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Analysis of the Essential Oils from Five VietnameseLitsea Species (Lauraceae)Le C. Sona, Do N. Daib, Tran D. Thangc, Duong D. Huyena & Isiaka A. Ogunwanded

a Institute of Ecology and Biological Resources, Vietnam Academy of Science and Technology,18-Hoang Quoc Viet, Cau Giay, Hanoi, Vietnamb Faculty of Biology, Vinh University, 182-Le Duan, Vinh City, Nghean Province, Vietnamc Faculty of Chemistry, Vinh University, 182-Le Duan, Vinh City, Nghean Province, Vietnamd Natural Products Research Unit, Department of Chemistry, Faculty of Science, Lagos StateUniversity, Badagry Expressway, Ojo, P. M. B. 0001, Lasu Post Office, Ojo, Lagos, NigeriaPublished online: 23 Dec 2014.

To cite this article: Le C. Son, Do N. Dai, Tran D. Thang, Duong D. Huyen & Isiaka A. Ogunwande (2014) Analysis of theEssential Oils from Five Vietnamese Litsea Species (Lauraceae), Journal of Essential Oil Bearing Plants, 17:5, 960-971, DOI:10.1080/0972060X.2014.935068

To link to this article: http://dx.doi.org/10.1080/0972060X.2014.935068

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Analysis of the Essential Oils from Five Vietnamese Litsea Species (Lauraceae)

Le C. Son 1, Do N. Dai 2, Tran D. Thang 3*, Duong D. Huyen1 , and Isiaka A. Ogunwande 4*

1 Institute of Ecology and Biological Resources, Vietnam Academy of Scienceand Technology, 18-Hoang Quoc Viet, Cau Giay, Hanoi, Vietnam

2 Faculty of Biology, Vinh University, 182-Le Duan, Vinh City, Nghean Province, Vietnam3 Faculty of Chemistry, Vinh University, 182-Le Duan, Vinh City, Nghean Province, Vietnam4 Natural Products Research Unit, Department of Chemistry, Faculty of Science, Lagos StateUniversity, Badagry Expressway, Ojo, P. M. B. 0001, Lasu Post Office, Ojo, Lagos, Nigeria

Abstract: In this paper, the compositional pattern of essential oils from five plants in the genus Litsea(Lauraceae family) cultivated in Vietnam, were being reported. The air-dried plant materials were hydrodistilledand the oils were analysed by Gas chromatography-flame ionization detector (GC-FID) and Gas chromatographycoupled with Mass spectrometry (GC-MS). The leaf oil of Litsea helferi Hook.f., was rich in limonene (17.5%), β-caryophyllene (14.2 %), bicyclogermacrene (13.1 %), bicycloelemene (12.4 %) and α-phellandrene(8.0 %). The main constituents of Litsea ferruginea Liou., leaf oil were dominated by monoterpenes comprisingof sabinene (34.5 %), α-pinene (10.1 %), γ-terpinene (7.8 %), limonene (6.9 %) and terpinen-4-ol (6.6 %). Thequantitative significant constituents of the leaf oil of Litsea verticillata Hance were also monoterpene compoundsrepresented by linalool (23.4 %), α-pinene (26.1%) and β-pinene (11.7 %). In addition, the monoterpenehydrocarbons (E)-β-ocimene (57.4 %), along with α-pinene (7.8 %) and β-pinene (7.3 %) were the mainconstituents in the leaf oil of Litsea glutinosa (Lour.) C. B. Rob. The main compounds in the leaf, stem, fruitsand roots oils of Litsea cubeba (Lours.) Pers., were (Z)-citral (32.9-66.1 %), sabinene (1.4-14.2 %), limonene(7.0-13.6 %) and linalool (1.9-9.5 %). The chemical compositions of L. helferi, L. ferruginea and L. verticillataare being reported for the first time.

