Essential Oil Composition and Antibacterial Activity of Juniperus oxycedrus ssp. macrocarpa (S. et Sm.) BalI. Growing in Oum El Bouaghi (Semi- arid area), Algeria

Leaf essential oils of Juniperus oxycedrus (Cupressaceae) wild grown in the region of Oum El Bouaghi (semiarid area) in Algeria have been analysed by gas chromatography/mass spectrometry (GC/MS). Fifty seven compounds were identified in the leave oils. The leaf oils were mainly composed of 5-Tetradecen-1-ol, acetate, (Z)-(12.9%) ç-Muurolene (9.1%), α-Cadinol (5.1%) (ñ)-Cadinene (3.9 %) and some other compounds which were only present in minor amounts. Aims: This study aimed to extracted, identification of essential oils of Leaves of J. oxycedrus L. ssp. macrocarpa (S. et Sm.) BalI. growing in Oum El Bouaghi (semiarid area), Algeria and evaluation of their antibacterial capacity. Results: The GC/MS analysis of the Leaves of J. oxycedrus L. (yielded 0, 36 %) permitted the identification of fifty seven components. The composition and percentage of the compounds are listed by their order of retention times. The main constituents of the essential oil were composed of 5-Tetradecen-1-ol, acetate, (Z) (12.9%) ç-Murolene (9.1%), α-Cadinol (5.1%) (ñ)-Cadinene (3.9 %) and some other compounds were only present in minor amounts. The antimicrobial activity of the Original Research Article Basa et al.; JPRI, 34(20A): 46-52, 2022; Article no.JPRI.84225 2 essential oils were evaluated by the disc diffusion method and tested against Gram-negative Escherichia coli, Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus bacteria. Results showed that Staphylococcus aureus was the highly resistant to the essential oil. Conclusion: The results of analysis of the components Leaves of Juniperus oxycedrus ssp. macrocarpa (S. et Sm.) BalI. growing in Oum El Bouaghi (semiarid area), Algeria, permitted the identification of fifty seven components. The essential oil showed that the essential oil of juniperus oxycedrus has the potential to act as an antibacterial agent.


INTRODUCTION
Essential oils are aromatic oily liquids, volatile, characterized by a strong odour, rarely coloured, and generally less dense than water. They can be synthesized by all plant organs (flowers, buds, seeds, leaves, twigs, bark, herbs, wood, fruits and roots) and therefore extracted from these parts, where they are stored in secretory cells, cavities, canals, epidermic cells orglandular trichomes [1]. Essential oils have a complex composition, containing from a dozen to several hundred components. The great majority of components identified in essential oils includes terpenes (oxygenated or not), with monoterpenes and sesquiterpenes prevailing. Nevertheless, allyl-and propenylphenols (phenylpropanoids) are also important components of some essential oils [2]. The wellknown families rich in essential oil are Apiaceae, Asteraceae, Hypericaceae, Lamiaceae, Lauraceae, Myrtaceae, Pinaceae, Piperaceae, Rutaceae, Santalaceae, Zingiberaceae, Zygophyllaceae and Cupressaceae [3,4].
The genus Juniperus (Cupressaceae) consists of approximately 67 species and 28 varieties. The genus is divided into three sections: Caryocedrus Edlicher (with only one species); Juniperus (syn: Oxycedrus Spach with 12 species) and Sabina (Miller) Spach (with 55 species) [5]. The genus Juniperus (Cupressaceae) is represented in the flora of Algeria by five species, namely J. Oxycedrus L.,J. Sabina L., J. thurifera L., J. phoenica L. and J. communis L., [6]. Juniperus oxycedrus is a shrub or small tree growing wild in stony places of the Mediterranean and Near East countries [7]. In folk medicine J. oxycedrus is used for the treatment of various diseases, such as hyperglycaemia, obesity, tuberculosis, bronchitis and pneumonia [8]. Leaves and stems of J. oxycedrus ssp. macrocarpa have been found to reduce the blood pressure of normotensive rats, to inhibit the response to histamine, serotonin and acetylcholine, and to exhibit significant antiinflammatory activity [9].

Extraction
Essential oils were obtained by hydrodistillation of 100g of dried leaves using a Clevenger-type apparatus for 3 h. The oil was dried over anhydrous sodium sulphate and stored in sealed vials protected from the light at −4 °C before analyses. The oil sample was subsequently analyzed by GC-MS.

Gas chromatography/mass spectrumetry (GC/MS)
The oil was analyzed by GC/MS using an Agilent 5973EI mass selective detector coupled with an Agilent GC6890A gas chromatograph, equipped with a cross-linked 5% PH ME siloxane HP-5MS capillary column (30m x 0.32mm, film's thicknesses 0.25μm). Operating conditions: The carrier gas flow was 1.6 ml He/min, column pressure was 100 Kpa. The injector and detector temperatures were 220°C and 250°C respectively. The column temperature was held at 60°C for 1 min, then raised from 60°C to 200°C at 10°C/min and held there for 5 min and from 200°C to 240°C at 10°C /min and held there for 6 min. The program was run in the splitless mode with a mass range of 50-400 u, and the scan interval was 0.5 s. Detector voltage was set at 1.5 kV (Table 1).

Identification of components
Identification of oil components was achieved on the basis of their retention times Rt, and by comparison of their mass spectral fragmentation patterns with those reported in the literature and stored on the MS library (NIST database). The concentration of the identified compounds was computed from the GC peak total area without any correction factor.
The composition of leaf essential oils of section Juniperus is generally much simpler and dominated by simple monoterpenes, in contrast to the essential oils of section Sabina, were oxygenated monoterpenes (e.g. camphor) and sesquiterpenes (e.g. cadinols, cedrol) are the major constituents [9,10].
It is renowned that the genotype, organ, season of collection, and geographic position Climatic conditions the applied extraction technique have a considerable effect on the composition [18,19].

Antibacterial Activity
Although some volatile compounds have proven to be effective against microorganisms, many studies are still based on the effect of the presence of some phenolic substances and oxygen compounds [1].The diffusion test was applied to three microorganisms including Grampositive, -negative bacteria. The results are summarized in Table 5

CONCLUSION
Our study by GC/MS on Juniperus oxycedrus leaves growing in Oum El Bouaghi (Algeria) showed the presence of 57 compounds of the essential oil. The antimicrobial activity of the essential oils was tested against 3 bacteria: Staphylococcus aureus, Escherichia coli and Pseudomonas aerugenosa through diffusion test disk. The anti bacterial activity showed that Staphylococcus aureus was highly resistant and the essential oil may represent potential source of antibacterial agents.

DISCLAIMER
The products used for this research are commonly and predominantly use products in our area of research and country. There is absolutely no conflict of interest between the authors and producers of the products because we do not intend to use these products as an avenue for any litigation but for the advancement of knowledge. Also, the research was not funded by the producing company rather it was funded by personal efforts of the authors.

NOTE
This research contributes to valuing effective natural resources and using them in the pharmaceutical industries in a scientific manner, as much as possible away, from the traditional methods common in many regions in the world.

CONSENT
It is not applicable.

ETHICAL APPROVAL
It is not applicable.