- Research article
- Open Access
Terpenes From the Root of Salvia hypoleuca Benth
© Saeidnia et al.; licensee BioMed Central Ltd. 2012
- Received: 17 June 2012
- Accepted: 20 October 2012
- Published: 24 October 2012
The genus Salvia, with nearly 900 species, is one of the largest members of Lamiaceae family. In the Flora of Iran, the genus Salvia is represented by 58 species of which 17 species are endemic. Salvia hypoleuca Benth., is one of these species growing wildly in northern and central parts of Iran. Salvia species are well known in folk medicine and widely used for therapeutic purposes. Literature review shows that there is no report on phytochemical investigation of the roots of S. hypoleuca.
The separation and purification process were carried out using various chromatographic methods. Structural elucidation was on the basis of NMR and MS data, in comparison with those reported in the literature. The isolated compounds were identified as sitosteryl oleate (1), β-sitosterol (2), stigmasterol (3), manool (4), 7α-acetoxy royleanone (5), ursolic acid (6), oleanolic acid (7), 3-epicorosolic acid (8), 3-epimaslinic acid (9) and coleonolic acid (10).
In the present study, three sterols, two diterpenes and five triterpenes were isolated from the ethyl acetate extract of the roots of S. hypoleuca. As the chemotaxonomic significance, some of the isolated compounds (1–7, 9) have not been previously reported from the species S. hypoleuca, while the triterpenes 8 and 10 are now documented from Salvia genus for the first time.
- Salvia hypoleuca
- Coleonolic acid
- 3-epimaslinic acid
- 3-epicorosolic acid
The genus Salvia L. (Lamiaceae), with more than 900 species throughout the world, is represented 58 species in Iran, 17 of which are endemic. Most of the species are used as herbal tea and flavoring agent by people and also used in traditional medicine as tonic, antirheumatoid, antimicrobial and carminative [1–3]. Flora Orientalis includes as many as 107 species of Salvia. Salvia hypoleuca Benth., is one of these species which growing wildly in northern and central parts of Iran .
Literature review show that various secondary metabolites such as terpenoids, phenolic acids , polyphenols, flavonoids [3, 6] and anthocyanins  have been reported from Salvia species. Limonene, α-pinene, β-pinene, 1,8-cineol, bicyclogermacrene, caryophyllene oxide and α-gurjunene are the main components of the essential oils of various species of Salvia growing wildly in Iran [8–11]. In the literature, there are several reports on phytochemical investigation of the above mentioned species.
Several sesterterpene lactones, isomeric epoxides, monolactone and hypoleuenoic acid have been reported from varies fractions of S. hypoleuca[12–14]. The main aromatic components of the essential oil of S. hypoleuca roots have been identified as hexadecanoic acid (27.4%) and viridiflorol (14.9%) , while germacrene D (15.1%) and β-caryophyllene (22.0%) identified as the major constituents during flowering stages . A great number of diterpenes exhibited interesting biological activities e.g. anti-tuberculous, antitumour, antibacterial, antileishmanial and antispasmolytic, and Salvia species are the excellent source of diterpenoids . In this study, we aim to report the isolation and identification of some sterols, diterpenoids and triterpenoids from the root extract of S. hypoleuca which have not been previously reported from this species.
Instruments and materials
1H-NMR and 13C-NMR spectra were recorded on a Brucker Avance 500 DRX spectrometer ® with tetramethylsilane as an internal standard and chemical shifts are given in δ (ppm). Multiple-pulse experiments (HSQC, HMBC and H-H COSY) were performed using the standard Bruker ® programs. Silicagel 60 F254 and Silicagel 60 RP-18 F254S pre-coated plates (Merck ®) were used for TLC. The spots were detected by spraying with anisaldehyde-H2SO4 reagent followed by heating.
The roots of Salvia hypoleuca Benth., were collected from Tehran province (near to Damavand city), Iran, at flowering stage in August 2008 and dried at room temperature. Voucher specimen was deposited at the Herbarium of Complex of Academic Center for Educational and Cultural Research under number ACECR-266.