Key words: Litsea glutinosa, Litsea helferi, Litsea ferruginea, Litsea verticillata, Litsea cubeba,essential oil composition

IntroductionLitsea is a genus of evergreen or deciduous trees

or shrubs belonging to the Laurel family,Lauraceae. The genus includes 200 to 400 speciesin tropical and subtropical areas of bothhemispheres. The flowers are from greenish towhite, greenish-yellow, yellowish. Theinflorescences are pseudo-umbels, flat-topped orrounded flower clusters, each pseudo-umbel withan involucre of four or six decussate bracts 1.

Literature information is relatively scarce on

the volatile and non-volatile constituents of L.helferi and L. ferruginea. However, L. verticillatais known to contained anti-HIV lignans andeudesmane sesquiterpenoid compounds 2-4,several sesquiterpenes 4 and butenolides 5.Extracts from various parts of L. glutinosa werefound to be antimicrobial 6-8, antioxidant, anti-inflammatory and wound healing effect 9,antinociceptive 10, aphrodisiac and possible maleanti-infertility activity 11, analgesic 12 and anti-diabetic 13. Phytochemical studies revealed the

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

*Corresponding author (Tran D. Thang; Isiaka A. Ogunwande)E-mail: < thangtd@vinhuni.edu.vn; isiaka.ogunwande@lasu.edu.ng > © 2014, Har Krishan Bhalla & Sons

TEOP 17 (5) 2014 pp 960 - 971 960

Received 15 August 2013; accepted in revised form 28 January 2014

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characterization of fatty acids 14, arabinoxylanwith immunomodulating properties 15, aporphinealkaloid 16, butanolides and benzoic acid 17,flavones glycosides 18, lignans 19 and gluco-pyranosides 20.

Previous essential oil constituents of L.glutinosa were found to be β-caryophyllene (22.8%), β-ocimene (7.2 %), phytol (6.9 %), β-pinene(6.8 %), α-pinene (5.9 %), caryophyllene oxide(5.9 %) from fresh leaves as well β-caryophyllene(23.2 %), β-ocimene (7.8 %), bicyclogermacrene(7.4 %), α-caryophyllene (5.9 %) and α-pinene(5.4 %) from dry sample 21. A steam distilled oilconsists mainly of (Z)-3-hexen-1-ol (43.9 %) and(E)-2-hexenal (7.4 %), while squalene (11.9 %),eucalyptol (9.9 %) and (+)-α-terpineol (5.4 %)were the main compounds of supercritical-CO2fluid extraction 22. (E)-β-ocimene (70.8 % and84.1 %, respectively) was the mainly compoundsin the unripe and ripe fruit oils from India 23. Theoil has effect on cardiovascular system 24.

Litsea cubeba (Lour.) Pers. is an evergreentropical shrub that is native to China and otherregions of East Asia. It is often referred to as MayChang, and has attracted much attention owingto its multiple functions. Extracts of L. cubebahave also been used in traditional Chinesemedicine for the treatment of a variety of ailments25. L. cubeba gives off an aromatic odour and themain constituents of its essential oils (citral) hasvarious industrial uses, including as a flavoring,in medicines and personal-care products, and asa botanical insecticide. Recently reports havedemonstrated the bioactivities of L. cubeba oils,which include antibacterial, antifungal, acaricidal,insecticidal, antioxidant, and anticancerproperties 26. Literatures pertaining to chemicalcompositions of L. cubeba oils have identified anumber of chemotypic forms such as cineole type,linalool type, neral/geranial type, linalool/citronella type etc. 27.

As part of our continued interest in the analysisof chemical constituents of Vietnamese flora 28-

29, we report herein compounds identified in theoils of five Litsea species. The plants are Litseahelferi Hook.f., Litsea ferruginea Liou., Litseaverticillata Hance, Litsea glutinosa (Lour.) C. B.Rob. and Litsea cubeba (Lours.) Pers.

