International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 22 Comparative analysis of essential oil components of two Daucus species from Algeria and their antimicrobial activity Meliani Nawel, Dib* Mohammed El Amine, Allali Hocine and Tabti Boufeldja Laboratoire des Substances Naturelles et Bioactives (LASNABIO) Département de Chimie, Faculté des Sciences, Université de Tlemcen, ALGERIAAvailable online at: www.isca.in Received 14th October 2012, revised 22nd October 2012, accepted 24th October 2012Abstract The essential oils obtained by hydrodistillation from leaves and stems of Algeria native, Daucus carota L. subsp. carota and Daucus carota L. subsp. gummifer were analyzed by GC and GC–MS. In total, 67 compounds, which accounted for more than 90.0 g/100g of the total composition of the oils, have been identified. -pinene (26.0-34.1 g/100g), sabinene (1.5-14.0 g/100g ), limonene (0.5-13.0 g/100g), -pinene (0.6-11.2 g/100g), myrcene (10.0-13.1 g/100g), terpinene-4-ol (2.4-4.9 g/100g), caryophyllene oxyde (0.8-6.0 g/100g), spathulenol (0.6-4.3 g/100g), p-cymene (3.3-4.4 g/100g) and isiospathulenol (0.2-3.8 g/100g) have been identified as the main components of both essential oils. The comparison of essential oil components of Daucus species between the present and previous work indicated that the composition of oils vary greatly with respect to the plant parts used, geographical locations and stage of development, mainly for the proportion of terpenoids and phenylpropanoids. In addition, we reported for the first time, the antibacterial activity of D. carota L. subsp. gummifer essential oil against eight bacteria. The most prominent inhibitory action of this essential oil was observed against Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus and Escherichia coli strains at a concentration of 2.5 mg/ml. The essential oil compositions of two Daucus samples can therefore be proposed as new potential sources natural for pharmaceutical industries. Keywords: D. carota L. subsp. gummifer; D. carota L. subsp. Carota; Essential oils; Antimicrobial activity; Comparative analysis. Introduction Essential oils as antimicrobial agents are recognized as safe natural substances to their users and for the environment and they have been considered at low risk for resistance development by pathogenic microorganisms1,2. Nowadays, the interests for essential oil extracted from aromatic plants are multiple and diversified. Based on their therapeutic properties and the chemical substances isolated from their volatile parts, it may allow further application in particular biological activities. Apiaceae represent one of the best-known plant families, widely distributed in temperate climate regions where they are often used as spices, vegetables or drugs owing to the presence of useful secondary metabolites such as essential oils. Essential oils of Apiaceae family have been widely used throughout history for their pharmacological activities, such as antibacterial, antifungal, antiviral, antiparasitic, insecticidal and antispasmodic5,6. Today they are being used in pharmaceutical, sanitary, cosmetic, agricultural and food industries. D. carota L. is an Apiaceae native from Europe, Asia, Africa and Macaronesia. Traditional medicine uses D. carota extracts for hepatic and renal insufficiency as well as for skin disorders, e.g. burns and furunculous. Various compositions of D. carota L. subsp. carota and D. carota L. subsp. gummifer essential oils have been reported, characterized by the occurrence of a main component, -pinene (up to 51%), sabinene (up to 60%), geranyl cetate (up to 76%), elemicin (up to 35%), Emethylisoeugenol (up 30%) and carotol (up to 66%)8-18 (table 2), but they didn’t give homogeneous results. These may be due to the genetic, plant parts used, geographical locations, season at the time of collection