Rubus rosifolius (Rosaceae) stem extract induces cell injury and apoptosis in human hepatoma cell line.

Rubus rosifolius, popularly known as "red mulberry", is a common medicinal plant in southern Brazil and is used as an antidiarrheal, analgesic, antimicrobial and antihypertensive, and to treat stomach diseases. The aim of this study was to analyze the R. rosifolius stem extract (RrSE) for possible in vitro cytotoxic and genotoxic effects, using the comet assay and the micronucleus test to assess genotoxicity, and flow cytometry to assess the impact on the cell cycle and apoptosis in HepG2/C3A cells, in addition to evaluating the expression of genes linked to the induction of DNA damage, cell cycle, apoptosis and metabolism of xenobiotics. The MTT assay observed no cytotoxic effects at concentrations between 0.01 and 100 µg/mL of the extract. However, genotoxic effects occurred in treatments with the extract from a 1 µg/mL concentration. Flow cytometry analysis revealed a significant increase in cells in the G2/M phase after treatment with 10 µg/mL, a decrease in cells in the G0/G1 phase in the treatment with 100 µg/mL, and a significant increase in total apoptotic cells. In the gene expression analysis, an increase in the CYP1A2 xenobiotics metabolizing gene expression was observed. Despite the promising pharmacological effects of R. rosifolius, the results revealed that the RrSE has genotoxic effect and induces apoptosis in HepG2/C3A cells, indicating danger in using this plant extract by humans.

New substances of Equisetum hyemale L. extracts and their in vivo antitumoral effect against oral squamous cell carcinoma.

ETHNOBOTANICAL RELEVANCE: Equisetum hyemale is used in traditional medicine as an anti-inflammatory, antioxidant, diuretic and anticancer agent. Recent studies have observed antiproliferative activity of this species in some tumor cell lines. AIM OF THE STUDY: The aim of this study was to evaluate the antiproliferative activity of the ethanol extract of E. hyemale and its partitions in oral squamous carcinoma cell lines, the death pathways induced by the most active partition, the acute toxicity and therapeutic activity, and the identification of the main compounds. MATERIALS AND METHODS: The ethanol crude extract was prepared from the stems of E. hyemale and partitions were obtained from this extract with n-hexane, dichloromethane and ethyl acetate. Cytotoxicity assays were performed using MTT on human oral tumor lines SCC-9, SCC4 and SCC-25, and normal primary fibroblasts. The main pathways of programmed cell death were analyzed. Acute toxicity in mice was performed using the most active partition, ethyl acetate. Antitumor activity was accessed in xenotransplants grafts of SCC-9 cells in Balb/nude mice. Phytochemical analysis was performed using UHPLC-MS/MS and dereplication was done using Global Natural Product Social Molecular Networking (GNPS) analysis. RESULTS: Ethanol extract, n-hexane and ethyl acetate partitions showed dose-dependent activity and selectivity towards oral tumor cells, with the ethyl acetate being the most bioactive. This medium polarity partition was shown to induce tumor cell death through apoptosis due to the presence of activated caspase 3/7, DNA fragmentation, chromatin condensation and phosphatidylserine exposure. The ethyl acetate partition also produced low toxicity in mice, provoking mild hepatic changes, but without causing necrosis and significantly reduced tumors volume and weight in xenotransplants of SCC-9 cells. Phytochemical analysis allowed identification of kaempferol glycosides and cinnamic acid derivatives previously described for E. hyemale. In addition it was possible to identify 6 new non-glycolyzed flavonoids 5-Hydroxy-3',4',7,8-tetramethoxyflavone (14), 5,4'-Dihydroxy-7,8,3'-trimethoxyflavone (15), 5,7-Dihydroxy-3',4'-dimethoxyflavone (16), 3',4,5,7-Tretramethoxyflavone (17), 5-Hydroxy-3'4',7-trimethoxyflavone (18), and 5,4'-Dihydroxy-3'-7'-dimethoxyflavone (19); besides 5 compounds already determined to be cytotoxic in other species, Isoferulic acid (1), Ferulic acid (2), Atractylenolide III (6), Dihydroxy-3',4'-dimethoxyflavone (16), and 5-Hydroxy-3'4 ',7-trimethoxyflavone (18). CONCLUSION: The results indicate that the E. hyemale extract and partitions inhibited 3 different cell lines of OSCC in a highly selective nontoxic way by inducing apoptosis of the cells. We identified 6 new non-glycosylated flavonoids and 5 other substances in this species.

