Genus Anemone Research Articles

Population genetic differentiation and structure of rare plant Anemone shikokiana based on genotyping-by-sequencing (GBS)

BackgroundAnemone shikokiana (Makino) Makino is a perennial herb of the genus Anemone in the family Ranunculaceae. Endemic to the Shandong Peninsula in China and Shikoku Island in Japan, it is a rare and endangered plant with a narrow, disjunct distribution. It is threatened with extinction and is in urgent need of conservation. Evaluating the genetic diversity of species, revealing the population genetic structure and gene flow, and inferring the population history are of great importance for species conservation, especially for rare and endangered plants.ResultsIn our study, 73 samples from eight wild populations in China were sequenced by Super-GBS, yielding a total of 40.59 G clean reads and 52,231 SNPs. Based on the obtained SNP data set, we evaluated the population genetic diversity, genetic structure, and gene flow of A. shikokiana. A low level of genetic diversity was found (He = 0.1925, Ho = 0.1422). The neighbor-joining (NJ) tree, principal component analysis and ADMIXTURE analysis suggested that these 73 A. shikokiana could be considered as two groups. Pairwise genetic differentiation coefficients (Fst) indicated that genetic differentiation was lower between adjacent populations and higher between geographically separated populations. The gene flow between Kunyu Mountain and Lao Mountain was very low. However, neither of the two regions showed evidence of Isolation by Distance.ConclusionsHere, we revealed the population genetic structure and gene flow of A. shikokiana from the Shandong Peninsula, China. This research provides valuable genetic resources for A. shikokiana and contributes to the scientific and effective conservation of the species.

Identification and quantification of flavonoids and hydroxycinnamic acids in yellow wood anemone (<i>Anemone ranunculoides</i> L.) by UHPLC-DAD-HESI/MS analysis

Introduction. Yellow wood anemone (Anemone ranunculoides L.) is a herbaceous perennial. This plant grows in the European parts of Russia, Ciscaucasia, Siberia, Central Europe and other regions. It has many different pharmacological activities: immunomodulatory, sedative, anti-inflammatory, antitoxic, diuretic, antibacterial, antioxidant, antitumor and antirheumatic activity. These various effects are due to their biologically active compounds, which include ephemeroids, protoanemonin, saponins, tannins, resins, ascorbic acid, ranunculin, oils lipids and triterpene glycosides. As to phenolic compounds, currently there is no sufficient information on flavonoids and hydroxycinnamic acids (HCAs) profiles in different species of genus Anemone. Because these groups of compounds are quite specific for yellow wood anemone, their detailed study seems to be relevant.Aim. The objective of this research is to identify and quantify flavonoids, HCAs, and their conjugates in Yellow wood anemone leaves, flowers and rhizomes with roots.Materials and methods. The identification was carried out by using ultra-high-performance liquid chromatography with diode array and electrospray ionization-mass spectrometric detection (UHPLC–DAD–ESI-MS) with quantification of individual compounds by external calibration method.Results and discussion. Among the 47 compounds, 30 flavonoids and 17 derivatives of HCAs were detected. Quercetin glycosides were found to be the major flavonoids in aerial parts whereas chalcone glycosides – in underground parts. The major HCA in rhizomes with roots was feruloyltartaric acid (1.18 mg/g) whereas chlorogenic acid was predominant in leaves and flowers (8.68 mg/g and 2.62 mg/g accordingly). The total content of phenolic compounds was estimated at 60 mg/g on a dry weight basis.Conclusion. As a result of the research a detailed profile of flavonoids and HCAs acids of anemon was described. The data obtained can serve to identify this species in the standardization of medicinal plant materials and taxonomic studies.

Chemical fingerprinting of three Anemone species and an adulteration study to detect cross mixing of medicinal plants by HPLC-HR-ESI-MS/MS method

The adulteration of plant raw materials used for the preparation of herbal drugs with foreign plant material is one of the important issues in the quality control of herbal products. Chemical fingerprinting is a well-known approach for the characterization of secondary metabolites associated with the plant species and can be used for quality control of plant material. The current study centred on the development of chemical fingerprinting of three medicinal plants of genus Anemone including A. obtusiloba, A. falconeri and A tetrasepala through identification of their metabolites using LC-ESI-QTOF-MS/MS analysis. Thirty compounds were identified by using high-resolution positive and negative electrospray- ionization (ESI) modes and MS/MS analysis. The identified compounds belong to diterpenoids, alkaloids, phenols, flavonoids and other classes and their distribution among the analysed species was studied using different statistical tools. Moreover, an LC-HR-ESI-MS/MS method was developed to detect the cross mixing of A. obtusiloba with Ziziphus jujuba. Seven chromatographically differentiative peaks confined to A. obtusiloba were selected to detect its contamination in adulterated samples. The method was able to detect as low as 20% mixing of A. obtusiloba in Z. jujuba. This study can play a significant role to manage the quality control of herbal medicines and to identify lead natural products of these plants.