and analytical methods However, only a few studies have been reported on the antimicrobial activity of the investigated essential oils10,12,16. As far as we were concerned, the chemical composition of D. carota L. subsp.carota and D. carota subsp. gummifer essential oils was never reported in Algeria. The aim of the present study was to investigate the qualitative and quantitative chemical composition (compared with the results reported in the previous studies) and to assess the in vitro antimicrobial activities of essential oils obtained from aerial parts of D. carota L. subsp.carota and D. carota L. subsp. gummifer. To the best of our knowledge, this is the first study to compare the chemical composition of these both subspecies and evaluate the antimicrobial activity of D. carota L. subsp. gummifer from Algeria. Material and MethodsPlant material and Oil isolation: The aerial parts (stems and leaves) of D. carota L. subsp. carota (DCSC) and D. carota L. subsp. gummifer (DCSG) were harvested from Bensekrane forest area (at about 30 km north west of Tlemcen - Algeria) in International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 23 July, 2009, and from Saf-Saf village at about 5 km north-Est of Tlemcen – Algeria, respectively. The plant material was botanically identified by Pr. Noury Benabadji (Laboratory of Ecology and Ecosystem Management of University of Tlemcen Algeria). The fresh aerial parts were submitted to hydrodistillation from 5 h using a Clevenger-type apparatus according to the European Pharmacopoeia19. Gas chromatography: GC analyses were carried out using a Perkin Elmer Clarus 600 GC apparatus (Walhton, MA, USA) equipped with a single injector and two flame ionization detectors (FID). The apparatus was used for simultaneous sampling to two fused-silica capillary columns (60 m x 0.22 mm, film thickness 0.25 m) with different stationary phases: Rtx-1 (polydimethylsiloxane) and Rtx-Wax (polyethylene glycol). Temperature program: 60 to 230°C at 2°C.min-1 and then held isothermal 230°C (30 min). Carrier gas: helium (1 mL.min-1). Injector and detector temperatures were held at 280°C. Split injection was conducted with a ratio split of 1:80. Injected volume: 0.1 L. Gas chromatography-mass spectrometry: The oils and the fractions obtained by CC were investigated using a Perkin Elmer TurboMass quadrupole analyzer, directly coupled to a Perkin Elmer Autosystem XL equipped with two fused-silica capillary columns (60 m x 0.22 mm, film thickness 0.25 m), Rtx-1 (polydimethylsiloxane) and Rtx-Wax (polyethylene glycol). Other GC conditions 146 were the same as described above. Ion source temperature: 150°C; energy ionization: 70 eV; electron ionization mass spectra were acquired with a mass range of 35-350 Da; scan mass: 1s. Oil injected volume: 0.1 L, fraction injected volume: 0.2 L. Component identification and quantification: Identification of the components was based i. on the comparison of their GC retention indices (RI) on non polar and polar columns, determined relative to the retention time of a series of n-alkanes with linear interpolation, with those of authentic compounds or literature data20; and ii. on computer matching with commercial mass spectral libraries20,21 and comparison of spectra with those of our personal library. The quantification of the collective essential oil components was carried out using peak normalization including response factors (RFs) with internal standard22. Component quantification was expressed in g/100g. Tridecane was introduced in all sample oils at same concentration (0.7 g/100 g) as internal standard. Test microorganisms: Essential oils were tested against eight microorganisms, including Gram positive Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalisATCC 29212, Listeria monocytogenes ATCC 7644, Bacillus cereus ATCC 9634, Gram