Perfil de flavonoides e avaliação do potencial antioxidante e citotóxico de Bauhinia purpurea (Fabaceae) da região amazônica

Bauhinia purpurea (Fabaceae). Plants of the Bauhinia species (Fabaceae) are used in the treatment of several infections and diseases, especially diabetes. In this paper, with an aim to contribute to the knowledge of the chemistry and therapeutic potential of the Amazonian species of the genus Bauhinia, flavonoid profiles in the leaves and branches of Bauhinia purpurea collected in Manaus and São Gabriel da Cachoeira/AM were analyzed by HPLC-DAD and ESIMS. Differences in the flavonoid profiles were detected along with the presence of rutin and isoquercitrin in the leaves collected in Manaus. All samples showed antioxidant activity, however, no cytotoxic activity against cell lines MDA-MB435 (breast), HCT-8 (colon), and SF-295 (glioblastoma) could be detected.

Intraspecific variability of Holostylis reniformis: concentration of lignans, as determined by maceration and supercritical fluid extraction (SFE-CO2), as a function of plant provenance and plant parts

Maceration and supercritical fluid extraction were used to prepare extracts from parts of plants (Holostylis reniformis) collected in two different regions of Brazil. ¹H NMR, HPLC-DAD-ESI/MS, HPLC-DAD, GC-MS, and chemometric techniques were used to analyse lignans in the extracts and showed that yields of SFE-CO2 were less than or equal to those of hexane maceration extracts. These analyses, in conjunction with the concentrations of aliphatic hydrocarbons, fatty acids and their methyl and ethyl derivatives in the extracts, also allowed the chemical composition of parts and provenance of the plant to be differentiated.

Phenolic composition and antioxidant properties of Brazilian honeys

The antioxidant activities and polyphenolic levels of "assa peixe," "cambara," and "morrão de candeia" Brazilian honeys were investigated. Phenolic extracts of 11 honeys were evaluated spectrophotometrically to determine their total phenolic and flavonoid contents, and their antioxidant activities were measured using DPPH, ABTS, and FRAP assays. High-performance liquid chromatography coupled with diode array detection was applied to determine the phenolic composition of the honey extracts. The presence of fourteen phenolic compounds was established (eleven phenolic acids and three flavonoids), as well as HMF and abscisic acid. Principal component analysis was applied to classify the honey samples according to their floral origins.

DETECTION AND QUANTIFICATION OF PHYTOCHEMICAL MARKERS OF Ilex paraguariensis BY LIQUID CHROMATOGRAPHY

Ilex paraguariensis (yerba-mate) is used as a beverage, and its extract requires adequate quality control methods in order to guarantee quality and safe use. Strategies to develop and optimize a chromatographic method to quantify theobromine, caffeine, and chlorogenic acid in I. paraguariensis extracts were evaluated by applying a quality by design (QbD) model and ultra high-performance liquid chromatography (UHPLC). The presence of these three phytochemical markers in the extracts was evaluated using UHPLC-MS and was confirmed by the chromatographic bands in the total ion current traces (m/z of 181.1 [M+H]+, 195.0 [M+H]+, and 353.0 [M−H]−, respectively). The developed method was then transferred to a high-performance liquid chromatography (HPLC) platform, and the three phytochemical markers were used as external standards in the validation of a method for analyses of these compounds in extracts using a diode array detector (DAD). The validated method was applied to quantify the chlorogenic acid, caffeine, and theobromine in the samples. HPLC-DAD chromatographic fingerprinting was also used in a multivariate approach to process the entire data and to separate the I. paraguariensis extracts into two groups. The developed method is very useful for qualifying and quantifying I. paraguariensis extracts.

Conversion of hydroxycinnamic acids into volatile phenols in a synthetic medium and in red wine by Dekkera bruxellensis

The conversion of p-coumaric acid, ferulic acid, and caffeic acid into 4-ethylphenol, 4-ethylguaiacol and 4-ethylcatechol was studied in Dekkera bruxellensis ISA 1791 under defined conditions in a synthetic medium and in a red wine. Liquid chromatography (HPLC-DAD) was used to quantify the phenolic acids, and gas chromatography (GC) coupled to a FID detector was used to quantify volatile phenols using a novel analytical methodology that does not require sample derivatization. Identification was achieved by gas chromatography-mass detection (GC-MS). The results show that phenolic acids concentration decreases while volatile phenols concentration increases. The proportion of caffeic acid taken up by Dekkera bruxellensis is lower than that for p-coumaric or ferulic acid; therefore less 4-ethylcatechol is formed. More important, 4-ethylcathecol synthesis by Dekkera bruxellensis in wine has never been demonstrated so far. These results contribute decisively to a better understanding of the origin of the volatile phenols in wines. The accumulation of these compounds in wine is nowadays regarded as one of the key factors of quality control.

