Beyond Traditional Use of Alchemilla vulgaris: Genoprotective and Antitumor Activity In Vitro.

Alchemilla vulgaris L. (lady's mantle) was used for centuries in Europe and Balkan countries for treatments of numerous conditions and diseases of the reproductive system, yet some of the biological activities of lady's mantle have been poorly studied and neglected. The present study aimed to estimate the potential of A. vulgaris ethanolic extract from Southeast Serbia to prevent and suppress tumor development in vitro, validated by antioxidant, genoprotective, and cytotoxic properties. A total of 45 compounds were detected by UHPLC-HRMS analysis in A. vulgaris ethanolic extract. Measurement of antioxidant activity revealed the significant potential of the tested extract to scavenge free radicals. In addition, the analysis of micronuclei showed an in vitro protective effect on chromosome aberrations in peripheral human lymphocytes. A. vulgaris extract strongly suppressed the growth of human cell lines derived from different types of tumors (MCF-7, A375, A549, and HCT116). The observed antitumor effect is realized through the blockade of cell division, caspase-dependent apoptosis, and autophagic cell death. Our study has shown that Alchemilla vulgaris L. is a valuable source of bioactive compounds able to protect the subcellular structure from damage, thus preventing tumorigenesis as well as suppressing tumor cell growth.

In Silico and In Vitro Studies of Alchemilla viridiflora Rothm-Polyphenols' Potential for Inhibition of SARS-CoV-2 Internalization.

Since the outbreak of the COVID-19 pandemic, it has been obvious that virus infection poses a serious threat to human health on a global scale. Certain plants, particularly those rich in polyphenols, have been found to be effective antiviral agents. The effectiveness of Alchemilla viridiflora Rothm. (Rosaceae) methanol extract to prevent contact between virus spike (S)-glycoprotein and angiotensin-converting enzyme 2 (ACE2) and neuropilin-1 (NRP1) receptors was investigated. In vitro results revealed that the tested samples inhibited 50% of virus-receptor binding interactions in doses of 0.18 and 0.22 mg/mL for NRP1 and ACE2, respectively. Molecular docking studies revealed that the compounds from A. viridiflora ellagitannins class had a higher affinity for binding with S-glycoprotein whilst flavonoid compounds more significantly interacted with the NRP1 receptor. Quercetin 3-(6″-ferulylglucoside) and pentagalloylglucose were two compounds with the highest exhibited interfering potential for selected target receptors, with binding energies of -8.035 (S-glycoprotein) and -7.685 kcal/mol (NRP1), respectively. Furthermore, computational studies on other SARS-CoV-2 strains resulting from mutations in the original wild strain (V483A, N501Y-K417N-E484K, N501Y, N439K, L452R-T478K, K417N, G476S, F456L, E484K) revealed that virus internalization activity was maintained, but with different single compound contributions.

Alchemilla viridiflora Rothm.: the potent natural inhibitor of angiotensin I-converting enzyme.

Alchemilla viridiflora Rothm., Rosaceae is a herbaceous plant widespread in central Greece, Bulgaria, North Macedonia and Serbia with Kosovo. Liquid chromatography-mass spectrometry analysis leads to the identification of 20 compounds in methanol extract, mainly ellagitannins and flavonoid glycosides. Given that various plant extracts have traditionally been used to treat hypertension and that some of the analyzed methanol extract constituents have beneficial cardiovascular effects, we hypothesized that some of these effects are achieved by inhibiting angiotensin I-converting enzyme (ACE). The dose-dependent ACE inhibitory activities of A. viridiflora and miquelianin were observed with an IC50 of 2.51 ± 0.00 µg/mL of A. viridiflora extract compared to the IC50 of 5.4139 ± 0.00 µM for miquelianin. The contribution of the single compounds to the tested activity was further analyzed through the in silico experimental approach. Computational docking results showed that tiliroside, ellagic acid pentose and galloyl-hexahydroxydiphenoyl-glucose exhibited even better binding affinity for the ACE active site than miquelianin, for which ACE activity was confirmed by an in vitro assay.

LC-ESI-MS/MS Identification of Biologically Active Phenolics in Different Extracts of Alchemilla acutiloba Opiz.

