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ISSN 2224-1698 Jagannath University Journal of Science, Volume-2, Number-I, January 2013

COMPARATIVE STUDIES ON PHYSICO-CHEMICAL PROPERTIES
AND GC-MS ANALYSIS OF ESSENTIAL OIL OF MYRISTICA
FRAGRANS
Shamsun Naher1, Mohammad Sayed Alam1, Md. Mizanur Rahman1 and
Shahin Aziz 2
1
1

2

Department of Chemistry, Jagannath University, Dhaka-1100, Bangladesh
Chemical Research Division, Bangladesh Council of Scientific and Industrial Research (BCSIR),
Dhaka-1205, Bangladesh

Abstract
Myristica fragrans Houtt fruit is locally known as jaiphal/nutmeg. It is one of the oldest and
widely used spice in the world. The essential oils from M. fragrans seeds are used in tonics. It also
shows antimicrobial, cytotoxic, anticancer, chemoprotective, antioxidant, antiinflammatory,
antithrombotic, hypolipidaemic, antiatherosclerotic activity. In the present study, essential oil were
extracted by steam distillation from the Indian and Sri Lankan nutmeg and investigated for their
chemical compositions by GC-MS. Total nine and ten compounds were identified in the essential
oil composition extracted from the Indian and Sri Lankan nutmeg, respectively. Among them
seven compounds was found common in both the Indian and Sri Lankan nutmeg essential oil e.g.
β-phellandrene, (1S)-6,6-dimethyl-2-methylene-bicyclo [3.1.1] heptane, (1R)-α-pinene, 4-methyl1-(1-methylethyl)-3-cyclohexen-1-ol, 1-methyl-4-(1-methylethyl) -1,4-cyclohexadiene, 1,2,3trimethoxy-5-(2-propenyl)-benzene and β-myrcene. On the other hand, 2,4-dimethyl-1,3dioxolane-2-methanol (12.462 %) and 2-methyl-5-(1methyethyl)-bicyclo[3,1,0] hex-2-ene (2.858
%) were found only in the essential oil of Indian nutmeg. Whereas, 4-hydroxy-4-methyl-2pentanone (12.244 %), (+)-4-carene (2.963 %) and 5-(2-propenyl)-1,3-benzodioxole (2.374%)
were identified only in the essential oil of Sri Lankan nutmeg. In both the Indian and Sri Lankan
nutmeg essential oil, β-phellandrene was found in highest concentration with 27.775 and 31.357
%, respectively. Among the common chemical constituents, the Indian nutmeg oil contains highest
percentage of each ingredient compared with that of Sri Lankan nutmeg oil except, βphellandrene. The specific gravity of the essential oil of Indian and Sri Lankan nutmeg were 0.88
and 0.87, whereas, the refractive index was found 1.4815 and 1.4805, respectively.
Key words: Myristica fragrans, physico-chemical properties, essential oil

1. Introduction
Myristica fragrans Houtt is a flowering vine in the family of myristiceae cultivated for its
fruit, which is usually used as a spice and seasoning. The local name of this plant is
nutmeg tree. Nutmeg is one of the oldest and widely used spice in the world. The nutmeg
spice is produced from its unripe but fully developed fruits. The fruit of this plant has the
characteristics aroma and flavor due to their chemical substances especially the volatile
Corresponding author: Mohammad Sayed Alam, E-mail: msalam@chem.jnu.ac.bd