ExperimentalPlant collection

Mature leaves of L. glutinosa, L. helferi, L.ferruginea, L. verticillata and L. cubeba werecollected from Bach Mã National Park, ThuaThiên - Hue Province, Vietnam, in August 2012.Voucher specimens LCS 263, LCS 261, LCS 264,LCS 832 and LCS 250, respectively, have beendeposited at the Botany Museum, VinhUniversity, Vietnam. Plant samples were air-driedprior to extraction.

Extraction of the oils0.5 kg of air-dried samples was shredded and

their oils were obtained by hydrodistillation for4 h at normal pressure, according to theVietnamese Pharmacopoeia 30. The plant samplesyielded a low content of essential oils: 0.95 %,0.30 %, 0.25 %, 0.25 % and 1.4 % (v/w; respecti-vely for L. glutinosa, L. helferi, L. ferruginea, L.verticillata and L. cubeba), calculated on a dryweight basis. All samples were light yellowcoloured.

Analysis of the oilsGas chromatography (GC) analysis was

performed on an Agilent Technologies HP 6890Plus Gas chromatograph equipped with a FID andfitted with HP-Wax and HP-5MS columns (both30 m X 0.25 mm, film thickness 0.25 μm, AgilentTechnology, Berkshire, United Kingdom). Theanalytical conditions were: carrier gas Hydrogen(1.0 mL/min), injector temperature (PTV-programmed temperature vaporization injection)250°C, detector temperature 260°C, columntemperature programmed from 40°C (2 min hold)to 220°C (10 min hold) at 4°C/min. Samples wereinjected by splitting and the split ratio was 10:1.The injection volume was 1.0 μL. Inlet pressurewas 6.1 kPa. Each sample was analyzed thrice.

An Agilent Technologies HP 6890N PlusChromatograph fitted with a fused silica capillaryHP-5 MS column (30 m x 0.25 mm, film thickness0.25 μm) and interfaced with a Mass SpectrometerHP 5973 MSD was used for the GC-MS analysis,under the same conditions as those used for GCanalysis. The conditions were the same asdescribed above with He (1.0 mL/min) as carrier

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gas. The MS conditions were as follows:ionization voltage 70 eV; emission current 40 mA;acquisitions scan mass range of 35-350 amu at asampling rate of 1.0 scan/s.

The identification of constituents wasperformed on the basis of retention indices (RI)determined by co-injection with reference to ahomologous series of n-alkanes, under identicalexperimental conditions. Further identificationwas performed by comparison of their massspectra with those from NIST 08 and Wiley 9th

Version and the home-made MS library built upfrom pure substances and components of knownessential oils, as well as by comparison of theirretention indices with literature values 31-32.

Cluster analysisThe Litsea species in this study were treated as

operational taxonomic units (OTUs). Thepercentage of composition of 15 major essentialoil components about e”5% was used to determinethe chemical relationship between the differentLitsea essential oil samples by cluster analysisusing the PAST software version 2.12 33.Euclidean and paired group were selected as ameasure of similarity for cluster definition.

Results and discussionTable 1 and Table 2 indicate the identities and

percentage composition of the compounds in thefive studied oil samples. Ubiquitous terpenehydrocarbons were the major fraction present in

L. glutinosa (85.9 %), L. helferi (86.1 %) and L.ferruginea (87.0 %). L. verticillata had terpenehydrocarbons (46.5 %) and oxygenatedderivatives (48.5 %). Oxygenated terpenes werethe major fractions in the leaf (50.9 %), stem(66.5 %), fruits (75.6 %) and roots (65.7 %) of L.cubeba. Monoterpenes were the dominantcompounds of L. glutinosa (85.7 %), L. ferruginea(85.4 %), L. verticillata (77.8 %) and L. cubeba(82.9-97.4 %). L. helferi contained an almostequal amount of monoterpenes (48.3 %) andsesquiterpenes (47.5 %).

Thirteen compounds namely α-pinene,myrcene, α-phellandrene, α-terpinene, limonene,γ-terpinene, linalool, terpinen-4-ol, β-caryophyllene, aromadendrene, α-humulene andspathulenol were present in all the oil samples.