Study of the physical and physicochemical characteristics of fruits of the licuri palm (Syagrus coronata (Mart.) Becc.) found in the Atlantic Forest of Minas Gerais, Brazil

AbstractThe Atlantic Forest has species of native fruits, consumed fresh and processed, which have an important contribution to food sovereignty of families that consume it. This study examined the physical and physicochemical characteristics, proximate composition, concentration of carotenoids, vitamin C, vitamin E and minerals in the pulp and kernels of fruits of licuri (Syagrus coronata (Mart.) Becc.). Titratable acidity was analyzed by volumetric neutralization, soluble solids by refractometry, proteins by the micro-Kjeldahl method, lipids by gravimetry using soxhlet, dietary fiber by non-enzymatic gravimetry, carotenoids and vitamin C by HPLC-DAD, vitamin E by HPLC-fluorescence, and minerals by ICP-AES. Pulp were a source of Zn (0.95 mg 100–1), a good source of fiber (6.15 g 100–1), excellent source of provitamin A (758.75 RAE 100–1), Cu (0.69 mg 100–1), Fe (3.81 mg 100–1), Mn (3.40 mg 100–1) and Mo (0.06 mg 100–1). The kernel were a source of Fe (3.36 mg 100–1) and excellent source of Mn (6.14 mg 100–1), Cu (0.97 mg 100–1) and Mo (0.07 mg 100–1). The nutritional value and wide availability of licuri fruit make it an important resource for reducing food insecurity and improving nutrition of the rural population and other individuals who have access to it.

Tanniinien ja proteiinien väliseen vuorovaikutuksen tutkimiseen käytettävät kemialliset menetelmät

Tanniinit ovat kasvien tuottamiin sekundaarimetaboliitteihin kuuluva polyfenolinen yhdisteryhmä. Maakasvit tuottavat aineenvaihdunnaassaan kahdenlaisia tanniineja: hydrolysoituvia tanniineja sekä kondensoituneita tanniineja eli proantosyanidiineja. Tanniinien on jo satojen vuosien ajan tiedetty saostavan proteiineja, mutta vasta nykyaikaisilla analyysitekniikoilla on onnistuttu tutkimaan saostureaktion aikana vallitsevia vuorovaikutuksia ja sidoksen muodostusta. Merkittävimpiä analyysitekniikoita tanniini‒proteiini -kompleksien tutkimuksessa ovat isoterminen titrauskalorimetria, massaspektrometria, turbidimetriset menetelmät sekä elektroforeettiset menetelmät. Tanniini‒proteiini -kompleksit muodostuvat olosuhteista riippuen joko kovalenttisesti tai ei-kovalenttisesti. Kovalenttiset kompleksit muodostuvat alkaalisissa ja hapettavissa olosuhteissa ja ne pysyvät todennäköisesti liukoisessa muodossa. Neutraaleissa tai heikosti happamissa olosuhteissa syntyy ei-kovalenttisia tanniini‒proteiini -komplekseja, jotka ovat herkästi saostuvia. Merkittävimipiä ei-kovalenttisia vuorovaikutuksia kompleksinmuodostuksessa ovat vetysitoutuminen ja hydrofobiset vuorovaikutukset. Liukoisten ja liukenemattomien kompleksien tutkimuksessa on usein turvauduttava eri analyysimenetelmiin, koska vain harva kokeellinen menetelmä soveltuu sekä liukoisille että saostuville analyyteille. Tämän tutkielman kirjallisessa osassa käsitellään lyhyesti tämänhetkinen käsitys ei-kovalenttisesta ja kovalenttisesta tanniini‒proteiini -kompleksinmuodostuksesta. Kirjallisessa osassa keskitytään niiden analyysimenetelmien käsittelyyn, joita on käytetty tanniini‒proteiini -vuorovaikutusten tutkimuksessa. Kokeellisessa osassa kehitettiin uusia menetelmiä tanniini‒proteiini -kompleksien tutkimiseen sekä yksi menetelmä proteiiniaffiniteettisten yhdisteiden tunnistamiseen kasviuutteesta. Ensimmäisessä kehitetyssä menetelmässä tanniini‒proteiini -komplekseja sekä reagoimatonta proteiinia analysoitiin UHPLC- ja HPLC-DAD-MS/MS-laitteistojen avulla, jolloin voitiin tarkkailla proteiinin massaspektrin muutoksia sekä eluutiokäyttäytymistä käänteisfaasiolosuhteissa ennen ja jälkeen kompleksin muodostuksen. Toisessa uudessa menetelmässä proteiiniin sitoutuneiden tanniinien osuus määritettiin käänteisesti kokonaisfenolipitoisuuden mittaukseen tarkoitetulla spektrofotometrisella menetelmällä. Lopuksi määritettiin kasviuutteen proteiiniin sitoutuvat yhdisteet vertaamalla alkuperäisen uutteen yhdistekoostumusta sellaisen uutteen yhdistekoostumukseen, josta proteiineihin sitoutuvat yhdisteet oli poistettu. Yhdisteiden määritykseen käytettiin UHPLC-DAD-MS/MS-laitteistoa.

