Determining the metabolic profile of some Achillea speci̇es by LC-MS-IT-TOF and LC-MS/MS. and investigation of their biological activities

In this pHd study, major secondary metabolites of 14 different Achillea species that belong to Asteracae family were determined by LC-MS/MS and LC-MS IT-TOF instruments. Also antioxidant and and anticancer properties of these species were investigated. The studied species (A. biebersteinii Afan, A. coarctata Poir., A. kotschyi subsp. kotschyi, A. lycaonica Boiss. et Heldr., A. millefolium L. subsp. millefolium L., A. schischkinii Sosn., A. setacea Waldst. et Kit.,A. sintenisii Hub.-Mor., A. wilhelmsii C. Koch subsp. wilhelmsii, A. teretifolia Willd., A. goniocephala Boiss. et Bal., A. nobilis L. subsp. neilreichii (Kerner) Formanek, A. spinulifolia Fenzl ex Boiss.ve A. monocephala Boiss. & Balansa) were taxonomically identified, dried and their overground and underground parts were seperated. The secondary metabolic profiles of the chloroform-methanol (1:1) extracts of the species were determined qualitatively by UHPLC-ESI-IT-TOF MS instrument. With this study 89 metabolites were identified in studied Achillea species. By developing and validating a comprehensive LC-MS/MS method, 37 phytochemicals of chloroform-methanol (1:1) and ethanol extracts of the species were analysed qualitativeley and quantitatively. In LC-MS/MS study, chlorogenic acid (55812.2 µg/g extract) in underground ethanol extract of A. kotschyi, quinic acid (38293.02 µg/g extract) in overground ethanol extract of A. schischkinii, fumaric acid (12986.93 µg/g extract) in underground chloroform-methanol extract of A. monocephala, malic acid (12945.44 µg/g extract) in overground chloroform-methanol extract of A. millefolium, rutin (11120.52 µg/g extract) in overground ethanol extract of A. coarctata, quercetin (9085.56 µg/g extract) in underground ethanol extract of A. teretifolia were found to be the richest analytes. Additionally, the antioxidant activities of the studied species were determined by using DPPH free radical scavenging assay, ABTS radical cation decolorization assay, β-carotene lipit peroxidation test system and CUPRAC cupper reduction capacity methods. For DPPH A. nobilis-underground (IC50: 12,23±0,24), for ABTS A. monocephala-underground (<10), A. nobilis-underground (<10) and A. kotschyi-underground (<10) and for CUPRAC A. nobilis and A. kotschyi-underground (3.675±0,130 and 3.547±0,039 respectively) extracts were the most active extracts for the studied methods. And for β-carotene lipid peroxidation test system all of the studied species were low or moderately active. Moreover, total phenolic content (A. nobilis-underground (282,97±3,14 µg pyrocatechol equivalent/mg extract)) and total flavonoid content (A. schischkinii-underground (35,36±0,54 µg quercetin eqivalent /mg extract)) of the studied species were investigated. According to the anticholinesterase activity results the highest activity in both butyrylcholinesterase and acetylcholinesterase enzyme inhibition was shown by overground extract of A. monocephala (39.73±1.51 and 19.60±0.44 respectively). Most importantly, the cytotoxic (anticancer) activities (on HeLa (Human Cervical Carcinoma Cell Line) of the chloroform-methanol extracts of the overground and underground parts of the 14 Achillea species were determined. Though most of the extracts were highly active, the highest cytotoxic activities were shown by A. goniocephala's (-0.009), A. millefolium's (-0.0059), A. wilhelmsii's (-0.0146), A. schischkinii's overground (-0.0034), A. spinulifolia's underground (-0.03) and overground (-0.02), extracts. Keywords : Achillea, seconder metabolite, LC-MS IT-TOF, LC-MS/MS, antioxidant, anticancer.