negative Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 10031. Biological screening: Antimicrobial activity was evaluated using a broth microdilution method. A dilution agar method was used to determine the Minimum Inhibitory Concentrations (MIC). Stock solutions were obtained by dissolving extracts in dimethylsulfoxide (DMSO 1%). Serial dilutions were made to obtain concentrations ranging from 0.2 to 10 mg/ml of the essential oil. Each mixture was added to Mueller–Hinton agar for bacteria23,24. The Petri dishes contained a sterile solution of DMSO and the culture medium, respectively and incubated at 37°C. Tetracycline served as positive control, while the solvent (10% aqueous DMSO) was used as a negative control. The experiments were performed in triplicate. According to the MIC values expressed in mg/mL, results were appreciated as follows: not sensitive (-) for value higher to 6 mg/mL, moderately sensitive (+) for value between 3.0 and 5.0 mg/mL and sensitive (++) for value to 2.5 mg/mL. Results and DiscussionEssential oil composition: GC and GC–MS analysis of DCSC and DCSG essential oils allowed to identify 67 components, 40 of which were common to both the two essential oils, which accounted for more than 90.0 g/100g of the oils. All components were identified by comparison of their EI–MS and GC-retention indices and mass spectra with those of our laboratory produced “Arômes” library, except three compounds which were identified by comparison with spectral data and retention indices from the literature (table -1). The essential oil yields, calculated from fresh material for DCSC and DCSG were 1.52 and 1.64 %, respectively (Table -1). The oil of DCSC made up the higher contribution of monoterpene hydrocarbons (83.2 g/100g), pinene (26.0 g/100g), sabinene (14.0 g/100g), limonene (13.0 g/100g), -pinene (11.2 g/100g), myrcene (10.0 g/100g), p-cymene (4.4 g/100g) and terpinene-4-ol (4.9 g/100g) were found as the characteristic components of this fraction in DCSC essential oils. However, this fraction was also the main one in the oil of DCSG (54.0 g/100g) with -pinene (34.1 g/100g), myrcene (13.1 g/100g) and p-cymene (3.3 g/100g) as the most abundant compounds. The oxygenated monoterpenes represented 8.6 g/100g of the total oil of DCSC. This fraction had a similar percentage (8.2 g/100g) in the essential oil of DCSG. In both cases, this fraction was dominated by terpinene-4-ol, which was more abundant in DCSC (4.9 g/100g) than that in DCSG (2.4 g/100g). The amount of oxygenated sesquiterpenes of DCSG (20.5 g/100g) was approximately tenfold as much as that of DCSC (2.0 g/100g). Caryophyllene oxyde (6.0 g/100g), spathulenol (4.3 g/100g) and isospathulenol (3.8 g/100g) were the main oxygenated sesquiterpenes in DCSG. The sesquiterpene hydrocarbons are present in small amounts and constitute about 6.4 and 1.2 g/100g of the oil composition in DCSC and DCSG, respectively. The content of the non-terpenic compounds of DCSG essential oil (1.0 g/100g) was more than that of DCSC (0.5 g/100g). This fraction was dominated by nonane (DCSG 0.5 g/100g) and 2-methyl butyl isovalerate (DCSG 0.3 g/100g, DCSC 0.2 g/100g). The phenylpropanoids compounds were the lowest component in International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 24 DCSC (0.2 g/100g). In, trans-methyl isoeugenol (0.1 g/100g) and elemicin (0.1 g/100g) were the only phenylpropanoids compounds. However, DCSC oil is characterized by the absence of phenylpropanoids compounds (table -1). Compared with previous studies: Compared the results obtained from our plant material of DCSC and DCSG with those previous reports on the same species from other localities8-18, it is interesting to point out that there are some qualitative and quantitative difference between the present work and those studies (table -2). Relative to the literature, it appears that