ANALYSIS OF ESSENTIAL OILS OF FLOWERS, RHIZOMES AND SEEDS OF ZINGIBER RUBENS ROXB

Trinh Dinh Chinh1 Hoang Thi Nhu Hanh2

1 2

Chemical Department, College of pedagogy, Hue university

Postgraduate student of College of pedagogy, Hue university 32 Le Loi street, Hue city, Viet Nam

Abstract The essential oils from flowers, rhizomes and seeds of Zingiber rubens Roxb were extracted. The essential oils were extracted using the hydrodistillation method and analysed by GC-MS. The yeild of volatile constituents (mL/100g) of flowers, rhizomes and seeds were 0.015, 0.075 and 0.094, respectively. The flowers essential oil contained trans-caryophyllene (22.38%), 2--pinene (19.22%) and bicyclogermacrene (10.88%) as major compound whereas the seeds essential oil were largely dominated by 2--pinene (33.74%), -pinene (11.79%) and trans--caryophyllene (11.61%). The essential oil of the rhizomes contained the major compounds (-) bornyl acetate (18.95%), 2--pinene (17.05%), endoborneol (11.35%) and linalool L (11.09%).

Introduction The pantropical Zingiberaceae is the largest family in the order Zingiberales with 53 genera and over 1300 species[2]. They are distributed throughout tropical Africa, Asia and America, with its greatest diversity in Southeast Asia. Of the 53 genera and 1300 species known in the world, at least 24 genera and 115 species are found in Viet Nam[1]. Zingiber rubens Roxb is classified under the genera of Zingiber. This plant is from the Indo-Malaysian region and has been found in many places throughout Southeast Asia. The paste of the rhizome (10 gm) is given after heating and is also applied to the head for giddiness. There were several previous studies of this plant. The study by Wimolpun Rungprom (1990) reported that the essential oil of rhizomes of Zingiber rubens Roxb,

obtained by steam distillation, gave the same bluish-green essential oil (0.40% of dried rhizomes and 0.28% of fresh rhizomes). Essential oil by gas chromatograph-mass spectrometer (GC-MS) showed that twelve compounds were the main components of the oil (84.05%). Four of these compounds were identified by computer as -pinene, conphene, -pinene and compaene. The residue from steam distillation was extracted with acetone by maceration. Five substances were obtained. Among them, two compounds, stigmasterol (C29H48O) and -sitosterol (C29H50O), were indentified. The ground dried roots of zingiber rubens Roxb were extracted with hexane, chloroform and methanol respectively. Three solid substances were obtained. The first subtances was believed to be the mixture of five compounds containing long chain aliphatic esters and long chain aliphatic hydrocarbons. The second substance was the mixture of three triterpenoids with molecular formular C30H50O. The third substance was believed to be sucrose[3]. The aim of this study was to analyze the volatile constituents of essential oils of Zingiber rubens Roxb from different parts of the plant namely flowers, rhizomes and seeds by gas chromatography and mass spectrometry. The study of these essential oils may provide interesting raw material for the application of this plant.

Experimental Plant material The fresh Zingiber rubens Roxb was collected at Binh Dien, Huong Tra, Thua Thien Hue in august, 2009. The plant was identified by Mr Cam, lecture of Hue college of Agriculture and Forestry, and transported to Natural compounds Laboratory of Hue college of pedagogy for essential oil extraction. Hydrodistillation 1 kg of flowers, 400 grams of rhizomes were chopped into small pieces, 160 grams of seeds were ground into pieces and subjected to hydrodistillation for 3-4 hours. The essential oils were isolated by extraction of the distillate with n-hexan and its yield was calculated. GC-MS analysis The essential oils were analyzed using GC model HP 6890 (America), MS model Agilent 5973 (America). The constituents were indentified by comparision with published mass spectra database Wiley 1998 and Nist 1998

