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Alchemilla Vulgaris Agg. (Lady's Mantle) from Central Balkan: Antioxidant, Anticancer and Enzyme Cite This: RSC Adv.,2019,9,37474 Inhibition Properties

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Alchemilla Vulgaris Agg. (Lady's Mantle) from Central Balkan: Antioxidant, Anticancer and Enzyme Cite This: RSC Adv.,2019,9,37474 Inhibition Properties RSC Advances View Article Online PAPER View Journal | View Issue Alchemilla vulgaris agg. (Lady's mantle) from central Balkan: antioxidant, anticancer and enzyme Cite this: RSC Adv.,2019,9,37474 inhibition properties a b c a Sanja Vlaisavljevic,´ * Sanja Jelaca,ˇ Gokhan¨ Zengin, Neda Mimica-Dukic,´ a a d Sanja Berezni,ˇ Milorad Miljic´ and Zora Dajic´ Stevanovic´ The current study was designed to evaluate the phytochemical profile and biological properties (antioxidant, enzyme inhibitory and cytotoxic activity) of methanolic, ethanolic, ethyl-acetate and water extracts of Alchemilla vulgaris from Southeast Serbia (central Balkan) which was traditionally used to alleviate and treat many diseases. Bioactive compounds were characterized by LC-MS/MS technique and biological properties were evaluated using antioxidant, enzyme inhibitory and cytotoxic assays. Twenty- six phenolic compounds were quantified in methanolic, ethanolic, ethyl acetate and water extracts, Received 9th October 2019 whereas the highest yields were found in ethyl-acetate (EA) extract (gallic acid, caffeic acid, catchin, Accepted 4th November 2019 Creative Commons Attribution-NonCommercial 3.0 Unported Licence. quercetin). This extract has also shown the greatest antioxidant, anticancer and inhibitory enzyme DOI: 10.1039/c9ra08231j activities which were demonstrated for the first time in this study. The obtained results indicated that rsc.li/rsc-advances Alchemilla vulgaris from South Serbia possesses high potential for pharmaceutical applications. Introduction According to Flora of Serbia (Genus Alchemilla, Gaji´c, 1972) name of the species – Alchemilla vulgaris L. is valid; the species Nature has been an inexhaustible source of bioactive is present in Serbia with three subspecies (subsp. montana, compounds which have been used since ancient times for subsp. pratensis and subsp. vulgaris).15 This article is licensed under a treatment of many diseases. Today more interest is being paid Among ethnobotanical reports on Alchemilla vulgaris for to understanding the complex role of plant nutrition where Southeast Europe and the territory of Balkan peninsula, it is to medical plants have particular importance. These plants mention a comprehenisve work of Tucakov (1997), and his include various types of plants for medicinal purposes. They are reports on adstringent and antidiarrheal, as well as effects Open Access Article. Published on 18 November 2019. Downloaded 9/28/2021 6:05:35 AM. considered as a rich source of bioactive ingredients. Serbia is against pruritus vulvae, bleeding from uterus and menstrual one of the world's most biodiverse centers, rich in medical pains. The most used folk names according to Simonovic (1959) plants, where some of them have not been determinated yet or are: “virak”, “vrkuta”, “rosanica”, “biserak”, “virkovina”, “viri´c” there is no information on their ingredients and bioactivity.13 and “gospinplaˇst” (it is interesting that the latter literally means Alchemilla vulgaris, an aggregate species, commonly known Lady's mantle).12,13 as Lady's mantle or lion's foot, is a perennial herbaceous plant It is 30 to 50 cm long half-rosette herbaceous plant with belonging to the rose family (Rosaceae). corrugated and lobed kidney-shaped to semicircular leaves and The status of the species Alchemilla vulgaris is still not the yellowish-green owers form clusters. The plant blooms completely resolved and there are a few opinions. However, from June through to September and can be found on road- most of them treat the taxon as an aggregate of microspecies, sides, grasslands bank, and mountain slopes. which were also considered as subspecies or varieties of the Aerial parts of the plant are oen used against gastrointes- species. tinal diseases and inammatory processes, as well as for improvement wound healing due to its strong antimicrobial and anti-inammatory effects.1 The plant is very popular espe- cially due to its great effect against women's problems and aDepartment of Chemistry, Biochemistry and Environmental Protection, Faculty of diseases such as broids, cysts, endometriosis, infertility, relief Sciences, University of Novi Sad, Trg Dositeja Obradovi´ca 3, 21000 Novi Sad, of menstrual problems and regulation of the menstrual cycle Serbia. E-mail: [email protected]; Fax: +381 21454065; Tel: +381 and reproductive and thyroid hormones balancing.2 Extract of 214852770 the plant exhibits high scavenge potential, especially towards bDepartment of Immunology, Institute for Biological Research “Siniˇsa Stankovi´c”, University of Belgrade, Serbia superoxide anion