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JOURNAL OF FOOD COMPOSITION AND ANALYSIS

Journal of Food Composition and Analysis 20 (2007) 52–56 www.elsevier.com/locate/jfca Short Communication Volatile components of the , and of wampee [ lansium (Lour.) Skeels]

Pratheung Chokepraserta,Ã, Albert Linton Charlesb, Kai-Hsin Suec, Tzou-Chi Huangc

aDepartment of Product Development, Faculty of Agro-Industry, Kasetsart University, Bangkok 10900, Thailand bDepartment of International Cooperation and Tropical Agriculture, National Pingtung University of Science and Technology, Pingtung 912, Taiwan cDepartment of Food Science, National Pingtung University of Science and Technology, Pingtung 912, Taiwan

Received 2 May 2005; received in revised form 22 June 2006; accepted 14 July 2006

Abstract

The volatile components of fruits, seeds and leaves from [Clausena lansium (Lour.) Skeel], obtained through headspace sampler, were analyzed by gas chromatography–mass spectrometry (GC–MS). The sesquiterpene fraction (28%) was the major component in the . Monoterpene (76–98%) was the dominant terpene in flesh, skin and and sabinene was the main component in leaf (14.9%), flesh (50.6%), skin (69.1%) and seed (83.6%). Other major components of wampee leaf were b-bisabolene (9.9%), b-caryophyllene (7.7%) and a-Zingiberene (6.5%); in the flesh, 3-cyclohexen-1-ol (15%), cyclohexene (6.5%), 1,4 cyclohexadiene (6.2%) and a-phellandrere (5%); in the skin, a-phellandrene (10.6%) and a-pinene (9.4%) and isosativene (1.4%); and in the seed, a-pinene (4.3%), a-phellandrene (3.0%), and myrcene (2.9%). r 2006 Elsevier Inc. All rights reserved.

Keywords: Wampee; Clausena lansium (Lour.) Skeel; GC–MS; Monoterpene; Sabinene; Headspace sampler

1. Introduction wampee, which is currently produced in Thailand as a sweet preserved . Wampee [Clausena lansium (Lour.) Skeels] is a minor The leaves, roots and fruits have been used as a folk member of the , and is a distant relative of citrus medicine in Taiwan (Li et al., 1991) and (Yang et al., fruit that originated in southern China. The Chinese 1988), for the treatment of certain dermatological diseases introduced it to the Nan Province in northern Thailand such as, for instance, acute and chronic viral hepatitis; the more than 100 years ago. Wampee has many vernacular fruit is used in the for influenza, colds and names and most are derived from the Chinese huang-p’i- abdominal colic pains (Stuart, 1977). The leaf decoction is kuo. In Thailand, it is called mafai jeen, but the common used as a hair wash to remove dandruff and preserve hair name is wampee. The fruit ripens from May to July; it colour (Perry, 1980). Recently, the extraction of seeds was tastes similar to grapefruit when ripe; resembles a found to possess antifungal and HIV reverse transcriptase- diminutive lemon, and is about 2.0 cm in diameter. It inhibitory activities (Ng et al., 2003). contains 1–3 seeds and the pulp is slightly acidic. When Previous studies on C. lansium included the character- fully ripe, it can be eaten with the peel. The pulp can be ization of dehydroindicolactone (Khan et al., 1983) and added to fruit cups, gelatines or other desserts, or made coumarins (Kumar et al., 1995) from the root bark; into pie, jam or jelly. Carbonated beverages resembling carbozole alkaloids from roots (Li et al., 1991); cinnam- champagne are made by fermenting the fruit with sugar amide derivatives from the seeds (Lin, 1989); and cyclic (Morton, 1987), but the most popular product is dried amines (Yang et al., 1988) and a triterpene alcohol from leaves. However, despite the many studies that have elucidated the non-volatile composition of C. lansium, ÃCorresponding author. Tel.: +66 15942158; fax: +66 53873063. there is little research on the volatile compounds respon- E-mail address: [email protected] (P. Chokeprasert). sible for the intense aroma of wampee, although Zhao et al.