Algerian DCSG essential oil exhibited chemical composition that differed from that of Spain origin, while was rather similar in comparison with that reported in the literature from Italy. The geranyl acetate (51.7-76.9%) was identified as major component in the spanish DCSG essential oil. Sabinene (26.8-60.6%) and -pinene (10.8-12.2%) were the major components of DCSGessential oil from Italy. However, -pinene (34.1 g/100g) and sabinene (13.1 g/100g) were the most important features of our essential oils. Other hand, the composition of DCSC essential oils (monoterpene hydrocarbons) was rather similar in comparison with those reported in the literature from Serbia, Poland and Lithuania16-18. Contrarily, the composition of DCSC essential oils was rather differed in comparison with those reported in the literature from Corsican, Turkey, Portugal, Italy and Tunisia10-14. Contrarily, the composition of DCSC essential oils was rather differed in comparison with those reported in the literature from Corsican, Turkey, Portugal, Italy and Tunisia10-14. Indeed, DCSC essential oils from Corse was largely dominated by (E)-methylisoeugenol (21.8-33.0%) and elemicin (11.4-16.3%). However, carotol (48.0-55.7%) and elemicin (31.5-35.3%) were the major components of DCSC essential oil from Tunisia. On one hand, some compounds, mainly including carotol, 11-H-himachal-4-en-1--ol, daucene and eudesm-7(11)-en-4-ol were present with higher amount (up 66%, 21%, 8% and 8%, respectively) in the essential oils of DCSC from Italy, Tunisia and Portugal11,12, whereas they were not identified in our essential oils (table -2). Generally, the observed differences in chemical composition of the various oils, when compared with those reported in previous studies, mainly for the proportion of terpenoids and phenylpropanoids could be due to a number of factors. Such factors may include differences in climatic conditions, plant parts used, geographical locations, season at the time of collection, stage of development and processing of plant materials before extraction of the oils. The phenomenon existed in many plants, and was reported by many previous studies, such as the genus Hypericum25. These showed that variations were sufficient to allow the distinction of different chemotypes that were the results of an adaptative process to particular ecologic conditions. Antibacterial activity: The antimicrobial activity of DCSC and DCSG essential oil was evaluated against 8 bacterial strains. The antibacterial effect is presented in table-3. The essential oil of DCSG shown higher activity against bacteria than the DCSCessential oil. Essential oil obtained from DCSG, exhibited activity against six bacterial strains, with MIC values ranging from 2.5 to 5 mg/ml. The most prominent inhibitory action of this essential oil was observed against three gram-positive bacterium (L. monocytogenes, B. cereus and S. aureus) and one gram-positive bacterium (E. coli) at 2.5 mg/ml.However, no inhibition was showed against B. subtilis and E. faecalis up to the value of 6 mg/mL. DCSC essential oil had, in most cases, moderate antibacterial activity, showing inhibitory effect at 3.8-5.1 mg/ml against B. cereus, L. monocytogenes and S. aureus(MIC = 3.8, 4.3 and 5.1 mg/ml, respectively) (table 3).It has been reported that the flowering umbel, and mature umbel oils from DCSC (collected in Poland), previously tested for inhibitory effect on microorganisms (S. aureus, B. subtilis, and C. albicans) were dominated by -pinene (17-42%) and sabinene (19-40%)16. On the other hand, oil sample, obtained from aerial parts of DCSC growing wild in Corsica, at the end of flowering stage, containing mainly (E)-methylisoeugenol (21.8%), -bisabolene (21.3%), elemicin (16.3%) and -pinene (15.9%) were reported as antimicrobial against the human enteropathogen Campylobacter10. The