Anthocyanins standards (cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside) isolation from freeze-dried açaí (Euterpe oleraceae Mart.) by HPLC

Availability of analytical standards is a critical aspect in developing methods for quantitative analysis of anthocyanins. The anthocyanins cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside were isolated from samples of freeze-dried açaí (Euterpe oleraceae Mart.), which is a round and purple well-known palm fruit in Brazil, and then used as standards. The isolation of the anthocyanins was performed by High Performance Liquid Chromatography (HPLC), using an adapted six-channel selection valve. The identification of anthocyanin pigments in açaí was based on mass spectrometric data for molecular ions and MS-MS product ions and on previous published data. After the collection procedure, standards with a high purity grade were obtained and an external standard curve of each anthocyanin was plotted.

Flavonóides de Calotropis procera R. Br. (Asclepiadaceae)

In this work we report the identification of two flavonol glycosides isolated from the leaves of Calotropis procera R. Br. (Asclepiadaceae), a plant species with large occurrence in Northwest of Brazil with some applications in folk medicine. Some proved pharmacological activities in this species could be attributed to the presence of flavonol glycosides. The extraction and isolation of flavonol glycosides was carried out firstly by a liquid-liquid partition, and then by elution of n-BuOH fraction with MeOH over a Sephadex LH-20 column. The identification of flavonol glycosides isorhamnetin-3-O-rutinoside (1), and isorhamnetin-3-O-robinobioside (2), was obtained by 1H and 13C NMR, one- and two-dimensional techniques.

Phenolic profile of Cynara scolymus L. and Silybum marianum(L.) Gaertn and related antimicrobial properties

Cynara scolymus L. (artichoke) and Silybum marianum (L.) Gaertn (milk thistle), belonging to the Asteraceae family, are medicinal plants vo.ith well-reported antioxidant and hepatoprotective effects. Widely consumed as infusions, these plants can also be found in several formulations to allow an easier consumption. The bioactivity of infusions, pills, and syrups based on artichoke and milk thistle was previously reported by our research group [1 ,2] and among the various phytochemicals present in these dietary supplements, phenolic compounds are pointed out as the most responsible for their beneficial properties. With the aim of studying the antimicrobial activity and possible relation vo.ith the phenolic composition, two different formulations of each plant were assessed (pills and syrups). The phenolic profiles were obtained by HPLC-DAD-ESIIMS, and the antimicrobial activity was performed with clinical isolates from hospitalized patients, namely Escherichia coli, Escherichia coli spectrum extended producer of P-lactarnases (ESBL), Proteus mirabilis, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus (MRSA). Vanillic acid (5.58 J.tg/g) and luteolin-7-0-glucoside (2.2 J.tg/g) were the most abundant compounds in artichoke syrup, that did not reveal antimicrobial activity against the studied strains, which could be due to their low concentrations. On the other hand, artichoke pills presented a prevalence of 5-0-caffeoylquinic (28.2 J.tg/g), 1,3-dicaffeoylquinic (24 J.tg/g), and 4-0-Caffeoylquinic acids (13.3 J.tg/g); revealing the capacity to inhibit MRSA vo.ith a MIC value of 1.9 mg!g. Regarding milk thistle, isorhamnetin-0-deoxyhexoside-0-hexoside, isorhamnetin-3-0-rutinoside, and isorhamnetin-0-deoxyhexoside-0-dihexoside were the major compounds detected in the syrup, in concentrations of 7.26, 5. 75, and 3.64 J.tg/g, respectively. This formulation proved to be able to inhibit the growth of E. coli, ESBL, MRSA and P. aeruginosa, with MIC values ranging from 0.2 to 1.3 mg!mL. Hydroxylated silibinin (1.565 J.!g/g) was the major flavonoid found in the pills, that revealed antimicrobial activity against ESBL, with a MIC value of 15 mg!mL, but did not inhibit the growth of the remaining bacteria None of the studied samples was able to inhibit P. mirabilis at the studied concentrations (1000 and 26.4 mg!mL for the syrups of artichoke and milk thistle, respectively; 150 mg/mL for both pills). Overall, the studied syrups and pills of artichoke and milk thistle revealed to be a good source of phenolic compounds, with some of these formulations revealing antimicrobial activity.