Chemical characterisation of wild populations of Thymus from different climatic regions in southeast Spain

Thymus is taxonomically a very complex genus with a high frequency of hybridisation and introgression among sympatric species. The variation in accumulation of leaf-surface flavonoids was investigated in 71 wild populations of Thymus front different putative hybrid swarm areas in Andalucia, Spain. Twenty-two flavones, five flavanones, two dihydroflavonols, a flavonol and two unknowns were detected by HPLC-DAD combined with LC-APCI-MS analysis. The majority of compounds were flavones with a lutelin-type substitution of the B-ring, in contrast to previous reports on Macedonian taxa, which predominantly accumulate flavones with apigenin-type substitution of the B-ring. Anatomical and morphometric studies, supported by cluster analysis, identified pure Thymus hyemalis and Thymus baeticus populations, and a large number of putative hybrids. Flavonoid variation was closely related to morphological variation in all populations and is suspected to be a result of genetic polymorphism. Principal component analysis identified the presence of species-specific and geographically linked chemotypes and putative hybrids with mixed morphological and chemical characteristics. Qualitative and quantitative flavonoid accumulation appears to be genetically regulated, while external factors play a secondary role. Flavonoid profiles can thus provide diagnostic markers for the taxonomy of Thymus and are also useful in detecting hybridising taxa. (C) 2007 Elsevier Ltd. All rights reserved.

A new type of Brazilian propolis: Prenylated benzophenones in propolis from Amazon and effects against cariogenic bacteria

Ethanol extracts of four propolis samples (E1-E4) from Manaus (Brazilian Amazon) were analysed by HPLC/DAD/ESI-MS/MS and GC/EIMS. The major constituents of E2 and E4 were analysed by NMR ((1)H and (13)C) and ESI/MS/MS. The main constituents of E2 and E4 are polyprenylated benzophenones: 7-epi-nemorosone, 7-epi-clusianone (major E4 constituents), xanthochymol and gambogenone (major E2 constituents), making up a chemical profile so far unreported for Brazilian propolis. Aristhophenone, methyl insigninone, 18-ethyloxy-17-hydroxy-17,18-dihydroscrobiculatone B, and derivatives of dimethyl weddellianone A and B, propolones, and a scrobiculatone derivative, were detected as minor constituents. Triterpenoids (beta-amyrins, beta-amyrenone, lupeol and lupenone) were ubiquitous and predominant in El and E3. The extracts E2 and E4 were highly active against the cariogenic bacteria Streptococcus mitis, Streptococcus mutans and Streptococcus salivarius. E2 was more active than E4, probably due to a higher content of 2-epi-nemorosone, while the latter was richer in di-hydroxylated compounds. (C) 2010 Elsevier Ltd. All rights reserved.

Coumarins and phenolic fingerprints of oak and Brazilian woods extracted by sugarcane spirit

A total of 25 sugarcane spirit extracts of six different Brazilian woods and oak, commonly used by cooperage industries for aging cachaca, were analyzed for the presence of 14 phenolic compounds (ellagic acid, gallic acid, vanillin, syringaldehyde, synapaldehyde, coniferaldehyde, vanillic acid, syringic acid, quercetin, trans-resveratrol, catechin, epicatechin, eugenol, and myricetin) and two coumarins (scopoletin and coumarin) by HPLC-DAD-fluorescence and HPLC-ESI-MS(n). Furthermore, an HPLC-DAD chromatographic fingerprint was build-up using chemometric analysis based on the chromatographic elution profiles of the extracts monitored at 280 nm. Major components identified and quantified in Brazilian wood extracts were coumarin, ellagic acid, and catechin, whereas oak extracts shown a major contribution of catechin, vanillic acid, and syringaldehyde. The main difference observed among oak and Brazilian woods remains in the concentration of coumarin, catechin, syringaldehyde, and coniferaldehyde. The chemometric analysis of the quantitative profile of the 14 phenolic compounds and two coumarins in the wood extracts provides a differentiation between the Brazilian wood and oak extracts. The chromatographic fingerprint treated by multivariate analysis revealed significant differences among Brazilian woods themselves and oak, clearly defining six groups of wood extracts: (i) oak extracts, (ii) jatoba extracts, (iii) cabreuva-parda extracts, (iv) amendoim extracts, (v) canela-sassafras extracts and (vi) pequi extracts.