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The effectiveness of the treatment may be due to the inhibition of prostaglandin synthesis in the mucosa and sub mucosa of the colon. to control flatulence. but found in sexual and general debility. oil. indigenous to India. 1959.. Finally.50 mm prior to the extraction process. The mixture of dirt free nutmeg powder (200 gm) and 600 mL distilled water were taken in a distillation flask (Clevenger’s apparatus. teas and soft drinks or mixed in milk and alcohol (Olaleye et al. collected from India and Sri Lanka. the dried nutmeg were crushed by Fritsch mortar grinder (Germany) for one hour and powder was sieved and collected with particle size 0.. JAOAC) and refluxed by electric heating mantle for four hours. dyspepsia. The raw samples were washed with water to remove dust materials and kept dried in air dry for 2 weeks. 2007) indicating enormous therapeutic potential of this plant. 2. fragrans are well known for their traditional medicinal uses.. 2005. stimulant. chutney and powder (Joshi et al. It is sometimes used as a stomachic. It is used as an antidiarrheal agent for patients with medullar carcinoma of the thyroid. Extraction of essential oil Steam distillation method was used to extract essential oil from the plant materials. antileukaemic effects and other biological activities (Latha et al. 2011). Nutmeg belongs to the order of Magnoliales which contains about 150 genera and more than 3000 species. It is also used as components of curry powder. and has a reputation as an emmenagogue and abortifacient (Green. Finally. carminative as well as for intestinal catarrh and colic. flatulence. to stimulate appetite. Yang et al. Recent reports on its antibacterial. 1996). Indonesia and Sri Lanka and now cultivated in many tropical countries of both hemispheres as well as in South Africa (Pal et al. The mixture of water vapor and essential oil of nutmeg were condensed by 70 . The oil of nutmeg is useful in the treatment of inflammation of the bladder and urinary tract. we have extracted essential oil from the fruits of M. candy.Shamsun Naher et al. impotence. fragrans (Nutmeg). halitosis. Panayotopoulos and Chisholm. Chirathaworn et al. M.25 and 0. Myristica species are natives of Moluccas.. insomnia and skin diseases. It is also used externally as a stimulant and ointment as a counter-irritant. In the present study. we also studied and compared the physico-chemical properties of essential oils extracted from aforesaid two different varieties.e. 1970).. and carried out the comparative studies of chemical composition using GC-MS analysis. pickle. nutmeg were collected from the local markets of Dhaka city (Chawkbazar) which are usually imported from India (kerala) and Sri Lanka. Materials and Methods The plant materials i. It has also been found useful as tonic for the heart and brain. antiviral. 2006). antidiabetic. It is used to produce nutmeg rind preserve.. 2006.

The spectrum of the unknown component was compared with the spectrum of the known component stored in the NIST library. After that the sample was injected by a micro syringe through a heated injection part which was vaporized and passed into the column. The retention time. samples were injected by splitting with the spilt ratio 10. The yield of percentage of essential oil was determined using the following formula described by Rao et al. helium carrier gas at constant pressure 90 KPa. The ether-extracted with ether was dried over anhydrous Na2SO4 and then filtered. Amount of essential oil recovered (g) Yield (%) = × 100 Amount of plant material used (g) Preparation of samples and GC-MS analysis Essential oil was diluted to 7% by chloroform (w/v) and analyzed by Electron Impact Ionization (EI) method on GC-17A gas chromatograph.Jagannath University Journal of Science. The ether was removed in vacuo and the remaining essential oil was kept in a dry vial. The signal was passed to a detector. molecular formula and composition percentage of the sample material was recorded. January 2013 water condenser and collected in a round bottom flask. Identification of the components Interpretation of mass spectroscopy (GC-MS) was conducted using data base of National Institute Standard and Technology (NIST) having more than 6200 patterns. range of linear temperature increase-100C per min. The GC-MS analysis of samples was carried out as follows: the flow rate of the carrier gas was adjusted to ensure reproducible retention time and to minimize the dirt of detector. coupled to a GC-MS 2010 plus mass spectrometer. molecular weight. injector temperature-2200C. column-initial temperature 400C. holding 5 min. The long tube of the column was tightly packed with solid particles. The carrier gas and sample component was then emerging from the column and passed through a detector. Essential oil makes a layer over water which was extracted and washed with analytical grade ether. The solid support was uniformly covered with a thin film of a high boiling liquid (the stationary phase). 71 .25mm. (2005). Volume-2. The mobile and stationary phases were then partitioned by the samples and it was separated into the individual components. Number-I. The amount of each component as concentration by the device and generates a signal which was registered electrically. The operating condition were as follows: fused silica capillary column (RTS-5MS) wtih diameter 30 cm and length 0. column packing was done with 10% diethylene glycol succinate on 100-120 mesh diatomic CAW.