Simultaneous UHPLC-MS Quantification of Catechins and Untargeted Metabolomic Profiling for Proof-of-Concept Authenticity Determination of Maytenus ssp. Samples.

Due to the widespread use of Maytenus ilicifolia leaves in the treatment of gastric ulcers, herbal medicines derived from such species are distributed by the national health system in Brazil. A related species, Maytenus aquifolium, is also used for the same disorders, and both are popularly known as Espinheira-santa. Due to their popular use, the quality and efficiency of the herbal medicines derived from these species is an important public health issue. The purpose of this study was to develop and test an analytical method that could quantify the content of catechin and epicatechin in dry Maytenus spp. leaves and simultaneously obtain their chemical profile to determine authenticity of the leaf samples. Ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-MS) was used to quantify these isomers, in the selected ion monitoring (SIM) mode, while simultaneously analyzing the extract in full-scan mode. This approach was successfully applied to the analysis of commercial and authentic samples of Maytenus spp. Fewer than half the samples presented the minimum epicatechin content of 2.8 mg per g of dry leaf mass, as specified in the 6th Brazilian Pharmacopoeia (2019) for M. ilicifolia. Furthermore, by using untargeted metabolomics, it was observed that the chemical profile of most the samples was not compatible with M. ilicifolia leaves, indicating the need for stricter quality control of this material. The method described herein could be used for this control; moreover, its concept could be adapted and used for an ample variety of medicinal plant products.

Antioxidant effect of chamomile tea on the salivary glands of streptozotocin-induced diabetic rats.

This study aimed to analyze oxidative stress and the activity of antioxidant enzymes in the salivary glands of streptozotocin (STZ)-induced diabetic rats with ad libitum consumption of chamomile tea in substitution of water for 21 days. Rats were divided in two control groups (untreated control and treated control) and two diabetic groups (untreated diabetic and treated diabetic). Superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) activities, total antioxidant status (TAS), and malondialdehyde (MDA) concentrations were determined. The chemical composition of the chamomile essential oil revealed 39 compounds, accounting for 93.5% of the total oils. The polyphenolic profile of the tea showed the presence of apigenin, luteolin, umbelliferone, and esculetin. SOD, GPx, CAT, and TAS levels were lower in the parotid (PA) diabetic glands, but treatment increased their concentration in both the submandibular (SM) and PA diabetic salivary glands. Increased MDA levels were observed in the PA diabetic glands, which were decreased by the consumption of chamomile tea with a reduction in hyperglycemia compared to that in untreated diabetic rats. However, the SM diabetic glands showed no difference in the MDA content. The consumption of chamomile tea prevented oxidative stress in the PA glands of diabetic rats, exhibiting hypoglycemic and antioxidant effects. Thus, chamomile tea could be a potential candidate for preventing oral complications in diabetes mellitus.

Bioactive compounds of parsley (Petroselinum crispum), chives (Allium schoenoprasum L) and their mixture (Brazilian cheiro-verde) as promising antioxidant and anti-cholesterol oxidation agents in a food system.

This study determined the bioactive composition and antioxidant potential of parsley, chives and their mixture (Brazilian cheiro-verde). Additionally, the effect of these herbs against cholesterol oxidation in grilled sardines (Sardinella brasiliensis) was also investigated. Ultra-high Performance Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (UHPLC-ESI-MS) analyses revealed the presence of phenolic acids (caffeic, chlorogenic, and ferulic acids) and flavonoids (apigenin, kaempferol, catechin) in the herbs. Higher levels of phenolics (2.10 ± 0.02 mg GAE/g) and carotenoids (205.95 ± 0.17 µg/g) were determined in parsley extracts. Moreover, parsley also presented higher antioxidant capacity by DPPH (59.21 ± 0.07 %) and ORAC (109.94 ± 18.7 µM TE/g) than the other herbs. In vivo analyses demonstrated that the herbs' extracts decreased the damage on Saccharomyces cerevisiae cells exposed to H2O2, except the chives extract at 10 µg/mL. Higher levels of cholesterol oxidation products (COPs) were determined after grilling. The total COPs increased from 61.8 ± 0.7 (raw fish) to 139.7 ± 10.1 µg/g (control). However, the addition of herbs effectively reduced cholesterol oxides formation, this effect was more pronounced in fish containing 4% parsley and 4% cheiro-verde. Promising results were found for cheiro-verde; however, it did not present synergic antioxidant effects.