Extraction and isolation process
Dried roots of S. hypoleuca (900 g) were cut into small pieces and extracted with ethyl acetate at room temperature by percolation method for 72 hours and 3 times. The solvent was evaporated by rotary evaporator. The ethyl acetate extract (2 g) was fractionated by silica gel column chromatography (CC) with hexane, hexane: chloroform (9:1, 5:5), ethyl acetate and methanol, to give seven fractions (A-G). Fraction A (88 mg) was subjected to silica gel CC with hexane: ethyl acetate (19:1) to obtain compound 1 (21 mg). Fraction B (200 mg) was submitted to silica gel CC with hexane: ethyl acetate (9:1) to give compound 2 and 3 (17 and 13 mg respectively). Fraction C (134 mg) was submitted to silica gel CC with hexane: ethyl acetate (19:1) to result in six fractions (C1-C6). Fraction C5 (14 mg) was chromatographed on silica gel CC with chloroform: ethyl acetate (19:1) to yield compound 4 (8 mg). Fraction D (126 mg) was fractionated on silica gel CC with hexane: ethyl acetate (19:1) to obtain six parts (D1-D6). Fraction D3 (27 mg) was separated on sephadex LH20 with methanol: ethyl acetate (7:3) to gain four fractions (D31-D34). Fraction D33 (10 mg) was subjected to reverse phase (RP) silica gel CC with methanol: water (8:2) to result in compound 5 (5 mg). Fraction F (624 mg) was fractionated on silica gel CC with chloroform: methanol (19:1) to yield three parts (F1-F3). Fraction F1 (204 mg) was chromatographed on silica gel CC with chloroform: ethyl acetate (8:2) to obtain nine fractions (F11-F19). Fraction F13 (30 mg) was subjected to sephadex LH20 with methanol to result in compound 6 and 7 (7 and 5 mg, respectively). Fraction F17 (8 mg) was submitted to sephadex LH20 with methanol to obtain compound 8 and 9 (3 and 2 mg, respectively). Fraction F2 (67 mg) was further isolated on RP silica gel CC with methanol: water (9:1) to give compound 10 (2 mg).
NMR data of the compound 4 in CDCl 3
1.00 (m, 1H)
1.76 (m, 1H)
1.36 (m, 1H)
1.55 (m, 1H)
1.17 (m, 1H)
C-4, C-5, C-18
1.36 (m, 1H)
1.08 (brd, J=12.3 Hz, 1H)
C-4, C-6, C-7, C-18, C-20
1.76 (m, 2H)
1.95 (m, 1H)
C-6, C-8, C-17
2.37 (brd, J=12.4 Hz, 1H)
C-5, C-6, C-8, C-9, C-17
1.55 (m, 1H)
C-8, C-10, C-17
1.48 (m, 1H)
1.55 (m, 1H)
C-9, C-10, C-12
1.27 (m, 1H)
1.76 (m, 1H)
5.92 (dd, J=17.3,10.7 Hz, 1H)
5.04 (d, J=10.6 Hz, 1H)
5.20 (d, J=17.3 Hz, 1H)
1.27 (s, 3H)
4.51 (s, 1H)
C-7, C-8, C-9
4.81 (s, 1H)
0.79 (s, 3H)
C-3, C-4, C-5
0.86 (s, 3H)
C-3, C-4, C-5, C-18
0.67 (s, 3H)
NMR data of the compound 5 in CDCl 3
1.20 (m, 1H)
2.72 (brd, J=13.0 Hz, 1H)
1.58 (m, 1H)
1.72 (dd, J=13.3,13.4 Hz, 1H)
1.21 (m, 1H)
1.47 (brd, J=12.7 Hz, 1H)
1.47 (brd, J=12.7 Hz, 1H)
C-4, C-7, C-10, C-18, C-20
1.60 (m, 1H)
1.93 (d, J=14.9 Hz, 1H)
C-5, C-7, C-8, C-10
5.92 (brs, 1H)
3.15 (m, 1H)
C-12, C-13, C-14, C-17
1.17 (d, J=7.0 Hz, 3H)
C-13, C-15, C-17
1.22 (d, J=7.0 Hz, 3H)
C-13, C-15, C-16
0.87 (s, 3H)
C-3, C-5, C-19
0.87 (s, 3H)
C-3, C-5, C-18
1.23 (s, 3H)
C-1, C-5, C-9, C-10
2.03 (s, 3H)
7.12 (s, 1H)
C-11, C-12, C-13
13 C-NMR data of the compounds 6–10
