FLAVOUR AND FRAGRANCE JOURNALFlavour Fragr. J. 2003; 18: 198–201Published online in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ffj.1181

Copyright © 2003 John Wiley & Sons, Ltd.

John Wiley & Sons, Ltd.

Constituents of the essential oils from Piper friedrichsthalii C.DC. and P. pseudolindenii C.DC. from Central AmericaESSENTIAL OILS FROM PIPER SPP.Roser Vila,1 Marisa Mundina,1 Felix Tomi,2 José F. Cicció,3 Mahabir P. Gupta,4 José Iglesias,1 Joseph Casanova2 and Salvador Cañigueral1*

1 Unitat de Farmacologia i Farmacognòsia, Facultat de Farmàcia, Universitat de Barcelona, Avda. Diagonal 643, E-08028 Barcelona, Spain2 Équipe ‘Chimie et Biomasse’, UMR CNRS 6134, Université de Corse, Route des Sanguinaires, F-20000 Ajaccio, France3 Centro de Investigaciones en Productos Naturales (CIPRONA), Escuela de Química, Universidad de Costa Rica, 2060 San José, Costa Rica4 Centro de Investigaciones Farmacognósticas de la Flora Panameña (CIFLORPAN), Facultad de Farmacia, Universidad de Panamá, Estafeta Universitaria, Panamá, Republic of Panamá, and Smithsonian Tropical Research Institute, Panamá, Republic of Panamá

Received 19 January 2002Revised 5 May 2002Accepted 25 May 2002

ABSTRACT: Essential oils obtained by hydrodistillation from the leaves and flower spikes of Piper friedrich-sthalii from Costa Rica and Panama, and leaves of P. pseudolindenii from Costa Rica were analysed by GC–FID, GC–MS and 13C-NMR. Monoterpenoids constituted the main fraction of the oils of P. friedrichsthalii from Costa Rica,particularly α-pinene, camphene, β-phellandrene, limonene and menthane derivatives, whereas the oils from leavesof P. friedrichsthalii from Panamá and of P. pseudolindenii from Costa Rica were characterized by a high sesquiter-pene content, especially 11-selinen-4α-ol, α-selinene, germacrene D and β-selinene in the former, and β-elemene,β-caryophyllene, germacrene D and α-humulene in the latter. Copyright © 2003 John Wiley & Sons, Ltd.

KEY WORDS: Piper friedrichsthalii; Piper pseudolindenii; Piperaceae; essential oil; GC–MS; 13C-NMR; α-pinene; α-selinene; 11-selinen-4α-ol; β-elemene; β-caryophyllene

Introduction

In the New World, the genus Piper contains at least500 different species.1 However, their classification isstill confusing, due to the great variation in vegetativecharacters or the use of inadequate botanical material.2

In particular, P. friedrichsthalii C.DC. has been dis-tinguished by Burger2 as a species different from P.lanceaefolium C.DC. However, in the last revision ofthe genus,3 P. friedrichsthalii has been reduced to asynonym under P. lanceaefolium, as it was consideredmerely to be a narrow-leaved form of this species.

The present work deals with the comparison of theessential oil from leaves and spikes of P. friedrichsthaliiC.DC. from Costa Rica and from the leaves of P.friedrichsthalii from Panama. In addition, it also reportson the composition of the essential oil from leaves ofP. pseudolindenii C.DC. from Costa Rica.

Experimental

Plant Material

Leaves and flower spikes of P. friedrichsthalii were col-lected in Pacayas, Cartago (Costa Rica) in September1996 and leaves of P. pseudolindenii in Turrialba,Cartago (Costa Rica) in the same month. The plant mate-rial was identified by L. J. Poveda (Escuela de CienciasAmbientales, UNA, Heredia). Voucher specimens weredeposited with the Herbarium of the University of CostaRica at the School of Biology under accession NosUSJ 56541 and USJ 62335, respectively. Leaves of P.friedrichsthalii from Panama were collected in the area ofFortuna, Quebrada Honda (Chiriqui) and identified by

* Correspondence to: S. Cañigueral, Unitat de Farmacologia i Farma-cognòsia, Facultat de Farmàcia, Avda. Diagonal 643, E-08028 Barcelona,Spain. E-mail: caniguer@farmacia.far.ub.esContract/grant sponsor: Generalitat de Catalunya, Spain.Contract/grant sponsor: Université de Corse, France.Contract/grant sponsor: Instituto de Cooperación Iberoamericana (ICI), Spain.Contract/grant sponsor: Vicerrectoría de Investigación, Universidad de CostaRica. 89 93600Contract/grant sponsor: Universidad de Panama.Contract/grant sponsor: Organization of American States.Contract/grant sponsor: CYTED

ESSENTIAL OILS OF PIPER SPP. 199

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18 198–201

Professor Mireya Correa (Director of the Herbarium ofthe University of Panama) in October 1998. A voucherspecimen was deposited in the Herbarium FLORPAN(University of Panama) under the accession No. 3107.

