7a-Hydroxyroyleanone is a bioactive diterpene isolated from leaves of Tetradenia riparia, active against tuberculosis and T. cruzi. In order to use this compound as starting material for the synthesis of compounds with enhanced bioactivity we aimed the dehydration of the 7-alpha-hydroxyl group of royleanone. The reaction performed in acetic acid and hydrochloric acid under reflux gave an unexpected chlorinated naphtoquinone, with aromatization and opening of one of the rings of the starting material, as major product, besides 6,7-dehydroroyleanone and a complex mixture of other products. The reaction products were purified by high-speed countercurrent chromatography with hexane-acetonitrile 1:1. Compounds were tested for their anti-leishmanial activity (L. amazonensis) pointing out a potential leishmanicidal agent which may favor the development of more effective therapies.

A new tetraglycosylated flavonoid derivative of kaempferol was isolated from the butanol extract of leaves of Platycyamus regnellii (Fabaceae) using high-speed countercurrent chromatography (HSCCC). The butanol extract (960.5 mg) was initially fractionated with the solvent system EtOAC-BuOH-H2O 2:8:10 (v/v/v) (elution in normal phase mode). The fractions containing the major tetraglycosylated flavonoid were further purified with the same solvent system by modifying the ratios to 3:7:10 (v/v/v) and alternating the elution to reversed phase mode. The structure of the isolated flavonoid (48.1 mg, 99.7% purity by ultra-high performance liquid chromatography with diode array detector (UHPLC-DAD)) was characterized by spectrometric and spectroscopic methods and identified as kaempferol 3-O-β-D-glucopiranosyl-(1®2)-α-L-rhamnopiranoside-(1®6)-α-L-rhamnopiranoside-7-O-α-L-rhamnopiranoside. Other minor flavonoids were also detected and tentatively identified using ultra-high performance liquid chromatography with diode array detector coupled to mass spectrometry using the electrospray ionization interface (UHPLC-DAD-ESI-MS/MS). By using the latter technique, a second tetraglycosylated flavonol with an O-methyl substituent (either rhamnetin, 3-O-methyl-quercetin or isorhamnetin) was detected, together with a triglycosylated kaempferol and diglycosylated flavonoids (either rhamnetin, 3-O-methyl-quercetin or isorhamnetin).

Abstract Choisya ternata Kunth, C. ternata var. sundance Kunth and the hybrid Choisya ‘Aztec-Pearl’ are three related species belonging to the Rutaceae family. Ethanol extracts were prepared from the leaves of these three species and evaluated in relation to their antioxidant activity using in vitro and ex vivo models. The ethanol extracts belonging to the three species produced a very high antioxidant profile as evidenced by the DPPH radical scavenging activity, the determination of total phenolics and flavonoid equivalent. The generation of reactive species of oxygen in leukocytes stimulated with LPS was dramatically reduced when the three ethanol extracts were used. The alkaloids anhydroevoxine and choisyine were isolated from the ethanol extract of C. ternata using HEMWat (4:6:5:5) as the solvent system by means of high-speed countercurrent chromatography. This was the first time quinoline alkaloids were isolated from this species using HSCCC. These compounds were also assayed for their capacity to inhibit the generation of ROS in leukocytes stimulated by LPS and the results also suggested that they are reactive oxygenase inhibitors.

ABSTRACT This work describes the isolation, by high-speed counter-current chromatography, of the diterpenes manool, jhanol and steviol and the benzaldehyde p-oxy-2-ethylhexyl benzaldehyde from the stilt roots hexane extract of the mangrove plant Rhizophora mangle L., Rhizophoraceae. For this, a non-aqueous biphasic solvent system composed of hexane–acetonitrile–methanol 1:1:0.5 (v/v/v) was applied. As far as we know, only steviol was previously isolated in Rhizophoraceae and this is the first time that p-oxy-2-ethylhexyl benzaldehyde is reported.

