Molecules 2007, 12, 149-154 molecules ISSN 1420-3049 http://www.mdpi.org Communication Comparative Chemical Analysis of the Essential Oil Constituents in the Bark, Heartwood and Fruits of Cryptocarya massoy (Oken) Kosterm. (Lauraceae) from Papua New Guinea Topul Rali 1, *, Stewart W. Wossa 2 and David N. Leach 3 1 2 3 Chemistry Department, University of Papua New Guinea, P.O. Box 320, University, Papua New Guinea Faculty of Science, University of Goroka, P.O. Box 1078, Goroka, Papua New Guinea; e-mail: wabowossa@yahoo.com Center for Phytochemistry and Pharmacology, Southern Cross University, P.O. Box 157, Lismore, NSW 2480, Australia; e-mail: dleach@scu.edu.au * Author to whom correspondence should be addressed; e-mail: rali.topul@upng.ac.pg Received: 8 November 2006; in revised form: 20 December 2006 /Accepted: 3 February 2007 / Published: 5 February 2007 Abstract: Exhaustive hydro-distillation of the bark, heartwood and fruits of Cryptocarya massoy (Lauraceae) afforded pale yellow-coloured oils in 0.7, 1.2 and 1.0 % yields, respectively. Detailed chemical evaluation of these distillates using GC/MS revealed the major components in the bark and the heartwood oils to be the C-10 (5,6-dihydro-6-pentyl2H-pyran-2-one) and C-12 (5,6-dihydro-6-heptyl-2H-pyran-2-one) massoia lactones, while the major fruit oil constituent was benzyl benzoate (68.3 %). The heartwood also contained trace amounts of the C-14 (5,6-dihydro-6-nonyl-2H-pyran-2-one) massoia lactone (1.4 %) and the saturated C-10 derivative δ-decalactone (2.5 %). Keywords: Cryptocarya massoy; Lauraceae; massoia lactones; 5,6-dihydro-6-pentyl-2Hpyran-2-one (C-10 massoia lactone); benzyl benzoate; 5,6-dihydro-6-heptyl-2H-pyran-2-one (C-12 massoia lactones); essential oil analysis; α,β-unsaturated δ-lactone; 5,6-dihydro-6nonyl-2H-pyran-2-one (C-14 massoia lactones); δ-decalactone. Molecules 2007, 12 150 Introduction The genus Cryptocarya of the family Lauraceae is comprised of more than 350 species distributed throughout the tropics, subtropics and the temperate regions of the world. Kosterman had established detailed taxonomic descriptions and classifications for this family [1], which has recently been the subject of much interest [2]. The growing wealth of phytochemical literature on the family Lauraceae further indicate their chemosystematics to be in agreement with the established classification schemes [3]. Several species from this genus have been used extensively as traditional medicines in a number of ethnobotanical practices [4,5] and detailed phytochemical and pharmacological studies have shown the chemical constituents to consist mostly of pyrones and styrylpyrones that exhibit anticancer, larvicidal and antifertility activities [6-8]. Information about the distribution of members of the family Lauraceae in Papua New Guinea (PNG) is scant, but pharmacological screenings of six species of Cryptocarya have indicated that they do exhibit biological activity [9]. No detailed phytochemical investigations were pursued, except for C. ainikini Kosterm. where a (+)-(5S)-δ-lactone previously identified in C. caloneura (Scheff.) Kosterm., was found to be one of the main constituents [10]. The massoia lactones (Figure 1) are 10, 12 and 14 carbon chain compounds, hereon referred to as the C-10, C-12 and C-14 massoia lactones, respectively, that possess characteristic α,β-unsaturated δlactone moieties They also present substitution at the C6 position of the α,β-unsaturated δ-lactone structures with chains of variable length containing five, seven or nine carbons. In nature these compounds are rare essential oil components that were first characterized by Abe in 1937 [11]. The report by Cavill and co-workers [12] on the essential oil composition in the bark oil of C. massoia and the formicine ants indicated that the constituents of the bark oil were the C-10 (5,6-dihydro-6-pentyl2H-pyran-2-one) massoia lactone, with traces of the C-12 (5,6-dihydro-6-heptyl-2H-pyran-2-one) and C-14 (5,6-dihydro-6-nonyl-2H-pyran-2-one) ones. The