Agriculture, active compounds investigation of cola herb (Artemisia abrotanum L.) recently introduced in Iraq.
Different extracts of medicinal plants have evoked interest as sources of natural products, their potential uses as alternative remedies for treatment of many infection diseases.  Artemisia is a genus of small herbs and shrubs found in northern temperature regions; it belongs to the important family compositae (Asteraceae).  Artemisia abrotanum its semi-evergreen subshrub in southern europe and hardy perennial in the eastern half of the United states and it reach from 3 to 5 feet in height. [3,4] A. abrotanum was traditionally considered as an antiseptic, astringent, emmenagogue, expectorant, febrifuge, stomachic, stimulant, tonic, anti-inflammatory, rermifuge, spasmolytic and used for treating upper respiratory tract disease. It's used against cancer, cough, fever, and tumors.  This type of Artemisia was introduced recently to Iraq for ornamental purposes without any study about adaptation this plant for environmental conditions of semi-arid of Iraq and no studies about active compounds and pharmacognostical characters of this plant, therefore, this study conducted to agriculture and investigation of active compounds of this plant.
MATERIALS AND METHODS
Plantation of Plant
The field experiment was conducted to evaluation the A. abrotanum plant adaptation of growing under semiarid conditions of Iraq during 2015-2016 growing season. The field experiment was conducted at the medicinal plants garden of College of Pharmacy of Al-Mustansiriyah University at Baghdad location. After preparation of soil to plantation, 2 months old seedlings with an average height of 10 cm were used. During growing season NPK fertilizer (18, 10, 16) was added 1 month after transplanting at 250 kg/ha. The plot area was 2 x 3 [m.sup.2], leaving 50 cm between plants and 50 cm between rows (Figure 1).
Collection of Plant Samples
Samples of plant collected at pre-flowering stage and 50 cm above soil level, samples were dried at room temperature 25[degrees]C in a laboratory room. The plant identification by national herbarium of the agriculture ministry.
Fresh specimens of A. abrotanum plant were used to study the morphological character of the plant such as the shape of leaves, stems, and margins.
Shade dried leaves were finely powdered and examined under microscope. Small quantity of the powder was placed on the slide which then was mounted two drops of chloral hydrate and covered with a cover slip and examined under microscope. Different cell components were observed and photography was done using digital camera.
The lower and upper epidermal layer of fresh leaf (in fragments) were mounted in chloral hydrate and observed under a microscope. Determination of the components of leaves (stomata, trichomes, and stomatal index) were carried out under a microscope. The stomatal index was carried out using the following equation. 
Stomatal index = [Number of Stomata/Number of Stomata+Number of epidermalcells]x100
The preliminary phytochemical study of ethanolic areal parts extract of plant was carried out by standard methods of phytochemical screening such as mayers, dragendroff's, borntrager's test, for alkaloids, and glycosides. The foam test, lead acetate test, ferric chloride test, alkaline test and Salkowski test were used for saponin, tannins, flavonoids, and terpenoids examination extracted using a clevenger - type apparatus.  The distillation was carried out with 50 g of plant material and 300 ml distilled water. Series of distillation, each lasting exactly 1 h respectively. [8-12]
Essential Oil Extraction
Essential oil was, from 1 h to 5 h. the essential oil yield percentage was measured, and the oil was collected, dried over by anhydrous sulfate [Na.sub.2][So.sub.4] and stored at -5[degrees]C in 2 ml vials for further analysis.
The extracted oil has been mixed with n-hexane, injected into gas-liquid chromatography using an auto-sample and the different compounds have been separated on an HP-INOWOAX (60 x 0.25 x 0.25 mm) capillary column. Helium was used as carrier gas (flow rate 1.5 m/[min.sup.-1]. The temperature program was 35-230[degrees]C (2.5[degrees]C/min.) in course of time (92 mm), injector temperature was 205[degrees]C and flame ionization detector used, area percentage were obtained using a PC programmer (maestro chromatograph data system), for identification of single compound internal and external standard substances have been used, the external standard was obtained from Oma company for chemical compounds.
