Chemical Composition of the Essential Oils of Two Alpinia Species from Hainan Island, China Peng Nana,b, Yaoming Huc, Jiayuan Zhaoa, Ying Fengb, and Yang Zhonga,* a Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China. Fax: 86-21-65642468. E-mail: [email protected] b Shanghai Center for Bioinformation Technology, Shanghai 201203, China c Center for Analysis and Measurement, Fudan University, Shanghai 200433, China * Author for correspondence and reprint requests Z. Naturforsch. 59c, 157Ð160 (2004); received July 24/August 29, 2003 The essential oils of two Alpinia species, i.e. A. hainanensis and A. katsumadai, from Hai- nan Island, China were analyzed by using GC-MS. The major constituents in the leaf oil of A. hainanensis were ocimene (27.4%), -pinene (10.1%), 9-octadecenoic acid (6.5%), n-hexadecanoic acid (5.8%), 9,12-octadecadienoic acid (5.4%), and terpinen (4.3%). The oil constituents obtained from the flowers of A. hainanensis were ocimene (39.8%), -pinene (17.7%), terpinene (5.5%), p-menth-1-en-ol (4.9%), caryophyllene (4.9%), and phellandrene (4.4%). In A. katsumadai, the major constituents in the leaf oil were p-menth-1-en-ol (22.0%), terpinen (19.0%), 4-carene (9.1%), 1,8-cineole (8.3%), and camphor (5.6%). The major constituents in the flower oil were p-menth-1-en-ol (21.3%), 1,8-cineole (20.2%), terpi- nen (12.6%), phellandrene (7.0%), 4-carene (6.4%), and -pinene (5.2%). Key words: Alpinia hainanensis and katsumadai, Essential Oil, GC-MS

Introduction binene, myrcene, and 1,8-cineole (Joseph et al., 2001), and A. zerumbet including terpinen-4-ol, The galangal genus of Alpinia with about 250 1,8-cineole, and -pinene (Ali et al., 2002) are in- species in the ginger family Zingiberaceae is dis- vestigated in detail. tributed mainly in China, India, East Indies, and Hainan Island (Hainan Province) is one of the Polynesia (Lemmon and Sherman, 1964; Lötschert major distribution regions of Alpinia plants in and Beese, 1983). The Alpinia plants have thick China. For example, the type specimens of A. katsu- fragrant rootstocks, resembling the scent of ginger, madai and A. hainanensis were collected from the from which the new shoots sprout in the spring, island. In recent years, about 80% of annual pro- and their leaves are lance-shaped with fringed bor- duction of Chinese A. katsumadai used as a fa- ders and they are produced on reedy stems. They mous Chinese traditional medicine were also from can grow up to 10 feet high with a 3-foot spread. Hainan Island. However, the chemical composi- Many Alpinia species are appreciated for their me- tion of essential oils of Alpinia plants in this region dicinal properties; they have a long history of use has not been reported yet. Therefore, the chemical in traditional medicine as a spasmolytic, hypoten- constituents of essential oils from leaves and flow- sive or diuretic due to their strong cardiovascular, ers of A. hainanensis and A. katsumadai in Hainan anti-emetic, anti-oxidant, anti-inflammatory, bac- Island were analyzed using GC-MS. teriostatic, or fungistatic effects in China, India and other regions (Jitoe et al., 1992; Habsah et al., 2000; Shin et al., 2002; Miyazawa and Hashimoto, Materials and Methods 2002; Ficker et al., 2003). Currently, the composi- Plant materials tions of essential oils from A. galanga including 1,8-cineole, -pinene, and camphor (Raina et al., The leaves and flowers of A. hainanensis and 2002; Mallavarapu et al., 2002), A. speciosa in- A. katsumadai used in this study were collected cluding limonene, 1,8-cineole, and terpinen-4-ol from Bawangling National Nature Reserve for (Zoghbi et al., 1999), A. purpurata including -pi- Black-crested Gibbon (18∞ 53Ј ~19∞ 30Ј N, 109∞ 0Ј nene, 1,8-cineole, and α-pinene (Zoghbi et al., ~ 109∞ 17Ј E), Changjiang County in Hainan Is- 1999), A. smithiae including -caryophyllene, sa- land in January 2003. The voucher specimens were