Hydrodistillation of L. glutinosa afforded oilwhose major constituent was (E)-β-ocimene (57.4%) with smaller quantities of α-pinene (7.3 %)and β-pinene (7.8 %). Monoterpene hydrocarbons(83.0 %) constitute the major oil fraction. Minorconstituent includes limonene (2.4 %), (Z)-β-ocimene (2.3 %) and β-caryophyllene (2.0 %) 23.β-Caryophyllene (27.2 %), bicyclogermacrene(18.2 %) and (E)-β-ocimene (13.4% ) were themain compounds of previous study 28; as well aslinalool (32.6 %), sabinene (13.4 %) and trans-verbenol (5.4 %) from another report from Vietnam29. The main compounds of sample analysed fromBangladesh were phytol (22.4 %), caryophyllene(21.5 %) and thujopsene (12.2 %) from the leaf

Table 1. Chemical composition of the studied oils of Litsea species from Vietnam

Compounds a RI RI Percent composition (%)(Cal.) (Lit.) L. L. L. L.

glutinosa helferi ferruginea verticillata

Tricyclene 926 921 - - - 0.1α-Thujene 930 924 0.1 0.3 1.9 -α-Pinene 939 932 7.8 2.4 10.1 26.1Camphene 953 946 1.1 - 0.3 1.3Verbenene 967 961 - - - 0.4Sabinene 976 969 - 2.0 34.5 0.4β-pinene 980 974 7.3 0.6 - 11.7Myrcene 990 988 1.0 1.2 5.3 0.6α-Phellandrene 1006 1002 1.7 8.0 0.3 0.1

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table 1. (continued).

Compounds a RI RI Percent composition (%)(Cal.) (Lit.) L. L. L. L.

glutinosa helferi ferruginea verticillata

Δ-3-Carene 1011 1008 - 0.4 - -α-Terpinene 1017 1014 0.1 0.4 5.4 Trρ-Cymene 1026 1020 - 4.3 0.2 0.4Limonene 1032 1024 2.4 17.5 6.9 0.81,8-Cineole 1034 1026 - - - 2.4(Z)-β-ocimene 1043 1044 2.3 - - -(E)-β-Ocimene 1052 1052 57.4 3.3 1.2 0.1γ-Terpinene 1061 1054 0.1 0.6 7.8 0.1α-Terpinolene 1090 1086 0.1 0.4 2.1 -trans-Sabinene hydrate 1097 1098 Tr - 0.3 0.1Linalool 1100 1095 0.3 0.5 0.5 23.4ρ-Mentha-1,4,5-triene 1110 1108 - - - 0.1cis-ρ-Menth-2-en-1-ol 1120 1118 - - 0.8 -allo-Ocimene 1128 1128 1.6 0.1 - -Camphor 1141 1141 - - - 0.6trans-Verbenol 1144 1140 - - - 2.3trans-Pinocarveol 1152 1135 - - - 1.7Pinocarvone 1165 1160 - - - 1.3Terpinen-4-ol 1179 1174 0.1 0.9 6.6 0.2α-Thujenal 1182 1182 - - - 1.7ρ-Cymen-8-ol 1183 1179 - - - 0.6α-Terpineol 1189 1186 0.1 0.2 - -Verbenone 1205 1204 - - - 0.7(E, E)-2,6-dimethyl- 1207 1208 0.2 0.2 - -3,5,7-Octatriene-2-oltrans-Carveol 1217 1215 - - 0.1 0.2Neral 1235 1235 - 0.5 - 0.1Carvone 1242 1239 - - - 0.1(E)-Citral 1270 1272 - 0.9 - -Safrole 1287 1285 0.3 - - -Bornyl acetate 1289 1287 - - 0.3 0.1Perilla alcohol 1295 1294 - - - 0.1Linalool propanoate 1324 1334 - - 0.2 -Bicycloelemene 1327 1338 - 12.4 2.7 0.3α-Cubebene 1351 1345 - 0.1 - -Eugenol 1356 1356 0.2 3.8 - -Citronellyl acetate 1356 1350 - - 0.3 -Neryl acetate 1362 1359 - - 0.3 -α-Copaene 1377 1374 0.9 0.3 - 0.2β-Cubebene 1388 1387 0.2 - 0.2 -β-Elemene 1397 1398 1.9 0.3 0.2 -Methyl eugenol 1407 1410 1.2 - - -

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table 1. (continued).