Systemic antioxidant and anti-inflammatory effects of yellow passion fruit bagasse extract during prostate cancer progression.

We evaluated the impact of yellow passion fruit (Passiflora edulis sp.) bagasse extract (PFBE) administration in systemic oxidative and inflammatory parameters in vivo, considering prostate cancer progression in transgenic mice (TRAMP). Piceatannol, scirpusin-B, dicaffeoylquinic acid, citric acid, and (+)-catechin were identified in PFBE, and the extract showed high in vitro antioxidant capacity. Some alterations in systemic parameters were verified during prostate cancer progression, as the increase in ALT and MDA levels, and SOD and GPx activities in the plasma. In the liver, higher MDA, TNF-α, and NF-κB levels, and GR and GPx activities were verified. Compared to their respective controls, the short- and long-term PFBE administration reduced MDA levels in the liver and plasma. The long-term treatment increased the catalase activity in the plasma, while the short-term treatment increased the hepatic SOD and catalase activities. Still, a reduction in hepatic TNF-α and NF-κB levels was verified after long-term treatment. PRACTICAL APPLICATIONS: Prostate cancer progression is associated with changes in systemic redox status and inflammation markers. Moreover, the intake of polyphenols with antioxidant properties, besides delaying prostate carcinogenesis, may improve the systemic antioxidant defenses and inflammatory response. In vitro studies pointed to a promising antioxidant and anti-inflammatory potential of yellow passion fruit bagasse. However, in vivo studies are scarce. Our results provided information about in vivo impacts of PFBE oral consumption on antioxidant defense and inflammation, indicating its potential as an adjuvant during the initial steps of prostate cancer.

The anticholesterol oxidation effects of garlic (Allium sativum L.) and leek (Allium ampeloprasum L.) in frozen fish burgers submitted to grilling.

This study determined the total phenolic content and antioxidant capacity of garlic (Allium sativum L.) and leek (Allium ampeloprasum L.), as well as evaluated their anticholesterol oxidation potential in fish burgers. The total phenolic contents were 1.1 ± 0.1 mg GAE/g FW to garlic and 1.3 ± 0.4 mg GAE/g FW for leek. Leek extract showed antioxidant activity index (1.3 ± 0.01) in DPPH and ß-carotene/linoleic acid assay (66.5 ± 1.6%); however, in ORAC assay, no statistic differences were observed (P > 0.05). Besides that, bioactive compounds of garlic and leek extracts were identified by ultra-high performance liquid chromatography-electrospray by ionization-mass spectrometry (UHPLC-ESI-MS). Fish burgers were prepared using different concentrations of leek and garlic and stored at -18 °C for 90 days. Thus, at days 0, 30, 60, and 90, the samples were grilled and analyzed as to their cholesterol and cholesterol oxidation products contents. Storage and grilling led to an increase in cholesterol oxidation products; however, addition of garlic and leek minimized cholesterol oxidation products formation. After 90 days, samples containing 3% leek + 0.5% garlic ware the most effective in inhibiting the cholesterol oxides formation during storage and showed the lowest increase in cholesterol oxidation products content (21.16%). Thus, the findings of this research indicate the potential application of garlic and leek as natural inhibitors of cholesterol oxidation in food. PRACTICAL APPLICATION: Garlic and leek have a set of bioactive compounds with a wide antioxidant capacity when used in meat foods such as fish burgers. Garlic and leek used as natural antioxidants perform well in the shelf life of fish burgers and can be substitutes for synthetic antioxidants in this type of product. The presence of both vegetables reduced the formation of prejudicial products to human health generated during the shelf life of the food.

Effect of aroeira (Schinus terebinthifolius Raddi) fruit against polyunsaturated fatty acids and cholesterol thermo-oxidation in model systems containing sardine oil (Sardinella brasiliensis).

