FLAVOUR AND FRAGRANCE JOURNAL Flavour Fragr. J. 2002; 17: 69–71 DOI: 10.1002/ffj.1044
Essential oils from aerial parts of Aristolochia gibertii Hook. ´ Horacio A. Priestap,1 Catalina M. van Baren,1 Paola Di Leo Lira,1 Hector J. Prado,1 Michael Neugebauer,2 2 1Ł Ralf Mayer and Arnaldo L. Bandoni 1
´ Catedra de Farmacognosia, IQUIMEFA (UBA-CONICET), Facultad de Farmacia y Bioqu´ımica, Universidad de Buenos Aires, Jun´ın 956 (1113), Buenos Aires, Argentina 2 Pharmazeutisches Institut der Universitaet Bonn, Kreuzbergweg 26, D-53115 Bonn, Germany
Received February 2001 Revised May 2001 Accepted July 2001
ABSTRACT: The leaf and stem volatile constituents of Aristolochia gibertii Hook., obtained by hydrodistillation, were analysed by GC and GC–MS. Monoterpenes and sesquiterpenes (20–21), amounting to 77.8–80.3% of the total oil, were identified in the leaf oils, bicyclogermacrene being the main component (38.1–42.2%). Components (28–35) were identified in the oils isolated from the stems, constituting 86.6–92.3% of the total volatiles. These oils were dominated by limonene (38.5–56.8%). The leaf and stem oils were also found to contain the sesquiterpene lactone, aristolactone, in varying amounts (1.6–9.9%). The olfactory profiles of these oils were characterized and found to have very interesting organoleptic qualities. Copyright 2001 John Wiley & Sons, Ltd. KEY WORDS: Aristolochia gibertii; Aristolochiaceae; essential oil; bicyclogermacrene; limonene; aristolactone
Introduction Aristolochia gibertii Hook. (Aristolochiaceae) is a woody vine which grows in the forests of south-eastern South America from Rio de Janeiro (Brazil) to Formosa and Corrientes (Argentina). In NE Argentina A. gibertii is popularly known as ‘mil hombres hembra’, ‘contrayerba’ and ‘patito’ and it is used medicinally in the treatment of swelling, stomach pain and other ailments.1 No previous phytochemical study on this species has been reported. In this investigation the composition of the essential oils of A. gibertii is presented.
Experimental
different years during the vegetative period of the plant, from May to July. Leaves from this single cultivated plant were gathered in May and June (1994) and immediately hydrodistilled (samples L-1 and L-2, respectively). Similarly, fresh stems from the same single plant were collected in July 1994, July 1996, June 1997 and July 1999, chopped into small pieces and immediately subjected to hydrodistillation (samples S-1, S-2, S-3 and S-4, respectively). Isolation of Oils
The essential oils were obtained by hydrodistillation from 200 to 1 kg (fresh weight) plant material in a Clevenger-type apparatus for 6–7 h.2
Plant Material
A whole plant of A. gibertii, growing in Paso de la Patria, Corrientes (Argentina) was collected and transplanted in Buenos Aires (Argentina) for this study (voucher herbarium specimen from Centro de Estudios Farmacol´ogicos y Bot´anicos (CEFYBO), No. BACP 3654). Plant material for essential oil analysis was collected in *Correspondence to: A. L. Bandoni, Libertad 1079, 2° piso (C 1012 AAU), Buenos Aires, Argentina. E-mail:
[email protected] Contract/grant sponsor: CYTED; Contract/grant number: IV.6. Contract/grant sponsor: UBACYT; Contract/grant number: TB-05.
Copyright 2001 John Wiley & Sons, Ltd.
