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Comparison between the essential oils of Santolina insularis (Genn. ex Fiori) Arrigoni and Santolina corsica Jord. et Fourr. from the island of Sardinia (Italy)
Journal of Ethnopharmacology 56 (1997) 201 – 208
Comparison between the essential oils of Santolina insularis (Genn. ex Fiori) Arrigoni and Santolina corsica Jord. et Fourr. from the island of Sardinia (Italy) Ferruccio Poli a,*, Leonardo Bonsignore b, Giuseppe Loy b, Gianni Sacchetti c, Mauro Ballero a a Istituto di Botanica ed Orto Botanico, Uni6ersita` di Cagliari, Via Fra Ignazio 13, I-09124 Cagliari, Italy Dipartimento Farmaco Chimico Tecnologico, Uni6ersita` di Cagliari, Via Ospedale 72, I-09124 Cagliari, Italy c Dipartimento di Biologia, Sezione di Botanica, Uni6ersita` di Ferrara, C. so Porta Mare 2, I-44100 Ferrara, Italy b
Received 13 May 1995; received in revised form 26 July 1996; accepted 20 February 1997
Abstract A comparative study of the essential oils of Santolina insularis and Santolina corsica has been carried out. The two specimens are found in Sardinia and were indiscriminately used in traditional medicine on the island. The essential oil was extracted by steam distillation of fresh aerial parts and analysed by GC, GC-MS and GC-FTIR. The analysis of the essential oils shows substantial qualitative and quantitative differences. Some of the components identified were present in both investigated species, others were characteristic of one species only. © 1997 Elsevier Science Ireland Ltd. Keywords: Essential oils; Santolina insularis; Santolina corsica; Sardinia
1. Introduction There are several shrubs widely growing in the Mediterranean region which belong to the genus Santolina (Asteraceae). Fiori (1927) described two varieties of the Sardinian species of this genus, Santolina chamaecyparissus L., S. chamaecyparissus L. var. insularis Genn. ex Fiori and S. * Corresponding author.
chamaecyparissus L. var. corsica Jord. et Fourr. Subsequently, Marchi and D’Amato (1973) and Marchi et al. (1979) in cytotaxonomic investigations pointed out caryological differences between the two Sardinian Santolina. S. chamaecyparissus var. insularis showed a hexaploid caryotype (2n= 54), while the S. corsica caryotype was tetraploid (2n= 36). Successively, with reference to morphological and caryological evidence, Arrigoni (1979) suggested that Santolina plants might be ascribed
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F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
Fig. 1. Simplified map of Sardinia and places of collection.
to seven different species in Italy. In particular, S. insularis (Genn. ex Fiori) Arrigoni and S. corsica Jord. et Fourr. are the two Santolina species reported on the island of Sardinia (Arrigoni, 1979). Recently, in ‘Flora d’Italia’, Pignatti (1982) confirmed the two different Santolina species in Sardinia and described their different distribution areas: S. corsica grows on Mt. Albo at Lula (North-West Sardinia) and in Corsica, whereas S. insularis is distributed on the Gennargentu (Central Sardinia) and Marganai-Linas massifs (South-West Sardinia). However, in ‘Flora Europaea’, Guinea and Tutin (1976) reported Santolina chamaecyparissus L. subsp. insularis Genn.
ex Fiori as the only ‘cotton lavender’ growing in the Central Mediterranean region. Recent ethnopharmacobotanical investigations on plants used in Sardinian folk medicine, pointed out the particular use of Santolina species as an intestinal vermifuge against horse strongyloidiasis and as a parasite repellent (Camarda, 1990; Ballero and Fresu, 1991; Ballero et al., 1994). Phytochemical studies on other South Mediterranean plants belonging to this genus, known for the pharmacological activity of their extracts, showed the presence of important bioactive compounds such as coumarins (Maqua et al., 1988), several acetylenic compounds (Christensen, 1992)
F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
essential oils (Baig et al., 1989; De Pascual et al., 1983; Villar et al., 1986) and flavonoids (Becchi and Carrier, 1980). The aim of this work is to evaluate possible phytochemical differences between the two Sardinian Santolina species.
