NMR and multivariate methods: Identification of chemical markers in extracts of pedra-ume-caá and their antiglycation, antioxidant, and enzymatic inhibition activities.

INTRODUCTION: In Brazil, the plant group popularly known as "pedra-ume-caá" is used in folk medicine for the treatment of diabetes, and its raw material is commonly sold. OBJECTIVE: The aim of the study was to apply a method for chemical identification of extracts of dry pedra-ume-caá leaves using HPLC-high-resolution mass spectrometry (HRMS) and NMR and develop a multivariate model with NMR data to authenticate commercial samples. In addition, to evaluate the biological activities of the extracts. MATERIALS AND METHODS: Dry extracts of Myrcia multiflora, Myrcia amazonica, Myrcia guianensis, Myrcia sylvatica, Eugenia punicifolia leaves, and 15 commercial samples (sold in Manaus and Belém, Brazil) were prepared by infusion. All the extracts were analysed using HPLC-high-resolution mass spectrometry (HRMS), NMR, principal component analysis (PCA), and hierarchical cluster analysis (HCA). The antidiabetic effect of extracts was evaluated according to enzymatic inhibition. Their content of total phenols, cell viability, and antioxidant and antiglycation activities were also determined. RESULTS: HPLC-HRMS and NMR analysis of these extracts permitted the identification of 17 compounds. 1H NMR data combined with multivariate analyses allowed us to conclude that catechin, myricitrin, quercitrin, and gallic and quinic acids are the main chemical markers of pedra-ume-caá species. These markers were identified in 15 commercial samples of pedra-ume-caá. Additionally, only the extracts of M. multiflora and E. punicifolia inhibited α-glucosidase. All the extracts inhibited the formation of advanced glycation end products (AGEs) and showed free-radical-scavenging activity. These extracts did not present cytotoxicity. CONCLUSION: This study revealed the chemical markers of matrices, and it was possible to differentiate the materials marketed as pedra-ume-caá. Moreover, this study corroborates the potential of these species for treating diabetes.

Hypoglycemic effect and toxicity of the dry extract of Eugenia biflora (L.) DC. leaves.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Eugenia biflora (Myrtaceae) are traditionally used by Amazonian populations for the control of diabetes. However, their chemical composition has not yet been described and pharmacological evidence has not been reported. OBJECTIVE: This study aimed to identify the chemical constituents and evaluate the hypoglycemic and toxic effect of the dry extract of the E. biflora leaves (DEEB). MATERIALS AND METHODS: DEEB, obtained by infusion, was analyzed using LC-HRMS and NMR, whose the catechin flavonoid was quantified using NMR. The antidiabetic effect of DEEB was evaluated according to its inhibition of the enzymes α-amylase and α-glucosidase, as well as the content of total phenols, free radical scavengingand antiglycation activities, and its in vitro cell viability. Oral maltose tolerance and chronic multiple dose tests (28 days) in streptozotocin-induced diabetic mice (STZ) were performed. The hypoglycemic effect and toxicity of this extract were evaluated in the multiple dose assay. Biochemical parameters, hemolysis, and levels of the thiobarbituric acid reactive species in the liver were investigated and histopathological analyses of the kidneys and liver were performed. RESULTS: Eight phenolic compounds were identified, with catechin (15.5 ± 1.7 mg g-1) being the majority compound and a possible chemical marker of DEEB. The extract showed inhibition activity of the enzyme α-glucosidase. Chronic administration of DEEB (50 mg/kg of body weight) reduced glucose levels in diabetic animals, similar to acarbose; however, DEEB (100 and 200 mg/kg bw) caused premature death of mice by D22 of the treatment. Our data indicate that one of the mechanisms of toxicity in DEEB may be related to the aggravation of oxidative stress in the liver. This histopathological study indicated that DEEB failed to minimize the progression of the toxicity of diabetes caused by STZ. CONCLUSIONS: This study demonstrated the hypoglycemic potential of E. biflora leaves. However, the prolonged use of this tea can be harmful to its users due to its considerable toxicity, which needs to be better investigated.

qNMR quantification of phenolic compounds in dry extract of Myrcia multiflora leaves and its antioxidant, anti-AGE, and enzymatic inhibition activities.