analysis of the light petroleum extract of the whole ant, on the other hand, indicated the presence of C-10 massoia lactone as the major constituent, correlating with its presence as a mode of chemical defense. This was further confirmed by the work of Lloyd and coworkers [13] investigating the chemistry of the mandibular gland secretion of the carpenter ant, Camponotus thoracicus fellah emery, another member of the formicine ants. Fleisher and Fleisher [14] also noted the occurrence of C-10 massoia lactone as a minor constituent in the volatile oils of Achillea fragrantissima. Brophy and co-workers [15] also reported the leaf oil of C. cunninghamii to contain all three series of the massoia lactones, with the C-14 massoia lactone being the major component. The characteristic α,β-unsaturated δ-lactone moiety found in the massoia lactones is also a common feature in a number of biologically active natural products such as (+)-goniothalamin, (-)callystatin A, canolide A and the kava lactones, either differing in the substituents at the C6 position or having a molecular structural architecture that maintains the α,β-unsaturated δ-lactone skeleton. Such interesting biological activities of this class of compounds have made their production a synthetic challenge [16-22], and biotechnological [23] and microbe-assisted conversion of various substrates into the desired massoia lactones have been described [24-27]. As part of an ongoing research program to identify and document the chemical constituents in the essential oils from endemic flora of PNG, we report herein a complete analysis of the essential oils obtained from the bark, heartwood and fruits of C. massoia from in this location. Molecules 2007, 12 151 Figure 1. Structures of the massoia lactones from C. massoia oil. O O O C-10 massoia lactone O O C-12 massoia lactone O O O δ-decalactone C-14 massoia lactone Results and Discussion Comparative data on the different volatile oil constituents in the bark, heartwood and fruits of C. massoia is presented in Table 1. The results indicate that C-10 massoia lactone is the major compound, present in similar amounts in the bark and heartwood and found in the fruit oil in trace amounts. This lactone is an important commercial oil, whose presence in the heartwood has not been reported previously. The heartwood oil also contained more C-12 massoia lactone, compared to the bark or fruit oils, and small amounts of C-14 massoia lactone, which was not found in the bark or fruit oils and is the main constituent of the leaf oil of Cryptocarya cunninghamii Meissner (Lauraceae) [15]. Benzyl benzoate is the main compound found in the fruit oil but it is present to a lesser extent in the bark and is absent in the heartwood oil. Table 1. A comparative analysis of the composition of the chemical constituents (area %) in the bark, heartwood and fruit oils of C. massoia. Chemical Component α-pinene β-pinene limonene linalool borneol α-cubebene α-copaene β-elemene β-caryophyllene α-humulene E,E-α-farnesene C-10 massoia lactone ledene β-bisabolene δ-cadinene cis-calamene δ-decalactone RI Bark Oil Heartwood Oil Fruit Oil 953 1000 1050 1110 1210 1375 1413 1420 1468 1505 1516 1520 1543 1555 1556 1566 1615 0.9 0.7 64.8 1.4 - 68.4 2.5 0.2 0.3 0.4 0.3 2.1 1.3 12.9 2.2 1.3 1.4 0.2 1.2 0.2 - Molecules 2007, 12 152 Table 1. Cont. Chemical Component caryophyllene oxide C-12 massoia lactone benzyl benzoate benzyl salicylate C-14 massoia lactone RI Bark Oil Heartwood Oil Fruit Oil 1647 1739 1838 1948 2068 17.4 13.4 - 27.7 1.4 2.2 0.2 68.3 1.8 - Conclusions The tree Cryptocarya massoia (Oken) Kostern (Lauraceae) is a species endemic to the island of New Guinea, from which the massoia lactones [11, 12] could be obtained from either Irian Jaya [15] or PNG. A comparative study of the bark, heartwood and fruit oils indicates the heartwood and bark oils to be the main natural sources of the C-10 and the C-12 massoia lactones. The C-14 massoia lactone and δ-decalactone were found to