turkjac Turkish Journal of Analytical Chemistry 2687-6698 Miraç OCAK 10.51435/turkjac.813224 Analytical Chemistry Analitik Kimya Antimicrobial, antioxidant, tyrosinase activities and volatile compounds of the Essential oil and Solvent Extract of Epilobium hirsutum L. growing in Turkey https://orcid.org/0000-0003-1248-7912 Kılıç Gözde Karadeniz Technical University https://orcid.org/0000-0002-3747-8855 Korkmaz Büşra Karadeniz Technical University https://orcid.org/0000-0002-9729-1013 Erik İshak Karadeniz Technical University https://orcid.org/0000-0002-8199-3336 Fandaklı Seda UNIVERSITY OF EURASIA https://orcid.org/0000-0002-5923-9302 Yaylı Sevde Seher Karadeniz Technical University https://orcid.org/0000-0003-2447-0763 Faiz Özlem Recep Tayyip Erdogan University https://orcid.org/0000-0003-1047-8350 Alpay Karaoğlu Şengül Recep Tayyip Erdogan University https://orcid.org/0000-0003-4174-3014 Yaylı Nurettin Karadeniz Technical University 12 29 2020 2 2 87 94 10 20 2020 11 21 2020 Copyright © 2019, Turkish Journal of Analytical Chemistry 2019 Turkish Journal of Analytical Chemistry

TThe essential oil (EO), solid phase micro extraction (SPME), and SPME of n-hexane extract of Epilobium hirsutum L. were analyzed by GC-FID/MS. A total of 35, 20, and 32 compounds were identified from E. hirsutum, accounting for 98.2% in HD, 97.6% in SPME, and 98.8% in SPME of n-hexane extract, respectively. The EO and SPME consisted mainly of alcohol (HD: 51.5% vs. SPME: 30.8%). Aromatic hydrocarbons (57.1%) was the major compounds for the SPME of n-hexane extract. Monoterpenes were the minor constituents for the EO (2.3%) and SPME (3.1%) of the E. hirsutum. (Z)-3-Hexene-1-ol (46.5%), (Z)-3-hexenyl acetate (18.2%), and cyclohexanone (38%) were major compounds for the EO, SPME, and SPME of n-hexane of the E. hirsutum, respectively. The comparative study gave that the total amount of monoterpenoids (6.3%) was found only in the EO of E. hirsutum. The antimicrobial activity of EO and solvent extracts (n-hexane, methanol, and water) of E. hirsutum were screened in vitro against nine microorganisms. The EO resulted the best activity (10 mm) against Escherichia coli. Antimicrobial activity for the methanol extract of E. hirsutum against the Pseudomonas aeruginosa, Enterococcus faecalis, and Mycobacterium smegmatis was found to be 12 mm, 12 mm, and 16 mm inhibition zone, respectively. Methanol extract gave moderate active against the Yersinia pseudotuberculosis, Staphylococcus aureus, and Bacillus cereus within 6-8 mm, respectively. Antioxidant activity of water extract were found to be 12.7647±0.0145 µm (CUPRAC) and 0.034 mg/mL (IC50, DPPH). IC50 values for the tyrosinase enzyme inhibitory activity for the methanol, and water extracts was determined to be 0.196±0.013 µg/mL and 0.160±0.09 µg/mL, respectively.