Myrcia multiflora (Lam.) DC. is often used in Brazilian folk medicine to control diabetes. Analysis using HPLC-HRMS and NMR of the dry extract from the infusion of leaves of this species revealed twelve phenolic compounds. Among these compounds, chlorogenic acid (1), 4-O-caffeoylquinic acid (2), corilagin (3), chebulagic acid (4), pedunculagin (5), quercetin-3-O-ß-2″-galloylglucoside (7), and kaempferol-3-O-rhamnoside (12) are described for the first time in this matrix. Furthermore, six compounds were quantified using qNMR. The compounds in the dry extracts are 3, 6 (myricetin-3-O-d-glucoside), 8 (myricitrin), 9 (hyperoside), 10 (guaijaverin) and 11 (quercitrin). These compounds may be considered chemical markers in this matrix. In addition, this extract presents activities of α-glucosidase inhibition (IC50 = 79.9 µg mL-1) and glycation in vitro (IC50 = 10.2 µg mL-1), in addition to antioxidant activity against DPPH and ABTS radicals (1,856.7 and 1,032.0 µmol TEq, respectively). This extract did not show significant cytotoxicity in human fibroblasts. Therefore, the enzymatic inhibition, anti-AGE (advanced glycation end-products) and antioxidant activities of Myrcia multiflora leaves corroborated its antidiabetic therapeutic potential and instigates future preclinical studies aimed at the treatment of diabetes mellitus and its complications.

Pedra-ume caá fruit: An Amazon cherry rich in phenolic compounds with antiglycant and antioxidant properties.

Species of Eugenia have been used as an antidiabetic natural source. Chemical, antioxidant and antiglycant screening of extracts from pedra-ume caá (Eugenia punicifolia) fruits were performed. 1H NMR assisted by non-supervised chemometric methods were employed for the evaluation of the chemical profiles which were distinguished according to the color of fruit maturation stages, as well as for pulp and seed fruit. Furthermore, 1H NMR fingerprint analysis of the crude extract allowed the identification of quercitrin and myricitrin, beside other nine compounds. The extracts of the yellow (YP) and green (GP) pulps presented higher antiglycant and antioxidant activities. Fresh juice from E. punicifolia was encapsulated in microcapsules produced with dextrose equivalent (DE) of 10, 20 or 30 as wall materials for the maintainment of their antioxidant and antiglycant properties. The more efficient retention of the bioactive compounds was found using the DE30. The Encapsulation Efficiency (EE) and the Retention Efficiency (RE) of this system was found around 89.7% and 97.6%, respectively. In addition, NMR spectra revealed the presence of flavonoids O-glycosylated (quercitrin and myricitrin) which might be related to the antiglycant and antioxidant activities. The YP presented larger content of quercitrin (117.6 ±â€¯0.4 mg per each 100 g of fresh fruit). Therefore, pedra-ume caá should be employed as an alternative nutraceutical source, as well as intherapeutic pourposes.

Chemical, Antioxidant, and Antimicrobial Evaluation of Essential Oils and an Anatomical Study of the Aerial Parts from Baccharis Species (Asteraceae).

The aim of this study was to evaluate the chemical, antioxidant, and antimicrobial activity of the essential oils as well as the anatomy of the aerial parts from Baccharis aracatubaensis, Baccharis burchellii, and Baccharis organensis owing to the therapeutic potential of Baccharis. The volatile constituents were analyzed using GC/MS, the antioxidant activity was evaluated by oxygen radical absorbance capacity (ORACFL ) and DPPH assays, and the antimicrobial activity by a microdilution technique. Of the 56 compounds identified, only seven (ß-caryophyllene, γ-muurolene, bicyclogermacrene, ß-germacrene, spathulenol, τ-muurolol, and α-cadinol) were common in the three specimens studied. Of these, γ-muurolene was found abundantly in B. aracatubaensis, while bicyclogermacrene was abundant in B. burchellii and B. organensis. The essential oils exhibited antioxidant activity in the ORACFL (>500.0 µmol TE g-1 ) and DPPH assays. However, they did not exhibit any antimicrobial activity. Secretory ducts and flagelliform glandular trichomes were observed in the anatomical study of all the Baccharis species studied.

Bacillus megaterium strains derived from water and soil exhibit differential responses to the herbicide mesotrione.

