growing wild in the Brezina area (southwest of Algeria, Wilaya El Bayadh). Thus, as a part of our investigation into Algerian medicinal plants [14-19], in this study, we investigate for the first time the chemical composition and antibacterial activity of the essential oils from the Saharan medicinal plant Brocchia cinerea.

Materials and Methods

Plant material

Brocchia cinerea Del. (Syn. Cotula cinerea Delile, Tanacetum cinereum (Delile) DC.) is a small annual herb with discoid capitula and homogamous hermaphrodite florets figure 1. It is widely distributed in the Sahara desert and represents one of the monotypic Anthemideae genera which are characteristic of the North African flora [20].

Figure 1: The medicinal plant Brocchia cinerea - Algeria Sahara

Aerial parts of Brocchia cinerea were collected in Brezina (Latitude: 33° 5′ 58″ N; Longitude: 1° 15′ 39″ E; Altitude: 1028m)) during the flowering period in February (2018). The plant was identified by Pr A. Marouf (Department of Biology, University Center Naama – Algeria) and a voucher specimen is kept in the Herbarium of POSL Laboratory, (UTMB and Algeria) under N° CA 07/10.

Isolation of the Essential Oil

The Brocchia cinerea essential oil (EO) was obtained from dry plant material (100 g) by hydro distillation using the Clevenger apparatus for 3 h, following the 3rd Edition of the European Pharmacopoeia cited by [21]. The obtained oil was dried over anhydrous sodium sulphate and stored in colored glass at 4 °C until analysis.

GC-MS Analysis

GC/MS analysis was performed on Shimadzu GC-17A gaschromatograph, interfaced with Shimadzu QP5000 mass spectrometer, operating at an electron impact of 70 eV with an ion source temperature at 250°C, scan a mass range of 40-400 m/z at a sampling rate of 0.5 scans/s. A 60 m ×0.25 mm fused silica nonpolar DB-5 capillary column, with a 0.3 μm film thickness was used. The operating conditions were: Temperature programmed as follows: 50°C for 2 min and then up to 240°C at 3°C/min, then to 300°C at 10°C/min, ending with a 10 min at 300°C., and injector and detector temperature were 240°C. The carrier gas was He (1.0 mL/min), injector, and detector temperature 240°C. Samples were injected by splitting and the split ratio of 1:5.

The EO component identification was confirmed by comparison of mass spectral fragmentation patterns with the computer library (NIST MS Library) and verified by comparison of their retention indices (determined relatively to the retention times of an n-alkanes homologous series, C-9 to C-24) of the identified compounds with the literature [15, 22-24]. The relative amounts of the individual components found in the oil are based on the peak areas obtained, without FID response factor corrections.

Antibacterial activity

The antibacterial activity of Brocchia cinerea essential oil was determined using the paper disc diffusion method according to our previous studies [15, 25, 26], against three-Gram negative bacteria (Escherichia coli (ATCC 25922), Klebsiella pneumonia (Isolated), Pseudomonas aereginosa (ATCC 27853)) and two-Gram positive bacteria (Enterococcus faecalis (ATCC 29212), Staphylococcus aureus (ATCC 25923)) which were obtained from Pasteur Institute (Algiers, Algeria). Antibacterial activities were evaluated by measuring the inhibition zone diameters in mm. Amoxicillin is included in the test as a reference [26, 27].

Results and Discussion

Chemical composition of the essential oil

Yellowish oil with a characteristic artemisias pleasant-smelling odor and sweet taste is obtained with a yield of 1.4% from Brocchia cinerea. The results of the chemical composition of the essential oil obtained by GC/MS analysis are presented in Table 1 and components are listed in order of their elution on the DB-5 capillary column. Twenty-nine compounds were identified representing 94.07% of the oil. The most important major components in the oil were trans-Thujone (36.11 %), Camphor (12.08 %), Santolina triene (11.25 %), 1,8 Cineol (6.98 %) and Cis-Verbenyl acetate (6.33 %), together reaching 72.75% of the total oil which is characterized by the presence of high content of oxygenated monoterpenes. Other compounds are present with low percentages between 1.5 and 2.5 % such as Terpinene-4-ol, Santolina alcohol, Camphene, α-Pinene, and β-Pinene.