2. Materials and methods
2.1. Plant material From at least five different plants of Santolina insularis and of Santolina corsica, the aerial parts were collected during full blossom (May) on the Marganai massif (Iglesias, Cagliari) and on the mountains at Lula (Nuoro) respectively (Fig. 1). The plants were identified following the keys reported in ‘Flora d’Italia’ (Pignatti, 1982) by M. Ballero, and a voucher specimen of each species was deposited in the Herbarium of the Institute of Botany and Botanical Garden, University of Cagliari (Santolina insularis: Cag no. 732; Santolina corsica: Cag no. 732/A).
2.2. Isolation of essential oils The fresh aerial parts of the plants (1500 g for each species) were distilled in a Clevenger-type apparatus for 5 h. The isolated oil was dried over anhydrous sodium sulphate and stored at 4°C.
2.3. Qualitati6e analysis Qualitative data were determined by gas chromatography (GC), gas chromatography mass spectrometry (GC-MS) and gas chromatography Fourier-Transform Infrared spectroscopy (GCTable 1 Chemical and physical characteristics of essential oils from S. insularis and S. corsica
Color Yield Boiling-point [n]20 D [a]D
Santolina insularis
Santolina corsica
Pale yellow 1.6% 95–106°C 1.4820 +5°6%
Yellow-green 0.92% 110–112°C 1.5122 −6°4%
203
FTIR). GC analysis was performed using a Carlo Erba gas chromatograph (model 5300) equipped with a flame ionisation detector and a spectra physical computing integrator. A fused silica capillary column Supelcowax 10 (30 m × 0.25 mm internal diameter, film thickness 0.25 mm) was employed. The temperature programme was 50°C for 10 min, then 250°C with a rise of 7°C/min. The carrier gas was nitrogen at a working flow rate of 2 ml/min, 100:1 split ratio and 1 ml sample. GC-MS analyses were performed on a Fisons mass spectrometer (model QMD 1000) combined with a Carlo Erba gas chromatograph (model 5300) using a fused silica capillary Carbowax (30 m× 0.25 mm internal diameter, film thickness 0.3 mm). Mass spectra were recorded at 70 eV. The carrier gas was helium at a working flow rate of 5 ml/min. GC-FTIR were performed employing an FTIR Perkin Elmer spectrophotometer (model System 2000) with an FT GC interface, connected with a Perkin Elmer chromatograph (model 8600). The working conditions were as reported for GC. The parameters of FTIR were the following: run time, 40 min; scans per slice, 2; GramSchmidt interferogram points, 25–75; peak threshold, 50.0; heated zone temperature, 200°C. A Varian FT unit 300 with CDCl3 as solvent was used for [1H]NMR.
2.4. Quantitati6e analysis GC quantitative analyses were based on computer calculated peak area normalisations without correction factors (Lorusso, 1983). Peaks obtained were compared with those of pure standards supplied by Extrasynthese (Merignac, France). GCFTIR was performed comparing the absorption of each peak of the Gram-Schmidt interferogram with those of pure samples inserted in the computer integrator as suggested by Herres (1987).
3. Results and discussion The two Santolina species investigated gave essential oils with significatively different characteristics (Table 1). S. insularis showed a yield
Fig. 2. Gas chromatogram of essential oil from the aerial parts of S. insularis plants.
204 F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
Fig. 3. Gas chromatogram of essential oil from the aerial parts of S. corsica plants.