The intense use of herbicides for weed control in agriculture causes selection pressure on soil microbiota and water ecosystems, possibly resulting in changes to microbial processes, such as biogeochemical cycles. These xenobiotics may increase the production of reactive oxygen species and consequently affect the survival of microorganisms, which need to develop strategies to adapt to these conditions and maintain their ecological functionality. This study analyzed the adaptive responses of bacterial isolates belonging to the same species, originating from two different environments (water and soil), and subjected to selection pressure by herbicides. The effects of herbicide Callisto and its active ingredient, mesotrione, induced different adaptation strategies on the cellular, enzymatic, and structural systems of two Bacillus megaterium isolates obtained from these environments. The lipid saturation patterns observed may have affected membrane permeability in response to this herbicide. Moreover, this may have led to different levels of responses involving superoxide dismutase and catalase activities, and enzyme polymorphisms. Due to these response systems, the strain isolated from water exhibited higher growth rates than did the soil strain, in evaluations made in oligotrophic culture media, which would be more like that found in semi-pristine aquatic environments. The influence of the intracellular oxidizing environments, which changed the mode of degradation of mesotrione in our experimental model and produced different metabolites, can also be observed in soil and water at sites related to agriculture. Since the different metabolites may present different levels of toxicity, we suggest that this fact should be considered in studies on the fate of agrochemicals in different environments.

Chemical characterization and antioxidant capacity of the araçá-pera (Psidium acutangulum): An exotic Amazon fruit.

The "araçá-pera" (Psidium acutangulum DC) is an exotic guava consumed by the Brazilian Amazon population. This paper describes 22 compounds from this fruit by UHPLC-HRMS and NMR methods, being one disaccharide, five monosaccharides, two organic acids, one trihydroxycinnamic acid glucopyranosyl, one tannine digalloyl glucopyranosyl, five triterpenoid acids, and six fatty acids. It also quantifies the level of ascorbic acid (AA) by HPLC-MS (74.32±1.23mg/100g of fresh fruit), and defines the chemical antioxidant activities by DPPH and ABTS+ assays (24.96±0.75, 90.57±0.63mg of vitamin C/100g fresh fruit, respectively), and a cell-based assays (76-100%). These results have shown that this exotic guava can be consumed as a nutraceutical ingredient, as well as be used in the production of functional foods in the Amazonian diet to prevent chronic and oxidative diseases.

Morpho-anatomical characteristics of Baccharis glaziovii in support of its pharmacobotany

Baccharis glaziovii Baker, Asteraceae, also known as carqueja or carqueja-arbustinho, is a native shrub of Brazil that reaches 0.5-2.5 m in height. It is a dioecious species that blossoms from September to December. This species has cladodes, which are winged stems that belong to the “carquejas” and are widely used indiscriminately by the population due to their gastric and diuretic properties. Carquejas are included in section Caulopterae and are difficult to identify even for taxonomists or Baccharis specialists. In the present study, a morpho-anatomical (cladodes and leaves) analysis of the medicinal plant was undertaken to improve its identification and add to the knowledge of section Caulopterae. Fragments of cladodes and leaves were collected and prepared in accordance with standard optical and scanning electron microscopy techniques. The morpho-anatomical characteristics found in B. glaziovii, include three-winged stems showing wings in a regular arrangement around the stem axis, short and petiolate leaves, flagelliform and simple non-glandular trichomes, concave-convex midrib, petioles with a concave shape and a slight projection on the adaxial face and convex with three projections on the abaxial surface, and calcium oxalate crystals in the form of raphides, styloids and pyramidal in the perimedullary region of the cladode, when evaluated as a whole, provide support for the quality control. .

Development and validation of an HPLC-MS/MS method for the early diagnosis of aspergillosis.

Invasive aspergillosis is an opportunistic infection that is mainly caused by Aspergillus fumigatus, which is known to produce several secondary metabolites, including gliotoxin, the most abundant metabolite produced during hyphal growth. The diagnosis of invasive aspergillosis is often made late in the infection because of the lack of reliable and feasible diagnostic techniques; therefore, early detection is critical to begin treatment and avoid more serious complications. The present work reports the development and validation of an HPLC-MS/MS method for the detection of gliotoxin in the serum of patients with suspected aspergillosis. Chromatographic separation was achieved using an XBridge C18 column (150 × 2.1 mm id; 5 mm particle size) maintained at 25 °C with the corresponding guard column (XBridge C18, 10 × 2.1 mm id, 5 mm particle size). The mobile phase was composed of a gradient of water and acetonitrile/water (95:5 v/v), both containing 1 mM ammonium formate with a flow rate of 0.45 mL min(-1). Data from the validation studies demonstrate that this new method is highly sensitive, selective, linear, precise, accurate and free from matrix interference. The developed method was successfully applied to samples from patients suspected of having aspergillosis. Therefore, the developed method has considerable potential as a diagnostic technique for aspergillosis.