Liquid chromatography electrospray ionization tandem mass spectrometric (LC-ESI-MS/MS) qualitative and quantitative analysis of different extracts from the aerial parts and roots of Alchemilla acutiloba led to the identification of phenolic acids and flavonoids. To the best of our knowledge, isorhamnetin 3-glucoside, kaempferol 3-rutinoside, narcissoside, naringenin 7-glucoside, 3-O-methylquercetin, naringenin, eriodictyol, rhamnetin, and isorhamnetin were described for the first time in Alchemilla genus. In addition, the antioxidant, anti-inflammatory and cytotoxic activity of all extracts were evaluated. The results clearly showed that among analyzed extracts, the butanol extract of the aerial parts exhibited the highest biological activity comparable with the positive controls used.

Protective effects of Aronia melanocarpa juices either alone or combined with extracts from Rosa canina or Alchemilla vulgaris in a rat model of indomethacin-induced gastric ulcers.

The aim of the study was to investigate the effects of four Aronia melanocarpa-based juices in a rat model of indomethacin-induced gastric ulceration. THE JUICES WERE: AM1 and AM2 (produced from aronia fruits at 20 °C and 60 °C, respectively), AMRC (a mixture of AM2 with Rosa canina extract) and AMAV (aronia juice with Alchemilla vulgaris). Male Wistar rats were used. Each of the juices (10 ml/kg) was administered for 10 days. Indomethacin (30 mg/kg) was injected subcutaneously and after 4 h, the effects were estimated. Indomethacin caused heavy destructions of the gastric mucosa, increased the expression of Bax and decreased the expression of Bcl-2, induced a certain increase in lipid peroxidation and a slight decrease in gastric PGE2 content. The pretreatment with the juices reduced the severity of indomethacin-induced gastric lesions and antagonized the effects of indomethacin on apoptosis and lipid peroxidation. The highest was the protective effect of AMAV, the juice with the highest polyphenolic content. The protective effect of Aronia melanocarpa-based juices against indomethacin-induced gastric lesions could be attributed to their polyphenolic contents. The mechanism involved to the highest extent in the protective effect of the juices was the inhibition of apoptosis.

[Prospects for the use of the herb lady's mantle in geriatrics.]

In the review are presented the relationship between a rich chemical composition with the breadth of the therapeutic activity of the Alchemilla vulgaris herb. The drugs, based on the Alchemilla vulgaris raw material are widely used medical application is prevention and maintenance in geriatrics, as a lymphotropic, anti-inflammatory, hypoglycemic, hypotensive, nootropic, hemorheological, wound-healing, antioxidant, antispasmodic and a mild diuretic remedy for prevention and maintenance therapy in case of the imbalance of the cardiovascular, digestive, nervous, endocrine, immune, excretory and bone systems.

Effect of herbal mixture composed of Alchemilla vulgaris and Mimosa on wound healing process.

Alchemilla vulgaris and Mimosa tenuiflora (Mimosa) have been used to treat cutaneous wounds as a traditional remedy due to their various biological activities. But, there are only a few studies about the effects of these herbs on wound healing. The purpose of this study is to investigate the wound healing effect of the herbal mixture, consisting of A. vulgaris and Mimosa, in mice and to determine the activity of the extract in vitro. In present study, application of an ointment containing the herbal mixture on the dorsal skin wounds of mice showed that the wound healing process was faster than treatment of Fusidic acid. Histological analysis demonstrated the herbal mixture promoted re-epithelialization, collagen synthesis, and especially the regeneration of skin appendages such as hair follicles. Immunohistochemical analysis revealed the herbal mixture improved angiogenesis and the stabilization of blood vessels, as well as accelerated the formation of granulation tissue. In addition, we demonstrated that herbal mixture enhanced the migration of HaCaT, fibroblasts, and HUVECs on a two-dimensional wound, and promoted the proliferation of macrophages and lymphatic vessels. Our results demonstrated that herbal mixture can promote the migration of keratinocytes, fibroblasts, and endothelial cells, and the proliferation of macrophages and lymphatic vessels. Furthermore, it showed that herbal mixture accelerates wound healing. Therefore, we suggest that herbal mixture may have a potential for therapeutic use for treatment and management of cutaneous wound.

Identification of a Collagenase-Inhibiting Flavonoid from Alchemilla vulgaris Using NMR-Based Metabolomics.