Brocchia cinerea collected in the region of Oued Souf ( Southeast Algeria) during two stages (flowering and fruiting), gave 0.0801% and 0.391% of essential oil (EO) respectively at the flowering and fruiting stage. The chemical study by GC/MS showed the presence of 22 chemical compounds in the flowering period with the dominance of 3-Carène (30.99%), Thujone (21.73%), Santolina triene (18.58%) and Camphor (6.21%). While 21 chemical compounds were obtained during the fruiting period with the dominance: Thujone (28.78%), 3-Carène (15.90%), Eucalyptol (15.13%), Santolina triene (13.38%) and Camphor (7.49%) [28]. The species from Morocco has Trans- Thujone (41.4%), cisverbenyl acetate (24.7%), 1,8-cineole (8.2%) and camphor (5.5%) as the major components [10], Other analyses indicate that EO is dominated by the existence of the iso-3-thujanol with 47.38%, followed by Santolina triene (11.67%) and of camphor (10.95%) [29]. However, the percentage of compounds differed substantially from that reported in species collected in Egypt, in which camphor (50%) and trans-Thujone (14.4%) were the main oil compounds [30].

Almost all of the components reported in our study were found in B. cinerea from Southeast Algeria, Morocco, and Egypt [10, 28-30] and in some species belonging to the tribe Anthemideae, confirming the chemotaxonomic relationships between Cotula and Anthemideae genera [10, 31,32]. We observed a qualitatively resembles in the chemical composition of our essential oils and that obtained from B. cinerea collected from other regions (Southeast Algeria, Morocco, and Egypt), but with some quantitative divergences, could be attributed to many factors such as vegetative phases of the plant, environmental and growing conditions (e.g. seasonal and geographical variations, soil composition [33].

Antibacterial activity.

Microorganisms have the genetic ability to transmit and acquire resistance to antibiotics and have become a major global healthcare problem in the 21st century [25]. Thus, one of the most efficient ways of finding new antibacterial compounds is collecting data on the use of medicinal plants in traditional pharmacopeia [1, 19].

Several studies have examined the antimicrobial activities of essential oils from Brocchia cinerea, Ghanmi et al [29] evaluated the anticandidal and antibacterial activity of the essential oil from Morocco, by using a panel of human pathogenic fungi (Candida albicans, C. krusei, C. glabrata and C. parapsilosis), authors demonstrate that the oil showed high anticandidal activity against all investigated strains with the inhibition zones against the tested Candida species ranging from 19.3 to 25.3 mm. Interesting results obtained of the antibacterial activity of essential oils against Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Micrococcus luteus. All bacterial strains were inhibited at 1/500 v/v concentration. In additional mold (Penicillium digitatum, Penicillium expansum, and Aspergillus niger) were less sensitive than bacteria and their growth was stopped at 1/250 v/v concentration [29]. The Escherichia coli strains and Staphylococcus aureus showed high sensitivity to the essential oil of this plant, until the concentration 1/8 (v/v) [28]. El Bouzidi et al. [10] demonstrated that the essential oil of B. cinerea presented a great activity against tested yeasts and the specie essences present a wide anticandidal activity spectrum.

We summarized in Table (2), the results of the in vitro antibacterial activity of the essential oils against pathogens causing urinary tract, lung, and gastrointestinal infection: Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aereginosa, and Staphylococcus aureus. The inhibition zone, measured in millimeters, including the diameter of the paper disk, was used as the criterion for measuring the antibacterial activity.

The Brocchia cinerea essential oil showed the best antibacterial activity, especially against Gram (+) bacteria Staphylococcus aureus, with zones of inhibition greater than 21.4 mm. One of the best known resistant Gram (-) bacteria, Klebsiella pneumonia seems to be more sensitive to the EO compared to the antibiotic Amoxicillin. Whereas, a moderate antibacterial effect against Enterococcus faecalis and Pseudomonas aereginosa was observed.

This antibacterial activity of the EO may be explained by its richness in oxygenated monoterpenes such as trans-Thujone (36.11%), Camphor (12.08 %) well known for their high and broadspectrum antimicrobial activity with the appreciable amount of 1,8-Cineol ( 6.98%) could also contribute to the antibacterial activity.

Table 1: Chemical composition of the Brocchia cinerea essential oil





Content  (%)


Santolina triene

























a- Terpinene










1,8 Cineol





Santolina alcohol




















trans- Thujone






























Carvacrol methy ether





Linalyl  acetate





Cis-Verbenyl acetate





Bornyl acetate










Neryl acetate





Geranyl acetate















Germacrene D





Caryophyllene oxide




Total Identified (%)                                        94.07

Table 2: Antibacterial activity of Brocchia cinerea essential oil



Inhibition Zone Diameters (mm)

Essential oil (Volume/disk (25µl))

Amoxicillin (15µg/ml)


Gram (+)

Enterococcus faecalis



Staphylococcus aureus



Gram (-)