Polyacetylenes from the leaves of Vernonia scorpioides (Asteraceae) and their antiproliferative and antiherpetic activities.

Polyacetylenes constitute an underexplored and unstable class of compounds that are found mainly in the Apiaceae, Araliaceae and Asteraceae families. Vernonia scorpioides (Lam.) Pers., Asteraceae is a lianous neotropical herb that usually grows in soils that have been deforested and are of poor quality. It is used in folk medicine for the treatment of several skin conditions. This study addresses the characterisation of eight polyacetylenes isolated from the leaves of V. scorpioides. Their structures were established on the basis of 1D and 2D NMR spectroscopy and MS analysis. Ab initio calculations including solvent effects were employed to aid the elucidation of the absolute configurations of the compounds. The in vitro antiproliferative and anti-herpetic activities of the polyacetylenes were determined. The isolated compounds presented no inhibitory effect against a human cell line of non-small cell lung cancer, but presented a mild non-selective in vitro antiviral activity, although their corresponding glycosides were inactive.

Pre-clinical anti-inflammatory aspects of a cuisine and medicinal millennial herb: Malva sylvestris L.

Malva sylvestris has been used since ancient times for its emollient, laxative and anti-inflammatory properties, being extensively used as salads, soups and teas. The preset study evaluated the topical anti-inflammatory action of M. sylvestris hydroalcoholic extract (HE) and its compounds in mice ear inflammation caused by 12-O-tetradecanoylphorbol-acetate in mice. The LC-MS analysis of the HE confirmed the presence of scopoletin, quercetin and malvidin 3-glucoside compounds in the HE of M. sylvestris. Topical application of the HE reduced ear oedema, polymorphonuclear cells influx (myeloperoxydase activity and histological analysis) and interleukin-1ß levels in the tissue. The topical application of the compound present in the HE, malvidin 3-glucoside was also able to inhibit ear oedema and leukocytes migration. The other tested compounds, scopoletin, quercetin and malvidin 3,5-glucoside were able to prevent the formation of oedema and cell infiltration, but with less effectiveness when compared to HE and malvidin 3-glucoside. Therefore, these results consistently support the notion that M. sylvestris leaves possesses topical anti-inflammatory activity, the compound malvidin 3-glucoside seems to be major responsible for this effect, with the participation of other anti-inflammatory compounds in the extract. Thus, as recommended by population, M. sylvestris can be used as a future treatment to skin disorders.

Sesquiterpene lactones from the leaves of Hedyosmum brasiliense (Chloranthaceae).

Hedyosmum brasiliense Miq. is an endemic aromatic arborescent shrub that is the only representative of the Chloranthaceae in Brazil. There have been few studies seeking to determine its chemical constituents and/or pharmacological effects. This work describes the isolation and identification of sesquiterpene lactones from the leaves, including guaianolides, elemanolides and a lindenanolide. These were tested against Mycobacterium tuberculosis, together with podoandin, onoseriolide and some other common phenolics. The structures of the isolated compounds were determined based on extensive analysis of 1D and 2D NMR spectroscopic and MS data, as well as comparison with published data. The compounds found were the guaianolides, 1,2-epoxy-10α-hydroxy-podoandin and 1-hydroxy-10,15-methylenepodoandin, the elemenolide 15-acetoxy-isogermafurenolide and the lindenanolide 8α/ß,9α-hydroxy-onoseriolide, along with the previously isolated guaianolide podoandin, the lindenanolide onoseriolide and the elemenolide 15-hydroxy-isogermafurenolide. The phenolic compounds isolated were scopoletin, vanillin, vanillic acid, protocatechuic aldehyde and ethyl caffeate. The isolated sesquiterpene lactones did not show anti-mycobacterial activity against isoniazid-sensitive M. tuberculosis cultures at concentrations of 1-30 µM.