0889-1575/$ - see front matter r 2006 Elsevier Inc. All rights reserved. doi:10.1016/j.jfca.2006.07.002 ARTICLE IN PRESS P. Chokeprasert et al. / Journal of Food Composition and Analysis 20 (2007) 52–56 53

(2004) analyzed the essential oil of the leaf, flower, 2.4. Qualitative and quantitative analyses sarcocarp and seed of C. lansium. The dominant consti- tuents identified were b-santalol, a-santalol, methyl santa- Most constituents were characterized by gas chromato- lol, bisabolol and ledol. The objective in this study graphy by comparison of their GC retention indices (RI) was to determine the headspace volatile components of with those found in literature (Sibanda et al., 2006; Davies, wampee fruits, seeds and leaves to reveal the volatile 1990). Further identification of volatile components was compounds that are responsible for its intense aromatic performed by matching their mass spectra with reference profile. spectra in the Wiley 275 Mass Spectral Library and the National Institute of Standard and Technology (NIST) 98 2. Materials and methods Mass Spectral Library (Revision D.01.00/Search pro- gramme v.1.6d) purchased from Agilent Technologies. 2.1. materials Quantitative analysis of each volatile component in percent was performed by peak area normalization measurement in The fruits, seeds and leaves of wampee [C. lansium triplicate. (Lour.) Skeels] were collected in July 2004 from the Horticultural Research Station, Department of Agricul- 3. Results and discussion ture, Nan Province, Thailand. The plant (Forest Herbar- ium No. BKF 135985) was identified and deposited In order to detect the ‘‘true’’ fragrance composition at the Forest Herbarium (BKF), National Park, Wildlife experienced by the consumers, a headspace sampling and Plant Conservation Department, Ministry of technique was performed in this experiment. Over 72 Natural Resources and Environment, Bangkok 10900 compounds were isolated and over 60 characterized from Thailand. All wampee fruits used were from the same their retention index, mass spectra and data from the batch. Fully ripe fruit were used in the study; ripeness is literature (Table 1). The volatile components of leaves, determined when the fruit turns yellow and has a thin, flesh, skin of fruit and seeds are summarized in Table 1. sometimes brittle skin, somewhat like paper. These components were found in different percentages in various parts of the plant. The majority of these 2.2. Sample preparation and headspace sampling components were found to belong to the hydrocarbon fraction, with percentages ranging from 50% in the leaves, An Agilent 7694 (Agilent Technologies Inc., Wilming- 77% in the flesh, to 96% in the skin and 99% in the seeds. ton, DE 19808, USA.) was used for headspace sampl- Among the components of the hydrocarbon fraction, the ing. Samples of 50 g were cut