0939Ð5075/2004/0300Ð0157 $ 06.00 ” 2004 Verlag der Zeitschrift für Naturforschung, Tübingen · http://www.znaturforsch.com · D 158 P. Nan et al. · Essential Oils of Alpinia Species from Hainan Island deposited at the MOE Lab for Biodiversity Sci- Table I. Chemical constituents of the essential oils from ence and Ecological Engineering, School of Life A. hainanensis and A. katsumadai in Hainan Island, China. Sciences, Fudan University. Compound A. hainanensis A. katsumadai Extraction of essential oils Leaf Flower Leaf Flower The fresh flowers (250 g) and dry leaves (100 g) 3-Hexenol 0.2 of A. hainanensis and A. katsumadai were sub- Hexanol 0.1 2-Heptanol tr 0.1 0.7 0.2 jected to steam distillation for 3 h using a Clev- Thujene 0.1 enger-type apparatus. The essential oils were col- Cyclofenchene tr 2.1 lected in a lighter than water oil graduated trap Camphene tr 1.4 1.2 0.7 and dried over anhydrous sodium sulfate. Phellandrene 2.1 4.4 0.7 7.0 -Pinene 10.1 17.7 2.7 5.2 Myrcene 1.3 1.3 GC-MS analysis 4-Carene tr 0.7 9.1 6.4 m-Cymene 1.0 1.8 The GC-MS analysis was performed on a com- Limonene 1.9 2.0 1.8 bined GC-MS instrument (Finnigan Voyager, San 1,8-Cineole tr 0.5 8.3 20.2 Jose, CA, USA) using a HP-5 fused silica gel capil- Ocimene 27.4 39.8 lary column (30 m length, 0.25 mm diameter, Terpinen 4.3 5.5 19.0 12.6 0.25 µm film thickness). A 1 µl aliquot of oil was Terpineol 0.2 0.4 2.7 1.1 p-Mentha-1,4-diene tr 0.6 4.2 3.4 injected into the column using a 10:1 split injec- Unidentified tr 0.1 tion, which temperature was set up at 250 ∞C. The Linalool 0.3 0.1 GC program was initiated by a column temper- 1-Menthyl-4-isopropyl- ature set at 60 ∞C for 2 min, increased to 250 ∞Cat 2-cyclohexenol 1.2 1.8 Dimethyl octatetraene tr 0.1 a rate of 10 ∞C/min, held for 10 min. Helium was Camphor 2.8 3.4 5.6 3.5 used as the carrier gas (1.0 ml/min). The mass Trimethyl norbornanol tr 0.1 0.1 0.1 spectrometer was operated in the 70 eV EI mode Borneol tr 0.1 0.2 0.2 with scanning from 41 to 450 amu at 0.5 s, and p-Menth-1-en-ol 3.0 4.9 22.0 21.3 Benzylacetone 0.7 tr mass source was set at 200 ∞C. The compounds 2-Isopropyl-5-methyl- were identified by matching their mass spectral 3-cyclohexenone 0.1 0.2 0.4 0.1 fragmentation patterns with those stored in the Nonenal tr 0.2 spectrometer database using the National Institute Unidentified 0.3 0.1 Unidentified 0.1 tr of Standards and Technology Mass Spectral data- Dodecane 0.4 tr base (NIST-MS, 1998). Copaene 0.1 Santalene tr 0.2 0.1 0.9 Results and Discussion Caryophyllene 3.0 4.9 2.3 1.7 Dimethyl-6- The steam distillation of the flowers and leaves methylpentenyl-2- of A. hainanensis and A. katsumadai yielded clear norpinene 1.1 0.5 Unidentified tr 0.1 and yellowish essential oils. They were about (Z)--Farnesene 0.3 0.2 0.08% v/w and 0.1% v/w in the flowers of A. hai- -Caryophyllene 2.1 1.9 0.6 0.6 nanensis and A. katsumadai, and about 0.14% v/w Cubebene 0.1 tr and 0.11% v/w in the leaves of A. hainanensis and Tetramethyl hexahydro- A. katsumadai, respectively. The chemical constit- benzocycloheptene 0.2 0.1 Germacrene D 1.5 2.1 1.5 1.8 uents identified by GC-MS in the essential oils of Eudesma-4,11-diene 0.7 0.1 leaves and flowers of A. hainanensis and A. katsu- Chamgrene 0.8 0.2 madai are listed in Table I. α-Farnesene 0.4 0.7 In A. hainanensis, a total of 34 and 36 com- Longipinene 0.8 0.6 Eudesma-3,7-diene 0.1 0.2 pounds (about 93.6% and 98.7% of the oils) were Cadina-1,4-diene tr 0.1 identified from leaves and flowers, respectively. Ledol 0.7 The major constituents identified in the leaf oil n-trans-Nerolidol 1.5 1.1 of A. hainanensis were ocimene (27.4%), -pinene Denderalasin 0.1 tr Spathulenol 0.4 0.1 tr (10.1%), 9-octadecenoic acid (6.5%), n-hexadeca- noic acid (5.8%), 9,12-octadecadienoic acid (5.4%), P. Nan et al. · Essential Oils of Alpinia Species from Hainan Island 159