Determination of flavonoids in stamen, gynoecium, and petals of Magnolia grandiflora L. and their associated antioxidant and hepatoprotection activities

Chromatographic analysis of flavonoids in ethyl acetate fractions of the stamen, gynoecium, and petal of Magnolia grandiflora L. by HPLC-PDA-MS/MS-ESI in the negative ionization mode was performed in this study. The results revealed the presence of eight flavonoids: apigenin 8-C-glucoside, luteolin 8-C-glucoside, quercetin 3-O-rutinoside, quercetin 3-O-galactoside, quercetin, 3-O-glucoside, kaempferol 3-O-rutinoside, isorhamnetin 3-O-glucoside, and isorhamnetin. Their quantification revealed that luteolin 8-C-glucoside is the major flavonoid and that the total phenolic content is concentrated primarily in the stamen. The antioxidant and hepatoprotective effects of ethanolic extract of the flower organs were evaluated against hepatotoxicity induced by CCl4, compared with the effects of silymarin.

Flavonoids, phenolic acids and abscisic acid in Australian and New Zealand Leptospermum honeys

Flavonoids, phenolic acids and abscisic acid of Australian and New Zealand Leptospermum honeys were analyzed by HPLC. Fifteen flavonoids were isolated in Australian jelly bush honey (Leptospermum polygalifolium), with an average content of 2.22 mg/100 g honey. Myricetin (3,5,7,3',4',5'-hexahydroxyflavone), luteolin (5,7,3',4'-tetrahydroxyflavone) and tricetin (5,7,3',4',5'-pentahydroxyflavone) were the main flavonoids identified. The mean content of total phenolic acids in jelly bush honey was 5.14 mg/100 g honey, with gallic and coumaric acids as the potential phenolic acids. Abscisic acid was quantified as twice the amount (11.6 mg/100 g honey) of the phenolic acids in this honey. The flavonoid profile mainly consisted of quercetin (3,5,7,3',4'-pentahydroxyflavone), isorhamnetin (3,5,7,4'-tetrahydroxyflavone 3'-methyl ethyl), chrysin (5,7-dihydroxyflavone), luteolin and an unknown flavanone in New Zealand manuka (Leptospermum scoparium) honey with an average content of total flavonoids of 3.06 mg/100 g honey. The content of total phenolic acids was up to 14.0 mg/100 g honey, with gallic acid as the main component. A substantial quantity (32.8 mg/100 g honey) of abscisic acid was present in manuka honey. These results showed that flavonoids and phenolic acids could be used for authenticating honey floral origins, and abscisic acid may aid in this authentication. (C) 2002 Published by Elsevier Science Ltd.

Saponinas triterpênicas de Tocoyena brasiliensis Mart. (Rubiaceae)

The present communication reports the isolation and identification of four triterpenoid saponins from the chloroform extract of the leaves of Tocoyena brasiliensis: 3-O-beta-D-quinovopyranosyl quinovic acid, 3-O-beta-D-quinovopyranosyl cincholic acid, 3-O-beta-D-glucopyranosyl quinovic acid and the 28-O-beta-D-glucopyranosyl ester derivative of quinovic acid as binary mixtures, respectively. From the ethanol extract a flavonoid identified as ramnazin-3-O-rutinoside was obtained. The structures of these compounds were assigned by data analysis of 1D and 2D NMR spectrometry and comparison with data recorded in the literature for these compounds.

Flavonoids in Astragalus corniculatus

Nine flavonoids were identified in aerial parts of Astragalus corniculatus Bieb. (Fabaceae) by liquid chromatography coupled with ionspray mass spectrometry in the tandem mode (LC/MS/MS) with negative ion detection. Vitexin, orientin and eriodictyol-7-O-glucoside are obtained for the first time in genus Astragalus L, and isorhamnetin-3-O-glucoside in the species.

Atividade anticolinesterásica e perfil químico de uma fração cromatográfica ativa do extrato etanólico das flores Bellis perennis L. (Asteraceae)

This work describes the isolation of an active flavonoid fraction and identification of isorhamnetin 3-O-β-D-(6''-acetyl)-galactopyranoside from flowers of B. perennis, and also the evaluation of anticholinesterase (AChE) activity of ethanolic extract from flowers (EEF) and the active fraction. The chemical structure of the flavonoid was defined on the basis of spectroscopic ¹H NMR, IR and UV data. EEF or flavonoid reduces AChE activity in vivo, while flavonoid also reduces AChE activity in vitro, showing a value of 1.49 mM for 50% inhibitory concentration (IC50), suggesting potential use as an insecticide or in the treatment of neurodegenerative diseases such as Alzheimer's disease.

