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 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 Spiranthera odoratissima A. St.-Hil., Rutaceae, known as “manacá” is a shrub native of the Brazilian Cerrado. Their leaves and roots are popularly used to treat rheumatism, infection and abdominal pain. This study analyzed the chemical composition of volatile oils from leaves of S. odoratissima and verified the seasonal variability of its chemical composition. The volatile oils were obtained by hydrodistillation using a Clevenger type apparatus and analyzed by gas chromatography coupled to mass spectrometry. The main chemical components found in samples of volatile oils were β-caryophyllene, bicyclogermacrene, δ-cadinene, amorphous-4,7(11)-diene, α-epi-muurolol, α-cadinol, α-muurolol and γ-cadinene. The hierarchical clustering identified three groups: the first was characterized by α-epi-muurolol, the second by amorphous-4,7(11)-diene and the third group was characterized by α-muurolol. The discriminant canonical analysis was used to differentiate between clusters on the basis of oil composition. The results suggest that the rainfall presented a relationship with the chemical composition of the volatile oil. This is the first study conducted on the seasonal behavior of the chemical constituents in volatile oil from leaves of S. odoratissima.

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 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.

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.

As sementes de guaraná (Paullinia cupana Kunth, Sapindaceae) são utilizadas como estimulante e os principais constituintes químicos são as metilxantinas e os taninos, tais como catequina e epicatequina. Este trabalho teve por objetivo investigar um método por cromatografia líquida de alta eficiência (CLAE) para quantificar, simultaneamente, cafeína, teofilina, teobromina, catequina e epicatequina em pó de guaraná. Os valores obtidos foram comparados com os resultados encontrados no método espectrofotométrico da Farmacopéia Brasileira IV ed. Pelo método cromatográfico, os teores de metilxantinas e taninos totais foram de 4,04% e 1,48%, respectivamente. Os teores de metilxantinas e taninos totais, obtidos pelo método espectrofotométrico, foram de 4,88% e 4,05%, respectivamente. Acredita-se que, nas condições experimentais empregadas, o método espectrofotométrico não foi específico, quantificando outras substâncias fenólicas presentes no guaraná. Portanto, sugere-se o emprego desse método cromatográfico no controle de qualidade do guaraná em pó, por ser preciso, sensível, específico e rápido.

The seeds of Paullinia cupana Kunth, Sapindaceae, are used as a stimulant and the main chemical constituents are the methylxanthines and tannins, as catechin and epicatechin. This study aimed to investigate a method by high performance liquid chromatography (HPLC) to quantify both caffeine, theophylline, theobromine, catechin and epicatechin in powdered of P. cupana. The values obtained were compared with the results of the spectrophotometric method of the Brazilian Pharmacopoeia IV ed. For the chromatographic method, the levels of methylxanthines and total tannins were 4.04% and 1.48%, respectively. The levels of methylxanthines and total tannins obtained by the spectrophotometric method, were 4.88% and 4.05%, respectively. It is believed that, under our experimental conditions, the spectrophotometric method was not specific and quantified others phenolic substances present in the P. cupana. Therefore we suggest the use of this chromatographic method in quality control of P. cupana powder, to be accurate, sensitive, specific and fast.

Memora nodosa (Silva Manso) Miers, Bignoniaceae, é uma planta do Cerrado utilizada popularmente como cicatrizante de úlceras e feridas externas (folhas e caules). O objetivo deste trabalho foi caracterizar morfologicamente essa espécie e fazer o estudo anatômico de suas folhas. O estudo morfológico foi realizado à vista desarmada e com o auxílio de microscópio estereoscópico. Para o estudo anatômico as folhas foram seccionadas e as lâminas histológicas confeccionadas de acordo com as técnicas convencionais. Memora nodosa é um arbusto com folhas opostas cruzadas, recompostas, bipinadas, imparipinadas, com foliólulos pequenos, lanceolados, verde-escuros e brilhantes. A inflorescência é racemosa com flores infundibuliformes e corola amarelo-ouro; glândulas nectaríferas são observadas no terço superior do cálice; androceu com quatro estames férteis, didínamos e um estaminódio reduzido; gineceu com estilete terminal, ovário súpero, assentado sobre um disco nectarífero. O fruto é do tipo cápsula septífraga achatada, com sementes aladas. Anatomicamente a lâmina do foliólulo é hipoestomática com estômatos predominantemente paracíticos. O parênquima paliçádico é pluriestratificado, ocupando cerca de dois terços do mesofilo dorsiventral. Tricomas glandulares e tectores são observados, em maior número, nas folhas jovens.