Table I. (cont.) The major constituents identified in the leaf oil Compound A. hainanensis A. katsumadai of A. katsumadai were p-menth-1-en-ol (22.0%), Leaf Flower Leaf Flower terpinen (19.0%), 4-carene (9.1%), 1,8-cineole (8.3%), camphor (5.6%). The major constituents Caryophyllene oxide 0.6 0.5 0.2 0.1 in the flower oil were p-menth-1-en-ol (21.3%), Unidentified tr 0.2 Dodecadienol acetate 0.1 tr 1,8-cineole (20.2%), terpinen (12.6%), phellan- Cadinol 0.8 drene (7.0%), 4-carene (6.4%), and -pinene Bisabolol 0.2 0.1 (5.2%). Unidentified 1.2 0.9 Of the major constituents of essential oils in Tetradecanioc acid 0.4 A. hainanensis and A. katsumadai, four constitu- Benzyl benzoate 0.3 tr Sinensal 0.5 ents, i.e. -pinene, 1,8-cineole, p-menth-1-en-ol, Hexahydrofarnesyl and ocimene, have strong antimicrobial activities acetone 0.5 0.2 (Juliani et al., 2002; Faleiro et al., 2003; Kim et al., Unidentified 0.4 0.7 2003) and 1,8-cineole has also cardiovascular ef- trans-Bergamotol 0.1 tr 9-Hexadecenoic acid 1.0 fects (Lahlou et al., 2002). However, the main n-Hexadecanoic acid 5.8 0.5 0.4 1.0 components were diversified, which led to dif- Myristamide 0.8 0.1 ferent effects in some Alpinia species (Zoghbi Isopropyl palmitate 0.9 et al., 1999; Raina et al., 2002). On the other hand, trans-Farnesyl actate 0.2 tr n-Heptadecanoic acid 0.4 the composition variations of essential oils in Alpi- Unidentified 0.6 0.2 nia species have chemotaxonomic implications. Phytol 0.5 0.1 Although some authors suggested that the four 9,12-Octadecadienoic Alpinia species, i.e. A. hainanensis, A. katsumadai, acid 5.4 9-Octadecenoic acid 6.5 A. henryi, and A. kainantensis, should be merged Unidentified 3.4 into one species A. hainanensis based on morpho- Hexadecanamide 4.0 tr 0.5 0.3 logical characters (Wu, 1997). Our study shows n-Tricosane 1.3 that there are significant differences between the 9-Octadecenamide 2.0 0.9 1.1 2.2 chemical constituents from essential oils of A. hai- Octadecanamide 1.8 n-Pentacosane 2.1 0.1 nanensis and A. katsumadai. It is obvious that fur- ther comparative studies of chemical constituents * Numbers represent the percentage of each constitu- of essential oils in Alpinia are necessary. ent in total essential oil. tr: Trace quantities (< 0.1%). Acknowledgements and terpinen (4.3%). The major constituents iden- We would like to thank Professor Suhua Shi of tified in the flower oil were ocimene (39.8%), Zhongshan University and Mr. Shaowei Chen of -pinene (17.7%), terpinene (5.5%), p-menth-1- Bawangling National Nature Reserve for Black- en-ol (4.9%), caryophyllene (4.9%), and phellan- crested Gibbon for their help in fieldwork. This drene (4.4%). study was partially supported by Chinese Nation- In another species, A. katsumadai, a total of 44 al Key Project for Basic Research (973) (2002 and 36 compounds (about 96.8% and 98.5%) were CB 512801) and Chinese PostDoc Foundation identified from leaves and flowers, respectively. (2003033270). 160 P. Nan et al. · Essential Oils of Alpinia Species from Hainan Island

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