This article was downloaded by: [SHERIFAT ABOABA]On: 12 March 2014, At: 02:46Publisher: Taylor & FrancisInforma Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House,37-41 Mortimer Street, London W1T 3JH, UK

Journal of Essential Oil Bearing PlantsPublication details, including instructions for authors and subscription information:http://www.tandfonline.com/loi/teop20

Chemical Composition, Toxicity and Antibacterialactivity of the Essential Oils of Garcinia mangostanafrom NigeriaSherifat Aboabaa, Akinsola Akandea & Guido Flaminiba Department of Chemistry, University of Ibadan, Ibadan, Nigeriab Dipartimento di Farmacia, Via Bonanno 33, 56126, Pisa, ItalyPublished online: 07 Mar 2014.

To cite this article: Sherifat Aboaba, Akinsola Akande & Guido Flamini (2014) Chemical Composition, Toxicity andAntibacterial activity of the Essential Oils of Garcinia mangostana from Nigeria, Journal of Essential Oil Bearing Plants, 17:1,78-86, DOI: 10.1080/0972060X.2014.884759

To link to this article: http://dx.doi.org/10.1080/0972060X.2014.884759

PLEASE SCROLL DOWN FOR ARTICLE

Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) containedin the publications on our platform. However, Taylor & Francis, our agents, and our licensors make norepresentations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of theContent. Any opinions and views expressed in this publication are the opinions and views of the authors, andare not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon andshould be independently verified with primary sources of information. Taylor and Francis shall not be liable forany losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoeveror howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use ofthe Content.

This article may be used for research, teaching, and private study purposes. Any substantial or systematicreproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in anyform to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions

Chemical Composition, Toxicity and Antibacterial activity ofthe Essential Oils of Garcinia mangostana from Nigeria

Sherifat Aboaba 1*, Akinsola Akande 1 and Guido Flamini 2

1 Department of Chemistry, University of Ibadan, Ibadan, Nigeria2 Dipartimento di Farmacia, Via Bonanno 33, 56126, Pisa, Italy

Abstract: The chemical composition of the essential oils obtained from the leaves and stem-bark ofGarcinia mangostana Linn. extracted by hydrodistillation were analyzed by GC and GC-MS. A total of 64 and101 constituents, representing 96.1 % and 97.5 % of the total oil were identified in the leaves and stem-barkessential oils, respectively. β-caryophyllene (17.3 % - 21.1 %) was the most abundant constituent common inboth oils. The essential oils were toxic to Artemia salina with LC50 value of 1.70 μg/ml and 5.15 μg/ml forleaves and stem bark oils, respectively. The antibacterial activities of the essential oils were carried out usingagar well diffusion method. Garcinia mangostana stem-bark essential oil was more active against the testedbacteria.

Key words: Garcinia mangostana Linn, β-caryophyllene, Lethal Concentration, antibacterialactivities, hydrodistillation.

IntroductionMedicinal plants play a crucial role in health

care needs of people around the world mostespecially in the developing countries. About 80% of the populations of most developingcountries depend on the use of traditionalmedicine derived from plants 1. The genusGarcinia mangostana Linn. belongs to the familyGuttiferae (Clusiaceae) 3, 4 is commonly knownas “mangoteen”. It is a tropical evergreen treethat grows from 7 to 25 m (20-80 ft) tall withcharacteristic taste, fragrance, visual qualities andnutrient richness 2. It is sometimes called“superfruit” and found in Thailand, Australia andsome other south-east Asian countries. From thefleshy pericarp, when prepared as an astringentmedicine, G. mangostana Linn. has beenemployed to cure dysentery, enteritis and chronicdiarrhea 5,6. People in the tropical rainforest of

some Southeast Asian nations like Indonesia,Malaysia, Sri Lanka, Philippines and Thailandhave used the pericarp (peel, rind, and hull) as atraditional medicine for the treatment of abdomi-nal pain, diarrhea, dysentery, infected wound andchronic ulcer 7.

