/ . Essent. Oil Res., 12, 19-23 (Jan/Feb 2000)
Composition of the Essential Oils from Galls and Aerial Parts of Pistacia lentiscus L.1 Ana Fernandez,* Ana Camacho, b Carlos Fernandez* and Joaquin Altarejosc* "Departamentos de Biologia Animal, Vegetal y Ecologia h Ciencias de la Salud c Quimica Inorgdnica y Orgdnica Facultad de Ciencias Experimentalesy de la Salud, Universidad dejaen, 23071 Jaen, Spain Pilar Perez Destilaciones Garcia de la Fuente-Universal Fragrances SA, Apdo. Correos no. 69, 18080 Granada, Spain Abstract The chemical composition of the essential oils obtained from the galls and the aerial parts of Pistacia lentiscus L. (Anacardiaceae) was analyzed by GC/MS. Among the 58 constituents identified (representing 82.8% of the oil composition) sesquiterpene hydrocarbons predominated (ca. 47%), J3-caryophyllene (13.1%), 6-cadinene (8.1%) and Germacrene D (6.8%) being the major components. Although galls and oils of the aerial parts were similar from a qualitative viewpoint, the latter oil was somewhat more complex (73 components identified, accounting for 85.4% of the oil) and the monoterpene hydrocarbons fraction (ca. 35%) was now dominant in the oil, a-pinene (13.0%), P-caryophyllene (6.9%), limonene and p-phellandrene (5.4%), {3-pinene (4.9%) and p-cymene (4.7%) being the main components. Key Word Index Pistacia lentiscus, Anacardiaceae, galls, essential oil composition, p-caryophyllene, a-pinene.
Introduction Pistacia lentiscus L. (Anacardiaceae) is an evergreen shrub or small tree native to the Mediterranean region and Middle East (1). It is commonly found in almost the whole Iberian Peninsula in dry open woods, pine forest and as a part of the scrub, and its flowering and fructification periods are during the spring and the autumn, respectively (2). The lentisco (Spanish common name) is valued because it is the source of mastic gum, an exudate which has been widely used in folk medicine to strengthen gums, deodorize breath, combat diarrhea and tuberculosis, etc. (2,3), and more recentiy to flavor alcoholic beverages, pharmaceuticals, chewing gum and traditional food (4,5). In addition, lentisco leaves and fruits have been used in a variety of ways by rural inhabitants of the areas where the plant grows wild (2,3,6). With regard to the composition of volatiles from different parts of the plant, the leaves (7-11), fruits (4,7,9) and mastic gum (7,12,13) have been the subject of previous research. However, in spite of all this work, nothing is known about the chemical composition of the galls, a teratologic structure •Address for correspondence x Part of this work was presented as a poster (Abstracts, p 37) at the Xllth International Conference on Aromas and Essential Oils, Seville, Spain, October 19-23, 1997 l04l-2905/00/0001-00l9$06.00/0—©2000 Allured Publishing Corp.
Received: October 1998 Revised: February 1999 Accepted: April 1999
20
FERNANDEZ ET AL.
Table I. Percentage composition 0 of the oils of Pistacia lentiscus Rl
a
839 841 852 921 931 943 971 975 989 1003 1009 1012 1013 1025 1028 1031 1042 1049 1054 1087 1097 1100 1105 1137 1144 1161 1165 1169 1178 1186 1192 1196 1202 1209 1219 1228 1231 1243 1247 1250 1290 1294 1319 1348 1354 1370 1375 1389 1391 1420
Compound
15
(E)-2-Hexenal (Z)-3-Hexenol Hexanol Tricyclene a-Pinene Camphene Sabinene p-Pinene Myrcene a-Phellandrene 8-3-Carene 1,4-Cineole a-Terpinene p-Cymene Limonene + p-Phellandrene (Z)-p-Ocimene (E)-p-Ocimene Isoamyl butyrate y-Terpinene Terpinolene + 2-Nonanone Linalool + a-Campholenic aldehyde Isoamyl isovalerate Isoamyl valerate trans-Pinocarveol p-Menthatriene Pinocarvone Borneol Carvotanacetone Terpinen-4-ol p-Cymen-8-ol a-Terpineol Myrtenal cis-Sabinol"1" Verbenone cis-Carveol (Z)-3-Hexenyl 2-methylbutyrate (Z)-3-Hexenyl isovalerate Cuminaldehyde Carvone Isoamyl hexanoate Bornyl acetate 2-Undecanone 2,4,6-Trimethylbenzaldehyde a-Cubebene 2,4,5-Trimethylbenzaldehyde (Z)-3-Hexenyl hexanoate a-Copaene p-Bourbonene P-Cubebene + p-Elemene p-Caryophyllene
A
d
Be
. t"
t 4.4 t 0.5 1.7 0.3 1.0 t
0.2 0.3 2.0 t t t 0.4 0.7f 0.19
0.2
0.2 t 1.8
1.9 0.2 t
t
0.1 0.3 0.9 2.7 0.3 2.2
1.8 0.1 1.5 13.1
L.