Compounds a RI RI Percent composition (%)(Cal.) (Lit.) L. L. L. L.

glutinosa helferi ferruginea verticillata

β-Caryophyllene 1419 1417 2.0 14.2 2.1 0.2trans-α-Begamotene 1435 1432 - 0.4 - -Aromadendrene 1441 1439 0.2 0.7 0.5 0.1α-Humulene 1454 1452 0.4 1.5 0.4 0.2δ-Gurjunene 1477 1475 - - 0.6 -Germacrene D 1485 1484 - - 1.0 0.2α-Amorphene 1485 1483 0.1 - - 0.6β-Selinene 1486 1489 - - 0.2 -cis-Cadina-1,4-diene 1496 1495 - 0.2 Tr 0.4Bicyclogermacrene 1500 1500 - 13.1 2.2 0.6β-Bisabolene 1506 1505 - - - 0.2γ-Cadinene 1514 1513 - - 0.4 -δ-Cadinene 1525 1522 - 0.6 0.5 0.6Eugenol acetate 1526 1521 0.7 - - -α-Calacorene 1542 1544 - - - 0.7Elemol 1550 1548 - 1.0 0.1 -Germacrene B 1561 1559 - 0.8 - -(E)-Nerolidol 1563 1561 - - 0.2 4.9Spathulenol 1578 1563 0.9 1.2 0.1 3.5Caryophyllene oxide 1583 1582 0.5 - - 2.2Globulol 1585 1590 - - - 0.2Viridiflorol 1593 1592 0.6 0.5 - -Guaiol 1601 1600 - - 0.4 -α-Cadinol 1654 1652 0.2 0.2 0.3 -Bulnesol 1672 1670 0.5 - 0.2 1.9(E,E)-Farnesol 1722 1722 - - - 0.2Benzyl benzoate 1760 1759 0.1 0.2 0.1 -Tetradecanoic acid 1762 1769 0.3 0.8 0.2 -(Z)-9-Octadecenamide b 2397 2397 0.3 2.3 1.2 -Total 95.2 99.3 99.2 95.0Monoterpene hydrocarbons 83.0 41.5 76.0 42.2Oxygenated monoterpenes 2.7 6.8 9.4 35.6Sesquiterpene hydrocarbons 5.9 44.6 11.0 4.3Oxygenated sesquiterpenes 2.7 2.9 1.3 12.9Non-terpenes 0.9 3.5 1.5 -

a Elution order on HP-5MS capillary columnRI (Cal.)= Retention indices on HP-5MS capillary columnRI (Lit.) = Literature Retention indices (see Experimental)b Tentative identification- Not identifiedTr, trace amounts < 0.1%;

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Table 2. Percentage composition of Litsea cubeba essential oils