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Table 2 Percentage of the components identified in the essential oils of S. insularis and S. corsica
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43
Compounds
S. insularis (%)
S. corsica (%)
a-Pinene Unknown Unknown Camphene Sabinene Limonene Cineole D-3-carene a-Terpinene 3,3,6-Trimethyl-1,5-heptadien-4-one Unknown Unknown g-Terpinene Bornyl acetate Phellandrene Camphor Unknown Unknown Unknown Terpinen-4-ol a-Terpineol Borneol Isobornyl acetate Geraniol a-Cubebene a-Copaene Unknown Unknown Unknown Unknown Guaiazulene Aromadendrene Carvacrol Thymol Unknown Caryophyllene Unknown 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene Unknown Muurolene Germacrene Unknown Cadinene
2.11 — — 8.47 1.27 1.58 9.01 0.50 0.42 21.18 — — 0.70 6.35 0.15 1.68 — — — 1.69 0.64 4.23
0.85 — — 1.29 0.83 0.92 1.85 — — 12.97 — — 0.62 0.70 — 18.53 — — — 0.45 0.75 7.41
0.31 0.20 0.18 — — — — 1.48 0.76 0.54 0.28 — 0.62 — 12.7 — 0.95 0.84 — 1.06
— 0.44 0.48 — — — — 0.93 5.55 0.61 — — 0.42 — 1.11 — 4.63 0.52 — 0.67
percentage of 1.6%, while S. corsica presented a yield of 0.92%. Furthermore the optical rotation of the two essential oils was found to be different; that of S. corsica was negative, as with other Santolina species, whereas that of S. insularis was positive.
The GC analysis of the essential oils showed substantial qualitative and quantitative differences; these principally concerned a-pinene (1), camphene (4), cineole (7), 3,3,6-trimethyl-1,5-heptadien-4-one (10), bornyl acetate (14), camphor (16), borneol (22), aromadendrene (32), 10-H-
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Table 3 Analytical and spectroscopical data of 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene Elemental Analysis for C15H24
FTIR (cm−1)a
[1H]NMR CDCl3 (d)b
[13C]NMR CDCl3 (ppm)b
GC-Mass (m/z)c
Calculated C% = 86.16 H% =11.84 Found C%87.98 H% =11.87
3050 2975 1530 1470 1380 1215 1195 890
0.70 =m,2H,2CH 0.77 =s,3H,1CH3 0.89 =d,3H,1CH3 1.03= s,3H,1CH3 1.90 = m,11H,CH2 4.55 =m,2H,CH2
166.30 98.95 48.12 47.95 45.08 44.20 43.38 36.11 33.50 30.43 30.00 29.76 25.70 21.08
204 189 175 161 147 133 119 105 91 79 69 55 41
B.p.=126 – 127°C, 10/mm; [n]20 D = 1.4998; l= 244mm. Vapour phase; b internal standard HMSDO; c instrument operating under EI (Electronic Impact) conditions (70 eV, 200 mA) and at ion source temperature of 200°C. a
cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene (38) and muurolene (40) (Figs. 2 and 3). GC-FTIR analysis allowed us to detect peaks not usually revealed with GC-MS. Table 2 shows the identified compounds and the percentage of each substance occurring in the two essential oils. In particular, a-pinene (1), camphene (4), cineole (7), 3,3,6-trimethyl-1,5-heptadien-4-one (10), bornyl acetate (14) and 10-H-cyclopropyl-1,1,7trimethyl-4-methylen-decahydro azulene (38) were more abundant in S. insularis. On the contrary camphor (16), borneol (22), aromadendrene (32) and muurolene (40) showed high values in S. corsica. Furthermore, D-3-carene (8), a-terpinene (9), phellandrene (15), geraniol (24) and thymol (34) were detected in the essential oil of S. insularis and not in that of S. corsica. Compound 38 was identified as 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene by [1H]NMR and FTIR. The analytical and spectroscopical data of this compound are shown in Table 3. This compound was found in both Santolina plants analysed but it was never identified in other plants of the same genus. In conclusion, the percentage yield of the essen-
tial oil of S. insularis was higher than that of S. corsica. Some identified compounds that had also been reported by other authors in studies on other Santolina species (Baig et al., 1989; De Pascual et al., 1983; Villar et al., 1986), were present in both essential oils examined though in different amounts. Furthermore, the detection and first isolation of 10-H-cyclopropyl-1,1,7-trimethyl-4methylen-decahydro azulene in the two examined species as well as the fact that the compound had never been isolated before in any other Santolina species, suggest the compound as a possible chemotaxonomical marker for Santolina species. Subject to the limitations of single bulk sample analyses as described here, the phytochemical data and the suggestions about qualitative and quantitative differences that emerged from this analysis support the assumption by Arrigoni (1979) who considered S. corsica and S. insularis as two new species on the basis of morphological and caryological observations. Finally, with the aim of carrying out a thorough study of Santolina plants in Sardinia, an investigation of the antibiotic and antiparasitic properties of the two different essential oils is currently in progress.