The results of field experiment of plant were referred to the adaptation of A. abrotanum to the environmental condition of Iraq (semi-arid condition). The adaptation of this plant may be belong to have this plant fine glandular hairs and wooly hairs which covered the leaves, that useful in preventing the water losing by transpiration and evaporation from leaves surface. Another reason of its adaptation may be belong to possibility of this plant to grow in clay-sandy soil of Iraq. Maybe belong to short, and crowded leaves of this plant, that lead to a reduction of water lose by both transpiration or evaporation from leaves surfaces. 
The results of macroscopical examination of the plant were showed that it was semi-erect, branched and green, up to 45 cm in height. Stems were angular rarely slender, green. The results were showed some typical xeromorphic features in the stem and leaf structure, which are more strongly. Microscopical examination of the leaf showed the presence of anomocytic stomata in which the guard cells are surrounded by epidermal cell that are not distinguishable from other epidermal cells as shown in Figure 2. Multicellular unbranched trichomes were represented. Furthermore, the microscopic examination was showed about 16.6% as stomatal index of upper surface while 25% for lower surface of leaves shown in Table 1.
The results of phytochemical screening were presented in Table 2 and these results were referred to the cola herb was contained different active compounds such as saponin, tannin, coumarin, and flavonoids.
The results of this study were referred to the percentage of volatile oil of this plant was reached to 2.4%. The values of specific gravity, oil density, and refractive index were reach to 0.98. 0.66 mg/ml and 1.5 mg degree, respectively. In addition, the results were obtained in Table 3 referred to 13 compounds of volatile oil of cola herb leaves. The most main compounds of volatile oil were soloinene (21.458%), limonene (14.368%), myrecen (13.634%), camphene (12.680%), thusene (4.163%), [alpha]-pinene (4.106%), [alpha]-terpinone (2.841%), trycyclene (1.541%), terpinolene (0.843%), [beta]-pinene (0.515%), [beta]-phellanderne (0.056%), and [beta]-ocimene (0.433%), respectively. These results were gave best indication for the possibility of this plant to grow under semi-arid conditions at middle region of Iraq and need more studies about active compounds and medical activity of these compounds. In addition, the results of quality and quantity of volatile oil of this plant encourage to increasing the different field experiment to obtain more quality and quantity of volatile oil.
The good adaptation of this plant for a semi-arid condition such as higher temperature with dry climate with very important active compounds of this plant lead to increasing the research or isolation of active compounds of this plant.
The authors would like to thank Al-Mustansiriyah University (www.uomustansiriyah.edu.iq) Baghdad - Iraq of its support in the present work.
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Ibrahim S Aljubory, Thamer Mouhi Jasiem, Sara K Baderden
Department of Pharmacognosy and Medicinal Plants, College of Pharmacy, Al-Mustansiriyah University, Baghdad, Iraq
Correspondence to: Thamer Mouhi Jasiem, E-mail: firstname.lastname@example.org
Received: October 01, 2016; Accepted: January 02, 2017
Table 1: Stomatal index Number of stomata Number of Stomatal index (%) epidermal cell Upper surface 2 10 16.6 Lower surface 4 12 2 Table 2: Results of phytochemical screening of cola herb Test Result Saponin + Tannin + Coumarin + Flavonoid + Table 3: Components of volatile oil of A. abrotanum L. which analysis by GLC Diagnosis RT Area Conservation (%) Trycyclene 5.696 504243 1.541 Thusene 7.833 1361801 4.163 [beta]-pinene 8.435 1343253 4.106 Camphone 8.975 4147914 12.68 [alpha]-pinene 10.048 178420 0.515 Soloinene 12.208 7010503 21.458 Myrecen 14.877 4459878 13.634 [alpha] terpinone 15.002 929273 2.841 Limonene 15.725 4700010 14.368 [beta]_phelladreene 16.578 18349 0.056 [beta]_ocimene 16.848 141527 0.433 Terpinolene 17.847 275681 0.843 [beta] thyjone 18.537 23317 0.071 GLC: Gas liquid chromatography, RT: Radiotherapy, A. abrotanum: Artemisia abrotanum
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|Title Annotation:||RESEARCH ARTICLE|
|Author:||Aljubory, Ibrahim S.; Jasiem, Thamer Mouhi; Baderden, Sara K.|
|Publication:||National Journal of Physiology, Pharmacy and Pharmacology|
|Date:||Apr 1, 2017|
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