Compounds a RI RI Percent composition (%)(Cal.) (Lit.) Leaf Stem Fruits Roots

α-Thujene 930 921 0.6 0.3 0.1 0.1α-Pinene 939 932 3.4 2.4 0.8 3.6Camphene 953 946 0.1 0.1 Tr 2.7Sabinene 976 969 14.2 10.2 2.1 1.4β-Pinene 980 976 - - 0.6 2.7Myrcene 990 988 2.0 3.3 2.0 2.4α-Phellandrene 1006 1002 0.1 0.1 Tr 0.1δ-3-Carene 1011 1008 - - - 0.3α-Terpinene 1017 1014 0.4 0.4 Tr 0.1Limonene 1032 1024 9.2 13.2 7.0 13.6(E)-β-Ocimene 1052 1052 0.3 Tr Tr Trγ-Terpinene 1061 1054 1.0 0.8 0.1 0.2α-Terpinolene 1090 1086 0.3 0.3 0.1 0.2trans-Sabinene hydrate 1097 1098 0.2 0.2 0.1 TrLinalool 1100 1095 9.5 2.4 2.9 1.9cis-ρ-Menth-2-en-1-ol 1121 1118 0.1 0.2 - -allo-Ocimene 1128 1128 0.4 - 7.9 -trans-Limonene oxide 1139 1137 - - - 0.1Camphor 1145 1141 - - - 2.1Borneol 1167 1165 - - - 0.1Terpinen-4-ol 1179 1174 4.0 5.3 - -α-Terpineol 1189 1186 1.8 2.1 1.1 -Citronellol 1223 1148 2.1 0.5 3.4 0.5Pulegone 1237 1233 - 1.4 - -(Z)-Citral (= Neral) 1238 1235 32.9 53.2 66.1 60.0Geraniol 1255 1249 - 0.8 0.6 -Geranial 1262 1264 0.1 0..3 0.2 0.1Linalool propaonaote 1324 1334 - - - 0.8Neric acid 1347 1347 0.1 - 0.9 0.4Geranic acid 1355 1355 0.2 0.4 0.5 -Eugenol 1356 1356 Tr - - -α-Copaene 1377 1374 0.1 - - -β-Elemene 1391 1398 0.1 - - -β-Caryophyllene 1419 1417 2.8 0.3 0.3 0.3α-Humulene 1454 1452 - Tr Tr Tr4-Dodecen-1-ol a 1460 1469 0.1 - - -Germacrene D 1485 1484 0.1 - Tr -β-Selinene 1486 1489 0.3 - - 0.4α-Muurolene 1500 1500 - - - TrBicyclogermacrene 1500 1500 Tr - - -(E, E)-α-Farnesene 1508 1505 0.1 - - -(E)-Nerolidol 1563 1561 Tr - - -Caryophyllene oxide 1583 1582 0.6 0.2 0.1 0.3

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and lauric acid (44.8 %), 2,7-dimethyl-3-octen-5-yne (28.7 %) and α-cubebene (6.8 %) from thefruit 34. The abundance (E)-β-ocimene makes theoil similar to the fruit oil sample from India 23.However, it differs considerably from otherprevious investigations where sesquiterpene 21, 28,aliphatic alcohols/aldehydes 22 and fatty acids 34

predominated as against the monoterpenecompounds. Notable compounds identified inprevious reports such as 1, 8-cineole, (Z)-3-hexen-1-ol, (E)-2-hexenal, α-cubebene, phytol,thujopsene, lauric acid, 2,7-dimethyl-3-octen-5-yne and squalene are conspicuously absent in thepresent oil sample. It is evident that two chemicalforms of essential oils of L. glutinosa exists sofar in Vietnam which are oils dominated bymonoterpene compounds 29 and oil rich insesquiterpene compounds 28.

The forty essential oil constituents of L. helferigave oil with almost equal amount ofmonoterpenes (48.3 %) and sesquiterpene (47.5%). The oxygenated terpenes are less common inthis oil (6.8 % and 2.9 % respectively). Also, the

main compounds were identified as limonene(17.5 %), β-caryophyllene (14.2 %), bicyclo-germacrene (13.1 %), bicycloelemene (12.4 %)and α-phellandrene (8.0 %). The minorcompounds in the oil were α-pinene (2.4 %),sabinene (2.0 %), (E)-β-ocimene (3.3%), β-caryophyllene (4.3 %), eugenol (3.8 %) and (Z)-9-octadecenamide (2.3 %).

The major monoterpene constituent of the leafoil of L. ferruginea consists of sabinene (34.5 %),α-pinene (10.1 %), γ-terpinene (7.8 %), limonene(6.9 %), terpinen-4-ol (6.6 %), α-terpinene (5.4%) and myrcene (5.3 %). In addition, the oil hasminor constituents such as α-terpinene (2.1 %),bicycloelemene (2.7 %), β-caryophyllene (2.1 %)and bicyclogermacrene (2.2 %). Sesquiterpenes(12.3 %) are less common in this oil.

