Eugenia punicifolia (Kunth) DC, known as “pedra ume-caá” or “pitanguinha do cerrado”, is popularly used to treat infectious diseases like inflammation, and diabetes. Therefore, this study aimed to develop and validate an analytical method by high-performance liquid chromatography with diode array detector (HPLC-DAD) for the seasonal quantification of gallic acid (GA) and ellagic acid (EA), and to co-validate the method for quantification from the ethanol extract and fractions of the E. punicifolia leaves. Chromatographic separations were performed with a mobile phase gradient of acetonitrile/methanol/water acidified with 0.2% formic acid, a flow of 0.8 mL min 1, detection at 254 nm, and a C18 column (250 × 4.6 mm, 5 µm). Method validation parameters were determined following Brazilian legislation. The HPLC-DAD analytical method developed for the quantification of GA and EA was validated and demonstrated to be selective, linear, precise, accurate, robust, and without matrix effect. Accordingly, it is helpful for the analysis of these tannins in crude ethanol extracts and fractions from E. punicifoila leaves. Thus, our results may be of great help for quality control assessment and standardization of raw materials containing Eugenia punicifolia.

Abstract The antinociceptive and anti-inflammatory activities of crude ethanolic extract of Celtis iguanaea leaves and their active fractions are reported. The oral treatment with crude ethanolic extract (CEE; 100, 300 or 1000 mg/Kg) inhibited the number of writhings in a dose-dependent manner. The intermediate dose also inhibited formalin-induced nociception in both phases. The oral treatment with dichloromethane fraction (DF; 9 mg/Kg) produced antinociceptive effect in both phases of formalin test; however, the treatment with ethyl acetate fraction (EAF; 16 mg/Kg) reduced pain only in the second phase of this test. The oral treatments with CEE (300 mg/Kg) or DF (9 mg/Kg) reduced the nociception induced by capsaicin and pre-treatment with naloxone did not change these effects. The oral administration of CEE (300 mg/Kg), DF (9 mg/Kg) or ethyl EAF (16 mg/Kg) reduced ear edema, leukocytes migration and myeloperoxidase activity. Furthermore, the oral treatment with CEE (300 mg/Kg) or EAF (16 mg/Kg) reduced the level of Tumor Necrosis Factor - Alpha (TNF-α) in the pleurisy test. In conclusion, the DF showed antinociceptive activity that involves the vanilloid system as well as anti-inflammatory effect and the EAF showed anti-inflammatory activity involving the reduction of TNF-α cytokine.

ABSTRACT In this work, we developed and validated a HPLC-PDA method for the quantification of hibalactone in Hydrocotyle umbellata L., Araliaceae, subterraneous parts extracts and optimized its ultrasound-assisted extraction. Chromatographic separations were carried out with an isocratic mobile phase of acetonitrile/methanol/water (10:65:25), a flow of 0.8 ml min−1, detection at 290 nm and C18 column (250 × 4.6 mm, 5 µm). The method validation parameters were determined according to Brazilian legislation. The optimization of the hibalactone ultrasound-assisted extraction was performed using Box–Behnken design and response surface methodology. The HPLC method for hibalactone quantification proved to be selective, linear, precise, accurate and robust, being useful for the analysis of hibalactone in H. umbellata subterraneous parts extracts. The optimal ultrasound-assisted extraction conditions were obtained with solid-to-liquid ratio of 1:5 g ml−1, ethanolic strength of 70% (v/v) and temperature of 65 °C. The results can provide support of the quality control and standardization of raw materials from H. umbellata.

