of Junipems communis L.

Transcription

1 RESEARCH REPORT J. Essent. Oil Res., 5, (Nov/Dec 1993) Chemical Investigation of the Leaf Oil of Junipems communis L. Pashalina S. Chatzopoulou and Stavros T. Katsiotis" Department of Pha~maceutics, School of Pharmacy Aristotle University of Thessaloniki PO Box 1589, Thessaloniki, G1-eece Received: March ABSTRACT: The essential oil obtained from the needles ofjuiziperus communis L. of Greek origin was subjected to GC/MS analysis. Fifty six compounds were identified accounting for more than 96% of the oil. The major component was a-pinene (41.25%), while the predominant minor constituents were sabinene (17.4%), limonene (4.2%), terpinen-4-01 (2.7%), myrcene (2.6%) and P-pinene (2.0%). KEY WORD INDEX: Juniperus communis, Cupressaceae, Juniper leaves, Greek mountain wild juniper, essential oil composition, a-pinene. 1NTRODUCTION:Junipei-us communis is awide spontaneous-spread essential oil plant in Greece; more often located in Thessalia, Peloponesus, Macedonia, Epirus and in certain islands. Its oil is of interest to the perfumery, cosmetics and pharmaceutical industries because of its aromatic and diuretic properties. Ill particular, there are monographs in the Austrian, French, Hungarian, Yugoslavian, Romanian and Swiss national Pharmacopoeias for juniper oil (1). After the 1960s, the interest of students of the juniper family turned from the berry oil to that of the needle oil. It is our belief that this new direction for the research m7as to either increase our knowledge on the chemical composition of the leaf oils of various species (2-5), or use the oils as genetic markers to better understand taxonomic interrelationships (6-8). Since the interest in juniper needle oil has been extensive, a systematic investigation was undertaken to determine the constituents present in an oil produced from Creek mountain juniper. MATERIALS AND METHODS: Plant Material-The plant material (leaves) samples were collected from wild growing Juniperus communis shrubby trees on the Olympus mountain at an altitude of 1300 m. In considering the changes in composition of the volatile terpenes during maturation, only foliage samples from the same tree were coinparedin order to avoid differences that might 'Address for correspondence /93/ $04.00/ Allured Publishing Coly. 603

2 604 JOURNAL OF ESSENTIAL OIL RESEARCH Table I. Chemical composition of the needle oil of Juniperus communis L. growing wild in North Greece Method of Components Percentage identification tricyclene a-t hujene a-pinene a-fenchene ,2,3 camphene ,3 sabinene P-pinene ,2,3 myrcene carene a-phellandrene 6-3-carene a-terpinene p-cymene P-phellandrene limonene 1,8-cineole (E)-p-ocimene y-terpinene trans-sabinene hydrate fenchone 3-octanol terpinolene linalool cis-sabinene hydrate octen-3-01 P-thujone trans-p-menth-2-en-1-01 cis-p-menth-2-en-l -ol camphor citronellal pinocarvone trans-carve01 borneol terpinen-4-01 a-terpineol citronellol linalyl acetate bornyl acetate citronellyl acetate a-copaene P-cubebene p-elemene P-caryophyllene a-humulene y-muurolene germacrene D a-muurolene y-cadinene

