Comparison of chemical composition and bioactivity of polar extracts Salvia fruticosa (Syn. salvia Triloba) plant growing in Bodrum and Marmara Island

Salvia (sage) species belonging to the Lamiaceae family are among the most studied plants due to their biochemical diversity which originates their different type variety in chemical structures. The genus Salvia is widely used in the pharmaceuticals/cosmetics and food industries, and their main biological activities include antibacterial, antifungal, antiviral, antioxidant, anticholinesterase, anti-inflammatory, and cytotoxic activities. The genus Salvia is represented by about 1000 species worldwide and over 110 species, half of them being endemic in Turkey. In this study, a medicinal plant, Salvia fruticosa Mill. (syn. Salvia triloba L.) which is called Anatolian sage, are collected from Bodrum and Marmara Island where grown wildly. Plants are dried in the open air under shade. The dried plants were grounded and extracted with polar solvents; methanol, aqueous ethanol (70%), water (infusion and decoction) separetely, and apolar solvent hexane for the comparison. The phenolic and flavonoid compounds of the prepared extracts were determined by LC-MS/MS, by qualitatively and quantitatively. Anticholinesterase activities of the extracts against acetylcholinesterase and butyrylcholinesterase enzymes were determined by the Ellman method. Cytotoxic effects of the extracts against MCF7 breast cancer cell lines, MDA-MB-231 breast cancer cell lines and on the CCD-1079Sk healthy skin fibroblasts were investigated. The LC-MS/MS analysis results of the all polar extracts showed the main phenolic compounds found in the plants are rosmarinic acid, syringic acid, caffeic acid, chlorogenic acid, and fumaric acid while flavonoids are hispidulin, nepetin, luteolin, nepetin-7-glucoside, luteolin-7- rutinoside. According to the anticholinesterase activity results, each of the plant extracts collected from Bodrum inhibited AChE (acetylcholinesterase) enzyme at 200 µg/mL concentration (61-69%) while the hexane extract (75.5%) and methanol extract (73.7%) prepared from Marmara Island collection showed higher inhibition against AChE enzyme. However, against the BChE (butyrylcholinesterase) enzyme, the Salvia extracts, specially infusion and decoction prepared from Bodrum plants showed very weak (5.8% and 14.2%, respectively) activity while the Salvia extracts prepared from Marmara Island collection, especially hexane (68.9%) and methanol (74.5%) extracts showed a higher inhibition against the BChE enzyme. However, the extracts of infusion and decoction showed a weak inhibition (13.4% and 10.7%, respectively) against BChE. The cytotoxicity of Salvia extracts collected from both locations Bodrum and Marmara Island were investigated at 6 different concentrations in the range of 31.25-1000 mg/mL on healthy skin fibroblast cells (CCD-1079Sk) and two human breast cancer cell lines (MCF-7 and MDA-MB-231). In fact, all the extracts showed very low cytotoxicity (almost not) up to 125 mg/mL concentration, after this concentration, the percentage of the viability of the healthy skin fibroblast cell lines (CCD-1079Sk) was found to be decreased as dose-dependant (70 - 40%) manner. As a result, Salvia extracts, which were prepared by collecting from both locations, showed a cytotoxic activity of 70-75% on both breast cancer cell lines at a concentration of 1000 mg/mL, while all the extracts exhibited a cytotoxicity as 30-35% at the lowest dose. It is noticeable that, among Salvia extracts, the most cytotoxic ones against two breast cancer lines were found to be methanol extract and followed by decoctions/infusions which are prepared in the sterilized water. This thesis work was supported by the Bezmialem Vakif University, Scientific Research Projects Unit (Project No: BAP-20210605). Keywords: Sage, Anatolian sage, Salvia triloba, Salvia fruticosa