ABSTRACT Chronic exposure to solar radiation could contribute to premature skin aging and skin cancer. Skin presents its own antioxidant defense, however when defenses are out of balance, reactive oxygen species could damage biological structures. In the present work, an oil-in-water photoprotective emulsion was developed and Bauhinia microstachya var. massambabensis Vaz, Fabaceae, extracts at 1% (obtained by extraction with different solvents) were added to this emulsion. In vitro and in vivo efficacy and safety of the formulations were evaluated. Spectrophotometric methods and in vivo Colipa test were performed to evaluated efficacy of the formulations, through sun protection factor (SPF) determination and UVA protection factor assessment. To the in vitro safety assessment HET-CAM, CAM-TBS and Red Blood Cell tests were performed. Results showed that both extracts contributed to a higher in vivo photoprotection (SPF 18) when compared to the formulation without extract (SPF 13), this result could be attributed to the antioxidant activity of the plant extracts that act by capturing reactive oxygen species. Concerning safety, all formulations were considered non-irritant according to in vitro tests. Formulations containing extracts could be considered efficient and safe for cosmetic use since they presented higher sun protection factor and passed the toxicity tests.

Lippia origanoides Kunth. Verbenaceae, is of great importance in the Brazilian traditional medicine. Because of it, this work had the purpose to contribute to the ethnopharmacological knowledge of L. origanoides through an ethnobotanical survey conducted within quilombola(maroon) communities of Oriximiná, Pará, Brazil. Among 254 plants cited in the survey, L. origanoides stood out among the ten most versatile species. The agreed main uses were to treat menstrual cramps, stomachache, and baby and postpartum colic. This could indicate a consensus of the informants to possible antispasmodic, anti-inflammatory and analgesic activities of L. origanoides.Therefore, anti-inflammatory and analgesic activities of L. origanoides extract (aerial parts) were assessed through thermal (hot plate) and chemical (formalin and acetic acid) models of nociception. A dose-dependent reduction in acetic acid-induced writhing was observed after treating mice with L. origanoidesextract. The same extract also inhibited significantly formalin-induced licking response and proved to have a central antinociceptive effect, in the hot plate test. This work demonstrates that L. origanoides is used specially by quilombola women from Oriximiná for disorders of the genitourinary system and that biological activities of this species could contribute to these uses. Furthermore, it was also observed antispasmodic, analgesic and antimicrobial uses of other species of the genus Lippia (Goniostachyum section), rich in thymol and carvacrol.

O raspador Acentroptera pulchella (Chrysomelidae, Hispinae) foi observado se alimentando de folhas de Aechmea blanchetiana (Bromeliaceae), mas não de forma indiscriminada. Este inseto aparentemente concentra seu ataque na metade superior das folhas intermediárias do tanque da bromélia. A fim de caracterizar a potencial influência de estratégias defensivas contra herbivoria no padrão alimentar intra-específico de A. pulchella, sobre folhas da bromélia A. blanchetiana, nós: i) quantificamos a aparente preferência intra-específica deste padrão alimentar, ii) comparamos folhas e partes distintas em relação à estratégias morfo-fisiológicas, mecânicas, nutricionais e químicas contra herbivoria e iii) avaliamos se algumas ou todas estas estratégias defensivas ajudariam a explicar o padrão alimentar observado. O raspador A. pulchella atacada preferencialmente a metade superior de folhas intermediárias e adultas de A. blanchetiana, mas não a metade inferior das mesmas folhas, as quais são mais resistentes à perfuração, mais espessas e apresentam menor teor de nitrogênio do que a parte superior. Ao mesmo tempo, porém, folhas jovens e tenras não foram atacadas. As metades atacadas também apresentaram maior teor de fenóis totais em comparação as não atacadas, indicando que os fenóis não atuam defensivamente neste caso. Outras estratégias morfo-fisiológicas e químicas contra herbivoria não variaram entre as partes ou folhas aqui avaliadas. O herbívoro estudado se alimenta preferencialmente de partes das folhas com maior teor de nitrogênio, evitando ao mesmo tempo outras com maior resistência à perfuração. Concluímos que o raspador A. pulchella apresenta claro padrão seletivo de forrageamento, onde o mesmo é direcionado mais por benefícios nutricionais do que estritamente por estratégias mecânicas ou químicas contra herbivoria.