ВЕСНЯНІ СУМЧАСТІ ГРИБИ (ASCOMYCOTA) УРОЧИЩА "ХОЛОДНИЙ ЯР"

Information about spring ascomycetous fungi on the territory of the nature tract "Kholodnyi Yar" is presented. As the result of the study 33 species of the ascomycetous fungi were identified, 19 of which are actually spring species and 14 – found in the spring. 15 species are registered for the nature tract "Kholodnyi Yar" for the first time. The largest number of species is represented by the family Morchellaceae (5 species), as well as by the families Pezizazeae (4 species) and Sclerotiniaceae (4 species). The ecological-trophic structure is dominated by species with a saprotrophic feeding strategy, which belong to xylotrophs, humus saprotrophs and herbophilic fungi. The group of xylotrophs was the predominant group of saprotrophic fungi in terms of the number of species (10 species). Humus saprotrophs numbered 9 species of the order Pezizales. In addition, representatives of the group of herbophilic (6 species), which use last year's remnants of herbaceous plants as a substrate, were quite common. Dumontinia tuberosa was the most common species among biotrophic species which parasitizes on the rhizomes of plants of the genus Anemone and forms fruiting bodies at the same time as the flowering period of the host plant. Symbiotrophs were represented by 3 species that are also capable of the saprotrophic feeding strategy inherent to humus saprophytes. A new location for Gyromitra slonevskii, Morchella crassipes and M. steppicola – species which were listed in the third edition of the Red Book of Ukraine – has been registered. Species included in the local red lists were also identified, namely Gyromitra gigas and Verpa bohemica (Kyiv region), Helvella acetabulum (Kherson region). The finding of Phylloscypha phyllogena is the second in Ukraine and the first on the territory of the Right-Bank Forest-Steppe.

Chemical constituents from the rhizome of Anemone hupehensis

A phytochemical investigation on the rhizome of Anemone hupehensis resulted in the isolation of thirteen compounds, including six neolignan glycosides (1-6), two phenylpropanoid glycosides (7, 8), a phenolic glycoside (9) and four triterpenoid saponins (10-13). The structures of the isolated compounds were elucidated on the basis of spectroscopic data. This is the first report of neolignan glycoside, phenylpropanoid glycoside, and phenolic glycoside from genus Anemone. This is also the first study to report compounds 1-5 and 7-9 from family Ranunculaceae. All the compounds, except 10 were isolated from A. hupehensis for the first time. The chemotaxonomic significance of the isolated compounds was discussed.

Chemical constituents from Anemone vitifolia Buch.-Ham. (Ranunculaceae)

A phytochemical investigation of Anemone vitifolia (Buch.-Ham.) led to the isolation of twelve compounds, including six lignans (1–6), four benzoic acid esters (7–10), one norsesquiterpenoid (11) and one lactone (12). The structures of these metabolites were established on the basis of detailed spectroscopic analysis, as well as comparisons with the data available in the literature. Among them, compounds 10 (dimethyl 2-(benzoyloxy)succinate) and 12 (4-hydroxy-5-methyl-γ-butyrolactone) were isolated for the first time as natural products. Compounds 1 and 11, compounds 2, 3, 5–7 and 9, and compounds 4 and 8 were isolated for the first time in family Ranunculaceae, genus Anemone and A. vitifolia respectively. The chemotaxonomic significance of the isolated compounds was discussed.

An overview of the genus Anemone L. (Ranunculaceae) in India

The present paper documents the genus Anemone L. (Ranunculaceae) from India. A total of 26 taxa (which includes 24 species, one subspecies and one variety) has been recorded from Indo Himalayas. Western Himalaya is endowed with 9 taxa, Eastern Himalaya with 8 taxa, while 9 are common to both the flanks. Three species are present in Northeast India which are common to IHR and one species has also been reported from Western Ghats. This genus constitutes flowering plants of high altitude and maximum diversity has been observed between 2000–3000 m asl. The genus possesses high medicinal potential and needs urgent assessment of taxonomic and conservation status for its sustainable utilization.

