Determination of bioactivity of Salvia ballsiana and Salvia huberi roots and resarch of chemical contents

The Salvia genus, an important member of the Lamiaceae family, has been used for different purposes since ancient times and has more than 900 species globally. The thesis's studying materials are the Salvia ballsiana (RECH. FIL.) HEDGE (endemic) and Salvia huberi HEDGE (endemic) belonging to the Salvia genus and growing in the eastern Anatolia. In this thesis, the antioxidant activity of petroleum ether, acetone, methanol, and water extracts of Salvia ballsiana and Salvia huberi roots was determined using β-carotene linoleic acid, DPPH free radical scavenging, ABTS cation radical scavenging, metal chelation, and CUPRAC assays. Besides, the in vitro anticholinesterase activity of the extracts against acetylcholinesterase and butyrylcholinesterase using the Ellman method, the in vitro tyrosinase inhibitory activity using the dopachrome method, the in vitro urease inhibitory activity using the indophenol method, the in vitro anti-inflammatory activity using the luminol (reactive oxygen species) assisted chemiluminescence method, and the in vitro cytotoxic activity against CaCo-2 colon cancer cell lines using MTT method were determined. In general, acetone, methanol, and water extracts of both species exhibited high activity in primary antioxidant activity tests. The acetone (EC50: 9.8 µg/mL) and methanol (EC50: 41.1 µg/mL) extracts of Salvia ballsiana and acetone (EC50: 29.2 µg/mL) extract of Salvia huberi displayed high cytotoxic activity. Besides, petroleum ether (IC50: 36.8 µg/mL) and acetone (IC50: 24.6 µg/mL) extracts of Salvia huberi showed anti-inflammatory activity. In general, all the root extracts had low anticholinesterase, tyrosinase, and urease inhibitory activity. Extracts possessing high antioxidant activity were fractionated. Ten compounds were purified using chromatographic methods. Betulinic acid (1), oleanolic acid (2), pisiferic acid (3), β-sitosterol 3-O-β-glycopyranoside (4), β-sitosterol (5), and ursolic acid (6) were isolated from the acetone and methanol extracts of Salvia ballsiana while β-sitosterol (7), tritetracontanoic acid (8), benzenepropanoic acid 3,5-bis (1,1-dimethylethyl) 4-hydroxy-octadecylester (octadecyl 3- (3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (9) and ursolic acid (10) from the acetone and methanol extracts of Salvia huberi. Spectroscopic methods were used to elucidate the structures of pure compounds (1-10). Besides, fingerprint studies were carried out in order to detect similarities and differences between species. For this purpose, acetone, methanol, and water extracts of Salvia ballsiana and Salvia huberi were analyzed using HPLC-DAD against 42 standard compounds. Twenty-three compounds were identified and quantified by the validated HPLC-DAD method. The chemical constituents, antioxidant, anticholinesterase, anti-inflammatory, tyrosinase inhibitory, urease inhibitory, and cytotoxic activities of all root extracts of Salvia ballsiana and Salvia huberi were examined for the first time in this thesis. Tritetracontanoic acid (8) and benzene propanoic acid 3,5-bis (1,1-dimethylethyl) 4-hydroxy-octadecylester (octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (9) were obtained from Salvia species for the first time, as well.