 Epilobium hirsutum essential oil GC-FID/MS SPME antimicrobial antioxidant tyrosinase activity Karadeniz Technical University [1] D.A. Baum, K.J. Sytsma, P.C. Hoch, A phylogenetic analysis of Epilobium (Onagraceae) based on nuclear ribosomal DNA sequences. Syst Bot, 19, 1994, 363-388. [2] D. Chamberlain, P.H. Raven, Epilobium L. Flora of Turkey and The East Aegean Islan, pp. 183-196, Edinburgh Universty Press, 1972. [3] N. Zeybek, U. Zeybek, Pharmaceutical botany, Ege University Faculty of Pharmacy Publication, Izmir, 1994. [4] T. Ege, The pharmacological and therapeutic potentials of Epilobium hirsutum L, Int. J Pharm Sci Rev Res, 57, 2019, 20-23. [5] S. Karakaya, I. Suntar, Ö.F. Yakinci, O. Sytar, S. Ceribasi, B. Dursunoglu, H. Ozbek, Z. Guvenalp, In vivo bioactivity assessment on Epilobium species: A particular focus on Epilobium angustifolium and its components on enzymes connected with the healing process, J Ethnopharmacol, 262, 2020, 113-207. [6] S. Granica, J.P. Piwowarski, M.E. Czerwińska, A.K. Kiss, Phytochemistry, pharmacology and traditional uses of different Epilobium species J. (Onagraceae): a review, J Ethnopharmacol, 156, 2014, 316–346. [7] L. Pirvu, V. Nicorescu, C. Hlevca, D.I. Udeanu, I. Nicorescu, Antimicrobial and synergistic activity of some whole and selective Epilobium hirsutum L. (great willowherb) extracts tested on standard and wild Staphylococcus aureus strains, Farmacia, 63, 2015, 690-695. [8] A. Vitalone, J. McColl, D. Thome, L.G. Costa, B. Tita, Characterization of the effect of Epilobium extracts on human cell proliferation, Pharmacology, 69, 2003, 79-87. [9] A. Sayik, A.S. Yusufoğlu, A. Leyla, G. Türker, B. Aydin, L. Arslan, Dna-binding, biological activities and chemical composition of wild growing Epilobium angustifolium L. extracts from Çanakkale, Turkey, J Turkish Chem Soc, 4, 2017, 811-840. [10] R. Jacquet, Methods of use for oenothein A and oenothein B from Epilobium species to treat inflammation or improve hormone balance in postmenopausal women, U S Pat Appl Publ, 2020, US 20200179426, A1 20200611. [11] S. Jürgenson, V. Matto, A. Raal, Vegetational variation of phenolic compounds in Epilobium angustifolium, Nat Prod Res 26, 2012, 1951- 1953. [12] I. Nitescu, D. Cucu, D. Lapa, O. Contz, Studies on a new method for photochemical screening using chromatography of medicinal plants, Farmacia, 42, 1994, 41-43. [13] D. Cando, D. Morcuende, M. Utrera, M. Estevez, Phenolic-rich extracts from Willowherb (Epilobium hirsutum L.) inhibit lipid oxidation but accelerate protein carbonylation and discoloration of beef patties, Eur Food Res Technol 238, 2014, 741-751. [14] I. Remmel, L. Vares, L. Toom, V. Matto, A. Raal, Phenolic compounds in five Epilobium species collected from Estonia Nat Prod Commun, 7, 2012, 1323-1324. [15] M. Pelc, O. Kosakowska, Z. Weglarz, J. Przybyl, A. Geszprych, Sterols and fatty acids in the seeds of evening primrose (Oenothera sp.) and willow herb (Epilobium sp.), Herba Pol, 51, 2005, 20-24. [16] M.A. Nawwar, M.S. Marzouk, W. Nigge, M. Linscheid, High-performance liquid chromatographic/electrospray ionization mass spectrometric screening for polyphenolic compounds of Epilobium hirsutum - the structure of the unique ellagitannin epilobamide-A, J Mass Spectrom, 32, 1997, 645-654. [17] J. De Pascual Teresa, B. Corrales, M. Grande, A review on presence of Oleanolic acid in Natural Products, Anales de Quimica, 75, 1979, 135-139. [18] S. Ivancheva, N. Manolova, J. Serkedjieva, V. Dimov, N. Ivanovska, Polyphenols from Bulgarian medicinal