acquired results confirm and supplement present findings about antibacterial characteristics of Apiaceae. ConclusionThe findings showed that both Daucus had a considerable variation in the essential oil composition and this study also demonstrated the occurrence of -pinene chemotype in D. carota ssp gummifer from Algeria. Both oils were the complex mixture of volatile compounds, most of which had antiseptic, anti-inflammatory and antimicrobial properties. Other hand, according to the antimicrobial results gram negative bacteria were the most resistant and the gram positive bacteria exhibited good sensitivity. However, further work is necessary to explore the efficacy of these essential oils as an antibacterial agents. AcknowledgmentsThe authors are grateful to Prof. Noury Benabadji (Botanical Laboratory, Biology Department, Aboubekr Belkaïd University) for the identification of the vegetable matter and are thankful to Professor Jean COSTA, Director of Laboratory of Chemistry of Natural Products, University of Corse, France for providing facilities for this work. 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S., Tchobo F., Yèhouénou B., Menut C. and Sohounhloué D., Chemical composition and Biological activities of the Essential oil extracted from the Fresh leaves of Chromolaena odorata (L. Robinson) growing in Benin, I. Res. J. Biological Sci., 1(3), 7-13 (2012) 4.Bakkali F., Averbeck S., Averbeck D. and Idaomar, M., Biological effects of essential oils – A review, Food Chem Toxicol., 46(2), 446-455 (2008)5.Olle M. and Bender I., The content of oils in umbelliferous crops and its formation, Agronomy Res., , 687-696. (2010)6.Teuscher E., Medicinal spices. 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Agric Food Chem., 55, 7332-7336 (2007) 11.Marzouki H., Khaldi A., Falconieri D., Piras A., Marongiu B., Molicotti P. and Zanetti S., Essential Oils of Daucus carota subsp. carota of Tunisia Obtained by Supercritical Carbon Dioxide Extraction, Natur Prod Commun., , 1955-1958 (2010) 12.Maxia A., Marongiu B., Piras A., Porcedda S., Tuveri E., Gonçalves M.J., Cavaleiro C. and Salgueiro L., Chemical characterization and biological activity of essential oils from Daucus carota L. subsp. carota growing wild on the Mediterranean coast and on the Atlantic coast, Fitoterapia, 80, 57-61 (2009)13.Gonny M., Bradesi P. and Casanova J., Identification of the components of the essential oil from wild Corsican Daucus carota L. using 13C-NMR spectroscopy, Flav Fragr j., 19, 424-433 (2004) 14.Özcan M.M. and Claude Chalchat J.C., Chemical composition of carrot seeds (Daucus carota L.) cultivated in Turkey: characterization of the seed oil and essential oil, Grasas y aceites., 58 (4), 359-365 (2007)15.Kula J., Izydorczyk K., Czajkowska A. and Bonikowski R., Chemical composition of carrot umbells oils from Daucus carota ssp. sativus cultivated in Poland, Flav Fragr J., 21, 667-669 (2006) 16.Staniszewska M., Kula J., Wieczorkiewicz M. and Kusewicz D., Essential oils of wild and cultivated carrots-the chemical composition and antimicrobial activity, J Essent Oil Res., 17, 579-583 (2005) 17.Sokovi M., Stojkovi D., Glamolija J., iri A., Risti M. and Grubiši D., Susceptibility of pathogenic bacteria and fungi to essential oils of wild Daucus carota, Pharmaceutical Biology., 47(1), 38-43 (2009) 18.Mockute D. and Nivinskiene O., The sabinene chemotype of essential oil of seeds of Daucus carota L. ssp. carotagrowing wild in Lithuania, J. Essent. Oil Res., 16, 277-281 (2004)19.Conseil de l’Europe. Pharmacopée Européenne, Maisonneuve S.A.: Sainte Ruffine., (1996)20.König W.A., Hochmuth D.H. and Joulain D., Terpenoids and Related Constituents of Essential Oils. Library of MassFinder 2.1 University of Hamburg, Institute of Organic Chemistry, Hamburg., (2001)21.Adams R.P., Identification of Essential Oil Components by Gas Chromatography/Quadrupole Mass Spectroscopy. 