The mass data of the compounds 1, 2, 3, 6 and 7 have been previously reported [27, 28]. The mass of other compounds are followed: Manool (4): EIMS (70eV) m/z: 290 [M]+ (8), 272 (40), 204 (20), 257 (58), 189 (28), 137 (100), 121 (48), 95 (67). 3-epicorosolic acid (8): 472 [M]+ (5), 248 (100), 223 (18), 203 (61), 189 (13), 133 (20), 119 (10). 3-epimaslinic acid (9): 472 [M]+ (4), 248 (100), 235 (9), 223 (12), 203 (54), 189 (15), 133 (28). coleonolic acid (10): m/z 470 [M] + (7), 452 (25), 264 (18), 206 (15), 201 (35), 159 (28), 146 (50), 105 (100).
β-sitosterol: 13C-NMR (125 MHz, CDCl3): δ C (from C-1 to C-29) 37.3, 31.7, 71.8, 42.3, 140.8, 121.7, 31.9, 31.9, 50.2, 36.5, 21.1, 39.8, 42.3, 56.8, 24.3, 28.3, 56.1, 11.9, 19.8, 36.2, 18.8, 34.0, 26.1, 45.8, 29.2, 19.0, 19.4, 23.1, 12.0.
Stigmasterol: 13C-NMR (125 MHz, CDCl3): δ C (from C-1 to C-29) 37.3, 31.7, 71.8, 42.2, 140.8, 121.7, 31.9, 31.9, 50.2, 36.4, 21.1, 39.7, 42.2, 56.9, 24.4, 28.9, 56.0, 12.0, 19.4, 40.5, 21.2, 138.3, 129.3, 51.6, 31.9, 19.0, 21.1, 25.4, 12.2.
Literature reviews show that Salvia species are important medicinal and food plants. About 200 triterpenoids have been isolated and identified from about 100 Salvia species and presented in a review article by Topcu . The oleanane, and ursane triterpenes display various pharmacological activities. These triterpenes can be considered as the lead compounds for the development of new multi-targeting bioactive agents . Both oleanolic and ursolic acid have been documented to protect liver against chemically induced injuries in laboratory animals via inhibition of toxicant activation and enhancement of immune systems. These two triterpenes have also been long-recognized as anti-inflammatory and anti-hyperlipidemic agents. Furthermore, anti-tumor activity has been noted from both non-toxic compounds .
Corosolic acid, a triterpenoid compound has been proved to have anti-diabetic effects on animal and human via enhancing glucose uptake in L6 myotubes and facilitating glucose transporters isoform 4 translocation in CHO/hIR cells. In addition, corosolic acid has been reported to inhibit the enzymatic activity of several non-receptor protein tyrosine phosphatases (PTPs) . The abietane diterpene 7 α-acetoxy-royleanone, containing quinone moiety in its structure, was demonstrated to possess cytotoxic activity on cancer cell lines and also alkylating properties using the nucleophile 4-(4-nitrobenzyl) pyridine . Among the reported antimicrobial labdane-type diterpenes, manool is the most active, since it furnished very promising MIC values for several tested bacteria that are closely associated with periodontitis .
According to chemotaxonomic significance, the isolated terpenes (manool (4), 7α-acetoxy-royleanone (5), ursolic acid (6), oleanolic acid (7), 3-epimaslinic acid (9)) were previously reported from other Salvia species such as S. sclarea, S. pubescens, S. lavandulifolia and S. officinalis. To the best of our knowledge, there is no report about the presence of the above mentioned compounds from S. hypoleuca. The triterpene 3-epicorosolic acid (8) and coleonolic acid (10) has not been reported from Salvia species, while some other genus of Lamiaceae such as Perilla frutescens and Coleus forskohlii contains these triterpenes.