Chemical constituent and chemotaxonomic study on the root of Anemone vitifolia Buch.-Ham

Thirty-four reported compounds were obtained from Anemone vitifolia Buch.-Ham., and were identified as triterpenoid saponins (1–28), flavonoid glycosides (29–32) and steroidal saponins (33–34). The structure of these compounds was determined by physicochemical constants and spectral analyses (NMR, MS, IR). Twelve compounds (7, 8, 10, 14, 20, 21, 23, 26–28, 30 and 32) were firstly identified in genus Anemone. Compounds 2, 4 and 14 can be considered as characteristic components of A. vitifolia. Finally, the chemotaxonomic significance of these compounds is discussed.

In Vitro Anti-proliferative Activity and Mechanism of Action of Anemone nemorosa.

Anemone nemorosa is part of the Ranunculaceae genus Anemone (order Ranunculales) which comprises more than 150 species. Various parts of the plant have been used for the treatment of numerous medical conditions such as headaches, tertian agues, rheumatic gout, leprosy, lethargy, eye inflammation as well as malignant and corroding ulcers. The Anemone plants have been found to contain various medicinal compounds with anti-cancer, immunomodulatory, anti-inflammatory, anti-oxidant and anti-microbial activities. To date there has been no reported evidence of its use in the treatment of cancer. However, due to the reported abundance of saponins which usually exert anti-cancer activity via cell cycle arrest and the induction of apoptosis, we investigated the mode of cell death induced by an aqueous A. nemorosa extract by using HeLa cervical cancer cells. Cisplatin was used as a positive control. With a 50% inhibitory concentration (IC50) of 20.33 ± 2.480 µg/mL, treatment with A. nemorosa yielded a delay in the early mitosis phase of the cell cycle. Apoptosis was confirmed through fluorescent staining with annexin V-FITC. Apoptosis was more evident with A. nemorosa treatment compared to the positive control after 24 and 48 h. Tetramethylrhodamine ethyl ester staining showed a decrease in mitochondrial membrane potential at 24 and 48 h. The results obtained imply that A. nemorosa may have potential anti-proliferative properties.

Transcriptomic analysis in Anemone flaccida rhizomes reveals ancillary pathway for triterpene saponins biosynthesis and differential responsiveness to phytohormones

Anemone flaccida Fr. Schmidt is a perennial medicinal herb that contains pentacyclic triterpenoid saponins as the major bioactive constituents. In China, the rhizomes are used as treatments for a variety of ailments including arthritis. However, yields of the saponins are low, and little is known about the plant's genetic background or phytohormonal responsiveness. Using one-quarter of the 454 pyrosequencing information from the Roche GS FLX Titanium platform, we performed a transcriptomic analysis to identify 157 genes putatively encoding 26 enzymes involved in the synthesis of the bioactive compounds. It was revealed that there are two biosynthetic pathways of triterpene saponins in A. flaccida. One pathway depends on β-amyrin synthase and is similar to that found in other plants. The second, subsidiary (“backburner”) pathway is catalyzed by camelliol C synthase and yields β-amyrin as minor byproduct. Both pathways used cytochrome P450-dependent monooxygenases (CYPs) and family 1 uridine diphosphate glycosyltransferases (UGTs) to modify the triterpenoid backbone. The expression of CYPs and UGTs were quite different in roots treated with the phytohormones methyl jasmonate, salicylic acid and indole-3-acetic acid. This study provides the first large-scale transcriptional dataset for the biosynthetic pathways of triterpene saponins and their phytohormonal responsiveness in the genus Anemone.

Род Anemone L. (Ranunculaceae) во флоре Таджикистана

Род Anemone L. (Ranunculaceae) во флоре Таджикистана

Morphological study of fruits of the genus Anemone L. (Ranunculaceae Juss.) for taxonomy

Results of comparative-morphological study of the genus Anemone L. are given. According to the modern data, this genus includes 16 subgenera, 19 sections, 5 subsections, 27 series and 118 species. We argumented the use in the taxonomy of Anemone ca. 25 carpologic characters (against 10 characters which were used earlier). In addition to fruits shape (ovoid, ellipsoid, globose, compressed, ribbed or winged), the peculiarities of their basal and apical parts (sessile or not, narrowed or dilated). The peculiarities of shape and size of stylodia and stigmas were clarificated.

