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.

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.

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.

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%).

Descreve-se o estudo morfo-anatômico e a análise de óleo essencial do fruto de Hennecartia omphalandra J. Poisson (Monimiaceae), uma planta tóxica. A ocorrência de feixes recurrentes e de um anel descontínuo de braquiesclereides no hipanto; assim como a presença de células oleíferas no mesocarpo, de "idioesclereídes" formando o endocarpo e de células com espessamento parietal em rede, constituindo a testa da semente, foram considerados caracteres importantes que podem auxiliar na sua morfo-diagnose. A análise do óleo essencial dos frutos revelou a presença de monoterpenos, sesquiterpenos e álcoois sesquiterpênicos, sendo este o grupo mais significativo. Os componentes majoritários identificados foram o biciclogermacreno (19,6%) e o β-felandreno (8,7%), além do germacreno D (5,3%), α-bulneseno (5,1%) e valerianol (4,9%).

The morphological and anatomical aspects of flower and fruit and the essential oil analysis of fruits of Hennecartia omphalandra J. Poisson (Monimiaceae), a toxic plant, are described in this paper. The presence of vascular recurrent bundles and of a ring of brachysclereids in the hypanthium; the oil cells in the mesocarp, the "idiosclereids" that constitutes the endocarp and cells with reticulated thick walls, shaping the seed coat were described as characters that may help in its diagnosis. The analysis of the essential oil of fruits showed the presence of monoterpenes, as well as sesquiterpenes hydrocarbons and alcohols, being the latter the most significant group. The major identified compounds were bicyclogermacrene (19.6%); β-felandrene (8.7%); germacrene D (5.3%); α - bulnesene (5.1%) and valerianol (4.9%).

Lippia lacunosa e L. rotundifolia são duas espécies brasileiras que formam um complexo de difícil delimitação taxonômica. A composição química do óleo essencial das folhas e flores dessas plantas foi investigada por cromatografia com fase gasosa (CG) e por cromatografia com fase gasosa acoplada à espectrometria de massas (CG-EM). Principais constituintes dos óleos essenciais de L. lacunosa (flores e folhas): mirceno (14,7% e 11,9%), mircenona (45,2% e 64,2%), Z-ocimenona (5,7% e 5,2%), e E-ocimenona (14,7% e 4,1%), respectivamente; L. rotundifolia (flores e folhas): a-pineno (8,7% e 1,8%), mirceno (5,1% e 3,6%), limoneno (26,0% e 7,9 %), cis-pinocanfona (4,5% e 3,1%) e mirtenal (22,3% e 16,7%), respectivamente. Os óleos essenciais de L. lacunosa apresentaram um forte e agradável aroma de manga, que foi relacionado à presença de mirceno e mircenona. Diferenças fundamentais na composição química de seus óleos essenciais podem representar uma poderosa ferramenta na classificação botânica das espécies.

Lippia lacunosa and L. rotundifolia (Verbenaceae) are two Brazilian species of complex taxonomic delimitation. The composition of the essential oils from leaves and flowers of these plants was investigated by gas chromatography (GC) and gas chromatography coupled with mass spectrometry (GC-MS) analysis. The major components of the essential oils of flowers and leaves of L. lacunosa were: myrcene (14.7% and 11.9%), myrcenone (45.2% and 64.2%), Z-ocimenone (5.7% and 5.2%), and E-ocimenone (14.7% and 4.1%), respectively; whereas in L. rotundifolia (flowers and leaves) were a-pinene (8.7% and 1.8%), myrcene (5.1% and 3.6%), limonene (26.0% and 7.9 %), cis-pinocamphone (4.5% and 3.1%) and myrtenal (22.3% and 16.7%), respectively. The essential oils from L. lacunosa exhibited a strong and pleasant mango aroma, which was related to the presence of myrcene and myrcenone. The marked differences in the chemical composition of their essential oils may represent a powerful tool for the botanical classification.

Quarenta e oito extratos brutos etanólicos e frações (em hexano, diclorometano, acetato de etila e n-butanol) de dez plantas brasileiras pertencentes às famílias Leguminosae, Monimiaceae e Verbenaceae; uma da Costa Rica (Verbenaceae) e uma da Argentina (Verbenaceae) foram ensaiados para verificação da atividade anti-micobacteriana contra Mycobacterium tuberculosis (ATCC-27294H37Rv), pelo teste do Alamar Blue, a uma concentração fixa de 100 µg/mL. Dentre os quarenta e oito extratos e frações estudados, sete mostraram-se ativos na concentração ensaiada - frações em hexano e diclorometano de folhas de Lantana trifolia, extrato em metanol:água, 1:1 de cascas de Vitex cooperi, frações em hexano e diclorometano de folhas de Lippia lacunosa e de Lippia rotundifolia, sendoque todas essas plantas pertencem à família Verbenaceae.

Forty eight ethanolic crude extracts and fractions (hexane, dichloromethane, ethyl acetate and n-butanol) from ten Brazilian plants (Leguminosae, Monimiaceae and Verbenaceae), 1 from Costa Rica (Verbenaceae) and 1 from Argentina (Verbenaceae) were screened for anti-mycobacterium activity against Mycobacterium tuberculosis (ATCC-27294H37Rv), by the Alamar Blue test, at a fixed concentration of 100 µg/mL. Out of the forty eight, seven were active at this concentration, corresponding to Lantana trifolia (hexane and dichloromethane extracts from leaves), Vitex cooperi (methanol:water, 1:1 extract from barks), Lippia lacunosa (hexane and dichloromethane extracts from leaves) and Lippia rotundifolia (hexane and dichloromethane extracts from leaves), all from the Verbenaceae family.

Proanthocyanidins from P. contorta leaves and from a commercial quebracho extract were isolated and characterized. Flavonoids, catechins and gallic acid were also identified in the extracts of P. contorta. Compounds were evaluated for their antioxidant properties and for their antiviral activity against an acyclovir-resistant herpes simplex virus type 1 strain. The low molecular weight phenolic derivatives and the proanthocyanidins from P. contorta showed the highest antioxidant activity. Purified proanthocyanidins from both P. contorta and quebracho showed the same maximum non toxic concentrations (25 µg/mL), with 82.2% and 100% of virus inhibition, respectively.