Medicinal used plants from India: analysis of the essential oils of Sphaeranthus indicus flowers, roots and stems with leaves

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1 Sc!entia Pharmaceutica (Sci. Pharm.) 71, (2003) 251 O Osterreichische Apotheker-Verlagsgesellschaft m. b.h., Wien, Printed in Ausia Medicinal used plants from Iia: analysis of the essential oils of Sphaeranthus iicus flowers, roots a stems with leaves L. Jirovetz*, G. Buchbauer a M. Shahabi Department of Pharmaceutical Chemisy, University of Vienna, Althansasse 14, A Vienna, Ausia P.M. Shafi a K.R. Rajeeve Department of Chemisy, Calicut University, Kerala , Iia Absact: The essential oils of flowers, roots a herb (stems with leaves) of Sphaeranthus iicus Kurz. (Asteraceae) from southern Iia were investigated by gas chromatographic-specoscopic (GC-FID a GC-MS) a olfactoric methods to identify those compous responsible for the characteristic odor as well as partly for the folk medicinal use of this plant. More than 95 volatiles were fou to be constituents of the three essential S. iicus oils with following composition of main components: flower oil: P- eudesmol (21.4%), 2,5-dimethoxy-p-cymene (16.2%), P-caryophyllene (7.8%), r-cadinol (7.2%). caryophyllene oxide (6.9%) a a-eudesmol (4.5%); root oil: 2,5-dimethoxy-p- cymene (28.3%), r-cadinol (25.3%), (2)-arteannuic alcohol (1 %), p-maaliene (3.9%) a caryophyllene oxide (3.1%); herb oil: 2,5-dimethoxy-p-cymene (27.0%), r-cadinol (12.5%), P-eudesmol (9.1 %), a-eudesmol (7.0%) a caroyphyllene oxide (4.7%). In addition, the odor impressions of the samples are described a the possible use of the essential Sphaeranthus iicus oils in medicinal, cosmetic a food flavoring discussed. Key Word Iex: Sphaeranthus iicus, Asteraceae, essential oils, flowers, roots, herb, odor, Iian folk medicine Inoduction: Sphaeranthu.~ iiczrs Kurz. (Asteraceae) is an important plant, used in Iian folk medicine especially for its antiseptic, antitussive a tonic effects14. While various solvent exacts were investigated for their biological active compous (mainly sesquiterpenoids)"'f the essential oils of S iicus plants a/or plant pans were studied very rarely 1.2. I I. I2

2 252 L. J i r o v e t z et al.: In continuation of our scientific work on analysis of essential oils used in aditional Iian medicine, also this research project was started to identify the volatiles of essential oils of different plant parts (flowers, roots a herb) of Sphaeranthus iicus a to discuss the constituents responsible for the characteristic aroma of these oils, their possible biological effects as well as applications in perfumery a cosmetics. To the best of our knowledge, no data on the composition of essential S. iicus oils from southern Iia are available until now. Experimental: Plant Material: The plants of Sphaeranthus iicus were collected on the Calicut University campus (Paddy field) in May 2000 a the plant material was identified by Dr.A.K. Pradeep, Department of Botany, Calicut University of Kerala. A voucher specimen is deposited in the specially maintained Herbarium of the Department of Chemisy at Calicut University (voucher-no. 35). The flowers, roots a herb of S. iicus were separated from the plant manually a dried at room-temperature for 14 days. After this, the three plant parts were powdered using an elecic grier each. The weights of powdered products were as follows: flowers: 650 g, roots: 250 g, herb: 1000 g. Essential Oil Exactions: The powdered S. iicus flowers, roots a herb were taken into a 10 L Erlenmeyer-flask a then 5 L of water was added, before steam-distillation for 3.5 hours each. The essential oils were exacted with diethyl ether as solvent a dried over anhydrous sodium phosphate. Yields were fou to be 0.31 g (0.05%) of an orange coloured essential flower oil, 0.30 g (2%) of a pale-yellow root oil a 0.44 (0.04%) g of a redorange herb oil. Olfactoric evaluations: The essential oils were diluted with dichloromethane, lop1 placed on a commercial odor-sip (Dragoco Co.) a the odor characterized by professional perfumers. Gas Chromatography:- GC analyses were carried out using a Shimadzu GC-14A with FID a the integrator C-R6A-Chromatopac a a Varian GC-3700 with FID a the integrator C- R1 B-Chromatopac (Shimadzu Co.). As columns a 30mx0.32mm boed unpoiar FSOT-RSL- 200 fused silica (film-thickness: 0.25pm, Biorad Co.) a a 30mx0.32mm boed polar Stabilwax (film-thickness: 0.50pm; Restek Co.) were used. Additional parameters as follows:

3 Medicinal used plants from Iia: analysis of the essential oils of Sphaeranthus Carrier gas: hydrogen; injector-temperature: 250 C; detector-temperature: 320 C; temp.- programs: 40 C/ 5min. to 280 C/ 5min. with a heating-rate of 6"Cl min.; quantification by %- peak-area-calculations (unpolar column); Some single components could be identified by co-injection of pure compous a correlation of their retention-times (using Kovats iices) with published datai3-16. Gas chromatography - Mass specomey:- The samples were analyzed by the GC-MS systems Shimadzu GC-17A with QP5000 a the data system Compaq-ProLinea (class5k- software), Hewlett-Packard GC-HP5890 with HP-5970MSD a PC-Pentium (Bohm Co., ChemStation-software) a Finnigan MAT GCQ with data system Gateway-2000-PS75 (Siemens Co., GCQ-software). Additional parameters as follows: Carrier gas: helium; injector-temperature: 250 C; interface-heating: 300 C; ion-source-heating: 200 C; EI-mode; scan-range: amu; other parameters see GC-FID-part. For compou identifications Wiley-, NBS- a NIST-library speca (on-line) as well as reference MS-speca data'5317'25 were used. Results a discussion: The essential Sphaerunthus iicus oils from southern Iia were olfactorically evaluated as follows: warm-spicy, dry-woody-medicinal, borneol-note, floral (direction of rose) a fruity side-note (flower oil); warm-herbal, dried fruits-like top-notes (direction of plums a apricot, cadinol-note), spicy (mixture of spices used for ready-to-cook soups), vetiver-, osmanthus-, rose- (both (3-phenylethyl alcohol-like) a lovage-notes (orterpineol-like), weak tobacco side-note (root oil) a warm-herbal, fresh-medicinal, fruityspice (like the root oil), vetiver-, osmanthus-, rose- a lovage-notes, tobacco side-note (sweet note when burned in a smoking-pipe) a aromatic-fatty notes in the backgrou (herb oil). Using gas chromatographic - specoscopic systems more than 95 components could be identified (see Table 1) as genuine constituents of the three essential S. iicus oils. As main compous (concenations higher than 3.094, calculated as %-peak-area of GC-FID analyses using an apolar column) of the essential oil of the flowers of S. iicus we fou P-eudesmol (21.4%), 2.5-dimethoxy-p-cymene ( 16.2%), P-caryophyllene (7.8%), s-cadinol (7.2%). caryophyllene oxide (6.9%) a a-eudesmol (4.5%). The root oil consists mainly of 23- dimethoxy-p-cymene (28.3%), r-cadinol (25.3%). (Z)-arteannuic alcohol ( 1 %), p-maaliene (3.9%) a caryophyllene oxide (3.1%) whereas in the herb oil 2.5-dimethoxy-p-cymene

4 254 L. J i r o v e t z et al.: (27.0%). r-cadinol ( 12.5%). p-eudesmol (9.1 %), u-eudesmol (7.0%) a caryophyllene oxide (4.7%) were fou. Table 1: Composition of the essential oils of the flowers (eof), roots (eor) a herb (eoh) of Sphaeranthus iicus from southern Iia. eor eoh 3-methyl butanol 2-methyl butanol 2-methyl but-3-en01 hexanal (Q-2-hexenal (2)-3-hexenol (E)-2-hexenol hexanol heptanal heptan-2-one a-t hujene a-pinene P-pinene heptanol 1 -octen methyl-5-hepten-2-one 6-methyl-5-hepten-2-01 (E)-3-hexenyl acetate myrcene (E)-2-hexenyl acetate hexanoic acid octanal para-cymene limonene (a-p-ocimene (E)-P-ocimene (2')-linalool oxide (furanoid) 1 -nonen-3-01 terpinolene linalool nonanal P-phenylethyl alcohol borne01 terpinen-4-01 verbenone a-terpineol cionel lo1 nerol ' t r

5 Medicinal used plants from Iia: analysis of the essential oils of Sphaeranthus neral geraniol geranial 7-epi-silphiperfol-5-ene eugenol a-cubebene neryl acetate geranyl acetate p-maaliene a-isocomene a-y langene a-copaene P-cubebene cyperene P-' isocomene p-elemene a-gurj unene P-bourbonene (2)-a-bergamotene a-cedrene a-himachalene p-caryophyl lenc aromaderene y-elemene 2,5-dimethoxy-p-cyn~ene (E)-a-bergamotene (E)-P-farnesene a-amorphene a-gualene a-humulene gennacrene D alloaromaderene (E,E)-a-farnesene a-muurolene (2)-a-bisabolene 6-cadinene p-selinene y-cadinene (2)-nerolidol elemol (E)-nerolidol spathulenol caryophyllene alcohol p-vetivene longifolene caryophyllene oxide a-cedrene epoxide guaiol globulol