plants with anti-infectious activity, Basic Life Sci 59, 1992, 717-728. [19] H.H. Barakat, S.A.M. Hussein, M.S. Marzouk, I. Merfort, M. Linscheid, M.A.M. Nawwar, Polyphenolic metabolites of Epilobium hirsutum, Phytochemistry, 46, 1997, 935-941. [20] B.H. Tóth, B. Blazics, Á. Kéry, Polyphenol composition and antioxidant capacity of Epilobium species. J Pharm Biomed, 49, 2009, 26-31. [21] A. Vitalone, F. Bordi, C. Baldazzi, G. Mazzanti, L. Saso, B. Tita, Anti-proliferative effect on a prostatic epithelial cell line (PZ-HPV-7) by Epilobium angustifolium L., Il Farmaco, 56, 2001, 483-489. [22] E. Ruszová, J. Cheel, S. Pávek, M. Moravcová, M. Hermannová, I. Matějková, J. Spilková, V. Velebný, and L. Kubala, Epilobium angustifolium extract demonstrates multiple effects on dermal fibroblasts in vitro and skin photo-protection in vivo, Gen Physiol Biophys, 32, 2013, 347-359. [23] V. Kaškonienė, M. Stankevičius, T. Drevinskas, I. Akuneca, P. Kaškonas, K.B. Survilienė , A. Maruška, O. Ragažinskienė, O. Kornyšova, V. Briedis, R. Ugenskienė, Evaluation of phytochemical composition of fresh and dried raw material of introduced Chamerion angustifolium L. using chromatographic, spectrophotometric and chemometric techniques, Phytochemistry, 115, 2015, 184-193. [24] H.C Onar, A. Yusufoglu, G. Turker, R. Yanardag, Elastase, tyrosinase and lipoxygenase inhibition and antioxidant activity of an aqueous extract from Epilobium angustifolium L. leave, J Med Plants Res, 6, 2012, 716-726. [25] B.T. Hevesi, P.J. Houghton, S. Habtemariam, Á. Kéry, Antioxidant and anti-inflammatory effect of Epilobium parviflorum Schreb, Phytother Res, 23, 2009, 719-724. [26] A.K. Kiss, A. Bazylko, A. Filipek, S. Granica, E. Jaszewska, U. Kiarszys, A. Kośmider, J. Piwowarski, Oenothein Bʼs contribution to the anti-inflammatory and antioxidant activity of Epilobium sp. Phytomedicine, 18, 2011, 557-560. [27] H. Juan, W. Sametz, A. Hiermann, Anti-inflammatory effects of a substance extracted from Epilobium angustifolium, Agents and Actions, 23, 1988, 106-107. [28] W.J. Bartfay, E. Bartfay, J.G. Johnson, Gram-negative and Gram-positive antibacterial properties of the whole plant extract of Willowherb (Epilobium angustifolium), Biol Res Nurs, 14,2012, 85-89. [29] L. Battinelli, B. Tita, M.G. Evandri, G. Mazzanti, Antimicrobial activity of Epilobium spp. Extracts, Il Farmaco, 56, 2001, 345–348. [30] I. Kosalec, N. Kopjar, D. Kremer, Antimicrobial activity of Willowherb (Epilobium angustifolium L.) leaves and flowers, Curr Drug Targets,14, 2013, 986-991. [31] Q. Zeng, J. Wu, P. Lin, Chemical composition and antimicrobial activity of the essential oil from Epilobium angustifolium. Chem Natl Compd, 52, 2016, 1113-1115. [32] S. Silló, E. Varga, Á. Belák, A. Maráz, Phytochemical and antimicrobial investigation of Epilobium angustifolium L., Acta Pharm Hung, 84, 2014, 105-110. [33] B. Tita, H. Abdel-Haq, A. Vitalone, G. Mazzanti, L. Saso, Analgesic properties of Epilobium angustifolium, evaluated by the hot plate test and the writhing test, Il Farmaco 56, 2001, 341–343. [34] M. Stolarczyk, M. Naruszewicz, A.K. Kiss, Extracts from Epilobium sp. herbs induce apoptosis in human hormone-dependent prostate cancer cells by activating the mitochondrial pathway, J Pharm Pharmacol, 65, 2013, 1044-1054. [35] F. Pourmorad, M.A. Ebrahimzadeh, M. Mahmoudi, S. Yasini, Antinociceptive activity of methanolic extract of Epilobium hirsutum, Pak J Biol Sci, 10, 2007, 2764-2767. [36] F. Vitali, G. Fonte, A. Saija, B. Tita, Inhibition of intestinal motility and