Escherichia  coli



Klebsiella pneumoniae



Pseudomonas aereginosa




In this work, we studied for the first time the chemical composition and antibacterial activity of the essential oil of the medicinal plant Brocchia cinerea from Brezina (Algerian Sahara). Chemical analysis of essential oil by GC/MS identified twenty-nine (94.07% ) compounds dominated by trans-Thujone (36.11 %), Camphor (12.08 %), Santolina triene (11.25 %), 1,8 Cineol (6.98 %) and Cis-Verbenyl acetate (6.33 %). The result obtained in this study shows that the essential oil has significant activity against Staphylococcus aureus and Klebsiella pneumonia, this, probably explains the use of this aromatic plant in traditional Saharan ethnopharmacopeae against some human diseases. Further experiments are planned to evaluate other biological activities and to determine the enantiomeric composition of this essential oil by chiral analysis.


We are thankful to MESRS and DGRST for the financial support given through this project.


  1. Cheriti A. Endemic Asteraceae from Algerian Sahara: Potential Medicinal Value and Chemical Diversities, in Asteraceae: Characteristics, Distribution, and Ecology, Millicent T (Ed.), Nova Publisher, New York, USA, ISBN: 978-1-53616-632-3:2020.
  2. Buckingham J, Cooper CM, Purchase R. Natural Products Desk Reference. CRC Taylor & Francis, NY. 2016.
  3. a) Wang M, Cao R, Zhang L, Yang X, Liu J, Xu M, et al. Remdesivir and chloroquine effectively inhibit the recentlyemerged novel coronavirus (2019-nCoV). Vitro, Cell Research, 2020;30:269–227. b) Yang P, Wang X. COVID-19: a new challenge for human beings. Cellular & Molecular Immunology. 2020. DOI: 10.1038/s41423-020-0407-x
  4. Dobignard A, Chatelain C. Index synonymique de la flore d’afrique du nord [Synonymic index of the flora of North Africa]. Editions des conservatoire et jardin botaniques, Genève. 2013.
  5. Ozenda, P. Flore et Végétation du Sahara [Flora and vegetation of the Sahara], CNRS, Paris. 2004.
  6. Cheriti A. Medicinal Plants of the Bechar Region, Ethnopharmacologie studies, CRSTRA Report, Algeria. 2000.
  7. a) Bellakhdar J. La pharmacopée marocaine traditionnelle. Médecine arabe ancienne et savoirs populaires [Moroccan traditional pharmacopoeia. Ancient Arabic medicine and popular knowledge]. IBIS Press. 1997. b) Ibn al-Baytar, Dhiya al-Din. Kitāb al-jāmiʿ li-mufradāt al-adwiya wa al-aghdhiya (Compendium on Simple Medicaments and Foods). Ed. Dar Kotob Elmia, Libanon. 1992. c) Leclerc L.  Traité des simples de ʻAbd Allāh ibn Aḥmad Ibn al-Bayṭār (Compendium on Simple of ʻAbd Allāh ibn Aḥmad Ibn al-Bayṭār). Traduction, Impr. Nationale, Paris. 1877.
  8. Belboukhari N, Cheriti A, Bouanini M. Chiral separation of flavanones from cotula cinerea by TLC methods. PhytoChem & BioSub J. 2012;6(1):1-5.
  9. Markouk M, Redwane A, Lazrek HB, Jana M, Benjama A. Antibacterial activity of Cotula cinerea extracts. Fitoterapia.  1999;70(3):314-316.
  10. El Bouzidi L, Abbad A, Fattarsi K, Hassani L, Leach D, Markouk M, et al. Chemical composition and anticandidal properties of the essential oil isolated from aerial parts of Cotula cinerea: a rare and threatened medicinal plant in Morocco. Nat Prod Commun. 2011;6(10):1491-1494.
  11. Markouk M, Lazrek HB, Larhsini M, Bekkouche K, Jana M. Preliminary screening of antiprotozoal activity of extracts from Cotula cinerea L. Therapie. 1999;54(6):763-765.
  12. Markouk M, Lazrek HB, Jana M. Analgesic effect of extracts from Cotula cinerea (L). Phytother Res. 1999;13(3):229-230.
  13. Larhsini M, Markouk M, Jaouhari JT, Bekkouche K, Lazrek HB, Jana M. The antipyretic activity of some Morroccan medicinal plants. Phytother Res. 2002;16 Suppl 1:S97-8.
  14. Oughilas A, Cheriti A, Reddy KH, Govender P. In vitro antioxydant activity and total phenolic content of extracts from the endemic argania spinosa (l.) skeels from Algerian Sahara. J Fundam Appl Sci. 2019;11(1):539-547.
