Chemical composition of Ambrette Essential Oil

Chemical topic of Ambrette Essential OilChemical composition of the crucial cover of ambrette (Abelmoschus moschatus Medik.) from subtropical orbit of north IndiaRam S. Verma*, Rajendra C. Padalia, Amit ChauhanABSTRACTAbelmoschus moschatus (Family Malvaceae), popularly know as ambrette or muskdana, is an important aromatic and medicinal plant of India. The plant is widely elegant in tropical countries for their musk-scented spills useful in perfumery and medicine. In this study, hydrodistilled ambrette author internal oil colour produced in subtropical region of north India was investigated using gas chromatography-flame ionisation detector (GC-FID) and GC- destiny spectrometry (GC-MS). A total of 27 constituents, re defering 90.4% of the total oil composition were identified. Major constituents of the oil were (2E,6E)-farnesyl ethanoate (58.0%), (Z)-oxacycloheptadec-8-en-2-one (12.1%), decyl acetate (4.8%), (2Z,6E)-farnesyl acetate (3.5%), (Z)-oxacyclopentadec-6-en-2-one ( 2.4%), dodecyl acetate (2.4%) and (2E,6Z)-farnesol (2.0%). Ambrettolide and its homologues, answerable for the characteristic musk-like odour, constitute 15.8% of oil composition.Keywords Abelmoschus moschatus, Malvaceae, ambrette seed, essential oil, (2E,6E)-farnesyl acetate1. IntroductionAbelmoschus moschatus Medik. (syn. Hibiscus abelmoschus (L.), commonly known as ambrette, is native to India (1). It is cultivated in tropical regions of Asia, Africa and South America for their seeds. The seeds sport been used as tonic, stimulant, carminative, diuretic, demulcent, and for stomachic property (2). The essential oil obtained by steam-distillation of ambrette seeds is a valuable material known for a rich, sweet, floral-musky, distinctly wine-like or brandy-like odor, which finds application in flavour and fragrance formulations (3). Moreover, the seed essential oil is used as anti-inflammatory, analgesic and antispasmodic drug. It is indicated against cramps and bowel disorders an d excessively useful in the itching caused by insect bites. The leaves and the fruits of the plant argon consumed in soups and the seeds atomic number 18 used as a spice (4). In addition to this, A. moschatus shows good antioxidant, antiproliferative and antimicrobial activities (5). The plant has been class as an herb of undefined safety by the Food and Drug Administration (FDA), and the extracts are classified as generally recognized as safe (GRAS) for their use in baked foods, candies, and alcoholic beverages (6). The chemical substance composition of essential oil and extracts of ambrette seed have been reported from different countries (7-16). The ambrette seed oil has a much smo another(prenominal) odor than synthetic musk compounds, and the major compounds responsible for the characteristic musky odor let in ambrettolide (Z)-7-hexadecen-16-olide and (Z)-5-tetradecen-14-olide (17).Despite a long history of uses in traditional medicines and in perfumery, information on A. mo schatus from subtropical region of India is meager. Therefore, in this study, volatile oil composition of the ambrette seed grown in north India (subtropical condition) has been investigated.2. Experimental2.1. Plant material and isolation of essential oilThe ambrette seeds were collected from experimental field of CSIR-Central Institute of medicinal and Aromatic Plants, Research Centre, Pantnagar (Uttarakhand) in the month of December (20092011). The experimental site is located between coordinates 29.02N, 79.31E and an altitude of 243 m in foothills of north India. isolation of the essential oil from ambrette seeds was carried out by hydrodistillation in a Clevengers type apparatus for 5 hours. Isolated oil was dried over anhydrous Na2SO4 and stored at 4C until further analyses.2.2. GC and GC-MS analysesGC analysis of the essential oil was carried out on a Nucon gas chromatograph model 5765 weaponed with DB-5 capillary column (30 m 0.25 mm internal diameter, film thickness 0.25 m) and flame ionization detector (FID). The oven column temperature ranged from 60230 C, programmed at 3 C/min, using H2 as carrier gas at 1.0 