".
t t t 0.2 13.0 0.3 3.3 4.9 0.8 0.7 0.1 t t 4.7 5.4 t t 0.2 0.2 1.0 0.5 0.3 t 0.2 0.3 0.1 0.1 0.1 2.8 0.8 2.2 0.3 0.2 t t 0.1 0.1 t t 0.4 0.4 1.6 0.7 0.2
0.2 1.1 0.1 1.5 6.9
PlSTACIA LENTISCUS
21
Table I. Continued Rl a 1429 1433 1443 1446 1453 1460 1472 1477 1480 1495 1500 1508 1514 1524 1534 1547 1553 1569 1583 1588 1612 1620 1630 1635 1646 1649 1659 1681 1751 1753
Compound 5
Ad
B8
Isoamyl benzoate1" Isoamyl octanoate Geranyl acetone Unknown a-Humulene Aromadendrene Unknown a-Amorphene Germacrene D Unknown a-Muurolene P-Bisabolene y-Cadinene 5-Cadinene Cadina-1,4-diene Elemicin Unknown (Z)-3-Hexenyl benzoate Spathulenol Caryophyllene oxide Unknown Unknown Unknown 10-epi-y-Eudesmol1' T-Muurolol Unknown a-Cadinol isomer* Bisabolol* Myristic acid Benzyl benzoate
0.5 0.1 0.2 0.6 3.0 1.2 1.2 3.3 6.8 1.2 3.4 0.6 1.1 8.1 0.7
0.5 0.3 0.2 0.2 1.6 0.7
0.2 0.5 1.3 0.4 0.6 1.2 3.5 3.8
4.3 0.6 0.5 0.2
2.0 2.3
2.6 0.4 0.9 4.1 0.1 0.1 0.6 0.1 1.4 3.7 1.2 0.5 0.9
3.6 0.8 3.7 0.9
0.2
"Temperature-programmed retention indices referred to n-alkanes, determined on DB-5 column according to reference 19; "Constituents listed in elution order from a 30 m DB-5 capillary column. Unidentified peaks with a presence in the oil of 0.5% or higher have been included. Mass spectra data of unknown compounds and those with a f are included in Table II; 'Percentage values less than 0.1% are denoted as t (traces); dA = Pistacia lentiscus galls; *B = Pistacia lentiscus aerial parts; 'Peak value corresponding to terpinolene alone; gPeak value corresponding to linalool alone. tentatively identified; *Correct isomer not determined
of this plant which appears in the leaves as a response to the parasitic action of the insect Aploneura ' lentisci (2). Therefore, here w e report on the oil composition of P. lentiscus galls (14), and also that from the aerial parts of the plant. Experimental Plant Material. The P. lentiscus samples were collected in October 1996 during the fructification period near the Cabra del Santo Cristo village (30S VG7783) in the Province of Jaen (Spain). Voucher specimens have been deposited at the Herbarium of the University ofJaen. The plant material consisted in the whole aerial parts formed by leaves and terminal stems (sample B in Table I) and galls (sample A in Table I). Air-dried aerial parts (135.4 g) were ground and hydrodistilled for 8 h using a Clevengertype apparatus (15). The oil was dried over anhydrous sodium sulfate and stored under refrigeration C-4°C) in the dark (335 mg, 0.25%). In the same manner the galls sample (56.8 g) yielded the
22
FERNANDEZ ET AL.
Table II. Mass spectra data of unknown and tentatively identified components of Pistacia lentiscus oil Rl a
Compound
m/z (rel. int.)