Experimental studies have revealed thatextracts from the plant have shown bioactiveproperties with potential therapeutic applications.A type of organic compound called “Mangostin”,(C24H26O6), a natural xanthonoid, and a varietyof other xanthones such as prenylated xanthonesisolated from various parts of the tree have beeninvestigated for biological activities includingantioxidant, anti-inflammatory, antibacterial,antifungal, antiviral, antitumoral and anti-canceractivities 7,8,9,10.

The brine shrimp lethality assay is a useful toolfor the preliminary assessment of toxicity 11,12,13.

ISSN Print: 0972-060XISSN Online: 0976-5026

*Corresponding author (Sherifat Aboaba)E-mail: < saboaba@gmail.com > © 2013, Har Krishan Bhalla & Sons

TEOP 17 (1) 2014 pp 78 - 86 78

Received 11 January 2013; accepted in revised form 04 June 2013

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

It is an assay based on the ability to kill labo-ratory-cultured Artemia spp. (brine shrimp). It hasbeen used for the detection of fungal toxins, plantextract toxicity, heavy metals, cyanobacteriatoxins, pesticides and cytotoxicity testing ofdental materials 14, 15. It can also be extrapolatedfor cell-line toxicity and antitumor activity 16.

Treatment of infectious diseases has becomeclinical problems as a result of indiscriminate useof antimicrobial drugs thereby increasing thenumbers of antibiotics resistant microorganisms17. Presently, there is increasing interest that gearstowards the development of new classes ofantibiotics from natural sources such as plantsso as to resolve the problems of drug resistancein pathogenic microorganisms 18.

This study evaluates the chemical constituents,toxicity and antimicrobial activities of essentialoils from the leaves and stem-bark of Garciniamangostana Linn. by GC-MS analysis, brineshrimp lethality test and antimicrobial assayrespectively.

Materials and methodsPlant Materials and Isolation of the Oils

Leaves and stem-bark of Garcinia mangostanawere collected from the plantation of theBotanical Garden, University of Ibadan, Ibadan,Nigeria and identified by Mr. Donatus of theBotanical Garden Herbarium. The fresh plant partsamples were air dried for 7 days and subse-quently pulverized separately and weighed.Pulverized leaves (250 g) and stem-bark (300 g)were subjected to hydrodistillation in an all glassClevenger distillation apparatus for 3.5 h inaccordance with British Pharmacopoeia method19. The essential oils thereby obtained as paleyellow were dried over anhydrous sodiumsulphate (Na2SO4) and refrigerated at 4°C untilfurther analysis was carried out.

Chemical Analysis and identification of theOils

Gas Chromatographic analysis was carried outon a HP-5890 Series II equipped with a HP-5capillary column (30 m x 0.25 mm, 0.25 μm filmthickness). The GC oven temperature wasprogrammed at 60°C (held for 10 min), heated to

220°C at 5°C/min. The injector and detectortemperatures were maintained at 250°C. Heliumwas used as carrier gas at flow rate of 2 ml/min.

The identification of components was carriedout by comparing the retention times of theconstituents with those of pure authentic samplesand by means of their linear retention indices(LRI) relative to the series of n-hydrocarbons.

GC-MS analyses were carried on a Varian CP-3800 gas chromatograph equipped with a HP-5capillary column (30 m x 0.25 mm; 0.25 μmcoating thickness) interfaced to a Varian Saturn2000 ion trap mass detector. The MS wasoperated in EI mode with ionization voltage 70eV. Operating conditions were: injector andtransfer line temperature, 220°C and 240°C,respectively, oven temperature programmed from60 to 240°C at 3°C/min. Helium was used ascarrier gas at flow rate of 2 ml/min.

Identification of the constituents was based ontheir retention indices (determined with referenceto a homologous series of normal alkanes), andby comparison of their mass spectra withpublished spectra and those of referencecompounds from NIST 98 Library 20,21,22,23,24.