mL/min, a split ratio of 135, an injection size of 0.03 L neat, and injector and detector temperatures were 220 C and 230 C, respectively for Nucon gas chromatograph model 5765. GC/MS analysis of the essential oil sample was carried out on a Clarus 680 GC interfaced with a Clarus SQ 8C vision spectrometer of PerkinElmer fitted with Elite-5 MS fused-silica capillary column (30 m 0.25 mm i.d., film thickness 0.25 m). The oven temperature program was from 60240 C, at 3 C/min, and programmed to 270 C at 5 C /min injector temperature was 250 C transfer line and source temperatures were 220 C injection size 0.03 L neat split ratio 150 carrier gas He at 1.0 mL/min ionization energy 70 eV mass scan range 40-450 amu. picture was achieved on the basis of retention index (RI, determined using a homologous series of n-alkanes, C8-C30 hydrocarbons), m ass spectra library search (NIST/EPA/NIH version 2.1 and Wiley registry of mass spectral data 7th edition) and by comparing the observed RI and mass spectral data with the literature (18,19). The relative amounts of individual components were calculated based on the relative % peak areas (FID response), without using a correction factor.2.3. Statistical analysisTo compare of the examined essential oil composition of ambrette seed from subtropics with the reported compositions from other regions, seven samples (1 present study and 2-7 other regions) (8,9,11,13,14) were treated as operational taxonomic units. The percentage of nine major components, representing composition up to 82.8-89.0% of ambrette essential oil (decyl acetate, dodecyl acetate, (E)--farnesene, (Z)-oxacyclopentadec-6-en-2-one, (2Z,6E)-farnesyl acetate, (2E,6E)-farnesyl acetate, (2E,6E)-farnesol, (Z)-oxacycloheptadec-8-en-2-one, and (E)-2,3-dihydrofarnesyl acetate) were used to determine the chemical relationship am ong the different essential oil samples by hierarchical cluster analysis using the average method (20). This package computes the hierarchical clustering of a multivariate dataset based on dissimilarities. The derived dendrogram depicts the grouping of chemical compositions as per their chemical constituents.3. Results and discussionThe essential oil yield and chemical composition of ambrette seeds observed in subtropics, north India is presented in Table 1. The seeds gave 0.12 0.01% (v/w) of essential oil on hydrodistillation. However, essential oil yield was 0.150.20% in ambrette seeds under easterly Indian conditions (12). The resulting essential oil was analysed using GC-FID and GC-MS techniques. Altogether, 27 constituents, representing 90.4% of the total oil composition were identified. Major constituents of the oil were (2E,6E)-farnesyl acetate (58.0%), (Z)-oxacycloheptadec-8-en-2-one (12.1%), decyl acetate (4.8%), (2Z,6E)-farnesyl acetate (3.5%), (Z)-oxacyclopentadec-6-en- 2-one (2.4%), dodecyl acetate (2.4%), (2E,6Z)-farnesol (2.0%), (Z)-oxacyclononadec-10-en-2-one (1.3%) and (E)-nerolidol (0.7%).The essential oil composition of ambrette seed has been investigated earlier from different countries and mainly five types of compositions are described. Garnero and Buil (1978) identified (2E,6E)-farnesol (39.0%) and (E,E)-farnesyl acetate (35.4%) as the major constituents of ambrette seed oil (13). slime et al (1999) reported two different compositions, viz. (E)-2,3-dihydrofarnesyl acetate (67.3%) type, and (E,E)-farnesyl acetate (35.5%) and (E)-2,3-dihydrofarnesyl acetate (32.9%) type for ambrette seed oil from Vietnamese (14). However, ambrette seed oils from Ecuador and China are reported to have (E,E)-farnesyl acetate (59.1% and 64.22%) and (Z)-oxacycloheptadec-8-en-2-one (7.8% and 14.9%) as major constituents (8,9). According to an earlier study from Odisha (eastern India), the main constituents of ambrette seed oil were (E,E)-farnesyl acetate (47.6 %), (E)--farnesene (9.6%) and (Z)-oxacycloheptadec-8-en-2-one (9.0%) (11). Moreover, to compare the examined essential oil composition with earlier reported compositions, the contents (%) of nine major components of different oils were subjected to the hierarchical cluster analysis. The derived dendrogram clearly demonstrate