radical. The results obtained in some studies cDepartment of Biology, Science Faculty, Selcuk University, Konya, Turkey indicate that presence of some phenolics, especially tannins, 3,4 dFaculty of Agriculture, University of Belgrade, Serbia may contribute to the antioxidant activity. Furthermore, Takir 37474 | RSC Adv.,2019,9, 37474–37483 This journal is © The Royal Society of Chemistry 2019 View Article Online Paper RSC Advances et al. (2015) demonstrated that methanolic extract of Alchemilla cholinesterase,41 anti-tyrosinase and anti-amylase activity of A. vulgaris has shown vasorelaxant and hypotensive effects.5 vulgaris extracts will be investigated in this study. The extracts of leaves and roots of Alchemilla vulgaris have These investigations were performed to verify whether there been reported to inhibit the orthopox viruses in Vero cell lines.6 are some other biological performances of Lady's mantle Bahadir et al., 2017 have performed in their study that extracts extracts which have been neglected so far, as there is not such of Alchemilla vulgaris demonstrated a hepatoprotective activity ethnopharmacological information. increasing the aminotransferase (ALT) level in carbon tetra- For best of our knowledge, the evaluation of Lady's mantle chloride induced mice serum.7 anticancer activity, especially on estrogen depended tumors, is In the Southeast Europe and the Balkan peninsula, the reported for the rst time. Summarily, the phytochemical Lady's mantle is very popular and acknowledged medicinal characterization in relation to antioxidant activity, enzyme plant, which is traditionally used in folk medicine and in range inhibitory effects and anticancer activity of different Alchemilla of herbal remedies. The herb is mostly used for preventing of vulgaris extracts are presented to clarify already known ethno- woman illnesses, such as menstrual problems, the oligomeno- pharmacological records, as well as to identify some novel rhea and dysmenorrheal, the menopausal complaints, for possible biological effects of the plant. regulation of vaginal secretion, polycystic ovaries and even for – infertility.8 10 Moreover, the Lady's mantle is used as diuretic, Material and methods anti-anemic and anti-diabetic, for wounds and ulcers, hernias and muscle atrophy,11 and it is readily used for wound healing, Plant material especially when used externally for ulcers, eczema and skin. Plant material was collected at the owering stage in There are also notes on use for mild and nonspeci c diarrhea midsummer 2017, from the site situated in south Serbia, the 12 and for slow metabolism. As reported for other regions, the Vlasina lake (E 22.36446000 N 42.73636300, 1234 m above sea most records refers to woman illnesses and disorders uses, in level). The habitat of the population is wet grassland of the Creative Commons Attribution-NonCommercial 3.0 Unported Licence. addition to astringent, antihemorrhagic, tonic, cardiotonic and Calthion type. Plant vouchers were deposited at Herbarium of 11 diuretic effects. In general, there are not many reports on Applied Botany Department of the Faculty of Agriculture, detailed phytochemical prole of Alchemilla and related bio- University of Belgrade, under voucher code 85000 BEO. The logical properties, especially focusing on anticancer properties collected material was air dried. and in particular of the estrogen depended cancer lines. The current study focuses on phytochemical composition assess- Extracts preparation ment, targeting the phenolic compounds, in different extracts of Alchemilla vulgaris collected from the central Balkan (South- Dry plant material was macerated for 24 h in 80% methanol (v/ east Serbia) region. The chosen populations originated from v), 70% ethanol (v/v), 70% ethylacetate (v/v) and distilled water This article is licensed under a sites where the plant material is collected from the wild for (dH2O) at room temperature using incubator shaker and l- Serbian and regional market needs. The main goal of the trated (IKA®). The supernatants were taken for drying. The present study was to verify the existing ethnobotanical data by obtained residues were dissolved in dimethyl sulfoxide (DMSO) À1 performing several antioxidant activity tests, as well as the to the concentration of 10 mg mL . The extracts (methanolic- Open Access Article. Published on 18 November 2019. Downloaded 9/28/2021 6:05:35 AM. anticancer and enzyme activity analyses. Evaluation of antioxi- MeOH, ethanolic-EtOH, ethylacetate-EA, water-H2O) were dant capacity has been also performed using different tests stored in dark at +4 C until further analyses. such as 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,20-azinobis-(3- ethylbenzothiazoline-6-sulfonic acid (ABTS), Cupric Reducing Quantication of bioactive compounds using liquid Antioxidant Capacity (CUPRAC), phosphomolibdenum, metal chromatography tandem-mass spectrometry (LC-MS/MS) chelating and Ferric Reducing Antioxidant
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