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Sci., , 99-104 (2012) 25.Hosni K., Msaâda K., Ben Taârit M., Ouchikh O., Kallel M. and Marzouk B., Essential oil composition of Hypericum perfoliatum L. and Hypericum tomentosum L. growing wild in Tunisia, Ind. Crops Prod., 27, 308-314 (2008) International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 26 Table -1 Essential oils of D. carota L. Subsp. carota (DCSC) and D. carota L. Subsp. gummifer (DCSG) from Algeria N° Components a RI a b Ri a c Ri p d DCSC e DCSG e Identification f 1 Nonane 906 908 907 - 0,5 RI, MS 2 Nonene 883 879 930 - 0,1 RI, MS 3 -Thujene 922 920 1019 0,9 - RI, MS 4 -Pinene 936 931 1016 26.0 34,1 RI, MS 5 Camphene 943 945 1062 1,8 0,8 RI, MS 6 Thuja-2,4(10)-diene 946 948 1117 0,2 - RI, MS 7 Sabinene 964 968 1117 14.0 1,5 RI, MS 8 -Pinene 978 970 1102 11,2 0,6 RI, MS 9 Myrcene 987 980 1152 10.0 13,1 RI, MS 10 -Phellandrene 997 999 1159 0,2 - RI, MS 11 3-methyl butyl isobutyrate 1002 1002 1170 0,1 - RI, MS 12 2-methyl butyl isobutyrate 1004 1007 1170 0,1 - RI, MS 13 -Terpinene 1008 1011 1267 0,9 - RI, MS 14 p-Cymene 1011 1015 1256 4,4 3,3 RI, MS 15 Limonene 1025 1022 1195 13.0 0,5 RI, MS 16 (Z)--Ocimene 1024 1027 1221 0,1 - RI, MS 17 (E)--Ocimene 1034 1037 1237 0,2 - RI, MS 18 -Terpinene 1047 1051 1233 0,1 0,1 RI, MS 19 Nonan-2-one 1070 1073 1392 0,1 0,1 RI, MS 20 Terpinolene 1078 1080 1274 0,2 - RI, MS 21 Linalool 1082 1085 1392 0,8 - RI, MS 22 2-methyl butyl isovalerate 1098 1090 1274 0,2 0,3 RI, MS 23 -Campholenal 1105 1093 1470 0,2 0,2 RI, MS 24 trans-Pinocarveol 1125 1124 1648 0,3 0,7 RI, MS 25 Pinocarvone 1136 1130 1547 0,3 1,3 RI, MS 26 Cryptone 1157 1158 1658 0,1 0,3 RI, MS 27 Terpinene-4-ol 1161 1165 1586 4,9 2,4 RI, MS 28 Myrtenal 1172 1171 1619 0,2 - RI, MS 29 -Terpineol 1176 1177 1685 0,4 0,2 RI, MS 30 Verbenone 1184 1182 1694 0,1 0,7 RI, MS 31 Carvone 1214 1216 1749 0,1 0,2 RI, MS 32 Cuminaldehyde 1217 1213 1778 0,2 0,2 RI, MS 33 Bornyl acetate 1269 1270 1515 0,9 1,8 RI, MS 34 -Terpenyl acetate 1334 1333 1676 - 0,1 RI, MS 35 Geranyl acetate 1362 1360 1749 0,1 0,1 RI, MS 36 -Ylangene 1375 1374 1470 0,1 0,2 RI, MS 37 -Copaene 1379 1371 1457 0,1 0,3 RI, MS 38 -Elemene 1389 1385 1550 tr - RI, MS 39 -Cedrene 1417 1410 1563 - 0,2 RI, MS 40 -Ylangene 1420 1413 1530 - - RI, MS 41 trans-Caryophyllene 1424 1426 1592 0,4 1,1 RI, MS 42 -Copaene 1430 1438 1602 - - RI, MS 43 trans--Bergamotene 1432 1431 1573 0,1 0,4 RI, MS 44 -Sesquisabinene 1435 1435 1637 0,1 1,2 RI, MS 45 (E)--Farnesene 1448 1449 1659 0,2 1,3 RI, MS 46 trans-methyl Isoeugenol 1463 1465 2170 0,1 - RI, MS 47 -muurolene 1473 1471 1667 0,1 0,2 RI, MS 48 Germacrene D 1480 1483 1663 - 0,2 RI, MS 49 -Humulene 1483 1487 1682 0,1 0,1 RI, MS International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 27 50 -Selinene 1483 1495 1703 - - RI, MS 51 Bicyclogermacrene 1494 1499 1683 - 0,1 RI, MS 52 -Bisabolene 1500 1494 1720 0,1 0,2 RI, MS 53 Cuparene 1504 1500 1824 - 0,6 RI, MS 54 -Cadinene 1515 1519 1762 0,1 0,1 RI, MS 55 Elemicin 1518 1527 2232 0,1 - RI, MS, Ref 56 E--Bisabolene 1531 1526 1733 - 0,2 RI, MS 57 -Caryophyllene oxide 1546 1550 2156 - 0,1 RI, MS, Ref 58 1,5 epoxy-Salvial-4(14)ene 1561 1563 1912 0,1 0,1 RI, MS 59 Spathulenol 1572 1568 2095 0,6 4,3 RI, MS 60 Caryophyllene oxyde 1576 1574 1937 0,8 6.0 RI, MS 61 Copaborneol 1592 1588 2154 - 1,9 RI, MS 62 Humulene epoxyde II 1601 1595 2009 0,1 2,1 RI, MS 63 Isospathulenol 1625 1615 2386 0,2 3,8 RI, MS, Ref 64 T-Muurolol 1634 1624 2138 0,1 0,4 RI, MS 65 -Cadinol 1645 1638 2227 0,1 0,7 RI, MS 66 (Z)--Santalol 1669 1667 2306 - 0,3 RI, MS 67 (E,Z)-Farnesol 1685 1680 2313 - 0,8 RI, MS Oil yield (%, w/w) 1,52 1,64 Total identification (g/100g) 95,9 90,1 Monoterpene hydrocarbons 83,2 54.0 Oxygenated monoterpenes 8,6 8,2 Sesquiterpene hydrocarbons 1,4 6,4 Oxygenated sesquiterpenes 