This paper describes the use of 1H NMR profiling and chemometrics in order to facilitate the selection of medicinal plants as potential sources of collagenase inhibitors. A total of 49 plants with reported ethnobotanical uses, such as the healing of wounds and burns, treatment of skin-related diseases, rheumatism, arthritis, and bone diseases, were initially chosen as potential candidates. The in vitro collagenase inhibitory activity of hydroalcoholic extracts of these plants was tested. Moreover, their phytochemical profiles were analyzed by 1H NMR and combined with the inhibitory activity data by an orthogonal partial least squares model. The results showed a correlation between the bioactivity and the concentration of phenolics, including flavonoids, phenylpropanoids, and tannins, in the extracts. Considering the eventual false-positive effect on the bioactivity given by tannins, a tannin removal procedure was performed on the most active extracts. After this procedure, Alchemilla vulgaris was the most persistently active, proving to owe its activity to compounds other than tannins. Thus, this plant was selected as the most promising and further investigated through bioassay-guided fractionation, which resulted in the isolation of a flavonoid, quercetin-3-O-ß-glucuronide, as confirmed by NMR and HRMS spectra. This compound showed not only a higher activity than other flavonoids with the same aglycone moiety, but was also higher than doxycycline (positive control), the only Federal Drug Administration-approved collagenase inhibitor. The approach employed in this study, namely the integration of metabolomics and bioactivity-guided fractionation, showed great potential as a tool for plant selection and identification of bioactive compounds in natural product research.

Protective effect of dietary Alchemilla mollis on UVB-irradiated premature skin aging through regulation of transcription factor NFATc1 and Nrf2/ARE pathways.

BACKGROUND: Alchemilla mollis (lady's mantle) is a common ingredient in skin care products. However, the protective mechanism of A. mollis against skin problems has not been elucidated. PURPOSE: This study was to investigate the effects of A. mollis ethanolic extract (AM) on UVB-irradiated normal human dermal fibroblasts (NHDF) and hairless mice. METHODS: The in vitro anti-photoaging effect of AM was performed in NHDFs. The antioxidant activities were assessed through DPPH, ABTS, and reactive oxygen species (ROS) assays. Matrix metalloproteinase 1 (MMP-1), IL-6, procollagen type I, and transforming growth factor-ß1 (TGF-ß1) were measured by kits. The protein levels of p-c-Jun, p-c-Fos, Nrf2, NQO-1, HO-1, nuclear NFATc1 and cytosolic p-NFATc1 were evaluated by western blotting. In in vivo, H&E and Masson's trichrome staining were carried out. Skin texture was analyzed using the roughness parameters. The expression of MMP-1, procollagen type I, TGF-ß1 and elastin were measured by western blot. RESULTS: AM included gallic acid as an active constituent. AM exhibited a strong antioxidant effect by inhibiting DPPH and ABTS free radicals, as well as ROS production. It was also found to upregulate transforming growth factor ß1, type I procollagen and elastin expression, and to downregulate matrix metalloproteinase-1 and interleukin-6 expression in AM-treated NHDFs under UVB irradiation. These effects were attributed to AP-1 and Nrf2/ARE signaling pathways. Significantly, it was demonstrated that AM regulated the UVB-induced NFATc1 dephosphorylation in nucleus. Based on dietary data, AM was effective for the prevention of wrinkle formation, skin thickening, water loss, and erythema in UVB-exposed mouse skin. CONCLUSION: Our data indicate that A. mollis provides protection from UVB exposure in both hairless mice skin in vivo and NHDFs in vitro. AM might therefore be useful as a cosmetic material and functional food for the prevention of UVB-induced human skin photoaging.

Antiviral Activity of Lady's Mantle (Alchemilla vulgaris L.) Extracts against Orthopoxviruses.

We studied toxicity and antiviral activity of bioactive substances extracted from the roots (ethylacetate extracts) and aerial parts (ethanol extracts) of lady's mantle (Alchemilla vilgaris L.). Plant extracts are characterized by low toxicity for continuous Vero cell culture, but inhibit the reproduction of orthopoxviruses (vaccinia virus and ectromelia virus) in these cells. Of all studied extracts, ethylacetate extract from lady's mantle roots characterized by the highest content of catechins in comparison with other samples demonstrated the highest activity in vitro towards the studied viruses (neutralization index for vaccinia and ectromelia viruses were 4.0 and 3.5 lg, respectively). The antiviral effect of Alchemilla vulgaris L. extracts was shown to be dose dependent.

Phenolic Composition, Anti-Inflammatory, Antioxidant, and Antimicrobial Activities of Alchemilla mollis (Buser) Rothm.