1202
cis-Sabinol
1429 1446
Isoamyl benzoate Unknown
1472
Unknown
1495
Unknown
1553
Unknown
1612
Unknown
1620
Unknown
1630
Unknown
1635
1 o-epi-y-Eudesmol
1649
Unknown
1659
a-Cadinol isomer
152 [M+](3), 134(28), 119(19), 109(20), 92(100), 91(87), 81(45), 79(37), 65(17), 41(41) [M+]?, 161(21), 133(5), 123(22), 105(100), 77(49), 70(55), 55(23), 41(17) 204[M+](27), 189(5), 161(100), 133(13), 119(45), 105(81), 91(34), 81(35), 69(12), 55(14), 41(27) 204[M+](32), 189(14), 161(100), 133(17), 119(30), 105(46), 91(23), 81(29), 69(7), 55(9), 41(18) 204[M+](21), 189(3), 161(100), 133(16), 119(35), 105(53), 91(45), 79(20), 67(9), 55(13), 41(25) 220[M+](10), 205(8), 202(10), 187(11), 177(15), 159(42), 146(30), 131(62), 123(83), 105(72), 91(87), 79(81), 67(44), 55(57), 41(100) 220[M+](6), 205(8), 202(7), 187(13), 177(14), 159(38), 145(33), 131(44), 119(32), 105(60), 91(64), 79(53), 67(75), 55(51), 41(100) 222[M+](11), 204(40), 189(17), 179(25), 161(42), 149(17), 135(33), 121(23), 109(100), 93(47), 81(51), 67(40), 55(44), 41(67) [M+]?, 204(42), 189(38), 179(19), 161(79), 147(52), 135(7), 119(100), 105(46), 95(28), 81(28), 69(21), 55(33), 43(40) 222 [M+](7), 204(67), 189(100), 161(87), 149(21), 133(49), 119(19), 107(35), 105(33), 91(46), 81(35), 67(19), 59(51), 43(37), 41(38) 222[M+](0.5), 204(29), 189(8), 177(6), 161(100), 147(8), 133(13), 119(64), 105(47), 95(43), 79(37), 67(19), 55(23), 43(72) 222[M+](4), 204(39), 189(9), 177(4), 161(40), 149(9), 137(14), 121(70), 105(33), 95(100), 79(39), 71(28), 55(23), 43(94)
Temperature-programmed retention indices (see Table I)
corresponding essential oil (49 mg, 0.09%). GC/MS: Analyses were done on a Hewlett-Packard gas chromatograph (model 5890) coupled to a Hewlett-Packard mass spectrometer (model 5970 B). Gas chromatograph unit: DB-5 30 m x 0.25 mm fused silica capillary column, film thickness 0.25 Jim; carrier gas He (30 cm/s); initial temperature 50°C; final temperature 220°C; final time 30 min; rate 3°C/min; "split mode" ratio 1:60; injector temperature 240C,C. Mass spectrometer unit: ion source temperature 230°C; ionizing voltage 70 eV; scan range from m/z 35 to m/z 300. Most components were identified with the use of a built-in NBS peak matching library search-system. Retention indices, authentic standards and/or data reported in the literature (16,17) were also used for further identification. Quantitative data were direcdy obtained from the total ion chromatogram, carried out under the conditions specified before. Results and Discussion
Table I shows the composition of the oils obtained from galls and aerial parts of P. lentiscus. Constituents are listed in order of their elution from a DB-5 capillary column and were identified by GC/MS analysis in combination with retention indices. The oil content in galls (0.09%) was lower than in the whole plant (0.25%) as it can be directly appreciated by the resinous-poorer aspect of this tipycal formations if compared, for instance, with those from the close related species P. terebinthus (18). Examination of the galls oil showed the presence of at least 80 components, 58 of which were identified (Table I) accounting for 83% of the oil. It was mainly composed of {3-caryophyllene (13.1%), 6-cadinene (8.1%) and Germacrene D (6.8%) and of somewhat lesser amounts of oc-pinene (4.4%), an a-cadinol isomer (4.3%), T-muurolol (3-8%), 10-epi-y-eudesmol (3.5%); the last three components identified
i
PlSTACIA LENTISCVS
23
according only t o their mass spectra. T h e oil w a s d o m i n a t e d b y s e s q u i t e r p e n e h y d r o c a r b o n s (ca. 47%), oxygenated sesquiterpenes (ca. 14%) and m o n o t e r p e n e h y d r o c a r b o n s (ca. 12%), the cadinane, caryophyllane, germacrane a n d p i n a n e b e i n g the most representative s k e l e t a found. Comparing this oil with that o b t a i n e d from t h e w h o l e aerial parts of t h e p l a n t (Table I, column B), w h i c h was collected o n the s a m e date, it can b e d e d u c e d that b o t h oils w e r e similar from a qualitative viewpoint. However, in this s e c o n d oil the major c o m p o n e n t s w e r e a - p i n e n e (13.0%), P-caryophyllene (6.9%), limonene plus J3-phellandrene (5.4%), P-pinene (4.9 %) and p - c y m e n e (4.7%), the m o n o t e r p e n e hydrocarbons fraction (ca. 35%) b e i n g n o w p r e d o m i n a n t in t h e oil v e r s u s the sesquiterpene hydrocarbons o n e (ca. 25%). F u t h e r m o r e , s o m e c o m p o u n d s found as m i n o r o r trace c o m p o n e n t s in the w h o l e plant oil, such as p-cymen-8-ol, p - m e n t h a t r i e n e , t r a n s - p i n o c a r v e o l , isoamyl isovalerate, verbenone, carvone and o t h e r s c o u l d n o t b e d e t e c t e d in the galls oil. In contrast, the galls contained 10-epi-y-eudesmol, 2,4,5-trimethylbenzaldehyde a n d myristic acid, w h i l e t h e w h o l e plant oil did not. References 1. T. G. Tutin, Pistacia L. In: Flora Europaea. Edits., T. G. Tutin, V. H. Heywood, N. A. Burges, D. M. Moore, D. H. Valentine, S. M. Walters and D. A. Webb, Vol 2, p 237, Cambridge University Press, Cambridge (1968). 2. P. Font Quer, Lentisco. In: PlantasMedicinales: El Dioscorides Renovado. p p 440-442, Labor, Barcelona (1978). 3. G. Lopez, Pistacia lentiscusL. In: La Guia Incafo de los Arbolesy Arbustos de la Peninsula Iberica. pp 714-715, Incafo, Madrid (1982). 4. S. G. Wyllie, J. J. Brophy, V. Sarafis and M. Hobbs, Volatile components of thefruit of Pistacia lentiscus. J. Food Sci., 55, 1325-1326 (1990). 5. B. M. Lawrence, Mastic gum oil. In: Essential Oils 1976-1978. Edit., Natural Flavor and Fragrance Materials "Perfumer & Flavorist", p 6, Allured Publ. Corp., Carol Stream, IL (1979). 6. A. M. Femandez-Ocana, I. Ortuno-Moya, A. I. Martos-Gilabert and C. Fernandez-Lopez, Sabery Utilizacion de Plantas en la Provincia defaen. Campana de 1993- Bol. Inst. Est. Giennenses, 161, 199-318 (1996). 7. M. H. Boelens and R. Jimenez, Chemical composition of the essential oils from gum and from various parts of Pistacia lentiscusL. Flav. Fragr. J., 6, 271-275 (1991). 8. V. Picci, A. Scotti, M. Mariani and E. Colombo, Composition of the volatile oil of Pistacia lentiscusL. In: Flavour Science and Technology. Edits., M. Martens, G. A. Dalen and H. Russwurm, p p 107-110, J. Wiley & Sons Ltd., New York (1987). 9. Z. Fleisher and A. Fleisher, Aromatic plants of the Holy Land and the Sinai. PartX. Volatiles of the Mastic treePistacia lentiscusL.). Essent. Oil Res., 4, 663-665 (1992). 10. H. L. DePooter, N. M. Schamp, E. A. Aboutabl, S. F. El Tohamy and S. L. Doss, Essential oils from the leaves of three Pistacia species grown in Egypt. Flav. Fragr. J., 6, 229-232 (1991). 11. V. Castola, A. Bighelli, F. Tomi and J. Casanova, Analyse de I'huile essentielle de Pistachier Lentisque de Corse parRMNdu carbone-13. Riv. Ital. EPPOS, 7, 559-563 (1996). 12. S. Katsiotis and N. G. Oikonomou, Qualitative and quantitative GLC analysis of the essential oil of Pistacia lentiscus (Mastix)from different districts of the Chios Island. Pharm. Delt., Epistem. Ekdosis, 10,17-28 (1984); Chem. Abstr., 103, 42382b (1985). 13. V. P. Papageorgiou, A. S. Mellidis andN. Argyriadou, The chemical composition ofthe essential oil of Mastic gum. J. Essent. Oil Res., 3, 107-110 (1991). 14. A. Fernandez, A. Camacho, C. Fernandez, J. Altarejos and P. Perez, Composition of an essential oil of Pistacia lentiscus galls. Paper No. 7, XLTth International Conference on Aromas and Essential Oils, Seville, Spain (1997). 15- R. P. Adams, Cedarwood oil-Analyses and properties. In: Modern Methods of Plant Analysis. Essential Oils and Waxes. Edits., H. F. Linskens and J. F. Jackson, p p 159-173, Springer-Verlag, Berlin (1991). 16. W. Jennings and T. Shibamoto, Qualitative Analysis of Flavor and Fragrance Volatiles by Glass Capillary Gas Chromatography. Academic Press, New York (1980). 17. R. P. Adams, Identification of Essential Oils by Ion Trap Mass Spectrometry. Academic Press, San Diego (1989). 18. P. Monaco, R. Caputo, G. Palumbo and L. Mangoni, Neutral triterpenes from the galls of Pistacia terebinthus. Phytochemistry, 12, 939-942 (1973). 19. H. Vandendool and P. D. Kratz, A generalization ofthe retention index system including Linear Temperature Programmed Gas-Liquid Partition Chromatography.]. Chromatogr., 1 1 , 463-471 (1963).
