Leaf essential oils of four Piper species from the State of Ceará - Northeast of Brazil

Abstracts

The essential oils, obtained by hydrodistillation, from leaf samples of four Piper species (P. arboreum, P. crassinervium, P. dilatatum and P. tuberculatum), harvested at the State of Ceará, were analyzed by GC-MS. The oil yields ranged from 0.03 to 0.11% (w/w), on fresh weight basis. The oils were characterized by mono- and sesquiterpenes, except the oil of P. arboreum to which only sesquiterpenes were identified. beta-Elemene (0.58-3.03%), (E)-caryophyllene (2.71-37.78%), germacrene D (3.43-11.81%), bicyclogermacrene (2.83 and 25.07%) and delta-cadinene (0.52-2.44%) were detected in all oil samples. The major identified monoterpenes were alpha-pinene (11.27%), beta-pinene (20.01%), 1,8-cineole (10.81%) and linalool (28.61%) for P. crassinervium and, alpha-phellandrene (22.53%) and delta-3-carene (10.20%) for P. dilatatum. The major sesquiterpenes were bicyclogermacrene (25.03 and 25.07%) for P. arboreum and P. dilatatum, (E)-nerolidol (11.12%) for P. arboreum, germacrene D (11.81%) for P. tuberculatum and, (E)-caryophylene (10.26 and 37.78%) for P. dilatatum and P. tuberculatum. To the best of our knowledge this is the first report about the chemical composition of the leaf essential oils of P. crassinervium and P. tuberculatum.

Piper arboreum; Piper crassinervium; Piper dilatatum; Piper tuberculatum; essential oils

Os óleos essenciais obtidos por hidrodestilação das folhas de quatro espécies de Piper (P. arboreum, P. crassinervium, P. dilatatum and P. tuberculatum), coletadas no estado do Ceará, foram analisados por CG-EM. Os rendimentos dos óleos, calculados sobre o peso do material fresco, variaram entre 0,03 - 0,11% (p/p). Os óleos foram caracterizados pela presença de mono- e sesquiterpenos, exceto o óleo de P. arboreum, no qual foram identificados apenas sesquiterpenos. beta-Elemeno (0,58-3,03%), (E)-cariofileno (2,71-37,78%), germacreno D (3,43-11,81%), biciclogermacreno (2,83-25,07%) e delta-cadineno (0,52-2,44%) foram detectados em todas as amostras analisadas. Os monoterpenos majoritários identificados foram alfa-pineno (11,27%), beta-pineno (20,01%), 1,8-cineol (10,81%) e linalol (28,61%) para P. crassinervium, e alfa-felandreno (22,53%) e delta-3-careno (10,20%) para P. dilatatum. Biciclogermacreno (25,03 e 25,07%) para P. arboreum e P. dilatatum, (E)-nerolidol (11,12%) para P. arboreum, germacreno D (11,81%) para P. tuberculatum e (E)-cariofileno (10,26 e 37,78%) para P. dilatatum e P. tuberculatum, foram os principais sesquiterpenos. Este trabalho descreve, pela primeira vez, a composição química dos óleos essenciais de P. crassinervium e P. tuberculatum.

ARTICLE

Leaf essential oils of four Piper species from the State of Ceará - Northeast of Brazil

Juliana B. Cysne^I; Kirley M. Canuto^I; Otília Deusdênia L. Pessoa^{*, I}; Edson P. Nunes^II; Edilberto R. Silveira^I

^IDepartamento de Química Orgânica e Inorgânica, Universidade Federal do Ceará, CP 12200, 60021-970 Fortaleza - CE, Brazil

^IIHerbário Prisco Bezerra, Departamento de Biologia, Universidade Federal do Ceará, 60451-970 Fortaleza - CE, Brazil

ABSTRACT

The essential oils, obtained by hydrodistillation, from leaf samples of four Piper species (P. arboreum, P. crassinervium, P. dilatatum and P. tuberculatum), harvested at the State of Ceará, were analyzed by GC-MS. The oil yields ranged from 0.03 to 0.11% (w/w), on fresh weight basis. The oils were characterized by mono- and sesquiterpenes, except the oil of P. arboreum to which only sesquiterpenes were identified. b-Elemene (0.58-3.03%), (E)-caryophyllene (2.71-37.78%), germacrene D (3.43-11.81%), bicyclogermacrene (2.83 and 25.07%) and d-cadinene (0.52-2.44%) were detected in all oil samples. The major identified monoterpenes were a-pinene (11.27%), b-pinene (20.01%), 1,8-cineole (10.81%) and linalool (28.61%) for P. crassinervium and, a-phellandrene (22.53%) and D-3-carene (10.20%) for P. dilatatum. The major sesquiterpenes were bicyclogermacrene (25.03 and 25.07%) for P. arboreum and P. dilatatum, (E)-nerolidol (11.12%) for P. arboreum, germacrene D (11.81%) for P. tuberculatum and, (E)-caryophylene (10.26 and 37.78%) for P. dilatatum and P. tuberculatum. To the best of our knowledge this is the first report about the chemical composition of the leaf essential oils of P. crassinervium and P. tuberculatum.