e.374%) were identified only in the essential oil composition of Sri Lankan nutmeg.2. Total nine and ten compounds were identified in the essential oil extracted from Indian and Sri Lankan nutmeg. Among them following seven compounds were found common (Figure 1) in the essential oil of Indian and Sri Lankan nutmeg.1]heptane (9. we also studied and compared the physico-chemical properties of essential oils extracted from the aforesaid two different varieties. 1methyl-4-(1-methylethyl)-1.4-dimethyl-1.462 %) and 2-methyl-5-(1methyethyl)-bicyclo[3.4-cyclohexadiene (4. 4-methyl-1-(1-methylethyl)-3-cyclohexen-1-ol. (1S)-6. sausages. 3. (1S)-6.g. Finally.4-dimethyl-1. respectively.841%). meats.244%). It is used to flavor many kinds of baked foods. vegetables.847%). Peak areas and retention times were measured by the electronic integrator. (+)-4-carene (2. GC-MS analyzed results which included retention time.597) and 1methyl-4-(1-methylethyl)-1. fragrans extracted from two varieties are presented in Table 1.963 %) and 5-(2-propenyl)-1.3-dioxolane-2-methanol (12. molecular formula.618-5.498-9. β-phellandrene were found in highest amount with 27. β-phellandrene.0]hex-2-ene (2. 4-hydroxy-4-methyl-2-pentanone (12.858 %) were found only in the essential oil composition of Indian nutmeg. In the essential oil of Indian and Sri Lankan nutmeg.1.3-trimethoxy-5-(2-propenyl)-benzene and β-myrcene.357 %. Essential oil compositions were identified by comparing retention time of the chromatogram peaks with those of standard compounds using data base of National Institute Standard and Technology (NIST).733%).3-benzodioxole (2. saucers. 4-methyl-1-(1-methylethyl)-3-cyclohexen-1-ol (5. On the other hand.Shamsun Naher et al.6-dimethyl-2-methylene-bicyclo[3. and beverages (Panayotopoulos and Chisholm. molecular weight and chemical composition of the essential oil of fruits of M.1.3-dioxolane-2-methanol (12. followed by (1R)-α-pinene (9.4-cyclohexadiene.894-7.244 %).1]heptane. Whereas. confections. The above mentioned variations in the chemical composition of the essential oil of Indian and Sri Lankan 72 .1. puddings. The chemical composition of essential oil extracted in the present study was found similar with some exceptions (Table 1). respectively. respectively.775 and 31. Results and Discussion Nutmeg has a characteristic pleasant fragrance and is slightly warm taste.6-dimethyl-2-methylenebicyclo[3.462 %) and 4-hydroxy-4-methyl-2-pentanone (12. 1970). The second highest chemical constituent of Indian and Sri Lankan nutmeg essential oil were 2. 2.461-9. (1R)-α-pinene. Among the common chemical constituents. the Indian nutmeg oil contains highest amount of each ingredient compared to that of Sri Lankan nutmeg oil except. 1. The present study reports the comparative studies on the chemical composition of essential oil which was extracted from the Indian and Sri Lankan nutmeg using GC-MS analysis. β-phellandrene.