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Acknowledgements Investigations supported by grants from CNR (Consiglio Nazionale delle Ricerche), MURST (Ministero dell’Universita` e della Ricerca Scientifica e Tecnologica) of Italy and INTERREG.
References Arrigoni, P.V. (1979) Le genre Santoline en Italie. Webbia 34, 257 – 264. Baig, M.A., Banthorpe, D.V. and Gutowski, J.A. (1989) Accumulation in cultivars of S. chamaecyparissus of a rare sesquiterpene with gasterop-repellent activity. Fitoterapia 60, 373 – 375. Ballero, M. and Fresu, I. (1991) Piante officinali impiegate in fitoterapia nel territorio del Marganai (Sardegna sud-occidentale). Fitoterapia 62, 524–531. Ballero, M., Bruni, A., Sacchetti, G. and Poli, F. (1994) Indagine etnofarmacobotanica del territorio di Arzana (Sardegna orientale). Annali di Botanica 52 (2), 489– 500. Becchi, M. and Carrier, M. (1980) 6-methoxy flavones of Santolina chamaecyparissus. Planta Medica 38, 267–268. Camarda, I. (1990) Ricerche etnobotaniche nel comune di Dorgali, Sardegna centro orientale. Bollettino della Societa` Sarda di Scienze Naturali 27, 147–204. Christensen, L.P. (1992) Acetylenes and related compounds in
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Anthemideae. Phytochemistry 31, 7 – 49. De Pascual, J.T., Vicente, S., Gonzales, M.S. and Bellido, I.S. (1983) Nerolidol-5,8-oxides from the essential oil Santolina oblongifolia. Phytochemistry 22, 2235 – 2238. Fiori, A. (1927) Nuo6a Flora Analitica d’Italia. Vol. 2, M. Ricci, Firenze, p. 652. Guinea, E. and Tutin, T. (1976) Santolina. In: T.G. Tutin, V.H. Heywood, N.A. Burges, D.M. Moore, D.H. Valentine, S.M. Walters and D.A. Webb (Eds.), Flora Europaea Vol. IV, Cambridge University Press, Cambridge. Herres, W. (1987) HRGC-FTIR Capillary Gas Chromatography Fourier-transform Infrared Spectrometry. Theory and Applications. A. Hutig, Verlag, Heidelberg. Lorusso, S. (1983) Spettrometria di massa nell’analisi degli alimenti. Chiriotti Editore, p. 22. Maqua, M.P., Vines, A.C.G., Caballero, E., Grande, M.C., Medarde, M. and Bellido, I.S. (1988) Components from Santolina rosmarinifolia, subspecies rosmarinifolia and canescens. Phytochemistry 27, 3664 – 3667. Marchi, P. and D’Amato, G. (1973) Numeri cromosomici per le flore italiane. Informatore Botanico Italiano 5, 93 – 100. Marchi, P., Capineri, R. and D’Amato, G. (1979) Il cariotipo di Santolina corsica Jordan et Fourreau (Compositae) provenienti dai pressi di Bastia (Corsica) ed altre osservazioni. Annali di Botanica 38, 1 – 13. Pignatti, S. (1982) Flora d’Italia. Vol. 3, Edagricole, Bologna, p. 65. Villar, A., Giner, R.M. and Rios, J.L. (1986) Chemical composition of Santolina chamaecyparissus ssp. squarrosa essential oil. Journal of Natural Products 49, 1143 – 1144.