This study evaluated the protective effect of ground aroeira (Schinus terebinthifolius Raddi) fruit addition against fatty acids and cholesterol oxidation in model systems containing sardine oil (Sardinella brasiliensis) during heating (150 and 180 °C). High temperatures reduced the amount of essential polyunsaturated fatty acids and caused the formation of oxidized products. Total cholesterol oxides content increased from 58.9 ± 0.26 to 577.5 ± 2.14 µg/g oil, after heating at 180 °C. However, aroeira significantly protected lipids from oxidation. Although the synthetic antioxidant applied as standard (butylated hydroxytoluene) showed greater results, it was used in the maximum concentration permitted by Brazilian legislation (0.01%), suggesting that aroeira fruit could be used as a natural antioxidant for the food industry. The protective effect of aroeira may be correlated to its antioxidant capacity and the presence of bioactive compounds which were identified by UHPLC-ESI-MS in the aroeira extract.

Aroeira fruit (Schinus terebinthifolius Raddi) as a natural antioxidant: Chemical constituents, bioactive compounds and in vitro and in vivo antioxidant capacity.

Schinus terebinthifolius Raddi fruit, known as Brazilian pepper or aroeira, is a natural source of bioactive compounds. However, studies about the antioxidant and nutritional contribution of this fruit in food systems are limited. Regarding the presence of bioactive compounds, flavonoids showed the highest level (10.33 ± 0.34 mg QE/g), and potential antioxidant components such biflavonoids were determined by ultra-high performance liquid chromatography/electrospray ionization mass spectrometry. The aroeira fruit extract showed antioxidant potential in DPPH (42.68 ± 0.05%), ORAC (43.40 ± 6.22 µM TE/g) and ß- carotene/linoleic acid (61.41 ± 5.30%) assays. Besides that, in vivo analyses demonstrated the ability of aroeira extracts to decrease the damage caused by oxidative stress promoted by H2O2 in Saccharomyces cerevisiae cells. Thus, the presence of phytochemicals with functional properties and the antioxidant capacity of aroeira fruit indicate its use as a potential natural antioxidant for the food industry.

Stability of hydroalcoholic extracts of two species of guaco; Mikania glomerata SPRENG. and Mikania laevigata SCHULTZ. (Asteraceae), by UHPLC-MS

It is important to study the stability of plant extracts used as active ingredients in phytotherapic medicine, as degradation of the active principles directly affects the efficacy and safety of these products. Therefore, a stability study of the hydroalcoholic extract of the species: Mikania glomerata and Mikania laevigata was conducted in order to determine the speed of degradation and shelf life of these extracts, which are incorporated in cough syrup in Brazil. Leaves of both species were dried in an oven or by lyophilization (freeze-dried). Hydroalcoholic extracts underwent both accelerated stability study of six months and long-term stability study for 12 months. Samples were stored at different temperatures and every three months were analysed by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) to monitor their chemical profile, quantifying coumarin and chlorogenic acid. For all conditions of the study, a reduction of the content of the chemical marker of this species, coumarin, greater than 5% was observed, so a shelf life of two years cannot be assigned to the hydroalcoholic extracts of these species as observed in commercial extracts.

Influence of environmental factors on the volatile composition of two Brazilian medicinal plants: Mikania laevigata and Mikania glomerata.

INTRODUCTION: Mikania laevigata Sch.Bip. ex Baker and Mikania glomerata Spreng. are medicinal plants popularly known as guaco, used for inflammatory diseases of the respiratory system, included in pharmaceutical formularies and often used without distinction. However, several studies show that the chemical composition varies between these species, as well as in plants are subjected to different environmental stresses. Few studies have been carried out with the volatile compounds of guaco, even less about the changes in volatile composition due to abiotic variation. OBJECTIVE: The aim of this work was to evaluate how volatile compounds vary according to the seasons and at different times of the day and if these compounds are influenced by the variations in the growth conditions such as: temperature, luminosity and water. METHODS: The headspace volatiles of the leaves were sampled by solid phase micro extraction and analyzed by gas chromatography-mass spectrometry. Untargeted metabolomic analysis of the resulting chromatograms and chemometrics was applied. The chemical profile of the volatiles of M. laevigata and M. glomerata were different; being clearly separated in the exploratory grouping analyzes (PCA), followed by analysis of variance of the marker compounds of both species. RESULTS: Only M. laevigata contained coumarin, considered to be the chemical marker of both species and to be responsible for the therapeutic activities. There was no significant difference between the morning and afternoon collections of either species. Coumarin, α-pinene and bicyclogermacrene were more intense in the volatiles of M. laevigata throughout the year and responsible for grouping the samples of this species. For M. glomerata, hexanal and 2-hexenal were responsible for grouping the samples and were more intense in all months. The growth conditions tested affected the intensity of specific compounds in the chromatograms. Some compounds were less intense with the increase of the temperature and in the plants subjected to full sunlight. However, certain volatile compounds-such as pinenes-were more intense in plants suffering drought. CONCLUSION: The variation in composition between species of guaco was greater than those observed in the seasonal and cultivation studies, indicating that these species cannot be used indistinctly.

