COMPARATIVE STUDY ON TANNINS, FLAVONOIDS, TERPENES AND MINERAL ELEMENTS OF SOME SALVIA SPECIES

Abstract

Salvia officinalis L., S. sclarea L., S. pratensis L. and S. nemorosa L. originated from Hungary and Transylvania were investigated. Tannin and flavonoid content of the S. species were determined and evaluated. Significant differences (p< 0.05) were found for tannin content between Hungarian and Transylvanian S. officinalis and for flavonoid content between Hungarian and Transylvanian S. sclarea. A qualitative comparative study was performed for terpenes from volatile oils by GC and GC-MS technique. The chromatogram of different S. species significantly differed from each other. The spectra of the same sage species originated from different places (e.g. S. officinalis from Hungary and Transylvania) were not qualitative different. Borneol and α-thujone were detected in each sage species whereas linalyl acetate, α-pinene and β-pinene were found to be present in three sage species. Element concentration of samples was determined by ICP-AES for Al, As, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Li, Mg, Mn, Mo, Na, P, Pb, S, Ti and Zn. The element contrentrations differ significantly between the species and within one species, greatly depend on the place of collection. The common feature of the species examined seems to be the relatively high chromium content. The element content of aqueous extract and the dissolution rate were also determined. INTRODUCTION S. officinalis L. (sage) grows wild in Southern Europe and Asia Minor. In Hungary the plant is cultivated similarly to S. sclarea L. (muscat sage). In Hungary several sages grow wild, e.g.: S. verticillata, S. aethiopis, S. glutinosa, S. nemorosa, S. pratensis, S. austriaca, S. nutans. From these S. nemorosa, S. pratensis and S. austriaca are only known as furrow-weeds and S. nutans is a protected plant. In this paper S. officinalis L., S. sclarea L., S. pratensis L. and S. nemorosa L. were investigated. The dried leaves of sage (Salviae officinalis folium) are registered as a medicinal drug for several indications (inflammatory diseases, common cold, gastric ulcer, state of exhaustion, nervosity) with numerous favourable effects (e.g. antiinflammatory, antifungal, antimicrobial) (Koga et al., 1999, Ozcan and Boyraz, 2000, Baricevic et al., 2001). The food, perfume and pharmaceutical industry use the volatile oil of sage (Aethorum salviae) (Then and Konszky, 1987). Several different investigations have been made on Salvia species (Lamiaceae) and it is still in the focus of interest today. The most effective bioactive components of sages are terpenoids formed and stored in Lamiaceae-type grandular hairs. S. officinalis. contains mainly monoterpenes (borneol, bornyl acetate, α-pinene, β-pinene, α-thujone, βthujone, eucalyptol, myrcene) (Bernath et al., 1991). S. sclarea is rich both in monoterpenes and diterpenes (linalool, sclareol, manool, salvipisone) (Ulubelen et al., 1994/a). Proc. XXVI IHC – Future for Medicinal and Aromatic Plants Eds. L.E. Craker et al. Acta Hort. 629, ISHS 2004 Publication supported by Can. Int. Dev. Agency (CIDA) 464 Diand triterpenes (Β-amyrin, germanicol, lupeol, loranthol) are present in S. pratensis (Anaya et al., 1989), while diterpenoids, e.g. nemorosin was found in S. nemorosa L. (Ulubelen et al., 1994/b). Each species contains a large amount of flavonoids (ThomasBarberan and Wollenweber, 1990; Areias et al., 2000) and several papers deal with tanning materials (e.g. caffeic acid, chlorogenic acid, ellagic acid, gallic acid) of sages (Lu et al., 1999; Lu and Foo, 2000). Only some investigations were made to determine the metal ion content of sages (Sullivan et al. 1987; Torshin et al., 1997), although a closer knowledge of the quality and quantity of the metal content would be essential, with special regard to the application of sage in medicine. For the study we collected different S. species from the same site in Hungary and also different S. species from the same site in Transyvania. Therefore, the differences between different S. species from the same site could be considered as taxonomical variances and the differences between same S. species from different site (Hungary and Transylvania) could be geological (or climate) divergences. MATERIALS AND METHODS Four Salvia species were examined: S. officinalis L. (No.:965/b ), S. sclarea L. (No.:8163), S. pratensis L. (No.:8725) and S. nemorosa L. (No.:8146). Plants were collected during the flowering period from the botanical garden in the vicinity of Budapest in Vacratot and from the botanical garden of the University of Medicine, in Tirgu Mures in 2000. For the extraction of the volatile compounds: 10 g of drug was allowed to stand in hexane (100 mL) in room temperature for 5 days and then was filtered. For tea preparation: 2 g of the drug was infused in 200 ml double distilled water. After cooling down, the mixture was filtered. The tannin content of the samples was determined according to the Hungarian Pharmacopoeia (Ph. Hg.VII, 1992) by spectrometry and calculated for pyrogallol as reference material. The total flavonoid content of the samples was determined according to the German Pharmacopoeia (DAB 10, 1996). After acidic hydrolyses of flavonoid glycosides, flavonoid aglycones were complexed with AlCl3, measured at 420 nm and the result were calculated in hyperoside equivalents. GC measurements of volatile oils were done with a FISON GC800 equipped with a capillar column (30m x 0.32mm, I.D:0.25m) packed with DB-1701 (OV-17). Temperature program: 3 min. at 60 C then heated by 8 C min. up to 230 C. GC-MS measurements were made with HP-5890 type instrument. The detection of the components was carried out with Eingine-5989 mass selective detector. Column: 30m x 0.32mm, packed with Supelco PTE-5. Temperature program: 2 min. at 60 C then heated by 10 C min. up to 300 C. The concentrations of elements of samples were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Type of instrument: Atom Scan 25 (Thermo Jarrell Ash). Sampling: the dry milled samples (0.5 g) or rotavapored extract (20 mL) were digested with a mixture of 5 mL HNO3 and 3 mL H2O2 in teflon vessels. After digestion the samples were diluted to 25 mL, from which the following elements were determined: Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, P, Pb, S, Ti, Zn. Mean values and standard deviations (SD) were calculated from the results. For comparison of the means one way analysis of variance (ANOVA) was used by GraphPAD software version 1.14 (1990). Significance limit was p < 0.05. RESULTS AND DISCUSSION Investigations showed that sages originated both from Hungary and Transylvania are rich in tanning materials (Table 1). S. officinalis L. shows the highest tannin content followed by S. sclarea, S. pratensis, then S. nemorosa. Almost the same trend can be seen in the flavonoid-, as in the tannin content. S. officinalis is the richest in flavonoids and S. nemorosa shows the lowest values. Significant difference (p< 0.05) was found for tannin content between Hungarian and Transylvanian S. officinalis and for flavonoid content