Memora nodosa (Silva Manso) Miers, Bignoniaceae, is a Brazilian Savannah plant, popularly used as external wound and ulcer healing (leaves and stems). The goal of this research was to perform the morphological study of M. nodosa species and the anatomical study of its leaves. The morphological study was done to the naked eye and with the aid of a stereoscopic microscope. For the anatomical study leaves were sectioned and histological slides made in accordance with conventional techniques. Memora nodosa is a shrub with decussate leaves recomposed, bipinnate, imparipinnate with small leaflets, lanceolate, dark green and shining. The inflorescence is racemose with infundibuliform flowers and gold yellow corolla; nectar glands are observed in the upper third calyx; androecium with four fertile stamens, didynamus and one staminode; gynoecium with terminal style, superior ovary, seated on a nectary disc. The fruit is flattened septifragal capsule with winged seeds. Anatomically, the blade leaflet is hypostomatic with predominantly paracytic stomata. The palisade parenchyma is pluriseriate, occupying about two thirds of the dorsiventral mesophyll. Glandular and non-glandular trichomes are observed in greater numbers in the young leaves.

Hyptidendron canum (Pohl ex Benth.) Harley, Lamiaceae, é utilizada popularmente como antimalárica, antiinflamatória, antiulcerativa, anti-hepatotóxica e anticancerígena. O objetivo deste trabalho foi realizar o estudo morfo-anatômico das folhas e caules e identificar as principais classes de metabólitos secundários presentes nas folhas de H. canum, dados ainda não descritos na literatura. As folhas e caules jovens coletados em Goiânia (GO) foram seccionados à mão livre e preparados para análise microscópica. Foram realizadas reações de identificação de metabólitos secundários do material dessecado e pulverizado. Preparou-se o extrato etanólico bruto, que posteriormente foi fracionado por partição líquido-líquido com hexano, clorofórmio e acetato de etila. As frações foram submetidas à análise cromatográfica em camada delgada (CCD). As lâminas foliares apresentam epiderme adaxial constituída por células poligonais com parede reta. Na epiderme abaxial observam-se células com parede reta a ondulada e estômatos diacíticos e anisocíticos. Tricomas tectores e glandulares estão presente em ambas as faces da lâmina foliar. O pecíolo apresenta aspecto canaletado, epiderme adaxial e abaxial unisseriada. O caule, em secção transversal possui contorno em geral quadrangular, com presença de tricomas tectores e glandulares. As reações e a CCD das folhas evidenciaram a presença de flavonóides, saponinas, terpenos e lignanas. Este trabalho contribuiu para um maior conhecimento da morfo-anatomia e das classes químicas presentes em H. canum.

Hyptidendron canum(Pohl ex Benth.) Harley, Lamiaceae, is popularly used as an antimalarial, anti-inflammatory, antiulcerative, antihepatotoxic and anticancer agent. The goal of this research was to perform the morphoanatomy study of H. canumleaves and stem and identify the main classes of secondary metabolites present in the of H. canumleaves. Such data have not been reported in the literature. The young leaves and stems were collected in Goiânia (GO), hand sectioned and prepared for microscope analysis. Reactions were performed for the identification of secondary metabolites of the dried and pulverized material. The crude ethanol extract was prepared and then fractioned by liquid-liquid partition with hexane, chloroform and ethyl acetate. Thin layer chromatography (TLC) analysis was performed on the fractions. The leaf blades presented adaxial epidermis constituted of polygonal cells with straight walls. On the abaxial epidermis cells with straight to wavy walls and diacytic and anisocytic stomates were noted. Non-glandular and glandular trichomes are present on both faces of the leaf blade. The petiole is grooved, and it presents single layered adaxial and abaxial epidermis. The cross section of the stem presents a generally quadrangular contour with the presence of non-glandular and glandular trichomes. The leaf reactions and TLC evidenced the presence of flavonoids, saponins, terpenes and lignanes. This works helps to increase knowledge of the morphoanatomy and the chemical classes present in H. canum.

This study evaluates the bioactivity of the crude ethanol extract and ethyl acetate, hexane and chloroform fractions obtained from Eugenia uniflora leaves using the hepatopancreas of Oreochromis niloticus L. as an experimental model. The ethanol extract and fractions were administered to the fish orally with their feed. Twenty-four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that the crude extract and the ethyl, chloroform and hexane fractions induced vasodilation, vascular congestion and toxicity due to the presence of eosinophilic granular cells, rodlet cells, some leukocytic infiltrate and rare focal necroses. The Nile tilapia proved to be a satisfactory model for screening plant products.

Hyptidendron canum (Pohl ex Benth.) Harley is a native tree of the Brazilian Savannah. The fish Oreochromis niloticus L. was used as an experimental model to determine the bioactivity of the crude ethanol extract as well as ethyl acetate, hexanic and chloroform fractions obtained from its leaves. The plant ethanol extract and fractions were administered to the fish orally with their feed. Twenty four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that both the crude ethanol extract and the fractions from H. canum induced vasoactive activity, causing vasodilation and vascular congestion, and the hexanic fraction also caused an apparent proliferation of capillaries. Hepatopancreas toxicity was evident through inflammatory processes. Pancreatic (chloroform fraction) and hepatic alterations, hemorrhagic spots and necroses were observed in fish treated with-ethanol extract and fractions. This study is the first description of the biologic action of the crude ethanol extract and the hexane, ethyl acetate and chloroform fractions in fish.