Physico-chemical analysis was performed according to the procedure described by Atti-Santos et al. respectively. The results of comparative studies of physico-chemical properties of essential oil of M.1. The specific gravity of the essential oil of Indian and Sri Lankan nutmeg were 0.915 and that of west Indian has been 0. It is well known that refractive index is an important parameter to evaluate the 73 . 2000 and Venturella. soil composition and climate differences.86 to 0.3-Trimethoxy-5(2-propenyl)-Benzene Figure 1. the refractive index of Indian and Sri Lankan nutmeg essential oil was found 1. respectively.4746 (Leela.87. 2008). while that of West Indian Nutmeg essential oil is ranging from 1. cultivation peculiarities. phenols and oxygenated compounds which are highly volatile at room temperature (Al-Shahhat.885 to 0. (2005).1.88 at 20˚ C. Previously. January 2013 nutmeg might be due to the genotype. Sarath-kumara (1985) have reported that the specific gravity of the East Indian Nutmeg oil ranges from 0. Volume-2. fragrans are presented in Table 2.4-cyclohexadiene 4-Methyl-1-(1-methylethyl)-3-cyclohexen-1-ol 4-Hydroxy-4-methyl -2-pentanone O 5-(2-Propenyl)-1.6-Dimethyl-2methylene-bicyclo [3. stage of maturity. The result shows that the smell of the essential oil from M. HO O H O (1S)-6.4729-1.88 and 0.4776-1.3 -benzodioxole O O O (+)-4-Carene β-Myrcene β-Phellandrene 1. Structure of identified compounds. The characteristic pungent odor and spicy taste of the essential oil nutmeg might be due to the presence of some low molecular weight compounds like alcohols. 2000). Number-I.4815 and 1. it has reported that the refractive index of the East Indian Nutmeg essential oil is ranging from 1. fragrans obtained was very strong which was turpentine like odor and the taste was spicy.4805.1] heptane OH 2.3dioxolane-2-methanol 2-Methyl-5-(1methyethyl) -bicyclo[3. However.0]hex-2-ene O O H OH (1R)-Pinene 1-Methyl-4-(1-methylethyl)1.2.4861. esters.Jagannath University Journal of Science.4-Dimethyl-1.

Table 1. cMolecular formula.4-Dimethyl-1. 11. 1550 208 C12H16O3 3.Pinene 4-Methyl-1-(1methylethyl)-3cyclohexen-1-ol 1-Methyl-4-(1methylethyl)-1. bMolecular weight.357 No RTa Name of the compound MWb MFc 1.618 8. 964 136 C10H16 2. 902 136 C10H16 2.682 3. 948 136 C10H16 9.753 4.3-Trimethoxy-5-(2propenyl)-Benzene β-Myrcene (+)-4-Carene 5-(2-Propenyl)-1.963 12.3dioxolane-2-methanol 2-Methyl-5(1methyethyl)bicyclo[3.858 - 5. especially the solid compounds (Ihsan.4cyclohexadiene 4-Hydroxy-4-methyl-2pentanone 1.Shamsun Naher et al.847 9. 1137 154 C10H18O 7.693 - 2.2.1. 1046 132 C6H12O3 12. 998 136 C10H16 5.374 a Retention time. 1327 β-Phellandrene (1S)-6.244 9.597 5. 845 116 C6H12O2 - 12.6-Dimethyl-2methylene-bicyclo[3.3benzodioxole 162 C10H10O2 - 2. quality of oils and fats and with the increase of refractive index value also increase the quality oils and vice versa.894 7.1] heptane 2. dEssential oli extracted from seeds of Indian nutmeg.498 6. dEssential oli extracted from seeds of Sri lankan nutmeg. The refractive index also related with the specific gravity of the oil and affected by the change in the concentrations of its components. Composition (%) Sample 1d Sample 2e 27.462 - 4. 1999).841 9. 958 919 136 136 C10H16 C10H16 2. 74 .0]hex-2-ene (1R)-α.775 31.501 2.1. Chemical constituents of the essential oil extracted from fruits of Indian and Sri Lankan nutmeg.894 10.461 3. 943 136 C10H16 9.