Identification of Components
The chromatographic analysis was performed on a Hewlett-Packard 5890 Series II gas chromatograph, equipped with two flame ionization detectors 255 ° C, two fused capillary columns of different polarity: methyl silicone and polyethylene glycol (HP-1 and HP-Wax, 50 m ð 0.25 mm, film thickness 0.25 (µm) were used simultaneously, injector 255 ° C, split ratio 1 : 150. Temperature program: from 90 ° C to 220 ° C at a rate of 3 ° C/min, using N2 as carrier gas at a working flow rate
70 H. A. PRIESTAP ET AL.
of 0.8 ml/min. Quantitative data were obtained from FID area values without considering the respective response factors. Mass spectra were obtained on a Perkin-Elmer Autosystem Gas Chromatograph coupled to a Perkin-Elmer Q-Mass 910 quadrupole selective detector at 70 eV. Analytical conditions: capillary columns with previously cited stationary phases: J&W DB-1, DB-Wax and J&W DB-5 (30 m ð 0.2 mm, film thickness 0.25 µm), helium as carrier gas (1 ml/min); temperature from 75 ° C (4 min) to 220 ° C at a rate of 3 ° C/min; split injector at 250 ° C; ratio 1 : 60. The essential oils components were identified by: (a) determination of their retention indices (RI) in relation to a homologous series of N-alkanes C8 –C20 ,
in three different stationary phases: polyethylene glycol (DB-Wax), methylsilicone (DB-1) and 5% phenyl–95% methylsilicone (DB-5); (b) by comparison of their mass spectra with MS data reported in the literature3,4 and with data stored in a library built up from authentic samples of reference oils.
Results and Discussion The essential oils isolated from the aerial parts of Aristolochia gibertii were obtained in a yield of 0.03% and 0.1% v/w for the leaves and stems, respectively. The analytical results of the isolated oils are shown in Table 1. All components present in a relative amount
Table 1. Percentage composition of the essential oils from leaves and stems of Aristolochia gibertii Compound
1 Tricyclene 2 ˛-Pinene 3 Camphene 4 Sabinene 5 ˇ-Pinene 6 Myrcene 7 -2-Carene 8 -3-Carene 9 p-Cymene 10 Limonene 11 ˇ-Phellandrene 12 1,8-Cineole 13 cis-Ocimene 14 trans-Ocimene 15 UnknownŁ 16 Bomeol 17 Isobomyl formate 18 Carvone 19 Bomyl acetate 20 υ-Elemene 21 ˛-Copaene 22 ˇ-Bourbonene 23 ˇ-Elemene 24 ˇ-Caryophyllene 25 Aromadendrene 26 ˛-Humulene 27 allo-Aromadendrene 28 Germacrene D 29 ˇ-Selinene 30 Bicyclogermacrene 31 -Cadinene 32 ˛-Calacorene 33 ˇ-Calacorene 34 trans-Nerolidol 35 Spathulenol 36 AristolactoneŁŁ Total identified Monoterpene hydrocarbons Oxygenated monoterpenes Sesquiterpene hydrocarbons Oxygenated sesquiterpenes Total sesquiterpenes
Kl1
917 929 942 965 971 989 981 1007 1021 1024 – – 1036 1046 1130 1163 1222 1229 1283 1339 1371 1379 1388 1413 1432 1444 1458 1475 1479 1491 1519 1530 1549 1561 1570 2060
Kl2
929 939 952 975 979 994 996 1009 1023 1039 – – 1051 1058 1136 1165 1238 1241 1290 1341 1374 1381 1391 1423 1440 1454 1460 1470 1481 1496 1526 1542 1561 1568 1574 2068
Kl3
1029 1037 1073 1118 1108 1153 1118 1144 1273 1206 1219 1223 1234 1252 1409 1728 1596 – 1602 – 1509 1535 1609 1616 – 1673 1674 1724 1732 1757 1776 1921 1956 – 2127 2633
Leaves
Stems
L-1
L-2
S-1
S-2
S-3
S-4
– tr – – tr tr – – – 0.4 – – – 0.1 – 0.1 tr – tr 1.0 0.4 0.5 2.5 2.3 5.7 – 0.6 6.0 – 42.2 0.9 – – tr 7.4 7.7 77.8 0.5 0.1 62.1 15.1 77.2
– t – – t – – – – 1.2 – – – 0.2 – 0.1 – – tr 1.1 0.3 0.5 2.7 2.2 4.5 tr tr 5.8 – 38.1 0.8 – – 0.1 12.8 9.9 80.3 1.4 0.1 56.0 22.8 78.8