The scraper Acentroptera pulchella (Chrysomelidae, Hispinae) feeds on the leaves of Aechmea blanchetiana (Bromeliaceae), but not indiscriminately. This insect apparently concentrates its attack on the apical region of the intermediate leaves from the bromeliad tank. To characterise the potential influence of leaf defensive strategies on the intraspecific feeding pattern of A. pulchella on the leaves of the bromeliad A. blanchetiana, we quantified the apparent intraspecific preference of this feeding pattern; compared distinct leaves and portions in relation to morpho-physiological, mechanical, nutritional and chemical anti-herbivore strategies; and investigated whether some or all these defensive strategies help explain the observed feeding pattern. A. pulchella preferentially attacked the mid-upper portions of mature and adult leaves of A. blanchetiana, but not the mid-lower portions of the same leaves, which are more resistant to punch, are thicker and present less nitrogen content than the upper portions. However, soft young leaves were not attacked. Attacked portions also presented higher total phenol content than did non-attacked portions, indicating that phenols do not act defensively. Other morpho-physiological and chemical anti-herbivore strategies evaluated did not differ between portions and leaves. This herbivore feeds preferentially on leaf portions that have higher nitrogen levels, avoiding others that present increased punch resistance. The scraper A. pulchella exhibits a clearly selective feeding pattern that is driven by nutritional benefits rather than strictly mechanical or chemical anti-herbivore strategies.

A cromatografia contracorrente (CCC) foi empregada como uma alternativa útil, rápida e econômica às técnicas cromatográficas convencionais na purificação de uma pterocapanoquinona sintética, LQB 118. A separação foi feita em duas etapas utilizando o sistema de solvente hexano-clorofórmio-metanol-água 2:1,5:5:2 em ambas as etapas. A purificação tradicional deste produto por cromatografia em coluna utilizando gel de sílica como fase estacionária requer a utilização de grande quantidade de solvente, sendo bastante demorada, além de permitir a adsorção da substância na coluna. O uso de ¹H RMN para o cálculo do K D da substância alvo foi proposto como uma alternativa às medidas por CLAE.

Countercurrent chromatography (CCC) was employed as a useful, fast and economic alternative to conventional chromatography techniques for the purification of a synthetic pterocarpanquinone, LQB-118. The separation was performed in a two-step CCC with the solvent system hexane-chloroform-methanol-water 2:1.5:5:2 in both steps. Traditional purification of these reaction products by silica gel column chromatography demanded a large amount of solvent and time, besides allowing the irreversible adsorption of the compound in the column. The use of ¹H NMR for the calculation of K D of target compound is proposed as an alternative for HPLC measurements.

The municipality of Oriximiná, Brazil, has 33 quilombola communities in remote areas, endowed with wide experience in the use of medicinal plants. An ethnobotanical survey was carried out in five of these communities. A free-listing method directed for the survey of species locally indicated against Tuberculosis and lung problems was also applied. Data were analyzed by quantitative techniques: saliency index and major use agreement. Thirty four informants related 254 ethnospecies. Among these, 43 were surveyed for possible antimycobacterial activity. As a result of those informations, ten species obtained from the ethnodirected approach (ETHNO) and eighteen species obtained from the random approach (RANDOM) were assayed against Mycobacterium tuberculosis by the microdilution method, using resazurin as an indicator of cell viability. The best results for antimycobacterial activity were obtained of some plants selected by the ethnopharmacological approach (50% ETHNO x 16,7% RANDOM). These results can be even more significant if we consider that the therapeutic success obtained among the quilombola practice is complex, being the use of some plants acting as fortifying agents, depurative, vomitory, purgative and bitter remedy, especially to infectious diseases, of great importance to the communities in the curing or recovering of health as a whole.