Abstract Trembleya parviflora (D. Don) Cogn., Melastomataceae, also known as "quaresmeira-branca", is a subshrub that is commonly used to treat verminosis, scabies, dermatoses, rheumatism, vaginal infections, ulcerations and wounds. The aim of this work was to perform a morphological study of T. parviflora, evaluate the composition and chemical variability of the volatile oils from the leaves, perform phytochemical screening of the powder from the leaves and to define parameters for quality control of the plant material. Macroscopic characterization of T. parviflora was carried out by naked eye in Serra dos Pireneus, Pirenópolis, Goiás for 12 months. Volatile oils were subjected to hydrodistillation with Clevenger apparatus and analyzed by gas chromatography-mass spectrometry. Phytochemical screening and ash and volatile compound content determination were performed by conventional techniques. T. parviflora has simple, oppositely crossed and petiolate leaves. The inflorescence of this plant is a cyme. The presence of coumarins, steroids, triterpenes, flavonoids and tannins was observed. The total ash content was 4.05 ± 0.02%; the insoluble ash content was 0.10 ± 0.03%; and the volatile compound content was 9.53 ± 0.02%. The major compounds present in the volatile oils were α-terpineol (2.7–16.5%), α-pinene (0.6–25.4%), β-pinene (2.7–23.1%), sabinene (1.2–14.1%), acetoxyeudesman-4-α-ol (0.6–6.3%) and 2,4a-8,8-tetramethyldecahydrocyclopropanaphtalene (2.4–24.4). Two clusters were identified: Cluster I represented the period with low levels of rainfall, and Cluster II represented the period with high levels of rainfall. This study provides data that can be applied for the quality control of powdered leaves and is the first description of the chemical composition and variability of the volatile oils from the leaves of T. parviflora.

ABSTRACT Campomanesia adamantium (Cambess.) O. Berg., Myrtaceae, is a plant popularly used for its anti-inflammatory, anti-diarrhoeal and urinary antiseptic activities. The aims of this study were to obtain the crude ethanolic extract and the hexane, dichloromethane, ethyl acetate, aqueous and concentrated aqueous tannin fractions from C. adamantium leaves, perform biomonitored fractionation to isolate and identify chemical compounds, study the chemical composition of the volatile oils of the leaves and flowers and test the antimicrobial activity of the ethanolic extract, fractions, isolated substances and volatile oils. Phytochemical screening and chromatographic and spectrometric techniques were used. Volatile oils were isolated by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. The antimicrobial activity was tested by a broth microdilution test. The component stictane-3,22-diol was isolated and identified from the hexane fraction, while valoneic and gallic acid were isolated and identified from the concentrated aqueous tannin fraction. The major constituents of the volatile oils of the leaves were verbenene (13.91%), β-funebrene (12.05%) and limonene (10.32%), while those of the volatile oils of the flowers were sabinene (20.45%), limonene (19.33%), α-thujene (8.86%) and methyl salicylate (8.66%). Antibacterial activity was verified for the hexane fraction, while antifungal activity was observed for the aqueous fraction and concentrated aqueous tannin fraction and for vanoleic acid. These results may justify the popular use of C. adamantium.

Abstract This paper describes the quantification of catechin in the spray-dried extract of Pimenta pseudocaryophyllus (Gomes) Landrum, Myrtaceae, citral chemotype using a validated HPLC-PDA method. The method employs a RP-18 column with acetonitrile:water-orthophosphoric acid 0.05% (gradient system) and UV detection at 210 nm. The method was demonstrated to be simple, sensitive, specific, linear, precise, accurate and robust. The response was linear over a range of 5–200 µg/ml (r > 0.999). The range of recoveries was 92.27–102.54%. The relative standard deviation values for intra- and inter-day precision studies were 4.30 and 3.78%, respectively. This assay can be readily utilized as quality control method for catechin in the dried extract of P. pseudocaryophyllus.

