A. mutica Chemical Composition of the Fruits Oil of Alpinia mutica Roxb. (Zingiberaceae) Hasnah Mohd Sirat,* Nor Farhida Mohd Khalid, Nor Akmalazura Jani and Norazah Basar, Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia; E-mail: hasnah@kimia.fs.utm.my Abstract The chemical composition of the young and mature fruit oils of Alpinia mutica were obtained by hydrodistillation. The oils were analyzed by capillary gas chromatography using flame ionization and mass spectrometric identification. The main constituents found in the oil were sesquiterpenoids (74.4–77.4%), monoterpenoids (17.0–19.6%), long chain hydrocarbon (0.5–0.6%), esters (0.9–1.2%), and ketone (0.5–2.8%). (E,E)-Farnesol was the major constituent (44.3–51.2%). Key Word Index Alpinia mutica, Zingiberaceae, essential oil composition, (E,E)-farnesol. Introduction Alpinia mutica Roxb. (syn. Languas mutica Merr.) is a perennial herb belonging to the family of Zingiberaceae (1). It is endemic to southern parts of Malaysia especially in the Johor area. The plants have been cultivated as ornamentals and the rhizomes have been used as a stomachic (2). The chemical components of the dried rhizomes showed the presence of phenolic compounds, such as 5,6-dehydrokawain, flavokawin B, and diarylheptanoid, 1,7-diphenyl-3-hydroxy-6-heptene5-one (3). The chemical composition of the fresh rhizome oil has also been reported (4), with camphor as the major constituent. Several phytochemicals from the fruits and rhizomes have been reported to show an inhibitory effect on platelet activating factor receptor binding (5), and an inhibitory effect on human platelet aggregation (6). To the best of the authors’ knowledge, there has been no report on the essential oil of the fruits. Therefore this paper describes the results of the GC and GC/MS analyses of the essential oil obtained from the young and the mature fruits of A. mutica. Experimental Plant material: Fresh fruits of Alpinia mutica were collected from Pontian, Johor, Malaysia. A voucher specimen, HS 135, has been deposited at the Herbarium of the Department of Biology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia. Oil isolation: The young (green color) and the matured (orange to red color) fresh fruits (305 g and 310 g, respectively) were submitted to hydrodistillation using a Clevenger-type ap- paratus for 8 h. Extraction of the distillate with diethyl ether, followed by drying over anhydrous magnesium sulphate and removal of the solvents yielded 0.29% and 0.52% (w/w) of colorless oil based on the fresh weight, respectively. Analysis: The quantitative GC analysis was performed on a Hewlett-Packard HP 5880A system equipped with a flame ionization detector, using a non-polar Ultra1 fused silica capillary column (25 m × 0.32 mm; 0.20 mm film thickness). Helium was used as carrier gas at a flow rate of 1 mL/min and 30 psi inlet pressure; split ratio 1:20. The oven temperature was programmed from 50°C for 5 min, 50–300°C at a rate of 4°C/min, and kept isothermal for 10 min. Both injector and detector temperatures were set at 250°C. The qualitative GC/MS analysis was carried out on a 5989A Hewlett-Packard instrument GC/MS system equipped with Wiley Library software. The capillary column and GC conditions were used as above. Significant operating parameters: ionization voltage, 70 eV; ion source temperature, 200°C; scan mass range, 40–350 m. The individual components of the oil were identified on the basis of their retention indices (RI) with reference to homologous series of C9–C24 n-alkanes, and by matching their 70 eV mass spectra with the data from the Wiley Library. Column chromatography: Column chromatography was performed on Merck silica gel 70–230 Mesh using, hexane and diethyl ether (9:1) as an eluent. Component identification: Sample components were identified by matching their mass spectra with those recorded in the MS Wiley Library and further supported by comparison *Address for correspondence Received: October 2007 Revised: December 2007 1041-2905/09/0005-0457$14.00/0—© 2009 Allured Business Media Vol. 21, September/October 2009 Accepted: July 2008 Journal of Essential Oil Research/457 Sirat et al. of their GC retention