secretion by extracts of Epilobium spp. in mice, J Ethnopharmacol, 107, 2006, 342–348. [37] E. Ehsan, M.R. Akhgar, A. Kariminik, Chemical constituents and antibacterial activity of the essential oil from Epilobium hirsutum, J Biodivers Environ Sci, 7, 2015, 338-344. [38] I. Erik, G. Kiliç, E. Öztürk, Ş.A. Karaoğlu, N. Yaylı, Chemical composition, antimicrobial and lipase enzyme activity of essential oil and solvent extracts from Serapias orientalis subs. orientalis, Turk J Chem, in press 2020. DOI: 10.3906/kim-2005-51. [39] G. Tosun, N. Kahriman, C. Albay, Ş.A. Karaoğlu, N. Yaylı, Antimicrobial activity and volatile constituents of the flower, leaf, and stem of Paeonia daurica grown in Turkey, Turk J Chem, 35, 2011, 145-153. [40] G. Renda, A. Özel, B. Barut, B. Korkmaz, N. Yaylı, The Volatile chemical compositions of the essential oil/spme and enzyme inhibitory and radical scavenging activities of solvent extracts and the essential oils from Coronilla orientalis Miller and C. varia L. grows in Turkey, Iran J Pharm Res, 18, 2019, 1831-1842. [41] B. Yaylı, G. Tosun, M. Karaköse, G. Renda, N. Yaylı, SPME/GC-MS analysis of volatile organic compounds from three Lamiaceae species (Nepeta conferta Hedge & Lamond, Origanum onites L. and Satureja cuneifolia Ten.) growing in Turkey, Asian J. Chem, 26, 2014, 2541-2544. [42] N. Yaylı, A. Yaşar, N. Yaylı, M. Albay, K. Coşkunçelebi, Essential oil analysis and antimicrobial activity of Paeonia mascula from Turkey, Nat. Prod. Commun, 3, 2008, 941-944. [43] R.P. Adams, Identification of essential oil components by gas chromatography/mass spectrometry, Allured publishing corporation Carol Stream, IL, 2007. [44] G.L. Woods, B.A. Brown-Elliott, P.S. Conville, E.P. Desmond, G.S. Hall, G. Lin, G.E. Pfyffer, J.C. Ridderhof, S.H. Siddiqi, R.J. Wallace Jr, N.G. Warren, F.G. Witebskyet, Susceptibility testing of mycobacteria, nocardiae, and other aerobic actinomycetes, Wayne (PA): Clinical and Laboratory Standards Institute, 2011. [45] A.L. Barry, Standards NCCLS document, Methods for determining bactericidal activity of antimicrobial agents: approved guideline. National Committee for Clinical Laboratory Standards Wayne, 1999. [46] R. Apak, K. Güçlü, B. Demirata, M. Özyürek, E.S. Çelik, B. Bektaşoğlu, I.K. Berker, D. Özyurt, Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay, Molecules, 12, 2007,1496-1547. [47] M.S. Blois, Antioxidant determinations by the use of a stable free radical, Nature, 181, 1958, 1199–1200. [48] R. Fu, Y. Zhang, Y. Guo, F. Chen, Antioxidant and tyrosinase inhibition activities of the ethanol-insoluble fraction of water extract of Sapium sebiferum (L.) Roxb. Leaves, S. Afr. J. Bot, 93, 2014, 98–104. [49] N.Y. Iskender, N. Yaylı, N. Yildrim, T.B. Cansu, S. Terzioglu, The volatile constituents of the flower, leaf, and stem of Verbascum wiedemannianum grown in Turkey, J Oleo Sci, 58, 2009, 117-121. [50] O. Üçüncü, N. Yaylı, A. Yaşar, S. Terzioğlu, N. Yaylı, Chemical composition of the essential oils from flower, leaf, and stem of Senecio trapezuntinus Boiss. grown in Turkey, Nat. Prod. Commun, 3, 2008, 925-928. [51] C.L. Cespedes, C. Balbontin, J. G. Avila, M. Dominguez, J. Alarcon, C. Paz,V. Burgos, L. Ortiz, I. Peñaloza-Castro, D.S. Seigler, I. Kubo, Inhibition on cholinesterase and tyrosinase by alkaloids and phenolics from Aristotelia chilensis leaves, Food Chem. Toxicol, 109, 2017, 984-995. [52] S. Momtaz, N.L. Basson, Inhibitory activities of mushroom tyrosine and DOPA oxidation by plant extracts, S Afr J Bot, 74, 2008, 577-582.