Olfactory perception threshold assessment of volatile acidity in Madeira wines

Madeira wine is a fortified wine with impact in the Madeira Island’s economy. Similarly to other wines, its acidity should be well controlled in order to ensure Madeira wine quality, mostly the volatile acidity. Due to Madeira wine complex flavour, it is crucial to get a better knowledge about the volatile acidity impact in its features, namely determine the perception limit of acetic acid and ethyl acetate, as both are the main contributors for volatile acidity. Firstly, the olfactory perception threshold of volatile acidity was assessed by a trained and an untrained panel, using 5 and 10 years-old Sercial and Malvasia wines. Moreover, the current work also presents the evolution of organic acids, acetic acid and ethyl acetate during 540 days of ageing of Madeira wines (Malvasia, Bual, Verdelho and Sercial), comparing the same wines aged by both traditional ageing processes: canteiro and estufagem. Other wine samples, aged in wood casks (canteiro) for at least 5 years, were also evaluated. HS-SPME followed by GC-MS analysis was used to determine ethyl acetate concentration and IEC-HPLC-DAD was used for the organic acids determination, including acetic acid. The results indicated that acetic acid and ethyl acetate olfactory perception threshold depends essentially on wine’s age. Concerning acetic acid, the untrained panel was in average 5.45 g/L (5 years-old) and 6.22 g/L (10 years-old). Training the expert panel to recognize acetic acid odour, the values decreased for 1.44 g/L (5 years-old) and 1.87 g/L (10 years-old), but still remained higher than the established volatile acidity legal limits. Ethyl acetate threshold was similar for both panels (in average 327.97 mg/L). Both compounds tend to increase exponentially with age, being more evident in sweet wines. Organic acids in young Madeira wines depend mostly on the nature of grape varieties, but this difference is minimized with wine ageing.

Desenvolvimento e validação de método analítico para determinação de WE010, WE014 e WE017

The synthetic guanylhydrazones WE010 (3,5-di-tert-butil-4-hidroxibenzaldehyde-guanylhydrazone), WE014 (4-bifenilcarboxialdehydeguanylhydrazone) and WE017 (3,4-diclorobenzaldehydeguanylhydrazone) showed high cytotoxic activity in terms of percentage inhibition of cancer cells growth. However, further progress in the development of these drug candidates requires precise and convenient methods for their qualitative and quantitative analyses. The aim of this study was to develop and validate High Performance Liquid Chromatography with diode-array detection (HPLC-DAD) and Ultra Fast Liquid Chromatography with diode-array detection (UFLC-DAD) methods suitable for as simultaneous as isolated determination of studied guanylhydrazones, based on the optimization of chromatographic parameters and obtaining reduced detection times. The chromatographic analyses of analytes by HPLC were performed on C18 ACE analytical column (150 mm x 4.6 mm), with a particle size of 5.0 μm. Among all the conditions assayed, the best results of separation were obtained with a mixture of methanol:water (60:40, v/v) as the mobile phase at a flow rate 1.5mL/min and pH of 3.5 adjusted at acetic acid. The UFLC method was developed by experimetal desing techniques in order to find optimal chromatographic analytical conditions, which were achieved on XR-ODS analytical column (50 mm x 3.0 mm), with a particle size of 2,2 μm, maintained at 25 ºC. The mobile phase was consisted of methanol:water (65:35, v/v) with 0.1% triethylamine (TEA) and pH of 3.5 adjusted at acetic acid, at a flow rate 0.5 mL/min. The procedure were validated following evaluating parameters such as specificity, linearity, limits of detection (LD) and quantification (LQ), precision, accuracy and robustness, giving results within the acceptable range. Although the UFLC method shows better sensitivity (lower values of LD and LQ), robustness (lower rates of relative standard deviation) and minimize spending time and solvent, both developed methods were adequately applied to the analysis of guanylhydrazones molecules, may be used in routine of quality control laboratories. Keywords: guanylhydrazones, HPLC/DAD, UFLC/DAD, validation of analitical method

Caracterização dos marcadores químicos e avaliação de atividades biológicas do extrato de Spondias mombim