6 (2)-arteannuic alcohol' humulene oxide khusimone cedrol (E)-arteannuic alcohol' y-eudesmol P-eudesmol t-cadinol a-cadinol a-eudesmol P-bisabolol a-guaiyl acetate (E,E)-farnesol khusimol y-eudesmyl acetate P-eudesmyl acetate a-eudesmyl acetate phytol 'in order of their elution 2concenations as %-peak-area calculated by GC-FID analysis '~ovats iices from unpolar FSOT-RSL column 4not detected 'ace compou (less than %) 'identified in accordance to cited literat~re'~.'~.~~.~' In accordance to these compositions, especially three points are remarkable: 1.) the relative high concenation of 2,5-dimethoxy-p-cymene in all three essential Sphaeranthus iicus oils from southern Iia. This aromatic monoterpene (syn. thymoquinol) was identified in e.g. oil samples of Monurada species, Eupatorium japonicum, Juniperus chinensis a Libocedrus de~urrens'~, but not in a Sphaeranthus species until now. 2.) the sesquiterpenes a- a P- eudesmol, known target compous of various Eucalyptus species27, were fou as free alcohols in such a significant concenation in S. iicus oils for the first time, while correspoing eudemanolides could be identified in different S. iicus exacts of previous works4. 3.) arteannuic alcohols were reported to be constituents of the essential oil of Artemisia annua2' as well as of the essential oil of Inula ~rithmoides~~, but no data are available that these sesquiterpene alcohols with cadinane sucture have been fou in a Sphaeranthus iicus sample. A comparison of analytical data with olfactoric ones, published el~ewhere~~-'~, allows following statements to the aroma impressions of essential flower, root a herb oils of Sphaerunthus iicus from southern Iia: warm-herbal aroma notes can be especially

7 Medicinal used plants from Iia: analysis of the essential oils of Sphaeranthus atibuted to 2,5-dimethoxy-p-cymene, dry-woody a spicy notes to cadinane a caryophyllane derivatives, fruity notes to minor aliphatic alcohols, such as hexanol, as well as some monoterpenes (e.g. geraniol), osmanthus- a rose-notes to 2-phenylethyl alcohol a the monoterpenes cionellol, geranial, geraniol, neral a nerol a lovage-like notes to a- terpineoi, vetiver notes of p-vetivene, khusimone, khusimol a P-bourbonene. An medicinal use of these essential S. iicus oils seems to be only possible as an antimicrobial medicament3';" for external application, because toxic effects a side-effects of the mainly sesquiterpenic constituents of the oils which are investigated very rarely until now. The same objection must be done on cosmetic applications as well as for the flavoring of food products. In summary, the essential oils of the flowers, roots a herb of Sphaeranthus iicus from southern Iia possess a very characteristic odor, showing some uncommon monoterpenes, such as 2,5-dimethoxy-p-cymene, a sesquiterpenes, such as arteannuic alcohols a eudesmols as main compous. These constituents as well as further lower alcohols a mono- a sesquiterpenes are responsible for the odor impressions. A possible use of these oils as medicaments, in cosmetic alor food products can be discussed again, after extensive investigations of the toxic potential of these identified sesquiterpenic components. Acknowledgements: We acknowledge the olfactoric evaluations by Mr. W. Hoppner a Mr. V. Hausrnann, perfumers of Dragoco Co., Vienna, Ausia, the botanical identification of the plant material by Dr.A.K. Pradeep, Department of Botany, Calicut University Kerala, Iia, the private correspoance with Dr.R. Bos, Department of Pharmacognosy, University of Groningen, The Netherlas, with Dr.F.-J. Hammerschmidt, Dragoco Co., Holzmien, Germany, a with Dr. D.D. Sawaikar, Division of Organic Chemisy, National Chemical Laboratory Pune, Iia for mass specal identifications of rare sesquiterpenes as well as the financial support (scholarship for M. Shahabi) by the University of Vienna. References: 1. Finnemore, H.; The Essential Oils, Ernest Benn Ltd., Loon, Great Britain, 861 (1926). 2. Gildemeister. E. a Hoffmann, F.: Die Atherischen Ole. Bd. VII, Akademie-Verlag, Berlin, Germany, 596 (1961). 3. Hiller, K. a Melzig, M.; Lexikon der Arzneipflanzen u Drogen, Bd. 2, Spekum Akademischer Verlag, Heidelberg, Germany, 296 (2000).

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