Reproductive ability of some species of the genus Anemone L. in conditions of culture

Reproductive ability of some species of the genus Anemone L. in conditions of culture

Nectar production in the pollen flower of Anemone nemorosa in comparison with other Ranunculaceae and Magnolia (Magnoliaceae)

The observation that the flowers of Anemone nemorosa offer nectar to pollinating bee-flies (Bombylius major) prompted this investigation into the site of nectar secretion and nectary tissue. To allow comparison on a broader basis, other nectar-secreting pollen flowers of the Ranunculaceae and Magnolia (Magnoliaceae) were included in the analysis. The contradictory information available on the function of the mouthparts of bee-flies during nectar and pollen feeding motivated us to investigate the proboscis structure in detail by SEM. Our investigations in Anemone nemorosa proved, for the first time, nectar secretion in the genus Anemone s.s. (i.e. other than the Pulsatilla group) and in addition, within the family, a new type of a carpellary nectary. The latter is an epithelial nectary involving the whole epidermis of the ovarian part of the carpel. The nectary of Anemone nemorosa resembles that of Magnolia (e.g. M. stellata), which we re-investigated. In both Anemone nemorosa and Magnolia stellata, nectar production is limited mainly to the female phase of the proterogynous flower. In this way, the attractiveness of the flower is also assured in the non-pollen presenting phase. Especially in Magnolia, with its numerous carpels arranged on a cone-like receptacle, the economic disadvantage of a choricarpous- compared to a coenocarpous-gynoecium is compensated for by nectar secretion by each carpel. When licking up the nectar droplets from the carpel surfaces, contact of the insect's body with each stigma may be achieved.

Two new glycosides from the whole plant of Anemone rivularis var. flore-minore

Phytochemical investigation on the whole plant of Anemone rivularis var. flore-minore led to the isolation of a new labdane-type diterpene glycoside (1) and a new trihydroxyfuranoid lignanoid glycoside (2), together with three known triterpene and triterpenoid glycosides (3–5). The structures of the two new compounds were elucidated as β-d-glucopyranosyl (13S)-13-hydroxy-7-oxo-labda-8,14-diene-18-oate (1) and (7S,7′R,8R,8′S)-7′-butoxy-7,9′-epoxy-4,4′,9-trihydroxy-3,3′-dimethoxylignane 9-O-β-d-glucopyranoside (2), on the basis of extensive spectral analysis and chemical evidence. Compound 1 is characterized by a glucose (Glc) esterified C-18 carboxyl group, which is a rarely encountered labdane-type diterpene glycoside in nature. The two new compounds (1 and 2) reported here are the first examples of diterpene glycoside and lignanoid glycoside found in the genus Anemone, and the known triterpene and triterpenoid glycosides (3–5) are identified for the first time from the title plant.

Evolution of the tetraploid Anemone multifida (2n = 32) and hexaploid A. baldensis (2n = 48) (Ranunculaceae) was accompanied by rDNA loci loss and intergenomic translocation: evidence for their common genome origin

In the genus Anemone two small groups of taxa occur with the highest ploidy levels 2n = 6x = 48, belonging to the closely related clades: the montane/alpine Baldensis clade and the more temperate Multifida clade. To understand the formation of polyploids within these groups, the evolution of allohexaploid A. baldensis (AABBDD, 2n = 6x = 48) from Europe and allotetraploid Anemone multifida (BBDD, 2n = 4x = 32) from America was analysed. Internal transcribed spacer and non-transcribed spacer sequences were used as molecular markers for phylogenetic analyses. Cytogenetic studies, including genomic in situ hybridization with genomic DNA of potential parental species as probe, fluorescence in situ hybridization with 5S and 18S rDNA as probes and 18S rDNA restriction analyses, were used to identify the parental origin of chromosomes and to study genomic changes following polyploidization. This study shows that A. multifida (BBDD, 2n= 4x = 32) and A. baldensis (AABBDD, 2n = 6x = 48) are allopolyploids originating from the crosses of diploid members of the Multifida (donor of the A and B subgenomes) and Baldensis groups (donor of the D subgenome). The A and B subgenomes are closely related to the genomes of A. sylvestris, A. virginiana and A. cylindrica, indicating that these species or their progeny might be the ancestral donors of the B subgenome of A. multifida and A and B subgenomes of A. baldensis. Both polyploids have undergone genomic changes such as interchromosomal translocation affecting B and D subgenomes and changes at rDNA sites. Anemone multifida has lost the 35S rDNA loci characteristic of the maternal donor (B subgenome) and maintained only the rDNA loci of the paternal donor (D subgenome). It is proposed that A. multifida and A. baldensis probably had a common ancestor and their evolution was facilitated by vegetation changes during the Quaternary, resulting in their present disjunctive distribution.