Hyperpigmentant activity of leaves and flowers extracts of Pyrostegia venusta on murine B16F10 melanoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Pyrostegia venusta is a native Brazilian plant which has a variety of uses in traditional folk medicine including the treatment of vitiligo. However, its effectiveness on melanogenesis is not yet elucidated. AIM OF THE STUDY: This study aimed to investigate the melanogenic activity of hydroalcoholic extracts from the leaves and flowers of P. venusta on murine B16F10 melanoma cells. MATERIALS AND METHODS: Different concentrations of the hydroalcoholic extracts of flowers and leaves of P. venusta were evaluated in trials of spontaneous melanin content (4 days), and cell viability by the MTT assay in murine B16F10 cells, and in the mushroom tyrosinase activity in vitro. RESULTS: Both extracts, leaves (0.1; 0.3; 1 and 3 µg/mL) and flowers (0.03 and 0.1 µg/mL) increased the melanin content in a concentration dependent manner after 4 days of incubation on melanoma cells. Leaves extract promoted enhancement of melanogenesis with maximum effect of 33.3±3% (3 µg/mL), and the flower extract increased in 23.4±3% (0.1 µg/mL). The cell viability test using MTT showed that in the same tested concentrations of both extracts no cell death was detected. Actually, either extract was not able to cause any change in the tyrosinase activity. HPLC analysis of P. venusta extracts found 0.09% and 1.08% of allantoin on leaves and flowers extracts, respectively. CONCLUSIONS: The leaves and flowers extracts of P. venusta stimulates B16F10 melanogenesis at very low concentrations. These findings support the folk medicinal use of P. venusta on the treatment of hypopigmentation diseases, such as vitiligo.

Development and validation of two methods based on high-performance liquid chromatography-tandem mass spectrometry for determining 1,2-benzopyrone, dihydrocoumarin, o-coumaric acid, syringaldehyde and kaurenoic acid in guaco extracts and pharmaceutical preparations.

In this study, two HPLC-ESI-MS/MS methods were developed and validated for the determination of 1,2-benzopyrone (COU), o-coumaric acid (OCA), kaurenoic acid (KAU), syringaldehyde (SYR), and dihydrocoumarin (DIH) in guaco extracts and pharmaceutical preparations (syrup and oral solution). The chromatographic separation was achieved using a C18 XBridge 150×2.1-mm (5-µm particle size) column maintained at 25°C. The mobile phases consisted of a gradient of water and acetonitrile containing 0.05% formic acid or 5 mM ammonium formate for the positive and negative ion modes, respectively. All of the calibration curves showed excellent coefficients of correlation (r≥0.9970) over the ranges of 1.25-400 ng/mL for coumarin, 10-600 ng/mL for dihydrocoumarin, 5-250 ng/mL for KAU, and 25-500 ng/mL for o-coumaric acid and syringaldehyde. The range of recovery was 96.3-103% with an RSD% of <4.85% for intraday and interday precision. The results indicate that the developed methods are fast, efficient, and sensitive for the quantification of the guaco metabolites in extracts and pharmaceutical forms while avoiding purification and derivatization steps.

Mikania glomerata Spreng. e M. laevigata Sch. Bip. ex Baker, Asteraceae: estudos agronômicos, genéticos, morfoanatômicos, químicos, farmacológicos, toxicológicos e uso nos programas de fitoterapia do Brasil / Mikania glomerata Spreng. e M. laevigata Sch. Bip. ex Baker, Asteraceae: agronomic, genetic, anatomical, chemical, pharmacological, toxicological studies and its use in herbal therapy programs in Brazil

No Brasil as espécies de Mikania glomerata Spreng. e M. laevigata Sch. Bip. ex Baker, Asteraceae, popularmente conhecidas como guaco, são amplamente utilizadas com evidências pré-clínicas para o tratamento de doenças onde estejam envolvidas suas ações broncodilatadoras, antialérgicas, antiasmáticas, anti-inflamatórias, antiulcerogênicas e relaxantes da musculatura lisa. Em virtude das propriedades terapêuticas atribuídas as essas espécies, ambas foram oficializadas na farmacopéia brasileira, e por serem bastante semelhantes, dificilmente são diferenciadas. Com o objetivo de contribuir com novas pesquisas, a presente revisão descreve, de forma sistematizada, sobre os aspectos farmacológicos, toxicológicos, químicos, agronômicos, morfoanatômicos, genéticos e o uso nos programas de fitoterapia do Brasil destas espécies. As principais diferenças e semelhanças entre as duas espécies são evidenciadas através da comparação das informações baseadas na literatura específica.