Keywords:Piper arboreum, Piper crassinervium, Piper dilatatum, Piper tuberculatum, essential oils

RESUMO

Os óleos essenciais obtidos por hidrodestilação das folhas de quatro espécies de Piper (P. arboreum, P. crassinervium, P. dilatatum and P. tuberculatum), coletadas no estado do Ceará, foram analisados por CG-EM. Os rendimentos dos óleos, calculados sobre o peso do material fresco, variaram entre 0,03 – 0,11% (p/p). Os óleos foram caracterizados pela presença de mono- e sesquiterpenos, exceto o óleo de P. arboreum, no qual foram identificados apenas sesquiterpenos. b-Elemeno (0,58-3,03%), (E)-cariofileno (2,71-37,78%), germacreno D (3,43-11,81%), biciclogermacreno (2,83-25,07%) e d-cadineno (0,52-2,44%) foram detectados em todas as amostras analisadas. Os monoterpenos majoritários identificados foram a-pineno (11,27%), b-pineno (20,01%), 1,8-cineol (10,81%) e linalol (28,61%) para P. crassinervium, e a-felandreno (22,53%) e D-3-careno (10,20%) para P. dilatatum. Biciclogermacreno (25,03 e 25,07%) para P. arboreum e P. dilatatum, (E)-nerolidol (11,12%) para P. arboreum, germacreno D (11,81%) para P. tuberculatum e (E)-cariofileno (10,26 e 37,78%) para P. dilatatum e P. tuberculatum, foram os principais sesquiterpenos. Este trabalho descreve, pela primeira vez, a composição química dos óleos essenciais de P. crassinervium e P. tuberculatum.

Introduction

The genus Piper (Piperaceae) has been recently revised.¹ With approximately 700 species, represented by herbs, shrubs and trees it is widely distributed in the tropical and subtropical regions of the terrestrial globe. Several plants of this genus are largely used in folk medicine in several parts of the world and have been reported to produce compounds with diverse biological and pharmacological properties.^2-8 Many Piper species are aromatic and as a consequence the chemical composition of the essential oils of several of them has been the subject of incessant studies, revealing a diverse range of volatile components, including monoterpenes, sesquiterpenes, arylpropanoids, aldehydes, ketones and long chain alcohols.^9-16

Pursuing the determination of the chemical composition of essential oils from aromatic and medicinal plants from Northeast of Brazil flora, especially from the State of Ceará, we are reporting now the results of the investigation of four Piper species: P. dilatatum L.C. Rich, P. arboreum Aubl., P. tuberculatum Jacq. and P. crassinervium Kunth. Although the non-volatile constitution of all four species have already been reported,^2,6,17-19 the essential oils of P. tuberculatum and P. crassinervium are being reported for the first time. During referee assessment of this paper a report of the essential oils from eleven different plant species, including P. crassinervium, came out in the literature.²⁰