Spondias mombin is a fruitful species dispersed in tropical regions of America, Africa and Asia. In Brazil, the species can be found mainly in the northern and northeastern regions. Scarce chemical and pharmacological studies have been reported for S. mombin and until this moment studies about chemical markers were not developed. In this context, the aims of this study were to characterize the chemical markers from S. mombin leaves and evaluate their anti-inflammatory, antioxidant and antiproliferative potentials. The chemical profile of the hydroethanolic extract from S. mombin leaves analyzed by HPLC-DAD, through a validated method, allowed the identification and quantification of ellagic acid and chlorogenic acid. This extract showed anti-inflammatory potential in acute peritonitis model induced by carrageenan. The hydroethanolic extract from S. mombin leaves was subjected to a liquid-liquid partition with the solvents: n-hexane, dichloromethane, ethyl acetate and n-butanol. Regarding the anti-inflammatory potential of the fractions obtained they were active; however, ethyl acetate fraction at 200 mg/kg showed highlighted results. The compounds ellagic acid and chlorogenic acid also inhibited the leukocyte migration to the site of inflammation at 2.5, 5 and 10 mg/kg. The hydroethanolic extract, fractions and the chemical markers showed significant antioxidant potential when evaluated in different assays: DPPH Free-Radical Scavenging, Superoxide Radical Scavenging, Hydroxyl Radicals Scavenging and Reducing Power. Taken together our results showed that hydroethanolic extract of S. mombin leaves has ellagic acid and chlorogenic acid as bioactive markers and it demonstrated antiinflammatory and antioxidant properties besides no cytotoxicity against 3T3 cells. It enables us to suggest S. mombin as an important species to develop herbal drugs

Estudo da funcionalidade de espécies comestíveis do semiárido nordestino e estratégias para sua utilização como ingredientes para fins alimentícios

The Brazilian caatinga is characterized by low annual rainfall and arid soils. Several cactaceae, either native or adapted species, grow in this semi-arid region, including the prickly pear (Opuntia fícus indica) and facheiro ((Philosocereus pachycladus Ritter) which produce underexploited edible fruits. In addition to these species, the algaroba is a leguminous with little studied technological applications and bioactive potential so far. Therefore, this research aims to investigate the physicochemical, bioactive and functional attributes of the prickly pear and facheiro fruit pulps and the algaroba flour. Specifically, this study approaches the physicochemical characterization, total phenolic compounds (TPC) and the betalain identification and quantification by HPLC-DAD-ESI-MS. It is also investigated the DPPH antioxidant capacity and the antienzymatic activities against alpha-amylase and alphaglucosidase of water and ethanolic extracts of these food material. In order to address their potential to be used as food ingredients, juice blends prepared with mixtures of cajá and prickly pear, biofilms with facheiro and cereal bars with algaroba flour were elaborated and analyzed. The prickly pear fruits presented low acidity and high sugar content when compared to facheiro. The Philosocereus pachycladus Ritter fruits had higher protein and ash content, but the algaroba flour was the species with higher protein and sugar content among all. The algaroba flour also presented outstanding food fiber content, which reveals its potentiality to be used as a natural intestinal regulator. The TPC of water and ethanol extracts ranged from 3.87 to 16.21 mg GAE/100g for algaroba flour, 79.24 to 110.20 GAE/ 100g for prickly pear and 412.23 to 539.14 mg GAE/100g for facheiro. The 70% (w/v) ethanol extract reached the highest DPPH antioxidant activity, which was linearly correlated to its high TPC content. In regard to the enzymatic inhibitory activities, the best performance was observed for the prickly pear extracts which presented a moderate inhibition for both investigated enzymes, but interestingly, no alpha-glucosidase inhibition was observed for facheiro extracts. This work shows, for the first time in the literature, the functional attributes of facheiro fruits, as well as the presence of betacianins and isobetanin in the pulp of this exotic fruit. When it comes to the food products developed here, the sensory attributes that better described the juice blend cajá-prickly pear were sweetness, acidity, color yellow-orange, body, turbidity and cajá flavor. The discriminative test applied for cereal bars produced with and without algaroba revealed that the texture was the only sensory attribute that differed (p<0.05) between these two samples. It was also observed that the addition of facheiro extracts did not influence the visual characteristics of the biofilms. Overall, this work unveils the physicochemical and bioactive attributes of these commercial and technologically underexploited species widely found in the Brazilian caatinga and presents alternatives for their rational use

Evaluation of anti-Mycobacterium tuberculosis activity of Campomanesia adamantium (Myrtaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)