Comparison between the essential oils of Santolina insularis (Genn. ex Fiori) Arrigoni and Santolina corsica Jord. et Fourr. from the island of Sardinia (Italy) Ferruccio Poli a,*, Leonardo Bonsignore b, Giuseppe Loy b, Gianni Sacchetti c, Mauro Ballero a a Istituto di Botanica ed Orto Botanico, Uni6ersita` di Cagliari, Via Fra Ignazio 13, I-09124 Cagliari, Italy Dipartimento Farmaco Chimico Tecnologico, Uni6ersita` di Cagliari, Via Ospedale 72, I-09124 Cagliari, Italy c Dipartimento di Biologia, Sezione di Botanica, Uni6ersita` di Ferrara, C. so Porta Mare 2, I-44100 Ferrara, Italy b
Received 13 May 1995; received in revised form 26 July 1996; accepted 20 February 1997
Abstract A comparative study of the essential oils of Santolina insularis and Santolina corsica has been carried out. The two specimens are found in Sardinia and were indiscriminately used in traditional medicine on the island. The essential oil was extracted by steam distillation of fresh aerial parts and analysed by GC, GC-MS and GC-FTIR. The analysis of the essential oils shows substantial qualitative and quantitative differences. Some of the components identified were present in both investigated species, others were characteristic of one species only. © 1997 Elsevier Science Ireland Ltd. Keywords: Essential oils; Santolina insularis; Santolina corsica; Sardinia
1. Introduction There are several shrubs widely growing in the Mediterranean region which belong to the genus Santolina (Asteraceae). Fiori (1927) described two varieties of the Sardinian species of this genus, Santolina chamaecyparissus L., S. chamaecyparissus L. var. insularis Genn. ex Fiori and S. * Corresponding author.
chamaecyparissus L. var. corsica Jord. et Fourr. Subsequently, Marchi and D’Amato (1973) and Marchi et al. (1979) in cytotaxonomic investigations pointed out caryological differences between the two Sardinian Santolina. S. chamaecyparissus var. insularis showed a hexaploid caryotype (2n= 54), while the S. corsica caryotype was tetraploid (2n= 36). Successively, with reference to morphological and caryological evidence, Arrigoni (1979) suggested that Santolina plants might be ascribed
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F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
Fig. 1. Simplified map of Sardinia and places of collection.
to seven different species in Italy. In particular, S. insularis (Genn. ex Fiori) Arrigoni and S. corsica Jord. et Fourr. are the two Santolina species reported on the island of Sardinia (Arrigoni, 1979). Recently, in ‘Flora d’Italia’, Pignatti (1982) confirmed the two different Santolina species in Sardinia and described their different distribution areas: S. corsica grows on Mt. Albo at Lula (North-West Sardinia) and in Corsica, whereas S. insularis is distributed on the Gennargentu (Central Sardinia) and Marganai-Linas massifs (South-West Sardinia). However, in ‘Flora Europaea’, Guinea and Tutin (1976) reported Santolina chamaecyparissus L. subsp. insularis Genn.
ex Fiori as the only ‘cotton lavender’ growing in the Central Mediterranean region. Recent ethnopharmacobotanical investigations on plants used in Sardinian folk medicine, pointed out the particular use of Santolina species as an intestinal vermifuge against horse strongyloidiasis and as a parasite repellent (Camarda, 1990; Ballero and Fresu, 1991; Ballero et al., 1994). Phytochemical studies on other South Mediterranean plants belonging to this genus, known for the pharmacological activity of their extracts, showed the presence of important bioactive compounds such as coumarins (Maqua et al., 1988), several acetylenic compounds (Christensen, 1992)
F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
essential oils (Baig et al., 1989; De Pascual et al., 1983; Villar et al., 1986) and flavonoids (Becchi and Carrier, 1980). The aim of this work is to evaluate possible phytochemical differences between the two Sardinian Santolina species.
2. Materials and methods
2.1. Plant material From at least five different plants of Santolina insularis and of Santolina corsica, the aerial parts were collected during full blossom (May) on the Marganai massif (Iglesias, Cagliari) and on the mountains at Lula (Nuoro) respectively (Fig. 1). The plants were identified following the keys reported in ‘Flora d’Italia’ (Pignatti, 1982) by M. Ballero, and a voucher specimen of each species was deposited in the Herbarium of the Institute of Botany and Botanical Garden, University of Cagliari (Santolina insularis: Cag no. 732; Santolina corsica: Cag no. 732/A).