and immediately crushed in a predominant compounds were found to be sesquiterpenes blender, then 1 g, of all samples used in this study in the leaves, and monoterpenes in the flesh, skin and seeds. were placed into 25 mL vials; they were then crimped A total of 39 components were identified in the head- and equilibrated for 20 min at 80 1C before head- space of leaves, amounting to 86% of the total volatiles. space sampling, following the method of Alasalvar et al. The sample was dominated by the sesquiterpenes (28%), (1999). with b-bisabolene, b-caryophyllene and a-Zingiberene (6.5%) as the main components. Monoterpenes were fewer 2.3. Gas chromatography–mass spectrometry (GC–MS) (22%), with sabinene (15%) as the main component. In addition to the hydrocarbons, an ester, 3-hexenyl 2- GC–MS was performed on an Agilent 6890 GC Plus methylbutanoate (0.19%), along with its alcohol, 3- equipped with a HP-5973 mass-selective detector (Agilent hexen-1-ol (0.17%) was characterized in the headspace of Technologies). A fuse silica capillary column, HP-5-MS, leaves. The compounds 3-hexen-1-ol and its acetate were with 5%-phenyl methylpolysiloxane as no-polar stationary often found in green leaves (Hatanaka, 1993). Sesquiter- phase (30 m 0.25 mm i.d. 0.25 mm film thickness, Agi- penes, b-caryophyllene (7.72%) and humulene (0.39%), lent Technologies) was utilized for analysis of volatiles and an ester (3-hexenyl 2-methylbutanoate, 0.19%) were obtained from wampee. The injection port temperature was released in response to the attack by the insect of 250 1C. The column temperature programme started at Spodoptera in cotton plantlets (Loughrin et al., 1994). 40 1C upon injection. The temperature was increased at a Camciuc et al. (1998) proposed that the biological activity rate of 3 1C/min to 100 1C, and then at a rate of 5 1C/min to of some of these compounds seems to support the 230 1C, and held there for 2 min. Purified helium gas at a hypothesis of their role in defence against insects. The flow rate of 1 mL/min was used as the GC carrier gas. monoterpenes and esters present in wampee leaves may act The mass spectrometer was operated in the electron as solvents and also have a synergic action with molecules impact (EI) mode with an electron energy of 70 eV; ion having irritant properties. Interestingly, significant amount source temperature, 230 1C; quadrupole temperature, of ar-curcumene (1.27%), a-zingiberene (6.52%) and b- 150 1C; mass range m/z 35–400; scan rate, 0.25 s/scan; bisabolene (9.88%) were characterized in the headspace of EM voltage, 1423 V; and the GC–MS transfer line was set leaves as well. Rani (1999) reported that the essential oil to 280 1C. from the rhizomes of Zingiber officinale Roacoe contains ARTICLE IN PRESS 54 P. Chokeprasert et al. / Journal of Food Composition and Analysis 20 (2007) 52–56