Isolation and Analysis of Essential Oils

Fresh plant material was submitted to hydrodistillationusing a Clevenger-type apparatus, giving essential oil yieldsof 0.3% from leaves and flower spikes of P. friedrich-sthalii and of 0.1% from the leaves of P. pseudolindenii.

Essential oils were analysed by GC–FID and GC–MSusing two fused silica capillary columns of differentstationary phases: Supelcowax™ 10 and methyl siliconeSE-30 (30 m × 0.2 mm i.d., 0.25 µm film thickness).GC–FID analysis were carried out on a Hewlett-Packard5890, using the following analytical conditions: carriergas, He (1 ml/min); split ratio, 1:60; injector temperature,250 °C; oven temperature programme, 80–220 °C at4 °C/min; detector temperature, 270 °C. Mass spectrawere obtained with a GC–MS computerized system (GCHewlett-Packard 5890 coupled to a mass selective detectorHewlett-Packard 5971A) using the same Supelcowax™10 and SE-30 capillary columns as above. Analyticalconditions were: carrier gas, He (1 ml/min); split ratio,1:60; injector temperature, 250 °C; interface temperature,290 °C; oven temperature programme, 80–220 °C at 4 °C/min. Identification of components was achieved fromtheir retention indices, determined with reference to ahomologous series of fatty acid methyl esters, and theirMS, which were compared with those stored in our ownlibrary and with literature data.4,5 The quantification ofthe components was performed on the basis of their GCpeak areas, without corrections for response factor.

The identity of major constituents was also confirmedby 13C-NMR. Spectra were recorded on a Bruker AC 200Fourier transform spectrometer, operating at 50.323 MHzfor 13C, in CDCl3, with all shifts referred to internal TMS.Analytical parameters were: pulse width, 3.2 µs; acquisi-tion time, 1.3 s for 32 K data table with spectral widthof 250 ppm, CPD mode decoupling and digital resolutionof 0.763 Hz/pt. The number of accumulated scans was10 000 for each sample of essential oil. Exponentialmultiplication of the free induction decay with a line broaden-ing of 1 Hz was used before Fourier transformation.6–8

Results and Discussion

Essential Oils from Leaves and Flower Spikes of P. friedrichsthalii from Costa Rica and Panama

The results obtained in the analysis of the essential oilfrom leaves and flower spikes of P. friedrichsthalii fromCosta Rica (Table 1) showed that differences between

them were mainly quantitative. In total, 46 componentswere identified, comprising more than 80% and 75% ofthe total oil from leaves and spikes, respectively. Mono-terpenes were shown to be the major group of constituents,while sesquiterpenes constituted a small percentage. Themain components of the leaf oil were the monoterpenesα-pinene (14.7%) and camphene (5.2%) and the sesquit-erpene germacrene D (7.0%). The main components in theoil from the flower spikes were the monoterpene hydro-carbons α-pinene (13.4%) and β-phellandrene (5.2%)and the oxygenated monoterpenes trans- and cis-p-menth-2-en-1-ol (7.0% and 5.1%, respectively). The main dif-ferences between the oils were the higher amounts ofoxygenated monoterpenes, especially menthane derivativessuch as p-menth-2-en-1-ol, piperitol and piperitone, anda minor presence of sesquiterpenes in the oil from theflower spikes. In both samples, the phenylpropanoidselemicin and eugenol were found in a low percentage (1%).

The analysis of the essential oil from the leaves ofP. friedrichsthalii from Panama (Table 1) revealedthat sesquiterpenes (79.0%) were the major group ofconstituents, especially the hydrocarbons α-selinene(12.0%), germacrene D (9.6%) and β-selinene (7.9%) andthe oxygenated 11-selinen-4α-ol (12.8%), spathulenol(4.3%), caryophyllene oxide (3.2%) and 6-E-nerolidol(2.8%). In contrast, monoterpenes constituted only a lowpercentage of the oil (0.8%), while phenylpropanoidswere not detected. In total, 42 different componentswere identified (83% of the total oil).

Comparison between the oils from the leaves fromboth localities showed some clear differences. In thesample from Costa Rica, monoterpenes proved to bethe main group of components, whereas in the oil fromPanama, sesquiterpenes constituted the major group.In addition, phenylpropanoids were not detected in thelatter, whereas elemicin (0.5%) and eugenol (0.5%) wereidentified in the sample from Costa Rica.