occur as minor constituents in the heartwood oil. The fruit oil, on the other hand, was found to be composed predominantly of benzyl benzoate. Acknowledgements The authors are grateful to Mr. Pius Piskaut of the University of Papua New Guinea Herbarium for the plant description and identification and the University of Papua New Guinea Research Council for a research grant and scholarship (to SW). The authors also thank Dr. Joseph J. Brophy for pointing out previous research work on massoia oil by Garnero and Joulain [28]. One of us (TR) would like to thank the people of Inawabui village for permission to access their traditional land to collect fresh samples of massoia bark and heartwood. Experimental Materials and Methods The bark and heartwood of C. massoia (Oken) Kosterm were collected from forests near Epa village, Central Province of PNG, in February 2003. The fruits were obtained from the same location during the fruiting season in August 2003. The fresh bark and heartwood were milled to a very fine homogeneous composition and the fruits were chopped into smaller pieces and ground to a fine powdery mixture. The individual plant parts were exhaustively hydrodistilled over an 8 hr period in an all-glass standard distillation set-up. The pure oils obtained were separated and dried over anhydrous magnesium sulphate to give pale yellow coloured oils in 0.70, 1.20 and 1.03 % yields from the bark, heartwood and fruits, respectively. The analyses of the oils were done using GC/MS on an Agilent 6890 Gas Chromatograph, equipped with a split/splitless injector and a 7963 Mass Selective Detector (MSD). Chromatography was performed on a BPX-5 capillary column (SGE, 50 m × 0.22 mm) terminated at the MSD operating as follows: transfer temperature: 310 °C; ionization 70 eV, source temperature: 230 °C; quadrupole temperature 150 °C and scanning a mass range 35-550 m/z. The injector temperature was 250 °C and the carrier gas was helium at 23.10 psi with an average velocity Molecules 2007, 12 153 to the MSD of 28 cm/sec. The column oven was programmed as follows: initial temperature: 50 °C; initial time 1.0 min; program rate 4 °C/min; final temperature 300 °C; final time 10 min. Identification of Oil Components The oil samples were injected in hexane. The individual compounds in the oil were identified by their retention indices relative to known compounds and further by comparison of their mass spectra with either the known compounds or published spectral data [29-31]. References and Notes 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. Hutchinson, J. Evolution and Phylogeny of Flowering Plants; Academic Press: London, 1969; p. 28 Rohwer, J.A. Lauraceae. In: Kubitzki, K., Rohwer, J.A. and Bittrich, V. Eds. The Families and Genera of Vascular Plants. Vol II. Flowering Plants – Dicotyledons; Springer-Verlag: Berlin, 1993; pp. 366 – 391. Gottlieb, O. R. Chemosystematics of the Lauraceae. Phytochemistry 1972, 11, 1537 – 1570. Zschocke, S.; van Staden, J. Cryptocarya species – substitute plants for Ocotea bullata? A pharmacological investigation in terms of cyclooxygenase-1 and –2 inhibition. J. Ethnopharmacol. 2000, 71, 473 – 478. Drewers, S.E.; Horn, M.H.; Mavis, S. Cryptocarya liebertiana and Ocotea bullata – Their phytochemical relationship. Phytochemistry 1997, 44, 437 – 440. Hawariah, A.L.P.; Stanslas, J.; Laily, D. Non-steroid receptor mediated antiproliferative activity of styrylpyrone derivative in human breast cancer cell lines. Anticancer Res. 1998, 18, 1739 – 1744. Hawariah, A.L.P.; Munawar, M.; Laily, D. Antifertility effect of Goniothalamin: A styrylpyrone isolated from Goniothalamus tapis Miqo. Asia Pacific J. Pharmacol. 