3 JOURNAL OF ESSENTIAL OIL RESEARCH 605 Table I. (cont.) Method of Components Percentage identification 8-cadinene calacorene* a-cadinene (E)-nerolidol P-caryophyllene oxide humulene oxide* T-cadinol a-cadinol * isomer not characterized Identification method: 1 = relative retention time, 2 = reference oils, 3 = mass spectra exist between trees. The relatively mall size of the trees investigated tended to minimize variations in composition due to the location of the foliage on the tree. Isolation and Yield of the Essential Oil-After separation from the berries and the branches, the needles were stored in freezer at -20 C. The oil percentage yield was determined by using the European Pharmacopoeia apparjtus (Clevenger-type) and after three replications the SD was f 0.15%. Prior to distillation, the needles were dried to 25% H,O (9). The plant material was kept in liquid N, for 30 min without being comminuted (10). Samples of 20 g of needles were water distilled with 340 ml of deionized water for 3 h at a &stillation rate of mumin (11). The lighter-than-water, limpid, colorless oil was diied over anhydrous Na,SO, and stored in sealed container under refrigeration (-20 C). GC-The oil sample was analyzed using a Hewlett Packard 5890 Series I1 gas chromatograph equipped with one injection port and a two-channel system of columns and respective flame ionization detectors connected to a chromatographic integrator (Hewlett Packard 3396 Series 11 Dual Channel). Two fused silica coluinns of different polarity were used: (a) Durabond-DB 1 and (b) DB-Wax, both of 60 m x 0.25 mm with a film thickness of 0.25 pn1 (J&W Scientific Inc., Rancho Cordova, California, USA). Oven temp: 45"-220 C (3.5"C/min); carrier gas: nitrogen, 140 Kpa; injection temp: 220 C; detector temp: 300 C. The percentage composition was con~puted from the GC peak areas without correction factors. GCMS-A GCIMS was also applied, using a 60 nix mm fused silica CP-Wax 52 CB column, film thickness 0.25 pm (Chrompack Nederland BV) and a Packard 438 A GC interhced with a Finnigan MAT Ion Trap Detector (Finnigan Mat, San Jose, California, USA). sofhvare version 3.0. Oven temp: 45"-240 C (3"C/min); carrier gas: helium (pressure 200 Kpa); splitting ratio 1:40; scan time, 1 sec. RESULTS AND DISCUSSION: The distillation experiments were performed by the European Pharmacopoeia apparatus. The average yield of the oil was 0.85% v/w, approximately midpoint between the previous yield values ( %) reported in literature (2,4,5-7). A list of the 56 compounds identified in the oil as well as their percentage and the method ofidentification is given in Table I. As can be seen, a-pinene (41.2%) was the predominant compound, while terpinen-4-01 (2.78%) and germacrene-d (1.83%) were the main oxygenated and sesquiterpenoid constituents respectively.

4 606 JOURNAL OF ESSENTIAL OIL RESEARCH The above results showed that the oil was rich in monoterpene hydrocarbons (77.18%). Even though there are 22 oxygenated conlpounds in the oil, their relative amount was quite low (7.5%). In contrast, the 17 sesquiterpenoids together totaled 11.6% of the oil. A rather complete analysis of the sesquiterpene fraction of the essential oil fromjrrniper leaves has been reported by Vernin et al. (E), and although these authors identified more than 50 components, no yuantitative data was presented. A co~nparison with the previously published data reveals that the a-pinene content of our oil is midway between the very high level found in an oil of N. American (3) and low altitude Norwegian origin (8) and the low values found in oils from France (5) or from high mountain areas of Nonvay (8). Similarly, the amount of sabinene found in our oil was between that found in oils of French (5) or high altitude Nonveigian origin (8), and that found for numerous samples of N. American origin (2). The monoterpene hydrocarbons of juniper needle oil have also been examined by Banthorpe et al. (IS), Looman and Baerheirn Svendsen (14) and Laakso et al. (15), although the latter two studies were headspace analyses. A further study on the monoterpene hydrocarbon content of a needle oil of ]. communis var, nana of Turkish origin has also been reported (16). Among the oxygenated constituents, teq~inen-4-01 which is reputed to be responsible for the diuretic properties of the oil (17), was found in 2.78% in our Greek juniper needle oil. This is lo we^ than the level reported for an oil of Romanian origin (18) in which the highest value for tei-pinen-4-ol(4.6%) was reported. Recently, terpinen-4-01 was also identified in Juniper branch (2.0%) and the wood oil (<0.1%) of tndian origin (19). ACKNOWLEDGMENTS: The authors are grateful to Dr. Wille, Quest Research Company Ltd., for comparison of some spectral data, and Mrs. A. Looman for valuable technical assistance. REFERENCES 1. Martindale, The Extra Phamacopoeia. 29th Ed., The Pharmaceutical Press. London (1989). 2. E. von Rudloff andv. K. Sood, Gas-liquidchromatography ofterpenes. Part XVIII. The volatile oil of the leaves ofjunipetrls communis L. Can. J. Chem.. 47,2081 (1969). 3. T. Shibamoto and W. G. Jennings, The volatile composition of the leaf oil of California Juniper (J. calfornica Carr.). VIIth International Congress of Essential Oils, sponsored by Japan Flavor and Fragrance Manufacturers Association, Kyoto, Japan (1977). 4. N. Gelsomini, V. Vidrich, P. Fusi and M. Michelozzi, Capillaty gas chromatography of the terpetzic fraction of Juniperus communis L. black, green, berrly and leaf extracts. J. High Resol. Chromatog. Commun., 10721, (1988). 5. G. Vernin, C. Boniface, J. Metzger, C. Ghiglione, A. Hammoud, K. N. Suon, D. Fraisse and C. Parkanyi, GCIMS Specma Bank Analysis of Juniperus cornmunis needles and berries. Phytochemistry, 27, (1988). 6. H. Horster, Variabilitat der Ole von Juniperus communis III. Die Olzusammcnsetzung won Individuen verschiedener Populationen und Wuchsformer. Planta Med., 26,45-51 (1974). 7. H. Horster. Vnriability ofthe oils ofjuniperus conununis L. The composition of the oils in leaves of dzflerent age. Planta bled., 23, (1973). 8. A. Baerheim Svendsen, J. J.C. Scheffer and A. Looman, A comparative study of the composition of the essential needle oils of Norwegian Low LandsJuniperaiad High Mountains Juniper. Sci. Pharm., 53, (1985). 9. P. Chatzopoulou, S. T. Katsiotis andp. P. Georgalzopoulos, Prelimina y study ofthecomposition of the Jutxiperus com7nzcnis L. essential oil. InjTuence of diyness on the yield. Acta of 4th Panhellenic Pharmaceutical Congress, Athens, pp , Penhelleic Association of Pharmacists (1988). 10. S. Katsiotis, G. Ktistis and N. Iconomou, Investigation of the simultaneous itzfluence of three dz9erentpartlo~eters on the essential oil yieldand qunlity of Mentha pipeiita. Pharln. Acta Helv., 60(8), (1985).