The current study was designed to evaluate the antioxidant, anti-inflammatory and antimicrobial activities of Alchemilla mollis (Buser) Rothm. (Rosaceae) aerial parts extracts. Chemical composition was analyzed by spectrophotometric and chromatographic (HPLC) techniques. The antioxidant properties assessed included DPPH· and ABTS·+ radical scavenging, ß-carotene-linoleic acid co-oxidation assay. Antimicrobial activity was evaluated with disc diffusion and micro dilution method. In order to evaluate toxicity of the extracts, with the sulforhodamine B colorimetric assay L929 cell line (mouse fibroblast) was used. The anti-inflammatory activities of the potent antioxidant extracts (methanol, 70% methanol, and water extracts) were determined by measuring the inhibitory effects on NO production and pro-inflammatory cytokine TNF-α levels in lipopolysaccharide stimulated RAW 264.7 cells. 70% methanol and water extracts which were found to be rich in phenolic compounds (184.79 and 172.60 mg GAE/g extract) showed higher antioxidant activity. Luteolin-7-O-glucoside was the main compound in the extracts. Ethyl acetate and 70% methanol extracts showed higher antibacterial activity against Staphylococcus aureus and Salmonella enteritidis with MIC value of 125 µg/ml. 70% methanol extract potentially inhibited the NO and TNF-α production (18.43 µm and 1556.22 pg/ml, respectively, 6 h).

Secondary Metabolites of Alchemilla persica Growing in Iran (East Azarbaijan).

Phytochemical investigations of Alchemilla persica Rothm. growing in Iran were performed taking into account both the volatile and polar constituents. The hydrodistilled essential oil was analysed by GC-MS that revealed the presence of alkanes (27.8%), diterpenoids (19.6%) and sesquiterpene hydrocarbons (17.1%) as the major constituents, while tannins and flavonol glycosides were identified as the most abundant constituents of the methanol extract by HPLC-MS. A. persica can be a valuable source of ellagitannins and polyphenols.

Anti-Helicobacter pylori Activity of Four Alchemilla Species (Rosaceae).

The present study evaluated the anti-Helicobacterpylori activity of Alchemilla glabra Neygenf. (A. sect. Alchemilla), A. monticola Opiz (A. sect. Plicatae S.E. Fröhner), A. fissa Günther & Schummel (A. sect. Calycinae (Buser) Buser) and A. viridiflora Rothm. (A. sect. Calycinae), and identified ellagic acid and quercetin-3-O-ß-glucoside. Anti-H. pylori activity was tested against ten clinical isolates and one reference strain (ATCC 43504). The methanol extracts were more active than the dichloromethane and cyclohexane extracts. The ranges of concentrations were between 4 µg/mL for methanol extracts of A. viridiflora, A. glabra and A. monticola, and 256 µg/mL for cyclohexane extracts of A. viridiflora, A. glabra and A. fissa. The best overall activity was obtained with A. monticola extracts. No significant difference was found in the ellagic acid contents of the methanol extracts of the tested Alchemilla species (0.2-0.3 mg/mL), and anti-H. pylori activity was similar (4-32 µg/mL). Ellagic acid exhibited strong activity at very low concentrations (0.125-0.5 µg/mL), while the second identified compound, quercetin-3-O-ß-D-glucoside, was also very active in concentration of 2-16 µg/mL.

Phytochemical analyses and effects of Alchemilla mollis (Buser) Rothm. and Alchemilla persica Rothm. in rat endometriosis model.

PURPOSE: The aim of the present study is to evaluate the treatment potential of Alchemilla mollis (Buser) Rothm. and Alchemilla persica Rothm. in the experimentally induced endometriosis model in rats. METHODS: Endometriosis was surgically induced in rats by autotransplanting endometrial tissue to abdominal wall. Thirty-six rats were randomly divided into six groups. The groups were orally treated with the methanol:water (80:20) extracts of aerial parts and roots of A. mollis and A. persica. Buserelin acetate (20 mg) was used as the reference drug. The phytochemical contents of the most active extracts were determined by high performance liquid chromatography. RESULTS: The cystic formation was determined to be significantly decreased with the aerial part extract of A. mollis. A reduction in the endometrioma was also determined for the aerial part extract of A. persica group. However, significant reduction on the levels of cytokine were recorded for the A. mollis aerial part extract group. Therefore, the phytochemical contents of the aerial part extracts of A. mollis. and A. persica were analyzed. CONCLUSION: The results of the present study revealed that the aerial part extracts of A. mollis and A. persica could be beneficial in the treatment of endometriosis.