Toxicity assayThe brine shrimp lethality test (BST) was

carried out, using Artemia salina eggs hatchedin sea water, to evaluate the level of toxicity inthe oils. Different concentrations (1000 ppm, 100ppm, 10 ppm stock solution each in triplicates)of the essential oil samples were prepared to testthe survival rate of the brine shrimps. 10 shrimpsin each of the triplicate stock solution of theextracted oils were introduced. A blank solutionwas also made consisting of DMSO and 10 brineshrimps in sea water. This serves as referencestandard. After 24 hours, the total number of deadshrimps was counted and the lethal concentrationat 50 % level was determined by the Finney probitcomputer progamme.

Antibacterial activitiesThe essential oils were tested for antibacterial

activity by disc and agar well diffusion methodsaccording to the National Committee for ClinicalLaboratory Standards (NCCLS). The essential

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 79

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

oils were tested against 6 clinical isolates ofStaphylococcus aureus, Proteus sp., Escherichiacoli, Staphylococcus epidermidis, Pseudomonasaeruginosa and Bacillus subtilis. Themicroorganisms were grown on Muller Hintonagar which was prepared following theManufacturer’s instruction and sterilized insideautoclave at 121°C for 15 min. The test bacteriawere seeded onto the agar surface by introducingstandardized inoculums (1-2 × 107 cfu/ml 0.5McFarland standards) onto the surface of sterileMuller Hinton agar plates, and a sterile glassspreader was used for even distribution of theinoculums.

Agar well diffusion method was also used forthe antibacterial assay of the essential oils. Thiswas done using a cork borer of diameter 9 mm tocut uniform wells on the surface of the agar. Theextracts were then applied in the crude form intothe various uniform wells using sterile plasticpipette for each test organism. Standard antibioticdiscs of chloramphenicol (30 μg), augmentin (30μg) and gentamycin (5 μg) were used as positivereference standards, while hexane was used asnegative reference standard. This is done toprovide a basis to evaluate the antibiotic effective-ness of the plant extracts. The set up was thenincubated for 24 hours at 37°C in the incubatorafter which the zones of inhibition (mm) weremeasured.

Results and discussionThe essential oils from Garcinia mangostana

Linn. were pale yellow in colour with percentageyield of 0.16 and 0.12 for the leaves and stembark tissues, respectively. A total of 64 and 101constituents representing 96.1 % and 97.5 % wereidentified in the essential oils of the leaves andstem-bark respectively. The leaves and stem barkessential oils consists of sesquiterpene hydro-carbons (82.9 to 87.9 %), oxygenated sesqui-terpenes (6.2 to 9.1 %), diterpenes (1.2 to 1.9 %)and non-terpene derivatives (0.8 to 3.6 %).

The major constituents identified in the leavesessential oil were β-caryophyllene (21.1 %), β-elemene (9.7 %), germacrene D (7.9 %), α-selinene (7.4 %), α-humelene (7.3 %), β-selinene(7.1 %) and γ-muurolene (5.0 %) while otherminor constituents were δ-cadinene (4.7 %),trans-γ-cadinene (2.7 %) and β-bourbonene (2.1%). The major constituents identified in the stembark essential oil were β-caryophyllene (17.3 %),β-selinene (10.7 %), trans-α-bergamotene (7.5%), α-selinene (6.4 %), α-humelene (6.4 %), δ-cadinene (5.3 %), β-elemene (5.3 %) andgermacrene D (5.0 %) while γ-muurolene (4.6%), trans-γ-cadinene (3.1 %), germacrene A (2.4%) and caryophyllene oxide (2.2 %) were alsopresent in significant quantity. β-Caryophyllene(the major constituents of the essential oils understudy) has been described as a new apoptosis

Table 1. Chemical constituents of the essential oils of Garcinia mangostana from Nigeria

Constituents LRI Leaves Stembark

(Z)-2-Octene 812 - 0.1Ethyl benzene 866 0.1 -Nonane 899 0.1 0.36-Methyl-5-hepten-2-one 986 tr 0.12-Pentyl furan 994 tr trMesitylene 996 tr trn-Decane 1000 tr 0.1Octanal 1002 - trLimonene 1031 tr -(Z)-β-Ocimene 1041 tr tr(E)-β-Ocimene 1051 tr trtrans-Decahydro naphthalene 1060 - trAcetophenone 1066 tr tr

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 80

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

table 1. (continued).