Development of Eighteen Microsatellite Markers in Anemone amurensis (Ranunculaceae) and Cross-Amplification in Congeneric Species

Polyploidy plays an important role in the evolution of plant genomes. To enable the investigation of the polyploidy events within the genus Anemone, we developed eighteen microsatellite markers from the hexaploid species A. amurensis (Ranunculaceae), and tested their transferability in five closely related species. The number of total alleles (NA) for each resulting locus varied from one to eight. The polymorphism information content (PIC) and Nei’s genetic diversity (NGD) for these microsatellites ranged from 0.00 to 0.71 and 0.00 to 0.91, respectively. For each population, the NA was one to seven, and the values of PIC and NGD varied from 0.00 to 0.84 and 0.00 to 0.95, respectively. In addition, most of these microsatellites can be amplified successfully in the congeneric species. These microsatellite primers provide us an opportunity to study the polyploid evolution in the genus Anemone.

Cytogenetic and phylogenetic studies of diploid and polyploid members of Tribe Anemoninae (Ranunculaceae)

The ancestry, phylogenetic differentiation and systematic classification of the worldwide-distributed genus Anemone have been debated for many years. In this paper 11 Anemone, three Pulsatilla species and Hepatica nobilis were subjected to detailed karyotype analysis with the aim of obtaining new cytogenetic data that will contribute to karyotype evolutionary studies of the tribe Anemoninae. The results are interpreted in a phylogenetic context, established from the intergenic nontranscribed spacer (NTS) of 5S rDNA and internal transcribed spacer (ITS) of 35S rDNA. One to three 35S and one to three 5S rDNA loci are present in diploid and polyploid taxa. The 35S rDNA loci are located terminally on the short arm of acrocentric chromosomes, while for 5S rDNA there is no preferential chromosomal position as it exhibits terminal, subterminal, interstitial or pericentromeric positions, and is located either on acrocentric or metacentric chromosomes. The karyotype of hexaploid A. baldensis (2n = 6x = 48) is presented for the first time, and A. sylvestris is proposed as one of its putative parental species. Chromosome fusion/translocation is proposed as the key mechanism involved in reduction of the basic chromosome number from 8 in the Anemone subgenus to 7 in the Anemonidium subgenus. The cytogenetic data obtained are mainly supported by ITS and NTS phylogeny. Diversification of the genus Anemone was accompanied by a large reduction of heterochromatin, from the Mediterranean anemones that have large amounts of heterochromatin to the New World anemones without any detectable heterochromatic blocks.

A synopsis of the genus Pulsatilla (Ranunculaceae) in Turkey

In this study, Pulsatilla Mill., which was considered a subgenus of the genus Anemone L. in Flora of Turkey, is accepted as a separate genus. Pulsatilla albana (Steven) Bercht. & J.Presl subsp. albana and P. violacea Rupr. subsp. violacea are declared as new records for the flora of Turkey. Anemone albana Steven subsp. armena (Boiss.) N.Busch, which was already known from Turkey, is accepted as a synonym of Pulsatilla violacea subsp. armena (Boiss.) Luferov. Descriptions, distribution maps, and identification keys are provided for these taxa in order to aid in the distinguishing of the genera Anemone and Pulsatilla as well as for species and subspecies of Pulsatilla in Turkey.