2.0 20,5 Phenylpropanoids 0,2 - Non-terpenic compounds 0,5 1.0 Order of elution is given on apolar column (Rtx-1)., Retention indices of literature on the apolar column (RI) reported from Konig et al., (2001).Retention indices on the apolar Rtx-1 column (RI). Retention indices on the polar Rtx-Wax column (RI). Algerien Daucus (concentrations in g/100g): D. carota L. subsp. carota (DCSC), D. carota L. subsp; gummifer (DCSG). RI: Retention Indices; MS: Mass Spectra in electronic impact mode; Ref.: compounds identified from literature data: Koning et al., 2001 (55, 57, 63) International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 28 Table -2 Main components of the essential oils of D. carota L. Subsp. carota and D. carota L. Subsp. gummifer from different origins previously reported Plant origin Italy Spain Tunisia (Sejnane) Tunisia Italy Portugal Subspecies Gummifer gummifer carota carota carota Carota References (A) (B) (C) (C) (D) (D) Extraction modes HD HD CO 2 CO 2 HD HD Major Components % % % % % % No Plant parts Leaves; Fruits Fruits Flowering umbels and umbels Flowering umbels and umbels Flowering umbels and umbels Flowering umbels and umbels 4 -Pinene 10.8-12.2 - - - - 13.0-37,9 7 Sabinene 26.8-60.6 4.42-11.13 12.0-14.5 - - 8 -Pinene 1.4-3.4 - - - - 2.3-3.5 9 Myrcene - - - - - - 15 Limonene 2.1-5.7 - - - - - 16 (Z)-Ocimene 0.4-5.0 - - - - - 18 -Terpinene - - - - - - 20 Terpinolene 0.6-4,7 - - - - - 21 Linalool - 3.97-5.18 - - - - 27 Terpinen-4-ol 4.8-5.4 - - - - - - Daucene - - - - - - 35 Geranyl acetate - 51,7-76,9 - - - 15.0-65.0 - (Z,Z)--farnesene - - - - - - - -selinene - - 7.4-8.6 - - - 48 Germacrene D 0.2-6.9 - - - - - 46 E-Methylisoeugenol - - - - 1.3-10.0 - 52 -Bisabolene - - - - 17.6-51.0 - 55 Elemicin - - - 31.5-35.3 5.2-6.4 0.3-6.4 - Carotol - - 3.5-5.2 48.0-55.7 2.4-25.1 - - 11-H-himachal-4-en-1--ol - - 12.7-17.4 - 9.0-21.6 21.6 - Eudesm-7(11)-en-4-ol - - 8.2-8.5 - - - Plant origin Turkey Corsican Corsican Serbia Poland Lithuania Subspecies carota Carota carota carota carota carota References (E) (F) (G) (H) (I) (J) Extraction modes HD HD HD HD HD HD Major Components % % % % % % No Plant parts Seed Aerial parts Aerial Parts All Organs Umbels Fruits 4 -Pinene - 15.9 24.9 7.05–51.23 41.0 16.0–24.5 7 Sabinene - - 3.7 2.68–36.69 18.0 28.2–37.5 8 -Pinene - - - - - - 9 Myrcene - - 3.5 3.04-7.18 7.0 2,5 15 Limonene - - - 1.79-9.59 5.0 2.3–4.0 16 (Z)-Ocimene - - - - - 18 -Terpinene - - - - - 2.9–6.0 20 Terpinolene - - - - - - 21 Linalool - - - - - - 27 Terpinen-4-ol - - - 1.22-3.48 5.0 4.6–7.5 - Daucene 8.74 - - - - - 35 Geranyl acetate - - - - - - - (Z,Z)--farnesene 5.86 - - - - - - -selinene - - - - - - 48 Germacrene D - - - - - - 46 E-Methylisoeugenol - 21.8 33.0 - - - 52 -Bisabolene - - - - - - 55 Elemicin - 16.3 11.4 - - - - Carotol 66.78 - - - - - - 11-H-himachal-4-en-1--ol - - - - - - - Eudesm-7(11)-en-4-ol - - - - - - Only the main components were reported; main components are classified by number corresponding to the table-1; Extraction mode: HD: Hydrodistillation; CO: Supercritic Carbone dioxide extract8,9,11,12,14,16,18 International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(1), 22-29, January (2013) Int. Res. J. Biological Sci. International Science Congress Association 29 Table -3 MIC (mg/mL) of D. carota L. Subsp. carota (DCSC) and D. carota L. Subsp. gummifer (DCSG) essential oils on bacterial strains Bacterial strains DCSC DCSG Gram-positive bacterium B. subtilis� 6.0 ( - ) � 6.0(-) L. monocytogenes 2.5 (++) 4.3 B. cereus 2.5(++)3.8 (+) S. aureus 2.5 (++) 5.1 (+) P. aeruginosa 5.0 (+) � 6.0 ( - ) E. faecalis � 6.0 ( - ) � 6.0 ( - ) Gram-negative bacterium K. pneumoniae 5.0 (+) � 6.0 ( - ) E. coli 2.5 (++) � 6.0 ( - ) Values are means of triplicate determinations, Essential oils are classified as (-) not active, (+) moderately active and (++) active.