Constituents LRI Leaves Stembark

1-Octanol 1071 - trcis-Linalool oxide (furanoid) 1075 tr trMethyl benzoate 1091 - tr2-Nonanone 1094 - trLinalool 1099 tr trNonanal 1103 tr 0.9(E,Z)-2,6-Nonadienal 1156 tr tr(E)-2-Nonenal 1160 tr -1,3-Dimethoxybenzene 1162 tr tr1-Nonanol 1172 - trNaphthalene 1180 - tr3-Decanone 1188 - trα-Terpineol 1190 - trMethyl salicylate 1192 tr trn-Dodecane 1200 - trDecanal 1204 - 0.2Nerol 1228 - trNeral 1241 - trGeraniol 1256 tr tr(E)-2-Decenal 1262 - trGeranial 1272 - trNonanoic acid 1280 - 0.32-Undecanone 1292 - trn-Tridecane 1300 - trUndecanal 1306 - tr(E,E)-2,4-Decadienal 1315 - trδ-Elemene 1339 0.1 trα-Cubebene 1351 0.6 0.31-Undecanol 1362 - trα-Ylangene 1372 0.4 0.4α-Copaene 1376 1.7 1.1β-Bourbonene 1384 2.1 1.1β-Cubebene 1390 - trβ-Elemene 1392 9.7 5.3Cyperene 1398 tr 0.2n-Tetradecane 1400 - tr(Z)-Caryophyllene 1405 tr trα-Cedrene 1409 - trα-Gurjunene 1410 tr -β-Ylangene 1414 tr -β-Caryophyllene 1418 21.1 17.3β-Copaene 1429 1.5 1.0trans-α-Bergamotene 1437 0.7 7.5α-Guaiene 1439 0.5 -

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 81

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

table 1. (continued).

Constituents LRI Leaves Stembark

Aromadendrene 1440 - Trα-neo-Clovene 1454 0.7 0.4α-Humulene 1455 7.3 6.4Alloaromadendrene 1461 tr 0.1cis-Muurola-4(14),5-diene 1463 tr trDrima-7,9(11)-diene 1469 tr trγ-Gurjunene 1474 0.4 0.3γ-Muurolene 1477 5.0 4.6Germacrene D 1480 7.9 5.0β-Selinene 1485 7.1 10.7α-Selinene 1494 7.4 6.4trans-β-Guaiene 1500 0.7 1.0Germacrene A 1503 2.9 2.4(E,E)-α-Farnesene 1508 0.2 0.5trans-γ-Cadinene 1513 2.7 3.17-epi-α-Selinene 1518 tr -δ-Cadinene 1524 4.7 5.3trans-Cadina-1(2),4-diene 1532 0.2 0.2α-Cadinene 1538 0.4 0.4α-Calacorene 1542 0.2 0.3Elemol 1550 - 0.1Germacrene B 1556 1.7 1.5(E)-Nerolidol 1565 0.5 1.2Spathulenol 1476 0.8 0.9Caryophyllene oxide 1581 1.9 2.2Globulol 1583 0.1 0.2Thujopsan-2-α-ol 1587 - 0.2n-Hexadecane 1600 - trHumulene epoxide 1606 0.3 0.5β-Oplopenone 1607 0.5 1.11-epi-Cubenol 1628 0.2 0.1β-Acorenol 1634 - 0.1T-Cadinol 1641 0.6 0.7α-Muurolol 1646 0.1 0.2Selin-11-en-4-α-ol 1653 1.2 1.7Intermedeol 1667 - tr(Z)-α-Santalol 1675 - trCadalene 1677 - trEudesma-4(15),7-dien-1-β-ol 1688 - trn-Heptadecane 1700 - tr14-Hydroxy-α-humulene 1714 - tr(E)-Nerolidol acetate 1717 - trMint sulfide 1741 - tr(E,Z)-Farnesol 1743 - tr

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 82

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

table 1. (continued).

