Variation in Essential Oil Composition of Iris Nigricans Dinsm. (Iridaceae) Endemic to Jordan at Different flowering Stages
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Arabian Journal of Chemistry (2012) xxx, xxx–xxx King Saud University Arabian Journal of Chemistry www.ksu.edu.sa www.sciencedirect.com ORIGINAL ARTICLE Variation in essential oil composition of Iris nigricans Dinsm. (Iridaceae) endemic to Jordan at different flowering stages Hala I. Al-Jaber * Department of Applied Sciences, Faculty of Engineering Technology, Al-Balqa Applied University, 15008, Amman 11134, Jordan Received 15 July 2011; accepted 7 January 2012 KEYWORDS Abstract Fresh aerial blooming parts of Iris nigricans Dinsm. (Iridaceae) were collected at differ- Iris nigricans; ent flowering stages. The essential oils obtained from the fresh aerial parts and from the rhizomes at Iridaceae; the post flowering stage were investigated by gas chromatography–mass spectrometry (GC/MS) Essential oil; analysis. The essential oil at the pre-flowering stage was characterized by a high proportion of oxy- Monoterpenoids; genated monoterpenes (40.93%), while the oils obtained during the full flowering and post flower- a-Pinene; ing stages were dominated by aliphatic hydrocarbons and their derivatives (81.28%, 79.52%, b-Pinene respectively). The oil of I. nigricans rhizomes was dominated by monoterpene hydrocarbons (35.47%) of the pinane skeleton (32.05%), represented by a- and b-pinenes (24.05% and 7.99%, respectively). In addition, rhizome’s aqueous and ethanolic extracts were investigated for their anti- microbial activity and were found to be inactive against Escherichia coli, Klebsiella pneumonia, Salmonella typhi, Staphylococcus aureus, Bacillus subtilis and Micrococcus luteus. ª 2012 King Saud University. Production and hosting by Elsevier B.V. All rights reserved. 1. Introduction of its species, many of which are grown in parks and gardens as ornamental plants due to their beautiful flowers (Baytop, Iridaceae, taking its name from the genus Iris, is an interesting 1984). It is a large family of perennial herbaceous plants with plant family from the viewpoint of the diverse floral structures rhizomes or corms comprising nearly 1750 species in 82 genera (Devoto and Medan, 2008). Plants belonging to this family are * Tel.: +962 6 4790333/4790359x108; fax: +962 6 4790350. distributed world-wide in tropical and temperate regions of the E-mail address: [email protected] world, with the highest diversity occurring in South Africa, fol- lowed by South America, Europe and temperate regions of 1878-5352 ª 2012 King Saud University. Production and hosting by Asia (Bahali, 2006). In Jordan, Iridaceae family is represented Elsevier B.V. All rights reserved. by four genera including Gladulus, Crocus, Gynandriris and Iris. There are 13 Iris species (Al-Eisawi, 1982) occurring wild Peer review under responsibility of King Saud University. in Jordan. Iris nigricans Dinsm. commonly known as ‘‘Black doi:10.1016/j.arabjc.2012.01.003 Iris’’ is endemic to Jordan. Plants of the Iridaceae family have been used in the tradi- Production and hosting by Elsevier tional medicines to treat cold, flu, malaria, toothache, bruises and burns (Lin et al., 2002). The use of several Iris plant Please cite this article in press as: Al-Jaber, H.I. Variation in essential oil composition of Iris nigricans Dinsm. (Iridaceae) endemic to Jordan at different flowering stages. Arabian Journal of Chemistry (2012), doi:10.1016/j.arabjc.2012.01.003 2 H.I. Al-Jaber rhizomes in traditional medicine dates back in time, especially coarsely powdered and then hydrodistilled separately using a in European communities. The dry rhizomes of several Iris Clevenger apparatus for 3 h. The extraction was repeated twice species, particularly those of Iris germanica L., were collec- and the obtained oils were pooled separately, dried over anhy- tively used as ingredients of the toothpowders. The drug Rhi- drous sodium sulfate (Na2SO4) and stored at 4 °C in amber zoma iridis, which is composed of small selected rhizome glass vials until analysis. species, enjoyed popularity due to its emetic, cathartic, diure- For the preparation of the ethanol and aqueous extracts, tic, stimulant, expectorant and errhine properties in addition each 10 g plant material was refluxed with 100 ml solvent to its use in relieving pain as masticatory for teething children (70% ethanol or water, respectively), kept overnight, filtered, (Rigano et al., 2006; Wollenweber et al., 2003). Traditional solvent evaporated and dissolved in DMSO (1 g extract in European healers recommended the aqueous extracts of the 10 ml DMSO). The DMSO solutions were then used in biolog- I. germanica roots as an enema or topically rubbing the oil ical tests at different concentrations. on arthritic limbs (Adams et al., 2009). In Iran, medicinal plants belonging to the genus Iris are known as ‘‘Irsa’’ and 2.3. GC–MS and GC–FID analysis were known to