ABSTRACT Trembleya phlogiformis DC., Melastomataceae, is a shrub whose leaves are used as a dye for dyeing wool and cotton. The present article aimed to carry out the morphological description of the species, to study the chemical composition of volatile oils from the leaves and flowers and the seasonal variability from the leaves during a year. Macroscopic characterization was carried out with the naked eye and with a stereoscopic microscope. Volatile oils were isolated by hydrodistillation in Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. The major components of the volatile oil of T. phlogiformis flowers were: n-heneicosane (33.5%), phytol (12.3%), n-tricosane (8.4%) and linoleic acid (6.1%). It was verified the existence of a large chemical variability of the volatile oils from the leaves of T. phlogiformis over the months, with the majority compound (oleic alcohol, ranging from 5.7 to 26.8%) present in all samples. A combination of Cluster Analysis and Principal Component Analysis showed the existence of three main clusters, probably related to the seasons. The results suggested that the volatile oils of T. phlogiformis leaves possess high chemical variability, probably related to variation associated with rainfall and the variation in the behavior of specimens throughout the year. This research provides insights for future studies on the volatile oils obtained from the T. phlogiformis leaves and flowers, mainly related to biological markers of applications monitored in the leaves and flowers of this species.

ABSTRACT A 33 Box–Behnken design and Response Surface Methodology were performed to evaluate the influence of extract feed rate, drying air inlet temperature and spray nozzle airflow rate on the process yield, stability parameters (moisture content and water activity) and on several physicomechanical properties of spray-dried rosemary extracts. Powder yield ranged from 17.1 to 74.96%. The spray-dried rosemary extracts showed moisture content and water activity below 5% and 0.5%, respectively, which indicate their chemical and microbiological stabilities. Even without using drying aids, some sets of experimental conditions rendered dried products with suitable flowability and compressibility characteristics for direct preparation of solid dosage forms. Analysis of variance and Response Surface Methodology proved that studied factors significantly affected most of the spray-dried rosemary extract quality indicators at different levels. The main processing parameter affecting the spray-dried rosemary extract characteristics was inlet temperature. The best combination of parameters used to obtain a reasonable yield of stable dry rosemary extracts with adequate technological properties for pharmaceutical purpose involves an extract feed rate of 2 ml/min, 80 °C inlet temperature and 40 l/min SA. The design of experiments approach is an interesting strategy for engineering spray-dried rosemary extracts with improved characteristics for pharmaceutical industrial purpose.

ABSTRACT Hyptis carpinifolia Benth., Lamiaceae, is a species known popularly as "rosmaninho" and "mata-pasto", and leaves are employed in Brazilian folk medicine to treat colds, flu, and rheumatism. The aim of this study was to perform a morphological description of H. carpinifolia and to evaluate the seasonal chemical variability of the leaf essential oils during 12 months. Macroscopic characterization of H. carpinifolia was carried out with the naked eye and with a stereoscopic microscope. Essential oils were isolated from leaves by hydrodistillation in Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. Major compounds were found to be 1,8-cineole (39.6-61.8%), trans-cadina-1(6),4-diene (2.8-17.5%), β-caryophyllene (4.4-10.0%), prenopsan-8-ol (4.2-9.6%) and β-pinene (2.9-5.3%). Results of essential oils compositions were processed by cluster analysis and principal component analysis. Data showed high variability in the concentration of the components. Besides, there was a seasonal variability of chemical composition, probably related mainly to the rainfall regime.

Trichogonia cinerea é endêmica no Brasil e ocorre em áreas de Cerrado e campo rupestre. Neste estudo, foram caracterizados os tricomas glandulares e não glandulares das partes aéreas dessa espécie, determinados os principais eventos no desenvolvimento dos primeiros e identificados os principais constituintes do óleo volátil produzido em seus órgãos aéreos. Folhas completamente expandidas, entrenós, flores, brácteas involucrais e ápices caulinares foram utilizados para a caracterização dos tricomas. Folhas, entrenós, flores e brácteas involucrais foram examinados em microscópio de luz e microscópio eletrônico de varredura, enquanto ápices caulinares foram examinados somente em microscópio de luz. Ramos, em fase reprodutiva, foram usados para a extração e a determinação da composição do óleo volátil. A espécie tem três tipos de tricomas glandulares, bisseriados vesiculosos, bisseriados pedunculados e unisseriados pluricelulares, que secretam óleos voláteis e compostos fenólicos. Os principais componentes identificados no óleo volátil foram 3,5-muuroladieno (39,56%) e hidroxitolueno butilado (13,07%).