Important differences between the composition of theleaf oils from P. friedrichsthalii and P. lanceaefoliumpreviously investigated by us9 were also found, thelatter being characterized by significant percentagesof sesquiterpenes (48%), mainly germacrene D and β-caryophyllene, and phenylpropanoids (36.3%), especiallyapiol isomers and elemicin. Moreover, the co-occurrenceof several menthane derivatives was established inthe oils of P. friedrichsthalii, but not in the oil of P.lanceaefolium.9 In conclusion, these results support themorphological classification established by Burger,2 inwhich both taxons were recognized as distinct species.

Essential Oil from Leaves of P. pseudolindenii from Costa Rica

The essential oil from the leaves of P. pseudolindeniifrom Costa Rica (Table 1) was characterized by a low

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 198–201

200 R. VILA ET AL.

Table 1. Chemical composition of essential oils ofPiper friedrichsthalii from Costa Rica and Panama andP. pseudolindenii from Costa Rica

Constituents RI P-1 P-2 P-3

%L %S %L %L

α-Pinene 117 14.7abcd 13.4abcd tabc 2.2abcd

Camphene 134 5.2abcd 4.5abcd tabc 0.1abc

β-Pinene 156 0.6abc 1.5abcd tabc 6.7abcd

Sabinene 164 — 0.1ab — —3-Carene 177 0.4abc 0.3abc — —Myrcene 182 2.3abcd 2.9abcd — 0.9abcd

α-Phellandrene 194 2.8abcd 3.7abcd — —α-Terpinene 200 1.9abcd 2.6abcd — —Limonene 207 4.4abcd 4.9abcd 0.1abc 0.5abcd

β-Phellandrene 212 4.4abcd 5.2abcd — 0.1ab

cis-2-Hexenal 214 — — — 0.3abc

cis-Ocimene 218 — — — 2.8abcd

trans-Ocimene 225 — — — 1.2abcd

γ-Terpinene 227 2.1abcd 2.6abcd — —p-Cymene 236 0.5abcd 0.3abcd — tab

Terpinolene 241 0.7abcd 0.9abcd — 0.1abc

2-Nonanone 300 — — — 0.1abc

Hexyl butyrate 311 — — — 1.8ad

α-Cubebene 326 — — 0.2abc 0.1abc

δ-Elemene 335 — — 0.2ac 1.3abc

α-Copaene 346 1.7abcd tab 3.3abcd 0.2abc

Camphor 358 0.2abcd 0.3abc — —α-Gurjunene 368 — — 0.1abc —Linalool 373 1.3abcd 3.0abcd 0.1ac 1.7abcd

trans-p-Menth-2-en-1-ol 383 3.8a 7.0a 0.2a —Bornyl acetate 394 0.3ab 0.6ab — 0.1ac

β-Gurjunene 396 0.3abc — 0.5ac —β-Elemene 400 — — 0.7abc 15.0abcd

2-Undecanone 401 — — — 0.1abc

Terpinen-4-ol 405 0.4abc 0.6ac — —β-Caryophyllene 404 1.4abcd 0.3abc 4.3abcd 11.8abcd

6,9-Guaiadiene 407 0.4abc 0.9abc — 0.3abc

cis-p-Menth-2-en-1-ol 411 2.8a 5.1a 0.1a —γ-Elemene 417 — — — 1.1ab

allo-Aromadendrene 418 — — 2.2abcd 0.2abc

γ-Gurjunene 431 1.0abc 2.1abc 1.8acd —α-Humulene 433 0.3abc — 1.4abcd 7.0abcd

cis-Piperitol 443 0.7abcd 1.4abcd tac —trans-Piperitol 443 1.2abcd 3.0abcd 0.1ac —α-Terpineol 444 0.7abcd 1.0acd 0.1ac 0.5ab

Borneol 446 2.2abcd 2.9abcd — —γ-Terpineol 451 0.7abc — — —γ-Muurolene 451 — — — 1.8acd

Germacrene D 456 7.1abcd 1.2abcd 9.6abcd 9.0abcd

δ-Guaiene 457 — — — 1.0ab

α-Selinene 458 — — 12.0abcd 0.7abc

Bornyl isovalerate 460 0.7a ta — —β-Selinene 463 — — 7.9abcd 0.3ab

Bicyclogermacrene 466 2.7abcd — 2.7abd 3.4abcd

δ-Cadinene 477 2.1abcd — 4.2abcd 1.5abcd

β-Bisabolene 478 — — — 1.7abcd

Calamenene 510 — — 0.1ab 0.3ac

Germacrene B 511 — — — 5.4abc

α-Ionone 520 — — — 0.1ab

α-Calacorene 543 — — 0.1ab —β-Ionone 560 — — — 0.1abc

Isocaryophyllene oxide 568 — — 0.1abc —Caryophyllene oxide 580 — — 3.2abcd 1.3abcd