2.2. Isolation of essential oils The fresh aerial parts of the plants (1500 g for each species) were distilled in a Clevenger-type apparatus for 5 h. The isolated oil was dried over anhydrous sodium sulphate and stored at 4°C.
2.3. Qualitati6e analysis Qualitative data were determined by gas chromatography (GC), gas chromatography mass spectrometry (GC-MS) and gas chromatography Fourier-Transform Infrared spectroscopy (GCTable 1 Chemical and physical characteristics of essential oils from S. insularis and S. corsica
Color Yield Boiling-point [n]20 D [a]D
Santolina insularis
Santolina corsica
Pale yellow 1.6% 95–106°C 1.4820 +5°6%
Yellow-green 0.92% 110–112°C 1.5122 −6°4%
203
FTIR). GC analysis was performed using a Carlo Erba gas chromatograph (model 5300) equipped with a flame ionisation detector and a spectra physical computing integrator. A fused silica capillary column Supelcowax 10 (30 m × 0.25 mm internal diameter, film thickness 0.25 mm) was employed. The temperature programme was 50°C for 10 min, then 250°C with a rise of 7°C/min. The carrier gas was nitrogen at a working flow rate of 2 ml/min, 100:1 split ratio and 1 ml sample. GC-MS analyses were performed on a Fisons mass spectrometer (model QMD 1000) combined with a Carlo Erba gas chromatograph (model 5300) using a fused silica capillary Carbowax (30 m× 0.25 mm internal diameter, film thickness 0.3 mm). Mass spectra were recorded at 70 eV. The carrier gas was helium at a working flow rate of 5 ml/min. GC-FTIR were performed employing an FTIR Perkin Elmer spectrophotometer (model System 2000) with an FT GC interface, connected with a Perkin Elmer chromatograph (model 8600). The working conditions were as reported for GC. The parameters of FTIR were the following: run time, 40 min; scans per slice, 2; GramSchmidt interferogram points, 25–75; peak threshold, 50.0; heated zone temperature, 200°C. A Varian FT unit 300 with CDCl3 as solvent was used for [1H]NMR.
2.4. Quantitati6e analysis GC quantitative analyses were based on computer calculated peak area normalisations without correction factors (Lorusso, 1983). Peaks obtained were compared with those of pure standards supplied by Extrasynthese (Merignac, France). GCFTIR was performed comparing the absorption of each peak of the Gram-Schmidt interferogram with those of pure samples inserted in the computer integrator as suggested by Herres (1987).
3. Results and discussion The two Santolina species investigated gave essential oils with significatively different characteristics (Table 1). S. insularis showed a yield
Fig. 2. Gas chromatogram of essential oil from the aerial parts of S. insularis plants.
204 F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
Fig. 3. Gas chromatogram of essential oil from the aerial parts of S. corsica plants.
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Table 2 Percentage of the components identified in the essential oils of S. insularis and S. corsica
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43
Compounds
S. insularis (%)
S. corsica (%)
a-Pinene Unknown Unknown Camphene Sabinene Limonene Cineole D-3-carene a-Terpinene 3,3,6-Trimethyl-1,5-heptadien-4-one Unknown Unknown g-Terpinene Bornyl acetate Phellandrene Camphor Unknown Unknown Unknown Terpinen-4-ol a-Terpineol Borneol Isobornyl acetate Geraniol a-Cubebene a-Copaene Unknown Unknown Unknown Unknown Guaiazulene Aromadendrene Carvacrol Thymol Unknown Caryophyllene Unknown 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene Unknown Muurolene Germacrene Unknown Cadinene
2.11 — — 8.47 1.27 1.58 9.01 0.50 0.42 21.18 — — 0.70 6.35 0.15 1.68 — — — 1.69 0.64 4.23
0.85 — — 1.29 0.83 0.92 1.85 — — 12.97 — — 0.62 0.70 — 18.53 — — — 0.45 0.75 7.41
0.31 0.20 0.18 — — — — 1.48 0.76 0.54 0.28 — 0.62 — 12.7 — 0.95 0.84 — 1.06
— 0.44 0.48 — — — — 0.93 5.55 0.61 — — 0.42 — 1.11 — 4.63 0.52 — 0.67
percentage of 1.6%, while S. corsica presented a yield of 0.92%. Furthermore the optical rotation of the two essential oils was found to be different; that of S. corsica was negative, as with other Santolina species, whereas that of S. insularis was positive.