Trichogonia cinerea is endemic to Brazil and occurs in areas of cerrado and campo rupestre. In this study, we characterized the glandular and non-glandular trichomes on the aerial parts of this species, determined the principal events in the development of the former, and identified the main constituents of the volatile oil produced in its aerial organs. Fully expanded leaves, internodes, florets, involucral bracts, and stem apices were used for the characterization of trichomes. Leaves, internodes, florets, and involucral bracts were examined by light microscopy and scanning electron microscopy, whereas stem apices were examined only by light microscopy. Branches in the reproductive phase were used for the extraction and determination of the composition of the volatile oil. The species has three types of glandular trichomes, biseriate vesicular, biseriate pedunculate, and multicellular uniseriate, which secrete volatile oils and phenolic compounds. The major components identified in the volatile oil were 3,5-muuroladiene (39.56%) and butylated hydroxytoluene (13.07%).

A presença de sistemas subterrâneos espessados em Asteraceae tem sido amplamente relatada na literatura. Devido à grande complexidade dos sistemas subterrâneos, que podem ser de natureza radicular, caulinar ou mista, as análises morfoanatômicas são importantes para assegurar a utilização da terminologia correta. Este estudo teve por objetivo descrever a morfoanatomia e ontogenia, investigar a presença de metabólitos secundários e avaliar os efeitos da sazonalidade no sistema subterrâneo de Chrysolaena simplex (Less.) Dematt. As amostras foram estudadas usando os protocolos padrão de anatomia vegetal, eletrônica de varredura, histoquímica e fitoquímica. O sistema subterrâneo de C. simplex foi identificado como rizóforo, que se originou do nó cotiledonar. Em indivíduos adultos, com rizóforos completamente desenvolvidos, as raízes primárias degeneram e formam sistemas radiculares adventícios. As gemas, nas porções subterrâneas, promovem o crescimento do rizóforo e formaram caules aéreos quando expostas à luz. Gotas lipofílicas foram evidenciadas nas células parenquimáticas do córtex e medula, endoderme e gemas. Frutanos tipo inulina foram evidenciados no eixo caulinar e gemas do rizóforo. A ocorrência de gemas, metabólitos secundários, frutanos e lipídeos armazenados no rizóforo, podem ser apontados como caracteres adaptativos.

The occurrence of thickened underground systems in Asteraceae is widely reported in the literature. Given the great complexity of underground systems, which may originate from roots, stems, or both, morpho-anatomical analyses are essential to ensure the use of correct terminology. The goals of this study were to describe the morpho-anatomy and ontogeny, investigate the occurrence of secondary metabolites and evaluate the effects of seasonality on the underground system of Chrysolaena simplex (Less.) Dematt. Samples were studied using standard protocols of plant anatomy, scanning electron microscopy, histochemical and phytochemical. The underground system of C. simplex was categorised as a rhizophore which started from cotyledonary node. In adult individuals, with rhizophores completely developed, the primary roots degenerated and adventitious radicular systems are formed. The buds in the subterranean portions promote the rhizophore growing, and form aerial stems when exposed to light. Lipophilic droplets were evident in the parenchymatous cells of the cortex and pith, endodermis and buds. Inulin-type fructans were observed in the stem axis and buds of the rhizophore. The presence of buds, secondary metabolites and the storage of fructans and lipids in the rhizophore can be seen as adaptive traits.

Abstract Hortia oreadica Groppo, Kallunki & Pirani, Rutaceae, known as “para-tudo”, “quina”, and “quina-do-campo”, is used in traditional medicine locally to treat stomach pain and fevers. The aims of this study were: analyze the chemical composition of essential oils from leaves, flowers and fruits of H. oreadica and verify the seasonal variation of the chemical components of essential oils from leaves. The essential oils were obtained by hydrodistillation using a Clevenger type apparatus and analyzed by GC/MS. The major components found in the samples of the essential oils were the amorpha-4,7(11)-diene (29.27% – flowers, 20.26% – fruits, 27.66–37.89% – leaves), bicyclogermacrene (23.28% – flowers, 20.64% – fruits, 14.71% to 31.37% – leaves). This work represents the first study of the chemical composition of essential oils from leaves, flowers and fruits and seasonal variation in the essential oils from leaves of H. oreadica.