dissimilarity based on the percentages of the constituents present among the different compositions (Figure 1). Thus, composition of the examined oil from subtropical northern India was closer to the oil composition reported from Ecuador (8). However, it was rather different from China (9) and eastern Indian (11) ambrette seed oils due to the content (%) of other constituents, viz. (E)--farnesene and decyl acetate.4. ConclusionsIn conclusions, the chemical composition of ambrette seed oil produced in subtropics was rich in (E,E)-farnesyl acetate (58.0%), and ambrettolide and its homologues (15.8%). The ambrette seed oil has a promising value for fragrance and fi xative purposes. Based on the results of this study, it can be said that ambrette can also produced good quality essential oil in the subtropical conditions of north India.AcknowledgementsCouncil of Scientific and Industrial Research (CSIR), New Delhi is thankfully acknowledged for the financial support to carrying out the work (Project BSC0203). Authors are also thankful to the Director, CSIR-Central Institute of Medicinal and Aromatic Plants for encouragement and the Central Chemical Facility (CSIR-CIMAP) for providing facility for GC and GC/MS analyses.ReferencesAnonymous, The wealth of India fresh materials National Institute of Science Communication, Council of Scientific and Industrial Research New Delhi, Vol. 5, pp 75-77 (1959).R. Sharma and A. Shahzad, Thidiazuran (TDZ) induced regeneration from cotyledonary node explant of Abelmoschus moschatus Medik. L. (A valuable medicinal plant). World J. Agric. Sci., 4(4), 449-452 (2008).S. Arctander, meat and flavor materials of nat ural origin Arctander Elizabeth, NJ, pp 58-60 (1960).De La Ripelle, H. F. (2006). Les hiscus, tradition et modernite. Phytotherapie, 3,136144.M.Z. Gul, L.M. Bhakshu, F. Ahmad, A.K. Kondapi, I.A. Qureshi and I.A. 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URL http//www.wessa.net/Table 1 Chemical composition of ambrette (Abelmoschus moschatus Medik.) seed essential oil from north IndiaS. no.CompoundaRIbRIc discipline (%)dS. no.CompoundaRIbRIcContent (%)d1-Pinene9339320.1 0.0915Decyl propanoate150215010.2 0.062-Pinene972974t16(E)-Nerolidol156015620.7 0.0636-Methyl-5-hepten-2-one9789810.1 0.0417(Z)-5-Dodecenyl acetate15881592*0.5 0.064-Terpinene10141014t18Dodecyl acetate160916072.4 0.255p-Cymene102210200.2 0.2519(2Z,6Z)-Farnesol169616980.1 0.006Limonene102610240.2 0.1620(2E,6Z)-Farnesol171317142.0 0.9371,8-Cineole102810260.2 0.2121(Z)-Oxacyclopentadec-6-en-2-one17192.4 2.438Linalool110010950.4 0.4622(2Z,6E)-Farnesyl acetate182218213.5 1.159Camphor11461141t23(2E,6E)-Farnesyl acetate1850184558.0 3.1310n-Decanol127012660.3 0.3524(2E,6E)-Farnesyl propanoate19190.4 0.1711Undecanal13041305t25(Z)-Oxacycloheptadec-8-en-2-one 1928192912.1 4.8812Decyl acetate140714074.8 0.9026(Z)-Oxacyclononadec-10-en-2-one21281.3 0.7913(E)--Farnesene145814540.2 0.3327Lino leic acid21292132t1410-Undecenol acetate149914980.1 0.10Total identified (%)90.4 6.25aMode of acknowledgment retention index (RI), mass spectral data (GCMS) RIb Experimental Retention Index (relative to n-alkane) RIc Retention Index from literature (18) dMean ( standard deviation) of three samples also known as (Z)-5-tetradecen-14-olide also known as (Z)-7-hexadecen-16-olide (= musk ambrette) *KI Kovat Index (19).Figure 1 Hierarchical cluster analysis of the essential oil compositions of ambrette (Abelmoschus moschatus Medik.) seed. 1 present study (2E,6E)-farnesyl acetate (58.0%), (Z)-oxacycloheptadec-8-en-2-one (12.1%) 2 (2E,6E)-farnesol (39.0%), (2E,6E)-farnesyl acetate (35.4%) (Garnero and Buil, 1978) 3 China (2E,6E)-farnesyl acetate (64.22%), (Z)-oxacycloheptadec-8-en-2-one (14.96%) (Tang et al., 1990) 4 Ecuador (2E,6E)-farnesyl acetate (59.1%), (Z)-oxacycloheptadec-8-en-2-one (7.8%) (Cravo et al., 1992) 5 Vietnam (E)-2,3-dihydrofarnesyl acetate (67.3%), (2E,6E)-farnesyl ace tate (14.9%) (Dung et al., 1999) 6 Vietnam (2E,6E)-farnesyl acetate (35.5%), (E)-2,3-dihydrofarnesyl acetate (32.9%) (Dung et al., 1999) 7 Eastern India (2E,6E)-farnesyl acetate (47.6%), (E)--farnesene (9.6%) (Rout et al., 2004).1