In conclusion, the results of this study indicated the presence of ten terpenes and sterols in the root extract of S. hypoleuca as: sitosteryl oleate (1), β-sitosterol (2), stigmasterol (3), manool (4), 7α-acetoxy royleanone (5), ursolic acid (6), oleanolic acid (7), 3-epicorosolic acid (8), 3-epimaslinic acid (9) and coleonolic acid (10). Some of the isolated compounds (1–7, 9) have not been previously reported from S. hypoleuca and the triterpenes 8 and 10 not reported from Salvia genus until now. The above mentioned compounds have been recognized as the biologically and pharmacologically active constituents from this medicinal and aromatic species of salvia.
This research was supported by Tehran University of Medical Sciences and Health Services grant (No. 11847). The authors wish to thank Mr. Yousef Ajani (Institute of Medicinal Plants, Jahade-Daneshgahi) for his help in collection and identification of the plant material.
- Hedge IC: Labiatae. Flora Iranica. Volume 151. Edited by: Rechinger KH. 1986, Graz: Akademische Druck-u Verlagsanstalt, 403-480.Google Scholar
- Saeidnia S, Gohari AR, Malmir M, Moradi-Afrapoli F, Ajani Y: Tryptophan and sterols from Salvia limbata. J Med Plants. 2011, 10: 41-47.Google Scholar
- Lu Y, Foo LY: Polyphenolics of Salvia - a review. Phytochemistry. 2002, 59: 117-140. 10.1016/S0031-9422(01)00415-0.View ArticlePubMedGoogle Scholar
- Tutin TG, Heywood VH, Burgess NA, Moore DM, Valentine DH, Walters SM, Webb DA: Salvia L. Flora Europa. Edited by: Hedge IC. 1972, Cambridge: Cambridge University Press, 188-192.Google Scholar
- Ulubelen A, Sönmez U, Topcu G, Johansson CB: An abietane diterpene and two phenolics from Salvia forskahlei. Phytochemistry. 1996, 42: 145-147. 10.1016/0031-9422(95)00888-8.View ArticlePubMedGoogle Scholar
- Gohari AR, Saeidnia S, Malmir M, Hadjiakhoondi A, Ajani Y: Flavones and rosmarinic acid from Salvia limbata. Nat Prod Res. 2010, 24: 1902-1906. 10.1080/14786411003766912.View ArticlePubMedGoogle Scholar
- Suzuki H, Sawada S, Watanabe K, Nagae S, Yamaguchi MA, Nakayama T, Nishino T: Identification and characterization of a novel anthocyanin malonyltransferase from scarlet sage (Salvia splendens) flowers: an enzyme that is phylogenetically separated from other anthocyanin acyltransferases. Plant J. 2004, 38: 994-1003. 10.1111/j.1365-313X.2004.02101.x.View ArticlePubMedGoogle Scholar
- Amiri H: Quantative and qualative changes of essential oil of Salvia bracteata Bank et Sol. in different growth stages. Daru. 2007, 15 (Suppl 2): 79-82.Google Scholar
- Sajjadi SE, Shahpiri Z: Chemical composition of the essential oil Salvia Limbata C.A. mey. Daru. 2004, 12 (Suppl 3): 94-97.Google Scholar
- Ghannadi A, Samsam-shariat SH, Moattar F: Volatile constituents of the flower of Salvia hydrangea DC. Ex Benth. Daru. 1999, 7 (Suppl 3): 23-25.Google Scholar
- Matloubi moghadam F, Amin GH, Safavi poorsohi E: Composition of stembark essential oil from Salvia macrosiphon Boiss. Daru. 2000, 8 (Suppl 1): 28-29.Google Scholar