RESUMO Memora nodosa, popularmente conhecida como "caroba", é amplamente encontrada em regiões de Cerrado no Brasil. Na Medicina Tradicional, as folhas e as raízes são utilizadas para o tratamento de úlceras externas e as raízes para dores abdominais. O presente trabalho objetivou avaliar o efeito do extrato etanólico das raízes da Memora nodosa (EMN) na mucosa gástrica de camundongos. No modelo de úlcera gástrica induzida por indometacina, o tratamento dos animais com EMN nas doses de 100, 300 e 1000 mg/kg, v.o., foi capaz de reduzir o índice de lesão. Nos modelos de úlcera gástrica induzida por etanol e por estresse o tratamento com EMN na dose de 300 mg/kg foi capaz de proteger a mucosa gástrica em 69 e 43%, respectivamente. Após sete dias de restrição alimentar, os animais tratados com EMN (300 mg/kg v.o.) mostrou uma redução no índice de lesão de 65% quando comparado ao grupo controle. A administração intraduodenal de EMN (300 mg/kg) não alterou os parâmetros de secreção ácida gástrica. O tratamento com EMN (300 mg/kg v.o.) não alterou os níveis de glutationa (GSH), mas foi capaz de aumentar o muco gástrico aderido, quando comparado ao grupo controle lesado. Esses resultados mostram que o EMN possui atividade gastroprotetora provavelmente devido ao aumento de muco gástrico aderido à mucosa gástrica.

ABSTRACT Memora nodosa is popularly known as "caroba" and widely found in the Cerrado regions of Brazil. In traditional medicine, the leaves and stems are used for the healing of external ulcer and the roots for abdominal pain. This study investigated the effect of ethanolic roots extract of Memora nodosa (EMN) on the gastric mucosa of mice. In the indomethacin induced gastric ulcer model, the treatments of the animals with EMN at doses of 100, 300 and 1000 mg/kg, p.o., markedly reduced the index of lesions. In the gastric ulcer models induced by ethanol and cold restraint-stress the previous treatment with EMN at dose of 300 mg/kg showed 69% and 43% of protection, respectively. Seven days after food-restriction, the animals treated with EMN (300 mg/kg p.o.) showed reduction in the index of lesion by 65% as compared to control group. The intraduodenal administration of EMN (300 mg/kg) did not alter the gastric acid secretion parameters. The treatment with EMN (300 mg/kg p.o.) did not alter glutathione levels (GSH), but showed an increase of adhered gastric mucus as compared to the control group with lesion. These results showed that EMN has gastroprotective activity probably due with an increase of adhered gastric mucus.

Os óleos essenciais de quatro espécies de Marsypianthes (Lamiaceae) foram investigados por meio de cromatografia gasosa e análise multivariada. Cada espécie foi representada por duas a sete populações, totalizando dezessete populações. β-Elemeno, (E)-cariofileno, α-humuleno, germacreno D, biciclogermacreno, δ-cadineno, espatulenol, óxido de cariofileno e globulol ocorreram em todas as amostras. As análises de componentes principais e de agrupamento hierárquico evidenciaram a presença de duas seções, uma contendo M. chamaedrys/M. montana (seção A) e a outra contendo M. burchellii (seção B). M. foliolosa apresentou maior complexidade, dividindo-se nas duas seções. Resultados similares foram obtidos de acordo com os esqueletos carbônicos biosssintéticos. Germacranos e biciclogermacranos preponderaram na seção A, enquanto aromadendranos e guaianos caracterizaram a seção B. A análise de redundância canônica mostrou que os agrupamentos não foram influenciados por variáveis edáficas dos locais de amostragem.