The GC analysis of the essential oils showed substantial qualitative and quantitative differences; these principally concerned a-pinene (1), camphene (4), cineole (7), 3,3,6-trimethyl-1,5-heptadien-4-one (10), bornyl acetate (14), camphor (16), borneol (22), aromadendrene (32), 10-H-
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Table 3 Analytical and spectroscopical data of 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene Elemental Analysis for C15H24
FTIR (cm−1)a
[1H]NMR CDCl3 (d)b
[13C]NMR CDCl3 (ppm)b
GC-Mass (m/z)c
Calculated C% = 86.16 H% =11.84 Found C%87.98 H% =11.87
3050 2975 1530 1470 1380 1215 1195 890
0.70 =m,2H,2CH 0.77 =s,3H,1CH3 0.89 =d,3H,1CH3 1.03= s,3H,1CH3 1.90 = m,11H,CH2 4.55 =m,2H,CH2
166.30 98.95 48.12 47.95 45.08 44.20 43.38 36.11 33.50 30.43 30.00 29.76 25.70 21.08
204 189 175 161 147 133 119 105 91 79 69 55 41
B.p.=126 – 127°C, 10/mm; [n]20 D = 1.4998; l= 244mm. Vapour phase; b internal standard HMSDO; c instrument operating under EI (Electronic Impact) conditions (70 eV, 200 mA) and at ion source temperature of 200°C. a
cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene (38) and muurolene (40) (Figs. 2 and 3). GC-FTIR analysis allowed us to detect peaks not usually revealed with GC-MS. Table 2 shows the identified compounds and the percentage of each substance occurring in the two essential oils. In particular, a-pinene (1), camphene (4), cineole (7), 3,3,6-trimethyl-1,5-heptadien-4-one (10), bornyl acetate (14) and 10-H-cyclopropyl-1,1,7trimethyl-4-methylen-decahydro azulene (38) were more abundant in S. insularis. On the contrary camphor (16), borneol (22), aromadendrene (32) and muurolene (40) showed high values in S. corsica. Furthermore, D-3-carene (8), a-terpinene (9), phellandrene (15), geraniol (24) and thymol (34) were detected in the essential oil of S. insularis and not in that of S. corsica. Compound 38 was identified as 10-H-cyclopropyl-1,1,7-trimethyl-4-methylen-decahydro azulene by [1H]NMR and FTIR. The analytical and spectroscopical data of this compound are shown in Table 3. This compound was found in both Santolina plants analysed but it was never identified in other plants of the same genus. In conclusion, the percentage yield of the essen-
tial oil of S. insularis was higher than that of S. corsica. Some identified compounds that had also been reported by other authors in studies on other Santolina species (Baig et al., 1989; De Pascual et al., 1983; Villar et al., 1986), were present in both essential oils examined though in different amounts. Furthermore, the detection and first isolation of 10-H-cyclopropyl-1,1,7-trimethyl-4methylen-decahydro azulene in the two examined species as well as the fact that the compound had never been isolated before in any other Santolina species, suggest the compound as a possible chemotaxonomical marker for Santolina species. Subject to the limitations of single bulk sample analyses as described here, the phytochemical data and the suggestions about qualitative and quantitative differences that emerged from this analysis support the assumption by Arrigoni (1979) who considered S. corsica and S. insularis as two new species on the basis of morphological and caryological observations. Finally, with the aim of carrying out a thorough study of Santolina plants in Sardinia, an investigation of the antibiotic and antiparasitic properties of the two different essential oils is currently in progress.
F. Poli et al. / Journal of Ethnopharmacology 56 (1997) 201–208
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Acknowledgements Investigations supported by grants from CNR (Consiglio Nazionale delle Ricerche), MURST (Ministero dell’Universita` e della Ricerca Scientifica e Tecnologica) of Italy and INTERREG.
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