- Rustaiyan A, Koussari S: Further sesterterpenes from Salvia hypoleuca. Phytochemistry. 1988, 27: 1767-1769. 10.1016/0031-9422(88)80440-0.View ArticleGoogle Scholar
- Rustaiyan A, Niknejad A, Nazarians L, Jakupovic J, Bohlmann F: Sesterterpenes from Salvia hypoleuca. Phytochemistry. 1982, 21: 1812-1813.View ArticleGoogle Scholar
- Ali MS, Ahmed W, Jassbi AR, Onocha PA: Hypoleuenoic acid: a trans- cinnamic acid derived secondary metabolite from Salvia hypoleuca (Lamiaceae). J Chem Soc Pak. 2005, 27: 316-319.Google Scholar
- Bigdeli M, Rustaiyan A, Nadimi M, Masoudi S: Composition of the essential oil from roots of Salvia hypoleuca Benth. from Iran. J Essent Oil Res. 2005, 17: 82-83. 10.1080/10412905.2005.9698837.View ArticleGoogle Scholar
- Rustaiyan A, Komeilizadeh H, Masoudiand S, Monfared A: Volatile constituents of three Salvia species grown wild in Iran. Flav Frag J. 1999, 14: 276-278. 10.1002/(SICI)1099-1026(199909/10)14:5<276::AID-FFJ825>3.0.CO;2-Y.View ArticleGoogle Scholar
- Atta-ur-Rahman: Studies in natural products chemistry. Elsevier B V. 2008, 35: 753-Google Scholar
- Julien-David D, Geoffroy P, Marchioni E, Raul F, Aoude-Werner D, Miesch M: Synthesis of highly pure (oxy) phytosterols and (oxy) phytosterol esters Part II. (Oxy)-sitosterol esters derived from oleic acid and from 9,10-dihydroxystearic acid. Steroids. 2008, 73: 1098-1109. 10.1016/j.steroids.2008.04.010.View ArticlePubMedGoogle Scholar
- Gohari AR, Saeidnia S, Shahverdi AR, Yassa N, Malmir M, Mollazade K, Naghinejad AR: Phytochemistry and antimicrobial compounds of Hymenocrater calycinus. Eur Asia J Bio Sci. 2009, 3: 64-68.View ArticleGoogle Scholar
- Nasiri M, Saeidnia S, Mashinchian-Moradi A, Gohari AR: Srerols from the red algae, Gracilaria salicornia and Hypnea flagelliformis, from Persian Gulf. Phcog Mag. 2011, 7: 97-100. 10.4103/0973-1296.80663.View ArticleGoogle Scholar
- Ulubelen A, Topcu G, Eris C, Sonmez U, Kartal M, Kurucu S, Bozok-Johansson C: Terpenoids from Salvia sclarea. Phytochemistry. 1994, 36: 971-974. 10.1016/S0031-9422(00)90474-6.View ArticlePubMedGoogle Scholar
- Rodriguez B: 1H and 13C NMR spectral assignments of some natural abietane diterpenoids. Mag Res Chem. 2003, 41: 741-746. 10.1002/mrc.1245.View ArticleGoogle Scholar
- Gohari AR, Saeidnia S, Hadjiakhoondi A, Abdoullahi M, Nezafati M: Isolation and Quantificative Analysis of Oleanolic acid from Satureja mutica Fisch. & C. A. Mey. J Med Plants. 2009, 8: 65-6934.Google Scholar
- Kojima H, Ogura H: Configurational Studies on Hydroxy Groups at C-2,3 and 23 or 24 of Oleanene and Ursene-type Triterpenes by NMR Spectroscopy. Phytochemistry. 1989, 28: 1703-1710. 10.1016/S0031-9422(00)97829-4.View ArticleGoogle Scholar
- Mahato SB, Kundu AP: 13C NMR Spectra of pentacyclic triterpenoids, a compilation and some salient features. Phytochemistry. 1994, 37: 1517-1575. 10.1016/S0031-9422(00)89569-2.View ArticleGoogle Scholar
- Raja Rao KV, Rao LJM, Prakasa Rao NS: An A-Ring Contracted Triterpenoid from Hyptis suaveolens. Phytochemistry. 1990, 29: 1326-1329. 10.1016/0031-9422(90)85456-P.View ArticleGoogle Scholar