Constituents LRI Leaves Stembark

n-Octadecane 1800 - trn-Nonadecane 1900 - trMethyl exadecanoate 1928 - trHexadecanoic acid 1959 - 1.6(3Z)-Cembrene 1965 1.2 1.9n-Eicosane 2000 - trn-Heneicosane 2100 0.6 -Kaurene 2034 - trClass of Terpene/Terpenoid PresentMonoterpene hydrocarbons tr trOxygenated monoterpenes tr trSesquiterpene hydrocarbons 87.9 82.9Oxygenated sesquiterpenes 6.2 9.1Diterpenes 1.2 1.9Non-terpene derivatives 0.8 3.6Total identified 96.1 97.5

tr = Trace quantity (< 0.1%)LRI. = Linear Retention Indices

inducing agent that acts, in a selective manner,against tumour cell-lines and not against normalcells. Eitan et al.25 recently reported an investi-gation on the biblical balm of Gilead(Commiphora gileadensis) for anticancerousactivity against tumor cell lines. β-Caryophyllenewas found to be a key component in the essentialoils extract which is known for its anti-inflammatory, local anesthetic, antifungal and asa potent cytotoxic compound over a wide rangeof cell lines.

The results of the brine shrimp lethality test ofthe essential oils are shown in Table 2. The resultswere analysed by the Finney probit computer

Table 2. Brine Shrimp lethality test of essential oils ofGarcinia mangostana leaves and stem-bark

Sample LC50 ( μμμμμg/ml) UCL LCL

Leaves 1.7014 21378 0.4678Stem-bark 5.1518 16.755 0.1745

UCL: Upper Confidence Limit.LCL: Lower Confidence Limit.

programme to determine the Lethal Concen-tration at 50 % level of toxicity (LC50). Theessential oils from the leaves and stem-bark haveLC50 values of 1.70 and 5.15 μg/ml, respectively.These values showed that the oils are highly toxic26. It is well known that natural products that aretoxic to Artemia species are medicinally activeand cytotoxic property by plant material has beenattributed to the presence of antitumour com-pounds 27.

The results of the antibacterial test of theessential oils are presented in Table 3. Theessential oils show selective levels of activitiesat varying degrees against all the clinical isolates

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 83

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

Table 3. Antibacterial activity of the essential oils fromthe leaves and stem bark of Garcinia mangostana

Test organisms with zone of inhibition (mm)Essential Staphylococcus taphylococcus Escherichia Pseudomonas Proteus Bacillus

Oils aureus epidermidis coli aeruginosa spp. subtilis

Leaves - - 48.2 - - -Stem-bark 18.0 20.0 9.0 - 14.0 21.3Standard controlAugmentin (30 μg) - - - - - -Chloramphenicol 7.0 10.0 20.0 - - 28(30 μg)Gentamycin (5 μg) - - 14.0 - - 16.0Hexane (100 %) - - - - - -

(-): No inhibition

viz: Staphylococcus epidermidis, Staphylococcusaureus, Escherichia coli, Proteus species,Bacillus subtilis, except Pseudomonas aerugi-nosa, from a moderate to high susceptibilities.The essential oil of G. mangostana leaves gavethe highest growth inhibitory activity (48.2 mm)and the only zone of inhibition againstEscherichia coli, (all other organisms showedresistance), while the least growth inhibitoryactivity (9 mm) was observed for essential oil ofG. mangostana stem bark against the same testorganism (E. coli).

Mazleena et al. 28 recently reported work on theantimicrobial properties of G. mangostana that

both ethanol and chloroform stem-bark extractswere ineffective against E. coli. However, fromthis study, the essential oil extract from the stem-bark was effective against E. coli, although witha low zone of inhibition. In addition, P.aeruginosa had been reported to show naturalresistance to many antibiotics 29,30,31,32.