Elemental characterisation of the medical plant Alchemilla velebitica.

Alchemilla, commonly called "lady's mantle", is a genus of herbaceous perennial plants belonging to the family Rosaceae. The species Alchemilla velebitica is found only in Southern Europe, like in the Croatian National Park Northern Velebit. Its benefits, such as a astringent and emmenagogue activity as well as wound healing are correlated to the organic compounds found in the plant, but also certain trace elements are known to reduce skin lesions, such as zinc, selenium, copper, manganese, silicon and lithium. Thus the objective of the present study was the elemental characterization of leaves, blossoms and roots of A. velebitica. After acidic microwave assisted digestion the concentrations of selected essential and trace elements were determined by inductively coupled plasma--atomic emission spectrometry and inductively coupled plasma--mass spectrometry. Other minor elements, such as Al, B, Cu, Fe, Mn, Sr and Zn are also found in leaves, blossoms and roots with contents in µg/kg range. The preparation of decoctions and the extraction yields of the elements of interest are calculated.

Anti-influenza activity of Alchemilla mollis extract: possible virucidal activity against influenza virus particles.

Influenza virus infection is a major public health problem that leads to significant morbidity and mortality. The emergence of resistance to the currently available anti-influenza agents has necessitated the development of new drugs with novel targets. Studying known ethno-medicinal plants is a promising approach for the discovery of new antiviral compounds. Alchemilla mollis is used in traditional medicine in Europe for different indications, including minimizing the symptoms of a sore throat. In this study, we found that A. mollis extract has anti-influenza activity, and investigated the mechanism underlying its inhibition of influenza virus replication. Plaque assays demonstrated that treatment of cells with A. mollis extract prior to infection did not inhibit influenza virus infection. However, plaque formation was markedly reduced when infected cells were overlaid with an agarose gel containing A. mollis extract. In addition, exposure of the virus to A. mollis extract prior to infection and treatment of cells during virus infection significantly suppressed plaque formation. Influenza virus-induced hemagglutination of chicken red blood cells was inhibited by A. mollis extract treatment. The inhibitory effect was observed against influenza A virus subtypes H1N1, H3N2, and H5N2. These findings suggest that A. mollis extract has virucidal or neutralizing activity against influenza virus particles. Furthermore, inhibitory effect of zanamivir synergistically increased after combination with A. mollis extract. Our results suggest that A. mollis extract has the potential to be developed as an antiinfluenza agent.

Phenolic constituents from Alchemilla vulgaris L. and Alchemilla mollis (Buser) Rothm. at different dates of harvest.

Acetone/water extracts from the leaves, including stalks, of Alchemilla vulgaris L. and A. mollis (Buser) Rothm. were investigated for their phenolic composition by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 24 and 27 compounds were detected for A. vulgaris and A. mollis, respectively. Pedunculagin and agrimoniin, as described in earlier reports for A. vulgaris, as well as other monomeric and oligomeric ellagitannins such as sanguiin H-10, castalagin/vescalagin, and galloyl-bis-hexahydroxydiphenoyl (HHDP) hexose constituted the major phenolic fraction of both plant species. Also, gallic and chlorogenic acids were found in both extracts. Interestingly, catechin and a procyanidin trimer were detected only in A. mollis. The flavonoid fraction comprised quercetin glucuronide as major compound in addition to several other quercetin glycosides. Most interestingly, a tentatively identified kaempferol glucuronide and a methylated quercetin glucuronide were exclusively found in A. mollis. Finally, the overall phenolic fingerprints of both Alchemilla species, harvested in May and August, i.e. at the beginning and the end of the flowering period, were compared. A general accumulation of phenolic constituents was observed later in the year, especially with regard to the ellagitannins.

Influence of nutrient medium composition on in vitro growth, polyphenolic content and antioxidant activity of Alchemilla mollis.