- Shahani S, Monsef-Esfahani HR, Saeidnia S, Saniee P, Siavoshi F, Foroumadi A, Samadi N, Gohari AR: Anti-Helicobacter pylori activity of the methanolic extract of Geum iranicum and its main compounds. Z Naturforsch. 2012, 67c: 172-180.View ArticleGoogle Scholar
- Gohari AR, Hadjiakhoondi A, Sadat-Ebrahimi SE, Saeidnia S, Shafiee A: Cytotoxic triterpenoids from Satureja macrantha C.A. Mey. Daru. 2005, 13 (4): 177-181.Google Scholar
- Topcu G: Bioactive triterpenoids from Salvia species. J Nat Prod. 2006, 69: 482-487. 10.1021/np0600402.View ArticlePubMedGoogle Scholar
- Jager S, Trojan H, Kopp T, Laszczyk MN, Scheffler A: Pentacyclic triterpen distribution in various plants-rich sources for a new group of multi-potent plant extracts. Molecules. 2009, 14: 2016-2031. 10.3390/molecules14062016.View ArticlePubMedGoogle Scholar
- Liu J: Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol. 1995, 49: 57-68. 10.1016/0378-8741(95)90032-2.View ArticlePubMedGoogle Scholar
- Shi L, Zhang W, Zhou YY, Zhang YN, Li JY, Hu LH, Li J: Corosolic acid stimulates glucose uptake via enhancing insulin receptor phosphorylation. Eur J Pharmacol. 2008, 584: 21-29. 10.1016/j.ejphar.2008.01.020.View ArticlePubMedGoogle Scholar
- Fronza M, Lamy E, Günther S, Heinzmann B, Laufer S, Merfort I: Abietane diterpenes induce cytotoxic effects in human pancreatic cancer cell line MIA PaCa-2 through different modes of action. Phytochemistry. 2012, 78: 107-19.View ArticlePubMedGoogle Scholar
- Souza AB, de Souza MGM, Moreira MA, Moreira MR, Furtado NAJC, Martins CHG, Bastos JK, dos Santos RA, Heleno VCG, Ambrosio SR, Veneziani RCS: Antimicrobial evaluation of diterpenes from Copaifera langsdorffii oleoresin against periodontal anaerobic bacteria. Molecules. 2011, 16: 9611-9619. 10.3390/molecules16119611.View ArticlePubMedGoogle Scholar
- Galicia MA, Esquivel B, Sanchez AA, Cardenas J, Ramamoorthy TP, Rodriguez-Hahn L: Abietane diterpenoids from Salvia pubescens. Phytochemistry. 1988, 27: 217-219. 10.1016/0031-9422(88)80617-4.View ArticleGoogle Scholar
- Passannantia S, Paternostroa M, Piozzi F: Triterpene acids from Salvia and Teucrium species. Phytochemistry. 1983, 22: 1044-1045. 10.1016/0031-9422(83)85059-6.View ArticleGoogle Scholar
- Brieskorn CH, Kapadia Z: Bestandteile von Salvia officinalis. Planta Med. 1980, 38: 86-90. 10.1055/s-2008-1074842.View ArticleGoogle Scholar
- Banno N, Akihisa T, Tokuda H, Yasukawa K, Higashihara H, Ukiya M, Watanabe K, Kimura Y, Hasegawa J, Nishino H: Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor- promoting effects. Biosci Biotech Biochem. 2004, 68: 85-90. 10.1271/bbb.68.85.View ArticleGoogle Scholar
- Roy R, Vishwakarma RA, Varma N, Tandon JS: Coleonolic acid, a rearranged ursane triterpenoid from Coleus forskohlii. Tetrahedron Lett. 1990, 31: 3467-3470. 10.1016/S0040-4039(00)97424-0.View ArticleGoogle Scholar
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