0.1 3.3 0.7 0.1 0.9 0.1 0.2 0.8 – 53.3 – – – 0.7 0.7 0.4 0.5 tr 0.1 0.4 0.3 0.4 1.7 2.4 0.8 0.2 0.2 4.1 – 7.6 0.9 – – 0.2 3.0 3.2 86.6 60.2 1.0 19.0 6.4 25.4
tr 1.9 0.6 tr 0.4 0.9 tr 1.0 0.1 38.5 – – 0.9 1.3 0.5 0.1 0.6 tr 0.1 0.3 0.8 0.8 2.8 3.7 1.7 tr 0.4 9.5 tr 16.5 1.7 – – 0.2 3.1 1.6 89.5 45.6 0.8 38.2 4.9 43.1
0.1 2.9 0.7 tr 0.7 1.3 0.2 0.8 – 56.8 – – – 0.8 0.7 0.1 0.9 – tr 0.2 0.2 0.3 1.8 3.0 0.7 0.2 0.2 4.0 – 10.7 0.8 – – 0.2 2.2 2.2 92.0 64.3 1.0 22.1 4.6 26.7
tr 2.9 0.8 tr 0.8 1.2 tr 0.7 t 54.3 tr tr tr 0.5 0.7 0.3 0.7 tr tr tr 0.3 0.4 1.5 2.7 0.4 0.2 0.3 5.1 tr 9.8 0.8 tr tr 0.3 2.5 5.8 92.3 61.2 1.0 21.5 8.6 30.3
Kov´ats indexes (KI) on: (1) DB-1; (2) DB-5 and (3) DB-WAX columns. Ł Unidentified (compound 15): m/z (rel. int.) 110 (17) [MC ], 96 (4), 95 (43), 93 (3), 81 (5), 69 (7), 68 (5), 67 (10), 55 (9), 53 (3), 45 (4), 43 (100), 42 (5). ŁŁ Aristolactone: m/z (rel. int.) 189 (20), 121 (32), 109 (27), 107 (55), 105 (28), 95 (100), 93 (31), 91 (20), 81 (31), 71 (28), 55 (39), 45 (39), 43 (66).
Copyright 2001 John Wiley & Sons, Ltd.
Flavour Fragr. J. 2002; 17: 69–71
ESSENTIAL OILS OF ARISTOLOCHIA GIBERTII
>1% have been identified. They were all mono- and sesquiterpenes, attaining 77.8–92.3% of the total oil. The leaf essential oils (L-1 and L-2) exhibit a similar composition pattern. Both show a very low content of monoterpenes (0.5% and 1.4%, respectively), the oils being mainly represented by sesquiterpenes (77.2% and 78.8%, respectively). Bicyclogermacrene is the main component (42.2% in L-1 and 38.1% in L-2 oils) in both oils, but other compounds could also be found in relative amounts higher than 5%: aromadendrene, germacrene D, spathulenol and aristolactone. Bicyclogermacrene was also found to be one of the main components in the leaf oils of A. elegans;5 however, this compound does not seem to be widely distributed in the Aristolochiaceae.6 The essential oils isolated from the stems also show a similar composition pattern (Table 1) but, in contrast to the leaf oils, they are characterized by a high limonene content (38.5–56.8%). Higher relative amounts of total monoterpenes are also observed (45.6–64.3%). The higher oil yields of the stems (ca. 1 ml/kg fresh weight) vs. those of the leaves (ca. 0.3 ml/kg fresh weight) may be assigned to the prevalent accumulation of limonene and other monoterpenes in the stems. This assertion can be confirmed by calculating the absolute value of limonene in stems [1 ml/kg ð 58.8% (average) D 0.6 ml/kg] in comparison with those in leaves [0.3 ml/kg ð 1% (average) D 0.003 ml/kg], while the total amount of sesquiterpenes in stems 1 ml/kg ð 31% D 0.3 ml/kg and leaves 0.3 ml/kg ð 78% D 0.23 ml/kg is nearly the same. This behaviour seems to be reflected in A. triangularis.7 A comparative study of the essential oils of the stems and leaves of A. gigantea 6 also showed a significant increase in the percentage of monoterpenes in the stem oil, although in this case such increase is probably due to a higher content of linalool and ˛-terpineol, rather than limonene. The slight quantitative differences found between the leaf oils and the stem oils can be partially explained by the diverse climatic conditions prevailing from year to year. All the samples analysed contained a significant amount of aristolactone, a sesquiterpene lactone first isolated from A. serpentaria and A. reticulata.8 – 11 Sesquiterpene lactones mostly occur in plants belonging to
Copyright 2001 John Wiley & Sons, Ltd.