Alchemilla mollis (Buser) Rothm. (Rosaceae) is a high-mountain medicinal plant growing in the Balkan Peninsula, with only one population in Bulgaria. Alchemilla plants (Lady's mantle) are commonly used in traditional medicine for treatment of many gynecological diseases. The commercial drugs "Herba Alchemillae" induce a rapid regeneration of skin epithelium and have styptic and anti-inflammatory actions. Because of the high content of phenolic compounds (tannins and flavonoids) and the ecological plasticity of the species, field cultivation or in vitro biomass production of A. mollis are possible alternatives to its collection from nature. Four MS based nutrient media differing in the concentration of the minerals and supplemented with alpha-naphthaleneacetic acid (NAA) and benzylaminopurine (BAP) were tested in order to examine their influence on the shoot multiplication effectiveness and the antioxidant activity of A. mollis, and also the possible relation between these parameters under the conditions of in vitro culture. The cultures grown for two months on these media differed significantly in their multiplication rates (p < 0.001), as well as in their morphological features--height, leaf color and root development. Methanol extracts of in vitro cultivated and ex vitro adapted and acclimated on Vitosha Mt. (1500 m a. s. l.) plants were analyzed for tannin and flavonoid content and for free radical scavenging activity. The contents of flavonoids and tannins in the in vitro cultures of A. mollis cultivated on the four tested media differed significantly (p < 0.05). The highest flavonoid content was found in the shoots cultivated on the control MS medium, as well as in the ex vitro adapted plants. The antioxidant activity of the in vitro cultures correlated positively with the concentrations of the PGRs in the respective media, and the ex vitro adapted plants had the highest antioxidant activity (IC50 13.1 +/- 1.9 microg/mL) commensurable with that of the commercial antioxidant butylated hydroxytoluene (BHT) used as a positive control, with an IC50 of 12.65 microg/mL.

Phenolic constituents from Alchemilla vulgaris L. and Alchemilla mollis (Buser) Rothm. at different dates of harvest.

Acetone/water extracts from the leaves, including stalks, of Alchemilla vulgaris L. and A. mollis (Buser) Rothm. were investigated for their phenolic composition by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 24 and 27 compounds were detected for A. vulgaris and A. mollis, respectively. Pedunculagin and agrimoniin, as described in earlier reports for A. vulgaris, as well as other monomeric and oligomeric ellagitannins such as sanguiin H-10, castalagin/vescalagin, and galloyl-bis-hexahydroxydiphenoyl (HHDP) hexose constituted the major phenolic fraction of both plant species. Also, gallic and chlorogenic acids were found in both extracts. Interestingly, catechin and a procyanidin trimer were detected only in A. mollis. The flavonoid fraction comprised quercetin glucuronide as major compound in addition to several other quercetin glycosides. Most interestingly, a tentatively identified kaempferol glucuronide and a methylated quercetin glucuronide were exclusively found in A. mollis. Finally, the overall phenolic fingerprints of both Alchemilla species, harvested in May and August, i.e. at the beginning and the end of the flowering period, were compared. A general accumulation of phenolic constituents was observed later in the year, especially with regard to the ellagitannins.

Flavonoid compounds identified in Alchemilla L. species collected in the north-eastern Black Sea region of Turkey.

This study identified flavonoid glycosides in species of the genus Alchemilla, A. procerrima, A. stricta, A. hirtipedicellata and A. sericata. A. procerrima is an endemic species for Turkey. After detailed investigation, flavonoid compounds of the species were identified for the first time. In this study, flavonoid compounds were determined by using two different chromatography techniques, TLC and HPLC. The following flavonoid compounds were identified from the Alchemilla species studied. They are as follows: orientin (luteolin-8-C-glucoside) Rf: 0,70, vitexin (apigenin-8-C-glucoside) Rf: 0,77 as flavone-C-glycoside, rutin (quercetin-3-O-rutinoside) Rf: 0,44, hyperoside (quercetin-3-O-galactoside) Rf: 0,65, isoquercetin (quercetin-3-O-glucopyranoside) Rf: 0,72, quercitrin (quercetin-3-O-rhamnoside) Rf: 0,84 as flavonol-O-glycoside. Three more folavonoids with Rf values of Rf1=0,36, Rf2=0,54 and Rf3=0,68 were also identified for the first time in this study. Rutin (quercetin-3-O-rutinoside) and the flavonoid glycoside, shown as Rf2 were found in all species. Quercitrin and isoquercetin were determined in all analysed species but A. procerrima. Hyperoside was identified in all species except for A. stricta. Vitexin was determined only in A.stricta. Orientin was determined in A. procerrima and A. stricta, but could not be determined in A. sericata and A. hirtpedicellata. Unknown flavonoid with Rf1 and Rf3 were determined outside of A. sericata. Description of these compounds in Turkish Alchemilla plants for the first time should be viewed as a discovery of an important chemosystematic feature.