Essential oils of four species of Marsypianthes (Lamiaceae) were investigated via gas chromatography and multivariate analysis. Each species was represented by two to seven populations, totaling seventeen populations. β-Elemene, (E)-caryophyllene, α-humulene, germacrene D, bicyclogermacrene, δ-cadinene, spathulenol, caryophyllene oxide, and globulol were found in all samples. Principal component and hierarchical cluster analyses revealed the presence of two sections, one containing M. chamaedrys/M. montana (section A) and the other M. burchellii (section B). M. foliolosa showed higher complexity, being divided in both sections. Similar results were obtained according to biosynthetic carbon skeletons. Germacranes and bicyclogermacranes predominated in section A, whereas aromadendranes, bourbonanes and guaianes characterized section B. Canonical redundancy analysis revealed that clusters were not influenced by edaphic factors in sampling sites.

Levantamentos etnobotânicos de plantas nativas do cerrado evidenciam que as folhas deCeltis iguanaea (Jacq.) Sargent (Cannabaceae), popularmente conhecidas como “esporão-de-galo” são usadas na medicina popular para dores no corpo, asma, cólicas, má-digestão, infecções urinárias, disfunções renais, como estimulante ou como diurético. O presente trabalho teve como objetivo avaliar as possíveis atividades citotóxica, genotóxica e antigenotóxica do extrato aquoso das folhas de C. Iguanaea (CALE) através do teste do micronúcleo em medula óssea de camundongos. Para avaliar a genotoxicidade de CALE, os camundongos foram tratados por via oral com três diferentes concentrações de extrato (100, 300 e 500 mg.kg−1). Para a investigação da atividade antigenotóxica, foram utilizadas as mesmas doses concomitantemente a uma única dose i.p. de Mitomicina C (MMC, 4mg.kg−1). Com exceção do controle negativo (apenas 24h), todas as outras doses tiveram as frequências de eritrócitos policromáticos micronucleados (EPCMN) avaliadas em 24 e 48 horas após o tratamento. A Genotoxicidade foi avaliada por meio da frequência de eritrócitos policromáticos micronucleados (MNPCE), enquanto que a citotoxicidade foi mensurada pela razão entre eritrócitos policromáticos e normocromáticos (PCE / NCE). Os resultados mostraram que CALE não apresentou uma redução significativa na razão PCE/NCE, nem um aumento considerável na frequência de EPCMN. No entanto, CALE reduziu a toxicidade na medula óssea (aumento na razão PCE/NCE) e diminuiu a frequência de micronúcleos induzidos pela MMC. Nos resultados obtidos, CALE não apresentou efeitos citotóxicos e genotóxicos, contudo apresentou ações antigenotóxicas e anticitotóxicas nas condições experimentais aplicadas neste estudo.

Ethnobotanical surveys of Cerrado native plants show that leaves of Celtis iguanaea (Jacq.) Sargent (Cannabaceae), popularly known in Brazil as “esporão de galo”, are used in folk medicine for body pain, asthma, cramps, poor digestion, urinary infection, kidney dysfunctions, as well as a stimulant and diuretic. This work aimed at evaluating possible C. iguanaea aqueous leaf extract (CALE) cytotoxicity, genotoxicity, and antigenotoxicity using the mouse bone marrow micronucleous test. To assess CALE genotoxicity, Swiss mice were orally treated with three different extract concentrations (100, 300, and 500 mgkg−1). To evaluate its antigenotoxicity, the same doses were used simultaneously with a single i.p. dose of mitomycin C (MMC, 4mg.kg−1). The frequencies of micronucleated polychromatic erythrocytes (MNPCE) were evaluated 24 h and 48 h after administration except for the negative control (24 h). Genotoxicity was evaluated using the frequency of micronucleated polychromatic erythrocytes (MNPCE), whereas cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). The results showed that CALE did not exhibit a significant reduction in the PCE/NCE ratio, neither a considerable increase in the frequency of MNPCE. Nonetheless, CALE reduced bone marrow toxicity (increased PCE/NCE ratio) and decreased the micronuclei frequency induced by MMC. We can conclude that CALE presented no cytotoxic and genotoxic effects, but showed antigenotoxic and anticytotoxic actions under the experimental conditions applied in this study.