AcknowledgementsThe authors are grateful to Mr. Abimbola O.

Adekanbi of Microbiology Department, Facultyof Science, University of Ibadan, for the anti-microbial assay.

References1. Cunningham, A.B. (1993). African Medicinal Plants: Setting Priorities at the interface between

Conservation and Primary Healthcare. People and Plants Working Paper 1, Paris.2. Primchanien, M., Nuttavut, K., Sineenart, K., Omboon, L., Narongchai, P. and Neelobol,

N. (2004). Antiproliferation, ant-oxidation and induction of apoptosis by Garcinia mangostana(mangosteen) on SKBR3 human breast cancer cell line. Journal of Ethnopharmacology; 90(1):161-6.

3. Phongpaichit, S., Rungjindama, N., Rukachaisirikul, V. and Saka-yaroj, J. (2006). Anti-microbial activity in cultures of endophytic fungi isolated from Garcinia species. FEMS Immunol.Med. Microbiol., 48: 367-372.

4. Mahabusakam, W. and Viriyacitra, P. (1987). Chemical constituents of Garcinia mangostana.J. Nat. Prod. 50 (5): 474-478.

5. Burkill, H.M. (1994). The useful plants of West Tropical Africa. (Edition 2). Vol. 2. FamiliesE-I. Royal Botanic Gardens Kew.

6. Drury, C.H. (1973). The useful plants of India: with notice of their chief medicinal value in

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 84

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

commerce, medicine and arts. Higginbotham and Co. Madras, Republic of India.7. Pedraza-Chaverri, J., Cardenas-Rodriguez, N., Orozco-Ibarra, M. and Perez-Rojas, J.M.

(2008). Medicinal properties of mangosteen (Garcinia mangostana). Food Chemistry andToxicology. 46: 3227-3239.

8. Jung, H., Su B., Keller, W., Mehta, R. and Kinghorn, A. (2006). “Antioxidant xanthonesfrom the pericarp of Garcinia mangostana (Mangosteen)”. J. Agric. Food Chem. 54(6): 2077-82.

9. Chen, L.-G., Yang, L.-L. and Wang, C.-C. (2007). Anti-inflammatory activity of mangostinsfrom Garcinia mangostana. J. Food and Chemical Toxicology. 46(2): 688-93.

10. Suksamrarn, S., Komutiban, O., Ratananukul, P., Chimnoi, N., Lartpornmatulee, N. andSuksamrarn, A. (2006). Cytotoxic prenylated xanthones from the young fruit of Garciniamangostana. Chemical & Pharmaceutical Bulletin. 54: 301-305.

11. Michael, A.S., Thompson, C.G. and Abramowitz, M. (1956). Artemia salina as a test organismfor bioassay. Science; 123:464.

12. Vanhaecke, P., Person, G., Claus, C. and Sorgeloos, P. (1981). Proposal for a short-term toxicitytest with Artemia nauplii. Ecotoxicol. Env. Safety. 5: 382-387.

13. Sleet, R.B. and Brendel, K. (1983). Improved methods for harvesting and counting synchro-nous populations of Artemia nauplii for use in development toxicology. Ecotoxicol. Env. Safety.7: 435-446.

14. Meyer, B.N., Ferrign, R.N., Putnam, J.E., Jacobson, L.B., Nicholas, D.E. and McLaughlin,J.L. (1982). Brine shrimp: A convenient general bioassay for active plant constituents. PlantaMedica. 45: 31-34.

15. Awal, M.A., Ainun, N., Hossain, M.S., Bari, M.R. and Haque, M.E. (2004). Brine shrimptoxicity of leaf and seed extracts of Cassia alata Linn. and their antibacterial potency. J. Med.Sci. 4(3): 188-193.

16. Anderson, J.E., Goetz, C.M., McLaughlin, J.L. and Suffness, M. (1991). A blind comparisonof simple bench-top bioassays and human tumor cell cytotoxicities as antitumor prescreens.Phytochemical microcladia Anal. 2: 107-111.