71
the Compositae,11 whereas their distribution in the Aristolochiaceae seems to be rather restricted.11,12 The olfactory profiles of these oils were characterized by a very strong, sweet and woody top note, slowly balsamic, powdery and worm core, and reminiscent of clary sage, davana and tobacco odours in the lasting undertones. It has a relatively short tenacity (not more than 1 day). However, the general performance of the oils appears to be very interesting for the fragrance industry, in view of its original olfactory profile. Acknowledgements—The authors are indebted to Dr Gordon L. Lange, University of Guelph, Ontario, Canada, for a sample of aristolactone. Part of this study was supported by CYTED (Project IV.6) and UBACYT TB-05.
References 1. Mart´ınez-Crovetto R. Las Plantas Utilizadas en Medicina Popular en el Noroeste de Corrientes (Rep´ublica Argentina), Miscel´anea No. 69. Fundaci´on Miguel Lillo: Tucum´an, Rep´ublica Argentina, 1981. 2. European Pharmacopoeia, 3rd edn. Maisonneuve SA: Sainte Ruffine, 1997. 3. Davies NW. Gas chromatographic retention indices of monoterpenes and sesquiterpenes on methyl silicone and Carbowax 20M phases. J. Chromatogr. 1990; 503: 1–24. 4. Adams RP. Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry. Allured: Carol Stream, IL, 1995. 5. Vila R, Mundina M, Muschietti L, Priestap HA, Bandoni AL, Adzet T, Ca˜nigueral S. Volatile constituents of leaves, roots and stems from Aristolochia elegans. Phytochemistry 1997; 46(6): 1127–1129. 6. Leitao GG, Lopes D, de Sousa Menezes F, Kaplan MAC, Craveiro AA, Alencar JW. Essential oils from Brazilian Aristolochia. J. Essent. Oil Res. 1991; 3: 403–408. 7. Priestap HA, Bandoni AL, Neugebauer M, R¨ucker G. Investigation of the essential oils from Aristolochia triangularis. J. Essent. Oil Res. 1990; 2: 95–98. 8. Stenlake JB, Williams WD. The chemistry of Aristolochia spp. J. Pharm. Pharmacol. 1954; 6: 1005–1013. 9. Steele JW, Stenlake JB, Williams WD. The chemistry of Aristolochia species: Part IV, The structure of Aristolactone. J. Chem. Soc. 1959; 3289–3299. 10. Martin-Smith M, de Mayo P, Smith SJ, Stenlake JB, Williams WD. Revised structure of aristolactone. Tetrahedron Lett. 1964; 2391. 11. Yoshioka H, Mabry TJ, Timmerman BN. Sesquiterpene Lactones. University of Tokyo Press: Tokyo, 1973. 12. Hegnauer RD. Chemotaxonomy der Pfflanzen, vol 8. Birkhauser Verlag: Basel, 1989.
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