Astringent Food Compounds and Their Interactions with Taste Properties

Astringency is traditionally thought to be induced by plant tannins in foods. Because of this current research concerning the mechanism of astringency is focused on tannin‐protein interactions and thus on precipitation, which may be perceived by mechanoreceptors. However, astringency is elicited by a wide range of different phenolic compounds, as well as, some non‐phenolic compounds in various foods. Many ellagitannins or smaller compounds that contribute to astringent properties do not interact with salivary proteins and may be directly perceived through some receptors. Generally, the higher degree of polymerization of proanthocyanidins can be associated with more intense astringency. However, the astringent properties of smaller phenolic compounds may not be directly predicted from the structure of a compound, although glycosylation has a significant role. The astringency of organic acids may be directly linked to the perception of sourness, and this increases along with decreasing pH. Astringency can be divided into different sub‐qualities, including even other qualities than traditional mouth‐drying, puckering or roughing sensations. Astringency is often accompanied by bitter or sour or both taste properties. The different sub‐qualities can be influenced by different astringent compounds. In general, the glycolysation of the phenolic compound results in more velvety and smooth mouthdrying astringency. Flavonol glycosides and other flavonoid compounds and ellagitannins contribute to this velvety mouthdrying astringency. Additionally, they often lack the bitter properties. Proanthocyanidins and phenolic acids elicit more puckering and roughing astringency with some additional bitter properties. Quercetin 3‐O‐rutinoside, along with other quercetin glycosides, is among the key astringent compounds in black tea and red currants. In foods, there are always various other additional attributes that are perceived at the same with astringency. Astringent compounds themselves may have other sensory characteristics, such as bitter or sour properties, or they may enhance or suppress other sensory properties. Components contributing to these other properties, such as sugars, may also have similar effects on astringent sensations. Food components eliciting sweetness or fattiness or some polymeric polysaccharides can be used to mask astringent subqualities. Astringency can generally be referred to as a negative contributor to the liking of various foods. On the other hand, perceptions of astringent properties can vary among individuals. Many genetic factors that influence perceptions of taste properties, such as variations in perceiving a bitter taste or variations in saliva, may also effect the perception of astringency. Individuals who are more sensitive to different sensations may notice the differences between astringent properties more clearly. This may not have effects on the overall perception of astringency. However, in many cases, the liking of astringent foods may need to be learned by repetitive exposure. Astringency is often among the key sensory properties forming the unique overall flavour of certain foods, and therefore it also influences whether or not a food is liked. In many cases, astringency may be an important sub‐property suppressed by other more abundant sensory properties, but it may still have a significant contribution to the overall flavour and thus consumer preferences. The results of the practical work of this thesis show that the astringent phenolic compounds are mostly located in the skin fractions of black currants, crowberries and bilberries (publications I–III). The skin fractions themselves are rather tasteless. However, the astringent phenolic compounds can be efficiently removed from these skin fractions by consecutive ethanol extractions. Berries contain a wide range of different flavonol glycosides, hydroxycinnamic acid derivatives and anthocyanins and some of them strongly contribute to the different astringent and bitterness properties. Sweetness and sourness are located in the juice fractions along with the majority of sugars and fruit acids. The sweet and sour properties of the juice may be used to mask the astringent and bitterness properties of the extracts. Enzymatic treatments increase the astringent properties and fermented flavour of the black currant juice and decrease sweetness and freshness due to the effects on chemical compositions (IV). Sourness and sweetness are positive contributors to the liking of crowberry and bilberry fractions, whereas bitterness is more negative (V). Some astringent properties in berries are clearly negative factors, whereas some may be more positive. The liking of berries is strongly influenced by various consumer background factors, such as motives and health concerns. The liking of berries and berry fractions may also be affected by genetic factors, such as variations in the gene hTAS2R38, which codes bitter taste receptors (V).