Um levantamento etnobotânico realizado em comunidades quilombolas de Oriximiná, Pará, destacou a saracuramirá (SAR), Ampelozizyphus amazonicus Ducke, com vasto uso popular no tratamento da malária, como tônica e depurativa. Por este motivo, o presente trabalho objetivou realizar um estudo etnofarmacognóstico da SAR nas respectivas comunidades. Por meio de uma análise etnobotânica quantitativa, foi verificado que SAR apresentou-se dentre as 10 espécies mais versáteis pela elevada importância relativa (1,3), dentre as cinco espécies com maior importância cultural pelo elevado índice de saliência (0,311) e a espécie com maior concordância de uso principal para malária (85,7%). Uma análise do índice de espuma e do índice de hemólise para SAR demonstra a presença de saponinas com elevado índice de espuma (833) e uma baixa atividade hemolítica (CH50 2,6 mg mL-1). Para realizar uma análise das agliconas das saponinas de SAR, a bebida preparada pelo método tradicional quilombola (BMT) foi hidrolisada e, após reação com diazometano, foi analisada por cromatografia gasosa. Dois sinais majoritários foram caracterizados por espectrometria de massas, um referente a um triterpeno de esqueleto damarânico, característico das saponinas da SAR, e outro referente ao éster metílico do ácido betulínico. Partindo das informações de uso popular da SAR, foi avaliada in vitro a atividade inibidora da acetilcolinesterase. Apesar de BMT não ter mostrado atividade neste ensaio, é possível supor que as indicações de uso desta planta pelos quilombolas como fortificante e contra malária podem estar relacionadas a uma possível atividade adaptógena e imunoestimulante, dada à presença das saponinas e do ácido betulínico em BMT.

In an ethnobotanical survey conducted within "Quilombola" communities of Oriximiná, Pará State, "saracuramirá" (SAR), Ampelozizyphus amazonicus Ducke, stood out as one of the most cited species with wide popular use, especially for malaria, as tonic and depurative. The aim of this paper was to carry out an ethnopharmacognostic study of SAR in these communities. Quantitative ethnobotanical analysis of the data showed that SAR stands out as one of the 10 most versatile species with a high relative importance (1,3); it remains among the five species with the greatest cultural importance, demonstrated by the high salience index (0,311), being the species with the highest major use agreement to malaria (85,7%). Analysis of the foam and hemolysis indexes of SAR shows the presence of saponins with high foam index (833) and low hemolytic activity (HD50 2,6 mg mL-1). In order to analyze SAR saponin aglycones, the drink was prepared by the traditional quilombola method (BMT), was hydrolyzed, and analyzed by gas chromatography after reaction with diazomethane. Two major peaks were characterized by mass spectrometry, one referring to a dammarane triterpene skeleton, characteristic from the SAR saponins, and the other identified as the methyl ester of betulinic acid. Owing to the popular usage of SAR, its in vitro acetylcholinesterase inhibitory activity was evaluated, but with negative results. However, it is possible to suggest that the indications of this plant as a tonic and for treating malaria may be related to an adaptogen and immunostimulant effect due to the presence of saponins and betulinic acid in BMT.

Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae, é uma planta utilizada na medicina popular brasileira contra úlceras e lesões de pele. Internamente é usada contra inflamações, febre, doenças renais e aterosclerose. Essa planta foi estudada com o objetivo de identificar os compostos bioativos majoritários que possam justificar seu uso terapêutico contra lesões de pele e arteriosclerose. A atividade antioxidante do extrato bruto etanólico, partições e os compostos majoritários isolados das raízes de S. cayennesis foi avaliada através de testes in vivo e in vitro. In vitro a atividade antioxidante foi avaliada pelo teste fotocolorimétrico do radical 2,2-difenil-1-picrilidrazil (DPPH•). In vivo, Saccharomyces cerevisiae, um modelo de célula eucariótica, foi utilizado tanto para avaliar a fototoxicidade quanto a capacidade antioxidante contra as espécies reativas de oxigênio (EROS) menadiona e peróxido de hidrogênio. O extrato, partições e os dois compostos majoritários isolados, verbascosídeo e ácido betulínico foram capazes de aumentar a sobrevivência e diminuir a peroxidação lipídica de S. cerevisiae contra EROS. Verbascosídeo apresentou atividade antimicrobiana moderada contra Streptococcus pyogenes, S.epidermidis e Staphylococcus aureus. O extrato etanólico e as partições testadas não apresentaram fototoxicidade, indicando que S. cayennensis é uma planta segura para o tratamento de lesões de pele e como possível ingrediente em cosméticos.