times, TLC and MS with those of reference compounds when the latter were available. The pure isolated compound, (E,E-farnesol) was identified by comparison with the authentic sample previously isolated from Amomum uliginosum (7) and literature value (8). Results and Discussion The fruits of A. mutica yielded ~ 0.30–0.50% oil. Fortyeight compounds representing more than 97% of the oil were identified. They are listed in Table I. Among these compounds were 17 sesquiterpenes (74.4–77.4%) and 23 monoterpenes (17.0–19.6%), together with four long chain hydrocarbons (0.5–0.6%), three esters (0.9–1.2%), and a ketone (0.5–2.8%). Both fruit oils consist of very similar constituents, with sesquiterpenes contributing the main components. The major sesquiterpene was (E,E)-farnesol (44.3–51.2%) followed by a-farnesene (7–8%). This showed that fruit oil of Alpinia mutica can be used as a source of farnesol. It is worth mentioning that the fruit oils are very different from the rhizome oil collected from cultivated plants in Universiti Putra Malaysia (4), which was rich in monoterpenes (81.7%) with camphor as the major constituent (35.6%). Acknowledgments The authors wish to thank Abd. Kadir A. Rahman and Ayob Jabal for the help of GC and GC/MS, respectively. The authors also would like to thank FELDA and MOSTI for financial support and Universiti Teknologi Malaysia for scholarships to Norazah Basar and Nor Akmalazura Jani. References 1. 2. 3. 4. 5. 6. 7. 8. R.E. Holttum, The Zingiberaceae of the Malay Peninsula. Gard. Bull. Sing., 1, 1–249 (1953). I. H. Burkill, A dictionary of the economic products of the Malay Peninsula. Vol II, p. 1333, Ministry of Agriculture and Co-operatives, Kuala Lumpur, Malaysia (1966). H.M. Sirat, A.A. Rahman, H. Itokawa and H. Morita, Constituents of two Alpinia species. Planta Med., 62, 188–189 (1996). H.M. Sirat and A.A. Rahman, Essential Oil Constituents of Alpinia mutica Roxb. J. Essent. Oil Res., 10, 83–84 (1998). I. Jantan, M. Pisar, H.M. Sirat, N. Basar, S. Jamil, J. Jalil and R. Ali, Inhibitory Effects of Compounds from Zingiberaceae Species on Platelet Activating Factor Receptor Binding. Pytotherap. Res., 18, 1005–1007 (2004). I. Jantan, S.M. Raweh, H.M. Sirat, S. Jamil, Y.H.M. Yasin, J. Jalil and J.A. Jamal, Inhibitory Effect of Compounds from Zingiberaceae Species on Human Platelet Aggregation. Phytomedicine, 15, 306–309 (2008). L.F. Hong, Isolation and Identification of chemical components from Amomum species. Unpublished results, B. Sc. Thesis, Universiti Teknologi Malaysia (1997). T.K. Devon and A.I. Scott, Handbook of Naturally Occurring Compounds. Vol. 1, p. 103, Academic Press, New York, NY (1975). 458/Journal of Essential Oil Research Table I. Chemical Compositiona of the Fruit Oils of Alpinia mutica Compoundsb a-pinene camphene b-pinene myrcene a-phellandrene p-cymene 1,8-cineole (Z)-b-ocimene (E)-b-ocimene g-terpinene terpinolene linalool a-fenchol camphor isoborneol borneol terpinen-4-ol a-terpineol 3-phenyl-2-butanone citronellol geraniol geranial bornyl acetate carvacrol a-cubebene geranyl acetate a-copaene b-elemene b-caryophyllene a-bergamotene* g-elemene a-humulene b-farnesene* ar-curcumene zingiberene a-farnesene* b-bisabolene b-sesquiphellandrene (E)-nerolidol caryophyllene oxide a-bisabolol (E,E)-farnesol (E,E)-farnesyl acetate docosane tricosane tetracosane pentacosane Total RIc Young Fruit Oil %d Mature Fruit Oil %d 936 948 974 983 995 1013 1018 1028 1040 1055 1081 1086 1097 1125 1140 1145 1160 1170 1199 1217 1236 1248 1253 1306 1344 1352 1374 1387 1418 1425 1427 1441 1446 1471 1480 1499 1503 1515 1543 1560 1668 1705 1809 0.1 0.1 0.7 0.2 1.1 0.5 3.7 0.1 3.0 0.1 0.1 2.2 0.1 0.1 0.6 1.5 0.5 1.5 2.8 2.3 0.8 0.2 0.2 0.1 0.1 0.4 0.4 0.2 3.1 1.8 4.6 1.1 1.5 7.0 0.2 4.3 3.2 0.6 2.3 44.3 0.6 0.4 0.1 0.1 98.9% 0.2 0.2 1.7 0.2 0.8 0.6 4.5 0.1 2.8 0.2 1.0 0.1 0.9 1.3 0.3 0.7 0.5 1.1 0.3 0.1 0.1 0.1 4.2 1.5 0.1 3.2 0.4 0.3 0.4 8.2 0.3 2.7 2.9 0.5 2.3 51.2 0.8 0.1 0.2 0.1 0.1 97.3% a Method of identiication: RI, MS, Lit; b Order of elution on Ultra 1 column; RI, retention index relative to C9–C24 n-alkanes; d %, percentage composition; * correct isomer not identiied. c Vol. 21, September/October 2009