In Brazil the species of Mikania glomerata Spreng. and M. laevigata Sch. Bip. ex Baker, Asteraceae popularly known as guaco, are widely used with preclinical evidence for the treatment of diseases whose actions involve bronchodilators, antiallergic, anti-asthmatic, anti-inflammatory, antiulcerogenic and smooth muscles relaxant. Due to the therapeutic properties attributed to these species, both were included in the official Brazilian Pharmacopoeia, and because of their similarity, they are hardly differentiated. In order to contribute to the new research, this review describes their several aspects: pharmacology, toxicology, chemical, agricultural, anatomical, genetics and the use in herbal medicine programs in Brazil, in a systematic way. The main differences and similarities between the two species are evidenced by comparing the information based on literature.

Sesquiterpene lactones from Vernonia scorpioides and their in vitro cytotoxicity.

Fresh leaves of Vernonia scorpioides are widely used in Brazil to treat a variety of skin disorders. Previous in vivo studies with extracts of this species had also demonstrated a high antitumor potential. This paper reports isolation of four sesquiterpene lactones (hirsutinolides and glaucolides), together with diacetylpiptocarphol, 8-acetyl-13-etoxypiptocarphol, luteolin, apigenin, and ethyl caffeate from fresh leaves and flowers of Vernonia scorpioides. The hypothesis that hirsutinolide 3 is formed during extraction was verified theoretically using Density Functional Theory. The effects of isolated compounds on in vitro tumor cells were investigated, as well as their genotoxicity by means of an in vitro comet assay. The results indicate that glaucolide 2 and hirsutinolide 4 are toxic to HeLa cells. These compounds were genotoxic in vitro, a property that appears to be related to the presence of their epoxy groups, which has been a more reliable indication of toxicity than substitution on C-13 or the presence of alpha,beta-unsaturated keto-groups. These results need to be replicated in vivo in order to ascertain their toxicity.

Alkaloids from the bark of Guatteria hispida and their evaluation as antioxidant and antimicrobial agents.

Phytochemical investigation of the bark of Guatteria hispida afforded three new alkaloids, 9-methoxy-O-methylmoschatoline (1), 9-methoxyisomoschatoline (2), and isocerasonine (3), along with 10 known alkaloids, 8-oxopseudopalmatine (4), O-methylmoschatoline (5), lysicamine (6), liriodenine (7), 10-methoxyliriodenine (8), nornuciferine (9), anonaine (10), xylopine (11), coreximine (12), and isocoreximine (13). The major compounds, 2, 6, 12, and 13, showed significant antioxidant capacity in the ORAC(FL) assay. Compounds 5, 6, and 7 were active against S. epidermidis and C. dubliniensis, with MIC values in the range 12.5-100 microg mL(-1).

A pyrimidine-beta-carboline and other alkaloids from Annona foetida with antileishmanial activity.

Bioassay-guided fractionation of the bark extract of Annona foetida afforded a new antileishmanial pyrimidine-beta-carboline alkaloid, N-hydroxyannomontine (1), together with the previously reported annomontine (2), O-methylmoschatoline (3), and liriodenine (4). The structure of compound 1 was established on the basis of extensive 1D and 2D NMR and MS analyses. This is the third reported pyrimidine-beta-carboline-type alkaloid and is particularly important for Annona genus chemotaxonomy. In addition, all compounds exhibit in vitro antileishmanial activity against promastigote forms of Leishmania braziliensis. Compounds 2 and 4 showed better activity than compounds 1 and 3 against L. braziliensis. Compound 2 was not active against L. guyanensis.

Trypanocidal activity of extracts and fractions of Bertholletia excelsa.

Crude extracts and fractions of Bertholletia excelsa stem barks were tested for trypanocidal activity. Acetone and methanol extracts showed significant in vitro trypanocidal activity against trypomastigote form of Trypanosoma cruzi since in the concentration of 500 microg/ml, the parasites were reduced in 100% and 90.3% respectively, whereas the triterpene betulinic acid pure isolated from hexane extract presented 75.4%.