Stachytarpheta cayennensis (Rich.) Vahl, Verbenaceae, plant extract, is a Brazilian medicinal plant externally used in folk medicine for purulent ulcers, skin lesions and internally for inflammations, fever, renal disorders and atherosclerosis. S. cayennesis was studied to identify potential bioactive compounds that may justify their therapeutic use against skin lesions and atherosclerosis. The antioxidant, antimicrobial and phototoxicity capacities of the crude ethanolic extract, fractions and isolated compounds from roots of S. cayennesis were evaluated through in vivo and in vitro tests. Strains of Saccharomyces cerevisiae, an eukaryotic cell model, were used to assess both the phototoxicity and the capacity to protect against the lethal oxidative stress caused by menadione and hydrogen peroxide. The extract, fractions and the two major isolated compounds, verbascoside and betulinic acid, of S. cayennensis were able to increase the tolerance and decrease the lipid peroxidation of S. cerevisiae to reactive oxygen species (ROS). The antioxidant activity was also evaluated by scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH•). Verbascoside exhibited a moderate antimicrobial activity against Streptococcus pyogenes, S.epidermidis and Staphylococcus aureus. Neither the ethanolic extract nor fractions showed phototoxicity, indicating that the S. cayennensis extract is safe for use in the treatment of skin lesions and as an active cosmetic ingredient.

Uma metodologia rápida e eficiente foi desenvolvida para separação de isômeros de isatinas utilizando a técnica de cromatografia contracorrente de alta velocidade. Os isômeros de derivados de isatinas foram preparados segundo a metodologia de Sandmeyer e separados com o sistema de solventes hexano:acetato de etila:etanol:água 1:0,5:0,5:1 (v/v/v/v).

A rapid and efficient method, using high-speed counter-current chromatography (HSCCC) technique, was developed for the separation of isomeric isatin derivatives, prepared following the Sandmeyer route. The biphasic solvent system composed of hexane:ethyl acetate:ethanol:water 1:0.5:0.5:1 (v/v/v/v) was used for all separations.

Com o objetivo de selecionar caracteres úteis na identificação de Polygonum acre Kunth var. aquatile Meisn., erva medicinal geralmente comercializada sob a forma de planta fragmentada, e para contribuir para o conhecimento botânico da espécie, descreveu-se sua anatomia e quantificaram-se os fenóis totais, taninos gálicos e condensados. O material foi processado segundo técnicas usuais de anatomia vegetal. Na análise anatômica, a comparação dos resultados com a bibliografia permitiu selecionar os seguintes caracteres como auxiliares à determinação de P. acre: 1) ausência de endoderme bem definida com estrias de Caspary conforme descrito para outras espécies do gênero Polygonum; 2) ócrea com numerosos tricomas pluricelulares e cavidades secretoras localizadas apenas na face abaxial da epiderme; 3) fibras presentes internamente ao xilema do caule; 4) feixe invertido na nervura principal; e 5) presença de pêlos lignificados no bordo da folha. As dosagens de fenóis totais e taninos demonstraram que a planta apresenta quantidade significativa de taninos condensados e que a princípio, os taninos gálicos não estão presentes na espécie.