Ledol 602 — — 1.0abd —6-E-Nerolidol 612 2.7abcd 1.2abcd 2.8abcd 2.5abd

Cubenol 623 0.4abc — 0.4abc 0.1ab

epi-Cubenol 624 0.4abc — 0.6acd 0.1ab

Globulol 634 — — 0.2abcd —Viridiflorol 636 0.3abc — — 0.4abc

Spathulenol 654 — — 4.3abcd 1.4abcd

T-Cadinol 672 1.1abc — — —

monoterpene content (17.0%), with α- and β-pinene(2.2% and 6.7%, respectively), and cis-ocimene (2.8%)the most important ones. Sesquiterpenes (70.2%)constituted the main fraction of the oil, especially β-elemene (15.0%) and β-caryophyllene (11.8%). It is inter-esting to note the co-occurrence of β-, δ- and γ-elemenes(15.0%, 1.3% and 1.1%, respectively), and of somegermacrane derivatives such as germacrene D (9.7%)and B (5.4%). Among the oxygenated sesquiterpenes,6-E-nerolidol (2.5%), α-cadinol (1.5%), spathulenol (1.4%)and caryophyllene oxide (1.3%) were also detected.In total, 50 components (89.7% of the total oil) wereidentified.

Acknowledgements— The authors are grateful to the Generalitat deCatalunya (Spain) and the Université de Corse (France), who gavefinancial support within the ACI Cooperation Program, as well as tothe Instituto de Cooperación Iberoamericana (ICI, Spain). We arealso indebted to the Vicerrectoría de Investigación, Universidad deCosta Rica (Project No. 809-93-600), and to the Universidad dePanama. CIFLORPAN acknowledges the support from Organization ofAmerican States. This study was performed within the framework ofthe Iberoamerican program CYTED (Subprogram X).

Constituents RI P-1 P-2 P-3

%L %S %L %L

Eugenol 674 0.5abc 0.5ac— —

T-Muurolol 679 — — 1.4ab 0.9abcd

Elemicin 689 0.5abc 0.5ac — —α-Elemol 690 — — 1.3abc —α-Cadinol 693 0.7abc — 2.4abcd 1.5abcd

α-Eudesmol 695 — — 0.2ab —Isospathulenol 705 — — — 0.9abc

11-Selinen-4-α-ol 709 — — 12.8abcd —Phytol 860 — — 0.1abc 0.2abc

Tricyclenee 206 0.1ac 0.1ac — —Nerale 350 — — — 0.1ac

Piperitonee 355 0.2ac 0.3ac — —E,E-α-Farnesenee 495 0.7ac — — —γ-Cadinenee 500 0.5ac — — 0.6ac

α-Muurolenee 520 — — 0.1ac —

Monoterpene hydrocarbons 40.1 43.0 0.2 14.6Oxygenated monoterpenes 15.2 25.2 0.6 2.4Sesquiterpene hydrocarbons 18.2 4.9 51.4 60.9Oxygenated sesquiterpenes 5.6 1.2 27.6 9.3Phenylpropanoids 1.0 1.0 — —Others — — 0.1 2.5

Total 80.1 75.3 83.0 89.7

P-1, P. friedrichsthalii from Costa Rica; P-2, P. friedrichsthalii from Panama; P-3, P. pseudolindenii.%L, essential oil from leaves; %S, essential oil from flower spikes.RI, retention indices in Supelcowax™10, except the last six constituents, which were only detected in SE-30.abcd Identification methods: a GC–MS; b retention index in Supelcowax™10; c retention index in SE-30; d H13

C-RMN; e Compounds only detected in SE-30.t, trace (≤0.05).

Table 1. Continued

ESSENTIAL OILS OF PIPER SPP. 201

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18 198–201

References

1. Tebbs MC. Bull. Br. Mus. Nat. Hist. (Bot.), 1989; 19: 117–158.

2. Burger WC. Fieldiana Bot., 1971; 35: 5–227.3. Tebbs MC. Bull. Nat. Hist. Mus. Lond. (Bot.), 1993; 23: 1–50.4. Central Institute for Nutrition and Food Research. Compilation

of Mass Spectra of Volatile Compounds in Food. TNO: Utrecht,1980.

5. McLafferty FW. Registry of Mass Spectral Data. Wiley: NewYork, 1993.

6. Tomi F, Bradesi P, Bighelli A, Casanova J. J. Magn. Res. Anal.,1995; 1: 25–34.

7. Bradesi P, Bighelli A, Tomi F, Casanova J. Can. J. Appl. Spectrosc.,1996; 41: 15–24.

8. Bradesi P, Bighelli A, Tomi F, Casanova J. Can. J. Appl. Spectrosc.,1996; 41: 41–50.

9. Mundina M, Vila R, Tomi F et al. Biochem. Syst. Ecol., 2001; 29:739–748.