17. Davis, J. (1994). Inactivation of Antibiotics and the Dissemination of Resistance Genes. Science.264: 375-382.

18. Cowan, M.M. (1999). Plant products as antimicrobial agents. Clinical microbiology reviews;12: 564-82.

19. British Pharmacopoeia. (1980). Vol. 2. H.M. Stationary office, London, 109.20. Adams, R.P. (1995). Identification of Essential Oil Components by Gas Chromatography/Mass

spectroscopy. Allured Publ. Corp., Carol Stream, IL.21. Swiger, A.A. and Silverstein, R.M. (1981). Monoterpenes. Aldrich Chem. Comp., Milwaukee.

WI.22. Massada, Y. (1976). Analysis of Essential Oils by Gas Chromatography and Mass Spectrometry.

John Wiley and Sons, New York.23. Jennings, W. and Shibamoto, T. (1980). Qualitative Analysis of Flavor and Fragrance Volatiles

by Glass Capillary Chromatography. Academic Press, New York.24. Davies, N.W. (1990). Gas Chromatographic retention indexes of monoterpenes and sesqui-

terpenes on methyl silicone and Carbowax 20M phases, J. Chromatogr. 503: 1-24.25. Eitan, A., Rivka, O., Nativ, D., Elaine, S., Tatiana, R., and Shimon, R. (2012). β-Caryophyllene,

a Compound Isolated from the Biblical Balm of Gilead (Commiphora gileadensis), Is a SelectiveApoptosis Inducer for Tumor Cell-Lines. Evidence-Based Complementary and AlternativeMedicine. Article ID 872394.

26. Moshi, M.J., Cosam, J.C., Mbwambo, Z.H., Kapingu, M. and Nkunya, M.H.H. (2004).

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 85

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4

Testing Beyond Ethnobotanical Claims: Brine Shrimp Lethality of Some Tanzania Plants. Pharma-ceutical Biology 42: 547-551.

27. Ara, J., Sultana, V., Ehteshamul-Haque, S. and Ahmad, Q.R.V.U. (1999). Cytotoxic activityof marine macroalgae on Artemia salina (brine shrimp). Phytotheraphy Res. 13: 304-307.

28. Mazleena, S.M., Sheikh Ahmad Izaddin, S.M.G., Nurulhuda, I. and Nora, M.B. (2011).Antimicrobial Properties of Mangosteen Stembark Extract. Empowering Science, Technologyand Innovation Towards a Better Tomorrow. UMTAS 2011. LSP 81: 582-585.

29. Hancock, R.E. (1998). Resistance mechanisms in Pseudomonas aeruginosa and other non-fermentative gram-negative bacteria. Clin. Intect. Dis. 27(Suppl 1): 593-599.

30. Ziha-Zarifi, I., Llanes, C., Kohler, T., Pechere, J.C. and Plesiat, P. (1999). In vivo emergenceof multidrug-resistant mutant of Pseudomonas aeruginosa over expressing the active effluxsystem MeXA-MeXB-Opru. Antimicrob Agents Chemother. 43(2): 287-291.

31. Brinkman, F.S., Bains, M. and Hancock, R.E. (2000). The amino terminus of P. aeruginosaouter membrane protein. OprF forms channels in lipid bilayer membranes: correlation withthree-dimensional model. J. Bacteriol. 182(18): 5251-5255.

32. Henwood, C.I., Livermore, D.M., James, D. and Warner, M. (2001). Antimicrobial suscepti-bility of P. aeruginosa: Results of a UK survey and evaluation of the British society for Anti-microbial Chemotherapy disc susceptibility test. J. Antimicrob. Chemother. 47(6): 789-799.

Sherifat Aboaba et al., / TEOP 17 (1) 2014 78 - 86 86

Dow

nloa

ded

by [

SHE

RIF

AT

AB

OA

BA

] at

02:

46 1

2 M

arch

201

4