Polygonum acre Kunth var. aquatile Meisn., a medicinal plant, was studied with the purpose of selecting useful characteristics for fragmented plant identification as well as to contribute to the botanical study of the species. Usual anatomical analysis was used and total phenolics, condensed and galic tannins were quantified. The comparison with bibliographic data allowed to select some characteristics to help the identification of P. acre: 1) absence of well defined endodermis with Casparian strips on the stem in accordance with others species of the Polygonum genus; 2) numerous multicellular trichomes on the ocrea and secretory cavities only in abaxial epidermis; 3) fibers on the interior of the xylem from the stem; 4) inverted bundle in midrib and 5) lignified protuberances in the edge of the leaf. The quantification of total phenolics and tannins from P. acre shows remarkable amounts of condensed tannins, but no hydrolysable tannins were detected.

Descreve-se a morfo-anatomia e a análise de óleo essencial das folhas de Hennecartia omphalandra J. Poisson (Monimiaceae), uma planta tóxica. A ocorrência de tricomas unicelulares, estômatos paracíticos; braquiesclereídes em grupos no pecíolo e na lâmina foliar, além de esclereídes vermiformes e ramiformes; sistema vascular em arco, com extremidades voltadas para o centro no pecíolo e na forma de arco atenuado na lâmina foliar, sendo o padrão de venação uma variação do tipo craspedódromo-semicraspedódromo, além de óleo essencial armazenado em idioblastos oleíferos não especializados no mesofilo, foram considerados caracteres importantes que podem auxiliar na sua morfo-diagnose. A análise do óleo essencial das folhas revelou a presença de monoterpenos, sesquiterpenos e álcoois sesquiterpênicos, sendo este o grupo mais significativo. Os componentes majoritários identificados foram β-felandreno (24,0%), seguido por germacreno D (10,5%), sabineno (9,3%), biciclogermacreno (8,4%) e β-elemeno (5,3%).

This paper describes the morphological and anatomical aspects and the essential oil analysis of the leaves from Hennecartia omphalandra J. Poisson (Monimiaceae), a toxic plant. The presence of trichomes formed by a single cell, paracitic stomata, groups of braquisclereids in the petiole and leaf blade; vermiform and ramiform sclereids, vascular system in arc softly invaginated at ends in the petiole, and in a flat arc in the middle vein, being the venation pattern a variation of craspedodromous-semicraspedodromous type, and essential oil stored in oil idioblasts in the mesophyll were the most important characters that can help the morphological diagnosis. The analysis of essential oil of the leaves showed the presence of monoterpenes, as well as sesquiterpenes hydrocarbons and alcohols, being the latter the most significant group. The major identified compounds were β-phelandrene (24.0%), germacrene D (10.5%), sabinene (9.3%), bicyclogermacrene (8.4%) and β-elemene (5.3%).

Trinta e seis extratos vegetais originários da Mata Atlântica foram testados quanto à sua atividade antimicobacteriana frente ao M. tuberculosis H37Rv e M. kansasii, utilizando o método REMA em concentrações seriadas de 100 a 0,20 µg/mL. Dentre os trinta e seis extratos testados, cinco mostraram atividade frente ao M. tuberculosis, e destes apenas, três mostraram atividade ao M. kansasii, que apresentou susceptibilidade a outros dez. O teste de citotoxicidade com células VERO foi realizado com os cinco extratos ativos frente ao M. tuberculosis em que identificou-se a não toxicidade em apenas um extrato (Peschiera affinis) na concentração de 100 µg/mL.

Thirty-six plant extracts from the brazilian Atlantic Forest were tested for their antimycobacterial activity against Mycobacterium tuberculosis H37Rv and M. kansasii, using the method REMA in seriate concentrations of 100 to 0.20 µg/mL. Among the thirty six extracts tested, five were active against M. tuberculosis, and three of these extracts also showed activity against M. kansasii. Cytotoxicity test with VERO cells was performed with the five extracts active against M. tuberculosis. Only the extract of Peschiera affinis was identified as non-toxic in the concentration of 100µg/mL.