Results and Discussion A total of fifty two components were indentified in the essential oils of flowers, rhizomes and seeds of Zingiber rubens Roxb. Among them, forty one compounds were indentified in essential oil of seeds, thirty eight compounds were indentified in essential oil of flowers and thirty nine compounds were indentified in essential oil of rhizomes. trans--caryophyllene (22.38%), 2--pinene (19.22%) and bicyclogermacrene (10.88%) were the major compounds of essential oil in flowers of Zingiber rubens Roxb. (-) Bornyl acetate (18.95%), endoborneol (11.35%), 2--pinene (17.05%) and linalool L (11.09%) were the major compounds of essential oil in rhizomes of Zingiber rubens Roxb. 2--pinene (33.74%), -pinene (11.79%) and trans--caryophyllene (11.61%) were the major compounds of essential oil in seeds of Zingiber rubens Roxb. -thujene, -terpinene, trans--ocimene were indentified in the essential oils of both flowers and seeds except rhizomes. Camphor, -elemene were indentified in the essential oils of both seeds and rhizomes except flowers. Linalool L, delta-cadinene were indentified in the essential oils of both flowers and rhizomes except seeds. Monoterpene hydrocarbons dominated the essential oil of seeds of Zingiber rubens Roxb with 61.03% whereas sesquiterpene hydrocarbons were the major type of compounds in the essential oil of flowers of Zingiber rubens Roxb with 46.21%. Oxygenated monoterpene derivatives were the major type of compounds present in the essential oil of rhizomes of Zingiber rubens Roxb with 53.02%. As a conclusion, major compounds in the essential oils depend on the part of plant studied. Chemical Compounds Monoterpene hydrocarbons Tricyclene -Thujene -Pinene Camphene Sabinene 2--pinene -Myrcene Percentage (%) Seeds 61.30 1.22 11.79 0.3 1.39 33.74 0.82 Flowers 31.24 0.17 5.63 0.45 1.23 19.22 0.58 Rhizomes 35.34 0.31 5.73 7.88 0.83 17.05 0.48

-Phellandrene -terpinene p-cymene Limonene trans--ocimene -terpinene -terpinolene Oxygenated monoterpene derivatives 1,8-cineole Linalool L Camphor Endo-borneol Terpinen-4-ol -terpineol Myrtenol Fenchyl acetate (-)-Bornyl acetat Sesquiterpene hydrocarbons -elemene -Gujurnene trans--caryophyllene -humulene Aromadendrene Germacrene B -cubebene -Chamigrene -Guaiene Valencene Ledene Bicyclogermacrene

0.46 0.95 2.16 1.68 0.29 6.34 0.16 3.84 0.36 0.17 0.15 2.04 0.69 0.43 14.87 0.3 11.61 1.21 0.69 -

0.14 0.5 1.16 1.09 0.81 0.26 16.38 4.76 7.88 0.75 0.37 1.19 1.09 0.34 46.21 0.6 22.38 2.58 2.07 0.69 0.39 10.88

0.15 2.37 0.22 0.32 53.02 7.42 11.09 0.81 11.35 0.87 2.27 0.26 18.95 5.19 0.66 0.65 0.19 0.19 0.27 0.7 0.12 0.17 0.4 0.33

Delta-guaiene -Bisabolene -Farnesene (+)-delta-selinene Neoisolongifolene Alloaromadendrene -selinene -selinene (-)-Isoledene Isolongifolene -amorphene Delta-cadinene Cycloheptane, 4-methylene-1-methyl-2-(2methyl-1-propen-1-yl)-1-vinyl Oxygenated sesquiterpene derivatives Santalol Caryophyllene oxide Miscellaneous compounds 1,2,3,5-tetramethyl benzene Acetic acid Total identified constituents Total non-identified constituents

0.39 0.17 0.5 2.7 0.24 2.46 0.47 0.29 0.18 83.18 16.82

0.43 1.09 2.6 0.32 0.55 0.53 0.87 0.23 93.83 6.17

0.19 0.83 0.25 0.24 93.55 6.45

70 60 50 40 30 20 10 0 Monoterpene Oxygenated Sesquiterpene Oxygenated Miscellaneous hydrcarbons monoterpene hydrocarbons sesquiterpene compounds hydrocarbons hydrocarbons Rhizomes Seeds Flowers

Figure 1. Chemical Compounds of the Essential oils of Zingiber rubens Roxb

(1)

(2)

(3)

(4)

(5)

(6)

The Major Chemical Constituents of Essential oils of flowers, rihzomes and seeds of Zingiber rubens Roxb (1) trans--caryophyllene ; (2) 2--pinene ; (3) -pinene ; (4) Endo-borneol ; (5) Bicyclogermacrene ; (6) Linalool L

References

1. Pham Hoang Ho (1993), Cay co Viet Nam quyen III, Mekong publisher, Montral. 2. W. John Kress, Linda M. Prince and Kyle J. Williams (2002), The Phylogeny and a new classification of the Gingers (Zingiberaceae): Evidence from molecular data, American Journal of Botany 89(11): 1682-1696, 2002. 3. Wimolpun Rungprom (1990), Chemical constituents of Zingiber rubens Roxb, Department of chemistry Faculty of science chulalongkorn University Thailand.