5 JOURNAL OF ESSENTIAL OIL RESEARCH 11. S. T. Katsiotis, Study ofdifferent parameters influeilcing the composition ofhydrodistilledsweet fennel oil. Flav. Fragr. J., 4, (1988). 12. G. Vernin, J. Metzger, K. N. Suon, D. Fraisse, C. Ghiglione, A. Haminoud andc. Parkanyi, GC/ MS Spectna Bank Analysis of Essential Oils and Aromas GC/MS (El-PCI) Data Bank Analysis of Ses~uiterpenic Cornpounds injuniperiveedle Oil. Application of Mass Fragmentomety SIM Technique. Lebensm. Wiss. u. Technol., 23,25-33 (1990). 13. D. V. Banthorpe, H. ff. S. Davies, C. Catford and S. R. Williams, Moi~oteryei~e patterns in Juniperus and Thuja species. Planta Med., 23, (1973). 14. A. Looman and A. Baerheim Svendsen, The needle essential oil of Norwegini~ MountainJulliper, Juniperus communis L. var. Saratilis Pall. Flav. Fragr. J., 7,23-26 (1992). 15. I. Laakso, R. Hiltunen, K. Antila and M. von Schantz, Application of headspace gas chromatography in essential oil analysis. X. Variation in the terpenes of Juniper leaves and bcrr-ics in some localities in Finland. Flav. Fragr. J., 2, (1987). 16. N. Tanker and E. Sarer, Gas liquid chromatographic researches on the volatile oils obtained fromleaues andfromfmit.r ofsome Junipemsspecies. J. Fac. Pharnl. Ankara, 5, (1975). 17. L. Janku, U. Hava and 0. Motl, Ein diuretisch wirksanwi- Stoff nus Wacholder (Junipems communis L.). Experientia, 13, (1957). 18. H. Horster, C. Cseclo andc. Racz, Gas chromatographicat~alysisofthevolatile oilfromjuniper leaves (Junipencs cov~munis L.) harvested in Romania. Revista Medicala, 20(1), (1974). 19. J. P. Bats, J. J. Moulines and G. Bourgeois, Chemical composition of an industrial essential oil: from Juniper branches (Junipems communis L.). Proc. 11th Inter. Coug. Essential Oils Fragr. Flav. (1989), New Delhi, by S. C. Bhattacharyya, N. Sen and K. L. Sethi, Oxford and IBTT Ltd., New Delhi, (1989).