357 %. Volume-2. (1R)-α-pinene. hypolipidaemic. fragrans is impressive.87.3dioxolane-2-methanol (12. Number-I. 1-methyl-4-(1-methylethyl)-1.1.2. antithrombotic.6-dimethyl-2-methylene-bicyclo[3.3trimethoxy-5-(2-propenyl)-benzene and β-myrcene. anticancer. Though the existing chemical and pharmacological literature on M. chemoprotective.775 and 31. more topics remain open to future investigation like.9 cms long and nearly as broad.1. the refractive index was found 1. 2. (+)-4-carene (2. whereas. β-phellandrene was found in highest concentration with 27. The essential oils from M. succulent. pendulous. antiinflammatory. 4-hydroxy-4methyl-2-pentanone (12. smooth. 6. In the present study. In the Indian and Sri Lankan nutmeg essential oil. 75 .88 and 0. β-phellandrene.244 %). The oil of nutmeg also have antimicrobial. bushy and evergreen. Whereas.4-dimethyl-1.1]heptane. cytotoxic.g. antioxidant. (1S)-6. respectively.3benzodioxole (2. January 2013 4.4805. essential oil were extracted by steam distillation from the Indian and Sri lankan nutmeg and investigated for their chemical composition by GC-MS.Jagannath University Journal of Science.0]hex-2-ene (2. On the other hand.858 %) were found only in the essential of Indian nutmeg. 9. respectively. Among them seven compounds were common e. The specific gravity of the essential oil of Indian and Sri Lankan nutmeg were 0. respectively. Total nine and ten compounds were identified in the essential oil extracted from the Indian and Sri Lankan nutmeg. fragrans seeds are used in tonics.12 m tall. fragrans obtained was very strong which was turpentine like odor and the taste was spicy. fragrans is a dioecious or monoecious tree. Among the common chemical constituents. 4-methyl-1-(1- methylethyl)-3-cyclohexen-1-ol.374%) were identified only in the essential oil of Sri Lankan nutmeg. antiatherosclerotic and behavioural effects.4-cyclohexadiene. β-phellandrene.963 %) and 5-(2-propenyl)-1.462 %) and 2-methyl-5-(1methyethyl)-bicyclo[3. The results physicochemical properties shows that the smell of the essential oil from M. Conclusion M. the Indian nutmeg oil contains highest percentage of each ingredient compared with that of Sri lankan nutmeg oil except. 1.4815 and 1. The fruit is a one-seeded fleshy drupe.

1Volume Soluble in 0. The Volatile Oils. Egypt.08Volume Soluble at any Volume Not Soluble Soluble at any Volume Soluble at any Volume Soluble at any Volume Soluble at any Volume Soluble at any Volume a Essential oli extracted from seeds of Indian nutmeg.. Rech J.87 1.. 76 .850 Not soluble Cloudy up to 9. Panseral M.D. Rotal L.A..R.A.8 volume 80% alcohol 90% alcohol 95% alcohol 100% alcohol Distilled water Chloroform CC14 Petroleum ether Diethyl ether n-hexane Soluble in 5. Pauletti G. Arabic House for Publishing and Distribution.. Physical properties Oil yield (%) Taste Organoleptic Odor properties Color Appearance at RT Specific gravity at 300c Refractive index [ήt c] Optical rotation[α]26 60% alcohol 70% alcohol Solubility Sample 1a Sample 2b 11. Agostini F. 2005. References Al-Shahhat N. pp. Brazilian Archives of Biology and Technology..21 Spicy Spicy Pungent Pungent Colorless Colorless Homogeneous.4815 1.C.. 48.0 volume Soluble in 0. and Moyna P. transparent liquid. Atti-Santos A. 2000.F..C.71 12.Shamsun Naher et al. lighter than water 0. Serafini L. Physico-chemical evaluation of Rosmarinus officinalis L.4805 +21. Comparative studies on physico-chemical properties of essential oil extracted from the fruits of Indian and Sri Lankan nutmeg.. 1035-1039. bEssential oli extracted from seeds of Sri lankan nutmeg.950 +21. essential oils. Rossatol M.88 0. Table 2.

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