have diuretic and expectorant properties at low doses and strong purgative and emetic properties at high About 1 ll aliquot of each oil sample, diluted to 5 llinGC doses. Also, ‘‘Irsa’’ was considered to be useful in the treat- grade n-hexane, was subjected to GC/MS analysis. The GC/ ment of pulmonary, liver and uterus diseases as well as in MS analysis was performed using Varian Chrompack the treatment of hemorrhoids (Ayatollahi et al., 2004). CP-3800 GC/MS/MS-200 (Saturn, Netherlands) equipped I. nigricans is an elegant endemic perennial herb, 20–30 cm with DP-5 (5% diphenyl, 95% dimethyl polysiloxane) GC cap- long with underground rhizomes. The leaves are 0.5–1.0 cm illary column (30 m · 0.25 mm i.d., 0.25 lm film thicknesses), wide, 10–15 cm long forming a tuft, curved, basal leaves, flat- with helium as a carrier gas (flow rate 0.9 ml/min). The actual tened, canal-like arranged in two opposite rows. The plant has temperature in MS source was 180 °C and the ionization volt- one green flower stalk rising from a rhizome and bearing a sin- age was 70 eV. The column temperature was kept at 60 °C for gle flower. Usually the leaves are much shorter than the stalk 1 min (isothermal), and programmed to 246 °C at a rate of of the flower. The flowers are 12–15 cm in diameter, 3 °C/min, and kept constant at 246 °C for 3 min (isothermal). black-dark lilac, glossy with transparent veins (Al-Eisawi, A hydrocarbon mixture of n-alkanes (C8–C20) was analyzed 1998). Recognized as the flower emblem of Jordan, this plant separately by GC/MS under the same chromatographic condi- is known to grow wild in marginal land spots and mountains tions using the same DP-5 column. of Amman, Madaba and Karak. The flowering period of this For the quantitative analysis (% area), a Hewlett–Packard plant is short, extending from April to May (Al-Eisawi, 1998). HP-8590 gas chromatograph equipped with a split–splitless Phytochemical investigations on this plant were very lim- injector (split ratio 1:50) and an FID detector was used. The ited, most of them dealt with determining the secondary column was an optima-5 (5% diphenyl, 95% dimethyl poly- metabolites of the rhizomes. Two new isoflavones named nig- siloxan) fused silica capillary column (30 m · 0.25 mm, 0.25 ricin and nigricanin were isolated from I. nigricans rhizomes film thickness). The temperature of the oven was increased at (Al-Khalil et al., 1994). In addition, the ethanol extracts of I. a rate of 10 °C/min from 60 to 250 °C and then held constant nigricans rhizomes afforded seven xanthones (Al-Khalil at 250 °C for 5 min. The temperatures of the injector and et al., 1995). A careful literature survey revealed that the essen- detector were maintained at 250 and 300 °C, respectively. tial oil of I. nigricans has not been studied before. Thus, this The relative peak areas of the oil components were measured current investigation was aimed to study the composition of and then used to calculate the concentration of the detected the essential oil of the aerial blooming parts of I. nigricans at compounds. Each sample was analyzed twice. different flowering stages and the composition of the oil of the rhizomes at the post flowering stage. In addition, the anti- 2.4. Identification of the components microbial activities of the ethanol and aqueous extracts ob- tained from the rhizomes were evaluated. The components of the essential oils at different flowering stages were identified using the built in libraries (Nist Co. and Wiley Co., USA) and by comparing their calculated reten- 2. Experimental tion indices relative to (C8–C20) n-alkanes literature values measured with columns of identical polarity (Adams, 2001), 2.1. Plant material or with authentic samples. The compounds, a- and b-pinenes, p-cymene, limonene, linalool (Fluka, Buchs, Switzerland) and The aerial blooming parts of I. nigricans at the pre-flowering, sabinene hydrate (Sigma–Aldrich, Buchs, Switzerland) were flowering, and post flowering stages and the rhizomes at the post used as reference substances in GC/MS analysis. GC-grade flowering stage were collected from Shafa-badran, Amman, hexane and analytical reagent grade anhydrous Na SO were during the period extending from March to April, 2011. A vou- 2 4 purchased from Scharlau (Barcelona, Spain) and UCB cher specimen (BAU/011/II 3010) has been deposited in the (Bruxelles, Belgium). Department of Applied Sciences, Faculty of Engineering Tech- nology, Al-Balqa Applied University, Amman, Jordan. 2.5. Antibacterial activity 2.2. Preparation of the volatile oils and extracts Overnight bacterial cultures of Gram-negative bacteria, Esch- erichia coli (ATCC 8739), Klebsiella pneumoniae (ATCC Each 340 g of fresh blooming aerial parts at different flowering 10031), Salmonella typhi (ATCC 6539) and Gram-positive stages and the rhizomes during the post flowering stage, was bacteria, Staphylococcus aureus (ATCC 6538P), Bacillus subtilis Please cite this article in press as: Al-Jaber, H.I.