Table 1 Volatile compounds identified in wampee using headspace sampler with HP-5MS non-polar column

No. Compounds RI % Relative area

Leaf Flesh Skin Seed ID

1 Ethanol t 2.46 ttMS 2 2-Propanone 3.02 tttMS 3 Propanal 1.63 tttMS 4 2-Methylfuran 1.10 tttMS 5 Butanal 8.61 tttMS 6 1-Pentene 1.89 tttMS 7 2-Ethylfuran 4.61 tttMS 8 Ethanone 0.20 tttMS 9 Acetic acid 0.94 2.65 0.08 0.03 MS 10 cis-2-Pentenol 0.71 tttMS

11 Hexanal 802 1.55 0.47 0.04 t MS, RI1 12 2-Hexenal 854 1.46 tttMS, RI1 13 3-Hexen-1-ol 857 0.17 tttMS, RI1 14 Styrene 921 0.13 tttMS, RI1 15 Tricyclene 928 tt0.03 t MS, RI1 16 a-Thujene 931 tt0.02 0.59 MS, RI1 17 a-Pinene 939 1.99 2.08 9.41 4.26 MS, RI1 18 Camphene 945 0.98 t 0.47 0.04 MS, RI1 19 Benzaldehyde 958 2.56 tt0.02 MS, RI1 20 b-Pinene 967 t 0.21 0.17 t MS, RI1 21 Sabinene 973 14.92 50.64 69.07 83.56 MS, RI1 22 6-Methyl-5-hepten-2-one 976 2.26 tttMS, RI1 23 Myrcene 993 1.10 1.70 3.15 2.94 MS, RI1 24 a-Phellandrene 1001 1.38 5.03 10.63 3.08 MS, RI1 25 3-Carene 1010 tt0.10 t MS, RI1 26 (+)4-Carene 1018 t 3.98 0.40 1.13 MS, RI1 27 Limonene 1026 t 0.21 ttMS, RI1 28 trans-b-Ocimene 1035 ttt0.02 MS, RI1 29 Benzeneacetaldehyde 1037 0.30 tttMS, RI1 30 1,3,6-Octatriene 1039 1.96 t 0.06 t MS, RI1 31 1,4-Cyclohexadiene 1043 t 6.19 0.32 t MS, RI1 32 g-Terpinene 1057 tt0.04 1.95 MS, RI1 33 Cyclohexene 1065 t 6.50 0.17 0.39 MS, RI1 34 2-Nonanone 1079 3.42 tt0.01 MS, RI1 35 Linalool 1086 2.25 t 0.16 t MS, RI1 36 (E)-4,8-dimethyl-1,3,7-nonatriene 1089 1.22 tttMS, RI1 37 3-Methyl-4-brendene 1095 t 0.13 ttMS, RI1 38 3-Cyclohexen-1-ol 1097 t 15.17 0.28 0.51 MS, RI1 39 2-Cyclohexen-1-one 1099 tt0.03 0.01 MS, RI1 40 3-Cyclohexen-1-methanol 1106 tt0.02 t MS, RI1 41 b-Fenchyl alcohol 1109 t 0.54 t 0.06 MS, RI1 42 Benzoic acid 1163 0.16 tttMS, RI1 43 cis-3-Hexenyl 2-methylbutanoate 1218 0.19 t 0.01 t MS, RI1 44 Bornyl acetate 1286 ttt0.01 MS, RI1 45 Geranyl acetate 1357 tt0.02 t MS, RI1 46 Copaene 1373 0.28 tttMS, RI1 47 b-Caryophllene 1417 7.72 tt0.55 MS, RI1 48 a-Bergamotene 1427 0.71 t 0.20 0.03 MS, RI1 49 (+)-Aromadendrene 1436 0.08 tttMS, RI1 50 Isosativene 1441 0.38 t 0.07 0.01 MS, RI1 51 b-Santalene 1444 tt0.02 t MS,RI1 52 a-Humulene 1447 0.39 t 0.02 0.03 MS, RI1 53 ar-Curcumene 1475 1.27 0.12 0.87 0.03 MS, RI1 54 Allaromadendrene 1478 tt0.10 t MS, RI1 55 a-Zingiberene 1486 6.52 tt0.06 MS, RI1 56 Bicyclogermacrene 1490 0.37 tt0.01 MS, RI1 57 a-Farnesene 1494 tt0.95 t MS, RI1 58 b-Bisabolene 1496 9.88 tt0.15 MS, RI1 59 b-Sesquiphellandrene 1512 0.70 t 0.30 t MS, RI1 60 d-Cadinene 1524 0.33 tttMS, RI1 ARTICLE IN PRESS P. Chokeprasert et al. / Journal of Food Composition and Analysis 20 (2007) 52–56 55

Table 1 (continued )

No. Compounds RI % Relative area

Leaf Flesh Skin Seed ID

Total monoterpenes 22.34 76.54 94.05 97.96 Total sesquiterpenes 27.69 0.12 2.22 0.85 Total alcohols 2.77 17.53 0.28 0.51 Total aldehydes 16.12 0.47 0.04 0.04 Total esters 0.19 0.00 0.03 0.01 Total ketones 8.90 0.00 0.03 0.02 Heterocyclics 5.72 0.00 0.00 0.00 Carboxylic acid 1.10 2.65 0.08 0.03 Hydrocarbons 1.35 0.13 0.00 0.00 Unidentified Total 86.18 97.44 96.73 99.42