  15. Boukhobza Z, Cheriti A, Kacimi El Hassani M, Lahreche MB. Comparison of chemical composition and antibacterial activity of mentha pulegium essential oil from two ecotypes (El Bayadh and Djelfa). J Fundam Appl Sci. 2019;11(3):1175-1187.
  16. Ghazi R, Boulenouar N, Cheriti A, Reddy K, Govender P. Bioguided fractionation of Citrullus colocynthis extracts and antifungal activity against Fusarium oxysporum f.sp. Albedinis. Curr Bio Comp. 2020;16(3):302-307. DOI: 10.2174/1573407214666181001124737
  17. Messaoudi R, Cheriti A, Bourmita Y. Bioassay-guided isolation of the major compound with antioxidant activity from the algerian medecinal plant Bubonium graveolens. Asian J Pharm Clin Res. 2018;11(11):424-426.
  18. Allaoui M, Cheriti A, Al-Gharabli S, Gherraf  N, Chebouat E, Dadamoussa B, Al-Lahham A. A Comparative Study of the Antibacterial Activity of Two Chenopodiaceae: Haloxylon scoparium (Pomel) and Traganum nudatum Del. Res J Pharma, Bio Chem Sci. 2014;5(5):85-89.
  19. Cheriti A. Limoniastrum feei from the Algerian Sahara: Ethnopharmacology, Phytochemistry and the Pharmacological Importance, in Plant sources and potential health benefits, edited by Iman Ryan. 2019:259-299. Nova Science Pub., N.York. USA.
  20. Greger H, Hofer O. Sesquiterpene-coumarin ethers and polyacetylenes from Brocchia cinerea. Phytochemistry. 1985;24(1):85-88.
  21. Bruneton J. Pharmacognosy, Phytochemistry, Medicinal Plants. 2nd edition, Ed. Tec & Doc: Paris. 1999.
  22. Adams RP. Identification of Essential Oil components by Gas Chromatography/Mass Spectrometry. 4th Ed., Allured publishing USA. 2007.
  23. Said ME-A, Bombarda I, Naubron J-V, Vanloot P, Jean M, Cheriti A, et al. Isolation of the major chiral compounds from Bubonium graveolens essential oil by HPLC and absolute configuration determination by VCD. Chirality. 2017;29(2):70–79.
  24. Cheriti A, Saad A, Belboukhari N, Ghezali S. The essential oil composition of Bubonium graveolens (Forssk.) Maire from the Algerian Sahara. Flavour and Fragrance Journal. 2007;22(4):286–288.  
  25. Berbaoui H, Cheriti A, Ould El Hadj-Khelil A. Répartition et polymorphisme des souches nosocomiales du genre Staphylococcus isolées dans la région de Bechar. PhytoChem & BioSub J. 2014;8(4):253-257.
  26. Fatehi N, Allaoui M, Berbaoui H, Cheriti A, Boulenouar N. Haloxylon Scoparium: An Ethnopharmacological Survey, Phytochemical Screening and Antibacterial Activity against Human Pathogens Causing Nosocomial Infection. PhytoChem & BioSub J. 2017;11(2):104.
  27. a) Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966;45(4):493-496. b) McFarland  J. Standardization of bacterial culture for disc diffusion assay, J Amer Med Assoc. 49, 1176.
  28. Chouikh A, Mayache B, Maazi MC, Hadef Y, Chefrour A. Chemical composition and antimicrobial activity of essential oils in xerophytic plant Cotula cinerea Del (Asteraceae) during two stages of development: flowering and fruiting. J Appl Pharma Sci. 2015;5(03):29-34.
  29. Boussoula E, Ghanmi M, Satrani B, Alaoui MB, Rhafouri R, Farah A, et al. Chemical quality, antibacterial and antifungal activities of Cotula cinerea essential oil from South Morocco. ESAIJ. 2016;12(5):209-216.
  30. Fournier G, Baghdadi H, Ahmed SS, Paris M. Contribution to the study of Cotula cinerea essential oil. Planta Med. 1989;55(6):580.
  31. Judzentiene A, Mockute D. The inflorescence and leaf essential oils of Tanacetum vulgare L. var. vulgare growing wild in Lithuania. Biochemical Systematics and Ecology. 2005;33(5):487-498.
  32. Habibi Z, Yousefi M, Shahriari F, Khalafi J, As’habi MA. Chemical composition of the essential oil of Tanacetum turcomanicum and T. canescens from Iran. Chemistry of Natural Compounds. 2009;45:93-95.
  33. Muller-Riebau FJ, Berger BM, Yegen O, Cakir C. Seasonal Variations in the Chemical Compositions of Essential Oils of Selected Aromatic Plants Growing Wild in Turkey. J Agric Food Chem. 1997;45:4821−4825.