Results and Discussion

The results for the leaf essential oil analysis of four Piper species are described in Table 1. A total of 52 volatile constituents were identified, accounting for 88.49 – 98.62% of the chemical composition of the correspondent oils. All oils showed some similarity in the qualitative composition, but they differ significantly from a quantitative point of view. The essential oils of P. arboreum and P. tuberculatum are characterized by the presence of sesquiterpenes, 88.49 and 81.07%, respectively; while a high percentage of monoterpenes was detected in the oil of P. crassinervium (74.55%). On the other hand, a similar contribution of monoterpenes (42.14%) and sesquiterpenes (54.19%) was observed to the oil of P. dilatatum. As can be seen from Table 1, monoterpenes were not detected in the essential oil of P. arboreum, to which the most significant constituents were the sesquiterpenes bicyclogermacrene (25.07%), (E)-nerolidol (11.12%) and (E)-caryophyllene (8.35%). Previous studies on leaf essential oils of what seems to be three varieties of P. arboreum have been reported and significant differences were found in their chemical compositions, although all them were dominated by sesquiterpenes.^10,11,15 The leaf essential oil of P. arboreum Aublet var. latifolium (C.DC) Yuncker from Rondônia, was characterized by an higher content of germacrene D (72.87%),¹⁰ while in the leaf oil of P. arboreum var. arboreum Yunck. from Rio de Janeiro, the most abundant constituents were g-eudesmol (14.61%) and a-eudesmol (12.21%), followed by bulnesol (8.13%) and (E)-caryophyllene (6.40%).¹⁵ On the other hand, in the study of P. arboreum var. latifolium oil from Panama, d-cadinene (25.80%) and a-copaene (7.40%) were the main compounds.¹¹ Comparison of those major constituents for all P. arboreum oils from different origins revealed that all components are present in the oil from Rio de Janeiro, just two of them (germacrene D and a-copaene) are present in the oil from Rondônia and, that the same compounds (germacrene D, (E)-caryophyllene, d-cadinene and a-copaene) are qualitatively but not quantitatively, present in both oils from Ceará and Panama. In the essential oil of P. dilatatum, from Ceará, a-phellandrene (22.53%) and D-3-carene (10.20%) were the major monoterpene hydrocarbons, while bicyclogermacrene (25.03%) and (E)-caryophyllene (10.26%) were the main sesquiterpene hydrocarbons. Earlier investigation of the leaf essential oil of P. dilatatum from Rio de Janeiro, showed that the monoterpenes myrcene (41.70%) and a-pinene (17.70%) were the major components.¹⁵ The major mono- and sesquiterpenes of the oil from Ceará are absent, or in very low percentage, in the oil from Rio de Janeiro. The same thing happens with the major monoterpenes of the oil from Rio de Janeiro. The oil of P. crassinervium, rich in monoterpenes, was characterized by a high content of linalool (28.61%), b-pinene (20.01%), a-pinene (11.27%) and 1,8-cineole (10.81%). Recently, the essential oil of P. crassivervium from Ecuador has been examined and was found to be rich in monoterpenes, including a-pinene (15.17%) and b-pinene (10.0%), in accordance with our analysis, but also rich in limonene (26.6%) and a-terpinene (7.79%),²⁰ not present in the oil from Ceará. All these facts would be maybe attributed to the edafoclimatic conditions related to the different environments in which the plants grown. The most significant constituents of the oil of P. tuberculatum were the sesquiterpenes (E)-caryophyllene (37.78%) and germacrene D (11.81%), while a-pinene (4.06%) and b-pinene (4.51%) were the main constituents of the monoterpene fraction. Unfortunately no other source of P. tuberculatum oil was found in the literature for comparison.

Of the 52 identified compounds, b-elemene, (E)-caryophyllene, germacrene D, bicyclogermacrene and d-cadinene were detected in all four oil samples, while a-pinene, b-pinene, myrcene, d-elemene, a-humulene, elemol, (E)-nerolidol and b-eudesmol were found in three of them. As can be observed from our findings, as well as from other analysis on essential oil from Piper species,^10-13,15,16 there is a notable tendency of those plants to biosynthesize sesquiterpenes, independent of their natural habitats.

Experimental

Plant material

The leaves of P. dilatatum (# 33.821), P. crassinervium (# 33.877) and P. arboreum (# 33.872) were harvested at the flowering stage from Guaramiranga Mountain (Ceará State, Brazil), in June 2004. P. tuberculatum (# 12985) was harvested from Horto de Plantas Medicinais Prof. Francisco José de Abreu Matos - Universidade Federal do Ceará, in July 2004. All species were harvested from 9 to 11 am. and the voucher specimens have been deposited at the Herbarium Prisco Bezerra (EAC), Universidade Federal do Ceará.

Isolation of the essential oils

Fresh leaf samples of all species were subjected to hydrodistillation for 2 hours in a Clevenger-type apparatus. The isolated essential oils were dried over anhydrous sodium sulfate and, after filtration, maintained under refrigeration before analysis. The yields (m/m) of the oils were calculated based on the fresh weight of the plant materials: P. dilatatum (600 g, 0.07%), P. crassinervium (290 g, 0.11%), P. arboreum (700 g, 0.07%) and P. tuberculatum (660 g, 0.03%).

GC-MS conditions

GC-MS analysis was carried out on a Hewlett-Packard Model 5971 GC/MS using a DB-5 fused silica capillary column (30 m x 0.25 mm i.d. x 0.25 µm film thickness); helium as the carrier gas, flow rate of 1 mL min^-1 and with split ratio 1:30. The injector temperature and detector temperature was 250 ºC and 200 ºC, respectively. The column temperature was programmed from 35 °C to 180 ºC at 4 ºC min^-1 and then from 180 °C to 250 ºC at 10 ºC min^-1. Mass spectra were recorded from 30 – 450 m/z.

Compound identification

The volatile components were identified by comparison of their 70 eV mass spectra with those of the spectrometer data base using the Wiley L-built library and other two computer libraries MS searches using retention indices as a preselection routine.^21,22 The identifications were confirmed by comparison of the fragmentation pattern and their retention indices with those reported in the literature.^23,24

Acknowledgements

The authors are grateful to Mrs. Olga Ramos (LPN/UFC) for recording the GC-MS spectra and the Brazilian Agencies CNPq, CAPES, FUNCAP and PRONEX for financial support.