The monoterpene compounds of L. verticillatawere identified as α-pinene (26.1 %), linalool(23.4 %) and β-pinene (11.7 %). Oxygenatedmono- and sesquiterpenes were pronounced (35.6% and 12.9 % respectively) in this oil. Its minorcompound included 1,8-cineole (2.4 %), trans-

table 2. (continued).

Compounds a RI RI Percent composition (%)(Cal.) (Lit.) Leaf Stem Fruits Roots

Isoaromadendrene epoxide 1597 1595 0.1 - - -(E, E)-Farnesol 1741 1742 0.1 - - -Tetradecanoic acid 1762 1765 0.3 - - -n-Hexadecanol 1883 1883 Tr - - -1,2-Benzenedicarboxylic acid 1917 1917 1.0 Tr 0.1 0.1n-Hexadecanoic acid 1970 1959 1.0 0.1 Tr 0.2(Z,Z)-9,12-Octadecadienoic acid 2066 2130 0.2 - Tr 0.3(Z)-9-octadecenamide 2397 2397 0.2 0.8 2.8 2.1Total 90.2 99.1 99.8 96.9Monoterpene hydrocarbons 32.0 31.1 20.7 27.4Oxygenated monoterpenes 51.1 66.8 75.8 65.8Sesquiterpene hydrocarbons 3.5 0.3 0.3 0.7Oxygenated sesquiterpenes 0.8 0.3 0.1 0.3Non-terpenes 2.8 0.9 2.9 2.7

a Elution order on HP-5MS capillary columnb Correct isomer not identifiedRI (Cal.)= Retention indices on HP-5MS capillary columnRI (Lit.) = Literature Retention indices (see Experimental)- Not identified

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verbenol (2.3 %), (E)-nerolidol (4.9 %), spathu-lenol (3.5 %) and caryophyllene oxide (2.2 %).

The authors are unaware of any previousanalysis of the essential oils of L. helferi, L.ferruginea and L. verticillata, as such the presentreport may represent the first of its kind.

In this study, essential oils extracted from theleaf, stem, fruit and root of L. cubeba byhydrodistillation were light yellow in colour. GC-MS analysis led to the identification of 39, 28,30 and 32 compounds respectively (Table 2). Themain component in all the samples was (Z)-citral(32.9-66.1 %) along with significant quantity ofsabinene (1.4-14.2 %), limonene (7.0-13.6 %) andlinalool (1.9-9.5 %). Terpinen-4-ol (4.0 and 5.3%, respectively) was an additional compound inthe leaf and stem oils, while allo-ocimene (7.9%) was present only in the fruit oil. The contentof geranial in all the samples was too low.Previous studies on L. cubeba from Vietnam have

Table 3. A summary of chemical constituents of Litsea cubeba from some other countries

Plant Biological Major constituents ReferencesParts activities

Leaves - 1, 8-Cineole (0.2-51.7 %), Bighelli, et al., 2005Linalool (0.4-91.1 %) andSabinene (0.4-8.1 %)

Stem barks - Citronellol (11.9-20.4 %) Choudhury, et al., 1998and Citronellal (7.7-10.0 %)

Fruits - Citronellal (44.8-77.2 %) and Choudhury, et al., 1998Citronellol (10.9-14.0 %)

Flowers - Sabinene (41.8-42.3 %), Choudhury, et al., 1998Citronellal (14.3-17.3 %),β-Phellandrene (7.7-9.0 %),α-Pinene (6.6-7.6 %) andβ-Pinene (5.8-6.1 %)

Part not known - Linalool (78.3 %) a, Nath, et al., 1996Citronellol (41.3 %) b andCitronellal (76.6 %) c