Flavonoids from Anemone tomentosa roots

The genus Anemone (Ranunculaceae) comprises about 150 species, of which 52 are distributed in northern China. As triterpenoid saponins are the main effective components of the genus Anemone, it has extensive biological activities, such as antitumor, antibacterial, and insect antifeeding properties. Anemone tomentosa (Maxim.) Pei is a perennial herb that is commonly called “Da Huo Cao” in China [1]. Its roots have been used as traditional Chinese medicine for the treatment of dysentery, malaria, infantile malnutrition, carbuncles, etc. [2]. Previous phytochemical studies of A. tomentosa have led to the isolation of coumarins [3], triterpenoids, and sterols [4, 5], but there are no records of the flavonoid constituents. Herein we communicate the additional isolation and structural elucidation of eight known flavonoids 1–8, of which 1, 4, and 6 were isolated from plants of the genus Anemone for the first time. Flavonoids were found first in A. tomentosa. Roots of A. tomentosa were collected in September 2007 from Ziwuling Mountain areas of Gansu Province (China) and identified by Prof. Xiaoqiang Guo, Department of Life Science, Longdong University. Ground air-dried roots (5 kg) of A. tomentosa were extracted at room temperature with 80% aq. EtOH (2 25 L). The residue (448 g) was partitioned between H2O and petroleum ether, CHCl3, EtOAc, and n-BuOH, successively. The EtOAc fraction (103 g) was chromatographed over silica gel with gradient elution by n-hexane:EtOAc (25:1 15:1) and CHCl3:MeOH (10:1 2:1) and monitoring by TLC, in order to produce thirteen subfractions (E1-E13). Subfractions E2 and E8 were applied to a Sephadex LH-20 column with elution by MeOH, and recrystallized to give compounds 1 (30 mg) and 4 (12 mg). The n-BuOH extract (92 g) was subjected to repeated silica gel column chromatography by elution with MeOH:CHCl3 (3:10 2:1) and monitoring by TLC, in order to produce sixteen subfractions (B1-B16). Subfractions B2, B4, B7, B8, B10, and B12 were repeatedly subjected to silica gel or Sephadex LH-20 column chromatography and preparative TLC, and recrystallized to afford compounds 2 (45 mg), 3 (19 mg), 5 (8 mg), 6 (21 mg), 7 (32 mg), and 8 (13 mg). Compound 1, C15H10O7, yellow powder (CH3COCH3), mp 310–312 C, positive response to Mg–HCl, AlCl3, and FeCl3–K3Fe(CN)6 tests, negative response to Molisch test. UV spectrum (MeOH, max, nm): 256, 302, 376. IR (KBr, , cm– 1): 3409 (OH), 1663 (C=O), 1612, 1563, 1520 (Ar). Spectral data (EI-MS, PMR, and 13C NMR) for 1 agreed with those reported in the literature for quercetin [6]. Compound 2, C21H20O12, yellow needles (MeOH), mp 221–223 C, positive response to Mg–HCl and Molisch tests. UV spectrum (MeOH, max, nm): 207, 252, 301, 354. IR (KBr, , cm–1): 3398 (OH), 1656 (C=O), 1603, 1560, 1493 (Ar). MS (FAB, m/z): 465 [M + H]+, 303 [M + H – Glc]+. PMR (500 MHz, CD3OD, , ppm, J/Hz): 12.61 (1H, s, HO-5), 11.24 (1H, s, HO-7), 9.71 (1H, s, HO-4 ), 9.21 (1H, s, HO-3 ), 7.53 (1H, d, J = 2.0, H-2 ), 7.41 (1H, dd, J = 2.0, 8.4, H-6 ), 6.91 (1H, d, J = 8.4, H-5 ), 6.37 (1H, d, J = 2.0, H-8), 6.24 (1H, d, J = 2.0, H-6), 5.31 (1H, d, J = 7.5, H-1 ), 3.99 (1H, dd, J = 2.1, 12.0, H-6 a), 3.76 (1H, dd, J = 6.1, 12.0, H-6 b), 3.61–3.35 (4H, m, H-2 -5 ). 13C NMR spectra data agreed with those reported for quercetin-3-O-D-glucopyranoside [7]. Compound 3, C21H20O12, yellow crystals (MeOH), mp 245–247 C, positive response to Mg–HCl and Molisch tests. MS (FAB, m/z): 465 [M + H]+, 303 [M + H – Glc]+. PMR (500 MHz, CD3OD, , ppm, J/Hz): 12.50 (1H, s, HO-5), 7.48 (1H, d, J = 2.1, H-2 ), 7.40 (1H, dd, J = 2.2, 8.5, H-6 ), 6.90 (1H, d, J = 8.6, H-5 ), 6.56 (1H, d, J = 2.0, H-8), 6.30 (1H, d, J = 2.0, H-6), 5.34 (1H, d, J = 7.4, H-1 ), 3.85 (1H, dd, J = 2.0, 11.9, H-6 a), 3.76 (1H, dd, J = 6.1, 11.9, H-6 b), 3.65–3.31 (4H, m, H-2 -5 ). 13C NMR spectral data agreed with those reported in the literature for quercetin-7-O-D-glucopyranoside [8].