Received: January 10, 2005

Published on the web: September 22, 2005

• 1. Parmar, V. S.; Jain, S. C.; Bisht, K. S.; Jain, R.; Taneja, P.; Jha, A.; Tyagi, O. D.; Prasad, A. K.; Wengel, J.; Olsen, C. E.; Boll, P. M.; Phytochemistry 1997, 46, 597.
• 2. Danelutte, A. P.; Lago, J. H. G.; Young, M. C. M.; Kato, M. J.; Phytochemistry 2003, 64, 555.
• 3. Stöhr, J. R.; Xiao, P. G.; Bauer, R; J. Ethnopharmacol. 2001, 75, 133.
• 4. Martins, R. C. C.; Lago, J. H. G.; Albuquerque, S.; Kato, M. J.; Phytochemistry 2003, 64, 667.
• 5. Rukachaisirikul, T.; Siriwattanakit, P.; Sukcharoenphol, K.; Wongvein, C.; Ruttanaweang, P.; Wongwattanavuch, P.; Suksamrarn, A.; J. Ethnopharmacol 2004, 93, 173.
• 6. Silva, R. V.; Navickiene, H. M. D.; Kato, M. J.; Bolzani, V. S.; Média, C. I.; Young, M. C. M.; Furlan, M.; Phytochemistry 2002, 59, 521.
• 7. Facundo, V. A.; Sá, A. L.; Silva, S. A. F.; Morais, S. M.; Matos, C. R. R.; Braz-Filho, R.; J. Braz. Chem. Soc 2004, 15, 140.
• 8. Lago, J. H. G.; Tanizaki, T. M.; Young M. C. M.; Guimarães, E. F.; Kato, M. J.; J. Braz. Chem. Soc 2005, 16, 153.
• 9. Rawat, A. K. S.; Tripathi, R. D.; Khan, A. J.; Balasubrahmanyam, V. R.; Biochem. Syst. Ecol 1989, 17, 35.
• 10. Machado, S. M. F.; Militão, J. S. L. T.; Facundo, V. A.; Ribeiro, A.; Moraes, S. M.; Machado, M. I. L.; J. Essent. Oil Res 1994, 6, 643.
• 11. Mundina, M.; Vila, R.; Tomi, F.; Gupta, M. P.; Adzet, T.; Casanova, J.; Cañigueral, S.; Phytochemistry 1998, 47, 1277.
• 12. Jirovetz, L.; Buchbauer, G.; Ngassoum, M. B.; Geissler, M.; J. Chromatogr. A 2002, 976, 265.
• 13. Mundina, M.; Vila, R.; Tomi, F.; Tomàs, X.; Cicció, J. F.; Adzet, T.; Casnova, J.; Cañigueral, S.; Biochem. Syst. Ecol. 2001, 29, 739.
• 14. Martins, A. P.; Salgueiro, L.; Vila, R.; Tomi, F.; Canigueral, S.; Casanova, J.; Cunha, A. P.; Adzet, T.; Phytochemistry 1998, 49, 2019.
• 15. Santos, P. R. D.; Moreira, D. L.; Guimarães, E. F.; Kaplan M. A. C.; Phytochemistry 2001, 58, 547.
• 16. Vila, R.; Begõna, M.; Tomi, F.; Casanova, J.; Ferro, E. A.; Cañigueral, S.; J. Ethnopharmacol 2001, 76, 105.
• 17. Terreaux, C.; Gupta, M. P.; Hostettmann, K.; Phytochemistry 1998, 49, 461.
• 18. Chaves, M. C. O.; Júnior, A. G. F.; Santos, B. V. O.; Fitoterapia 2003, 74, 181.
• 19. Capron, M. A.; Wiemer, D. F.; J. Nat. Prod. 1996, 59, 794.
• 20. Sacchetti, G.; Maietti, S.; Muzzoli, M.; Scaglianti, M.; Manfredini, S.; Radice, M.; Bruni, R.; Food Chem 2005, 91, 621.
• 21. Alencar, J. W.; Craveiro, A. A.; Matos, F. J. A.; J. Nat. Prod 1984, 47, 890.
• 22. Alencar, J. W.; Craveiro, A. A.; Matos, F. J. A.; Machado, M. I. L.; Quim. Nova 1990, 13, 282.
• 23. Stehagen, E.; Abrahamson, S.; McLafferty, F. W.; Registry of Mass Spectral Data Base, Government Printing Office: Washington DC, 1974.
• 24. Adams, R. P.; Identification of Essential Oil Components by Gas Chromatography/Quadrupole Mass Spectroscopy, Allured Publishing Corporation: Carol Stream, Illinois, USA, 2001.

Publication Dates

• Publication in this collection
  20 Jan 2006
• Date of issue
  Nov 2005

History

• Received
  10 Jan 2005