RI ¼ programmed temperature retention indices relative to the homologous series of n-alkanes (C5-C25); RI1 ¼ retention data in literature; t ¼ traces40.01%; ID ¼ identification method. ar-curcumene (20%), a-zingiberene (22%) and b-bisabo- that represents the major volatile component in the leaves lene (14%). He postulated that bisabolyl cation may be and seeds. The steam distillation procedure was applied in derived from farnesylpyrophosphate. Bisabolyl cation is their experiment to obtain the essential oils from the the penultimate precursor of ar-curcumene, a-zingiberene flowers, leaves, sarcocarps and seeds of C. lansium, and b-bisabolene; and two 1,2-hydrogen shifts lead to the respectively. High boiling point alcohols, such as b-santalol formation of a-zingiberene whereas one 1,2-hydrogen shift (35.2%), bisabolol (13.7%) and ledol (6.5%) in leaves, were leads to the formation of ar-curcumene. The chemical extracted under much higher temperature (boiling point of composition of the essential oil obtained from the rhizomes water) compared to the extraction conditions in this of Z. officinale Roscoe was characterized by the presence of experiment (80 1C). ar-curcumene (22.1%), zingiberene (11.7%), b-bisabolene Sabinene (15%, 51%, 69% and 84% in leaf, flesh, skin (11.2%) and cadina-1,4-diene (12.5%). It is reasonable that and seed, respectively) was the major headspace volatile in in (Kumar et al., 1995) the leaves are used as a wampee. Sabinene was found to be extremely abundant in substitute for curry leaf in cooking. the plant and the fresh fruit essential oil of Peucedanum Monoterpenes constituted the major part of the fra- verticillare(Fraternale et al., 2000). The emission of grance emitted from wampee flesh, skin and seed. The sabinene and limonene (trace amounts; see Table 1) was monoterpene hydrocarbon fraction (76%) dominated the two to three times higher in the middle of the light cycle flesh, with sabinene (50.6%) and 1,4-cyclohexadiene than it was in darkness in flowers and leaves of Brassica (6.2%) as the main components. The alcohol fraction napus in situ (Jackobsen et al., 1972). The essential oils represented 17.5% volatiles with 3-cyclohexen-1-ol (15%) from the leaves of Clausena anisata (Willd.) J.D. Hook ex as the major component. In the skins, 30 components could Benth, were isolated and found to contain mainly sabinene be identified, amounting to 76.7% of the total volatiles. (33.0%), germacrene-D (17.0%), Z-b-ocimene (6.0%), The sample was dominated by the monoterpenes (94%), germacrene-B (5.5%), (E)-b-ocimene (4.9%) and terpi- with sabinene (69%), a-phellendene (10.6%) and a-pinene nen-4-ol (4.7%) (Gundidza et al., 1994). (9.4%) as the main components. In the seeds, 25 The headspace composition of wampee samples gives a components could be identified, amounting to 99% of the better overall representation of the compounds detected by total volatiles. The sample was dominated by the mono- smell as compared with that of the steam distillate. The terpenes (98%), with sabinene (84%), a-pinene (4.3%), a- study successfully isolated and characterized the low- phellandrene (3.1%) and myrcene (2.9%) as the main temperature volatile aromatic compounds in wampee using components. the headspace sampling technique. a-phellandrene was abundant in all of the samples examined. a-pinene was more abundant in skin than in References seed and flesh, whereas 3-cyclohexen-1-ol was a distinctive component in the flesh. Sulphur-containing compounds Alasalvar, C., Grigor, J.M., Quantick, P.C., 1999. Method for the static could not be identified in any part of the samples examined. headspace analysis of carrot volatiles. Food Chemistry 65, 391–397. The composition of the volatile compounds of the leaves Camciuc, M., Bfssiere, J.M., Vilarem, G., Gaset, A., 1998. Volatile components in okra seed coat. Phytochemistry 48, 341–345. and seeds of C. lansium was distinctly different from that Davies, N.W., 1990. Gas chromatographic retention indices of mono- previously studied from Hainan Island, China (Zhao et al., terpenes and sesquiterpenes on methyl silicone and Carbowax 20 M 2004) by the absence of the b-santalol, bisabolol and ledol phases. Journal of Chromatography 503, 1–24. ARTICLE IN PRESS 56 P. Chokeprasert et al. / Journal of Food Composition and Analysis 20 (2007) 52–56

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