Part not known Antioxidant activity Limonene (26.3 %), Wang, et al., 2012and free radical followed by α-Citral (25.9 %),scavenging β-Citral (21.90 %), and

β-Pinene (6.2 %)Fruits Analgesic, Geranial (37.2 %), Chi-Jung, et al., 2012

anxiolytic, CNS Neral (28.3 %),and d-Limonene (22.9 %)

identified three chemotypes, viz- linalool type 35,1,8-cineole/sabinene/α-terpineol type 35 and neral/geranial/limonene type 36. The compo-sitionalpattern of neral/sabinene/limonene may representanother chemical form of the oil of L. cubeba.However, the major composition data was inagreement with the results from other studies thatcitral was the major component 37, 38, whilst theproportion vary widely, which demonstrates thevariability of the natural oil.

Table 3 represents the summary of majorchemical constituents of essential oils fromvarious parts of L. cubeba analysed fromdifferent countries 25. It could be seen differentchemical classes of L. cubeba oils exists whichmay be due to the nature of the plant,geographical areas, methods of extraction, plantparts, rearing condition and maturation of theharvested plant.

Fig 1 and 2 represent the cluster analysis and

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table 3. (continued).

Plant Biological Major constituents ReferencesParts activities

Leaves Antifungal β-cis-Ocimene (25.1 %), Wang, et al., 19993,7-Dimethyl-1,6-octadien-3-ol (16.8 %)and trans Nerolidol (13.89 %)

Stem barks Antibacterial Citral b (neral), β-Phellandrene, Wang & Liu, 2010and β-Terpinene,

Roots - d-Limonene (8.5 %), Hu, et al., 2011α-Citral (26.2 %),β-Citral (33.2 %)

Fruits Antimicrobial Geranial (39.2 %), Tsung-Shi, et al., 2013Neral (30.6 %)

Fruits Antioxidant Geranial (39.2 %), Xiao-Wei, et al., 2013Neral (30.6 %),Limonene (8.1 %) andLinalool (5.2 %)

Leaves - (E)-Citral (41.3 %), Ko, et al., 2009(Z)-Citral (30.1 %),Methylheptenone (5.6 %) Si, et al., 2012

Aerial parts - d-Limonene (0.7-5.0 %),Neral (34.2-37.4 %),Geranial (44.4-50.0 %)

Leaves - Sabinene (12.3 %), Chen-Lung, et al., 20101,8-Cineole (47.5 %),α-Terpinyl acetate (9.8 %)

Fruits Anticancer Limonene (11.1%), Chen-Lung, et al., 2010Neral (31.3 %) andGeranial (37.6 %)

Stem barks inhibitory effects on 1,8-Cineole (26.6 %) Muchtaridi & Anas, 2011locomotor activity and Citronellol (21.7 %)in mice

Fruits Antifungal Limonol (44.2 %), Yang, et al., 2010β-Linalool (8.8 %),1,8-Cineole (5.4 %)

the principal components analysis for the studiedoil samples. The cluster is in 2 groups namely(Fig 1). Cluster 1 which comprises of L. helferi,L. ferruginea, L.cubeba and L. verticillata.Cluster 2 is basically composed of L. glutinosa.The Principal component analysis is representedin Fig 2. L. ferruginea has the followingcompounds sabinene, terpinen-4-ol, myrcene, γ-terpinene and α-terpinene; L. verticillata and L.glutinosa have (E)-β-ocimene, linalool, β-pineneand α-pinene in common, while L. helferi has the

following compounds limonene, β-caryophyllene,bicycloelemene, α-phellandrene and bicyclo-germacrene in the analysis. However, L.cubebacomprised mainly of (Z)-citral.

AcknowledgmentsWe are grateful to Miss Onwordi Theresa for

the assistant in cluster and principal componentsanalysis. Mrs. Ogunwande Musilimat was alsoacknowledged for typesetting manuscript to oursatisfaction.

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Fig 1. Cluster analysis dendogram for components of the Litsea oil samples

Fig 2. Principal component analysis for Litsea oil samples

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