Extraction of polyphenols from processed black currant (Ribes nigrum L.) residues

The total phenol and anthocyanin contents of black currant pomace and black currant press residue (BPR) extracts, extracted with formic acid in methanol or with methanol/water/acetic acid, were studied. Anthocyanins and other phenols were identified by means of reversed phase HPLC, and differences between the two plant materials were monitored. In all BPR extracts, phenol levels, determined by the Folin-Ciocalteu method, were 8-9 times higher than in the pomace extracts. Acid hydrolysis liberated a much higher concentration of phenols from the pomace than from the black currant press residue. HPLC analysis revealed that delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside were the major anthocyanins and constituted the main phenol class (approximate to 90%) in both types of black currant tissues tested. However, anthocyanins were present in considerably lower amounts in the pomace than in the BPR. In accordance with the total phenol content, the antioxidant activity determined by scavenging of 2,2'-azinobis(3-ethylbenzothiazoline-6- sulfonic acid) radical cation, the ABTS(center dot+) assay, showed that BPR extracts prepared by solvent extraction exhibited significantly higher (7-10 times) radical scavenging activity than the pomace extracts, and BPR anthocyanins contributed significantly (74 and 77%) to the observed high radical scavenging capacity of the corresponding extracts.

Saponinas triterpênicas de Tocoyena brasiliensis Mart. (Rubiaceae)

The present communication reports the isolation and identification of four triterpenoid saponins from the chloroform extract of the leaves of Tocoyena brasiliensis: 3-O-β-D-quinovopyranosyl quinovic acid, 3-O-β-D- quinovopyranosyl cincholic acid, 3-O-β-D-glucopyranosyl quinovic acid and the 28-O-β-D-glucopyranosyl ester derivative of quinovic acid as binary mixtures, respectively. From the ethanol extract a flavonoid identified as ramnazin-3-O-rutinoside was obtained. The structures of these compounds were assigned by data analysis of ID and 2D NMR spectrometry and comparison with data recorded in the literature for these compounds.

New biflavonoid and other flavonoids from the leaves of Chimarrhis turbinata and their antioxidant activities

A new biflavonol, named chimarrhoside (1), and eight known flavonol glycosides (2-9), were isolated from the leaves of Chimarrhis turbinata. Their structures were established on the basis of 1D and 2D NMR experiments as quercetin-3-O-rutinoside (2), kaempferol-3-O-rutinoside (3), kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-β-D-galactopyranoside (4), quercetin-3-O-α-L-rhamnopyranosyl-(1→6)-β-D- galactopyranoside (5), 6-hydroxy-rutin (6), kaempferol-3-O-D-galactopyranoside (7), kaempferol-3-O-D-glucopyranoside (8) and kaempferol-3-O-α- Lrhamnopyranosyl-(1→6)-α-L-rhamnopyranosyl-(1→4) -β-D-glucopyranoside (9). In addition, catechin (10) and catechin-(4α→8)-catechin-procyanidin B-3) (11) were isolated. The crude extract, fractions and isolated compounds were evaluated for their antioxidative properties using an autographic assay based on β-carotene bleaching on TLC plates, and spectrophotometric detection by reduction of the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical. Flavonoids 2, 5, 6, 10 and 11 displayed strong free radical scavenging activity, when compared with the standards BHT and rutin. ©2005 Sociedade Brasileira de Química.

Constituintes micromoleculares de Aristolochia melastoma Manso: compostos nitrados

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Investigação eletroquímica e aplicação de eletrodos de carbono vítreo modificados por filmes de poli-L-lisina na determinação de antioxidantes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

A new flavonoid derivative from leaves of Oxandra Sessiliflora R. E. Fries

A fração em acetato de etila (EtOAc) obtida a partir da partição do extrato de etanol (EtOH) das folhas de O. sessiliflora R. E. Fries (Annonaceae) foi submetida a diversos procedimentos cromatográficos, incluindo cromatografia líquida de alta eficiência (HPLC), o que resultou no isolamento dos flavonóides: quercetina-3-O-α-L-ramnopiranosil-(1→4)-β-D-glucopiranosídeo (1), inédito na literatura, canferol-3-O-α-L-ramnopiranosil-(1→4)-β-D-glucopiranosídeo (2), rutina (3) e canferol-3-O-rutinosídeo (4). As estruturas foram definidas através da análise dos espectros de ressonância magnética nuclear (NMR) de ¹H e de 13C (1D e 2D) e espectrometria de massas.