1994, 9, 273 – 277. Hawariah, A.L.P.; Stanslas, J. In vitro response of human breast cancer cell lines to the growthinhibitory effects of styrylpyrone derivative (SPD) and assessment of its antiestrogenicity. Anticancer Res. 1998, 18, 4383 – 4386. Collins, D.J.; Culvenor, C.C.J.; Lamberton, J.A.; Loder, J.W.; Price, J.R. Plants for medicine: a chemical and pharmacological survey of plants of the Australian region. CSIRO Australia, 1990. Hlubucek, J.R.; Robertson, A.V. (+)-(5S)-δ-lactone of 5-hydroxy-7-phenylhepta-2,6-dienoic acid, a natural product from Cryptocarya caloneura (Scheff.) Kostermans. Aust. J. Chem. 1967, 20, 2199 – 2206. Abe, S. Massoy oil. J. Chem. Soc. Japan 1937, 58, 246 – 251. Cavill, G.W.K.; Clark, D.V.; Whitfield, F.B. Insect Venom, Attractants and Repellents. XI. Massoialactone from two species of Formicine ants, and some observations on constituents of the bark oil of Cryptocarya massoia. Aust. J. Chem. 1968, 21, 2819 – 2823. Lloyd, H.A.; Schmuff, N.R.; Hefetz, A. Chemistry of the male mandibular gland secretion of the carpenter ant, Camponotus thoracicus fellah emery. Comp. Biochem. Physiol. Part B. Biochem. Mol. Biol. 1984, 78, 687 – 689. Fleisher, Z.; Fleisher, A. Volatiles of Achillea fragrantissima (Forssk.) Sch. Bip. Aromatic Plants of the Holy Land and the Sinai. Part XI. J. Essent. Oil Res. 1993, 5, 211 – 214. Molecules 2007, 12 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 154 Brophy, J.J.; Goldsack, R.J.; Forster, P.I. The Leaf essential oil of Cryptocarya cunninghamii Meissner (Lauraceae). J. Essent. Oil Res., 1998, 10, 73 – 75. You, Z-W.; Wu, Y-Y.; Qing, F-L. Synthesis of gem-difluoromethylenated massoialactone by ring-closing metathesis. Tetrahedron Lett. 2004, 45, 9479 – 9481. Fournier, L.; Korcienski, P.; Pons, J-M. The β-lactone route to α,β-unsaturated δ-lactones. Total syntheses of (+)-goniothalamin and (-)-massoialactone. Tetrahedron 2004, 60, 1659 – 1663. Gupta, P.; Naidu, S.K.; Kumar, P. A practical enantioselective syntheses of massoialactone via hydrolytic kinetic resolution. Tetrahedron Lett. 2004, 45, 849 – 851. Ramachandran, P.V.; Reddy, M.V.R.; Brown, H.C. Asymmetric synthesis of goniothalamin, hexadecanolide, massoialactone and parasorbic acid via sequential allylboration-esterification ring-closing metathesis reaction. Tetrahedron Lett. 2000, 41, 583 – 586. Pais, G.C.G.; Fernandes, R.A.; Kumar. P. Asymmetric synthesis of (S)-Massoialactone. Tetrahedron 1999, 55, 13445 – 13450. Romeyke, Y.; Keller, M.; Grabley, S.; Hammann, P. Secondary metabolites by chemical screening-13- Enantioselective synthesis of δ-lactones from streptenola, a chiral building block from Streptomyces sp. Tetrahedron 1991, 47, 3335 – 3346. Pohmakotr, M.; Jarupan, P. A new syntheses of α,β-unsaturated γ- and δ-lactones via intramolecular acylation of α-sulfinylcarbanion. Tetrahedron Lett. 1985, 26, 2253 – 2256. Gil, B.; Christopher, D. Process for the biotechnological production of Delta-decalactone and Delta-dodecalactone. European Patent EP0899342, 1999. D’Arrigo, P.; Fuganti, C.; Fantoni, G.P.; Servi, S. Extractive biocatalysis: A powerful tool in selectivity control in yeast biotransformation. Tetrahedron 1998, 54, 15017 – 15026. Fuganti, C.; Fantoni, G.P.; Sarra, A.; Servi, S. Stereochemistry of Baker’s yeast mediated conversion of α,β-unsaturated δ-lactones in the goniothalamin series. Tetrahedron: Asymmetr. 1994, 5, 1135 – 1138. Shinobu, G.; Rumi, K.; Tsuyoshi, K. Process for the production of the δ-decalactone. Eur. Pat. EP0822259, 1998. Cardillo, R.; Fugati, C.; Babeni, M.; Allegro, G. Process for the preparation of saturated delta lactones by biohydrogenation of the corresponding unsaturated compounds by microorganisms. European Patent EP0577463, 1994. Garnero, J.; Joulain, D. Massoia essential oil. IXth International Congress of Essential Oils, Essential Oil Technical Paper Book 3, 1983; pp. 184 – 193. Joulain, D.; Konig, W.A. The Atlas of Spectral Data of Sesquiterpene Hydrocarbons; E.B. Verlag: Hamburg, Germany, 1998. Adams, R.P. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry; Allured Pub. Corp.: Carol Stream, IL, USA, 1995. Swigar, A.A.; Silverstein, R.M. Monoterpenes; Aldrich: Milwaukee, USA, 1981. Sample Availability: Available from the authors. © 2007 by MDPI (http://www.mdpi.org). Reproduction is permitted for noncommercial purposes.