Comparison of the Morphology, Anatomy, and Chemical Profile of Mikania glomerata and Mikania laevigata.

Mikania glomerata and Mikania laevigata are medicinal plants popularly used in Brazil for colds and asthma. Although several studies report differences in their composition, they are frequently used indistinctly, which may be due to misidentification of the species or the plant part used. Herein, the chemical, anatomical, and morphological profiles of both species were evaluated, seeking to differentiate them. Due to the pattern of secondary metabolites found in M. glomerata and M. laevigata, the chemical profile of the leaves was the characteristic that best differentiated the two species. Coumarin was present in all the leaf samples of M. laevigata but absent or in low concentration in leaf samples of M. glomerata. The stems presented coumarin in both species, albeit in different concentrations. The quantification of chlorogenic and dicaffeoylquinic acids was crucial for the separation of the two species, as higher concentrations are found in M. glomerata. The anatomic structure of the leaves, stem, and petioles of both species was conserved, even when exposed to different environments, but presented only subtle differences between species. The morphology of the leaves presented different characteristics for each species but altered when exposed to shade. Therefore, the chemical analysis together with the morphological characteristics of plants grown in full sunlight may be used for the correct identification of these species.

Comparative Study of Chemical Composition and Biological Activity of Yellow, Green, Brown, and Red Brazilian Propolis.

The chemical composition and biological activity of a sample of yellow propolis from Mato Grosso do Sul, Brazil (EEP-Y MS), were investigated for the first time and compared with green, brown, and red types of Brazilian propolis and with a sample of yellow propolis from Cuba. Overall, EEP-Y MS had different qualitative chemical profiles, as well as different cytotoxic and antimicrobial activities when compared to the other types of propolis assessed in this study and it is a different chemotype of Brazilian propolis. Absence of phenolic compounds and the presence of mixtures of aliphatic compounds in yellow propolis were determined by analysing (1)H-NMR spectra and fifteen terpenes were identified by GC-MS. EEP-Y MS showed cytotoxic activity against human tumour strain OVCAR-8 but was not active against Gram-negative or Gram-positive bacteria. Our results confirm the difficulty of establishing a uniform quality standard for propolis from diverse geographical origins. The most appropriate pharmacological applications of yellow types of propolis must be further investigated.

Unraveling the biosynthesis of pilocarpine in Pilocarpus microphyllus.

Pilocarpine is found exclusively in species of Pilocarpus and the presence of other imidazole alkaloids has been reported in several species of the genus. Pilocarpine has several important pharmaceutical applications. Although several imidazole alkaloids related to pilocarpine have been reported in the previous years, little is still known about its biosynthetic route. At most, histidine has been reported as the precursor of pilocarpine. Based on our own previous reports and in an experiment where pilocarpine and related alkaloids (pilosine, trachyllophiline and anhydropilosine) were supplied to P. microphyllus leaves and the alkaloid profile analyzed by UPLC-MS, we suggest a biosynthesis pathway for pilocarpine. Further experiments using labeled precursors associated with transcriptome data may allow us to understand the whole biosynthesis pathway and its genetic control.

Green tea extract supplementation induces the lipolytic pathway, attenuates obesity, and reduces low-grade inflammation in mice fed a high-fat diet.

The aim of this study was to evaluate the effects of green tea Camellia sinensis extract on proinflammatory molecules and lipolytic protein levels in adipose tissue of diet-induced obese mice. Animals were randomized into four groups: CW (chow diet and water); CG (chow diet and water + green tea extract); HW (high-fat diet and water); HG (high-fat diet and water + green tea extract). The mice were fed ad libitum with chow or high-fat diet and concomitantly supplemented (oral gavage) with 400 mg/kg body weight/day of green tea extract (CG and HG, resp.). The treatments were performed for eight weeks. UPLC showed that in 10 mg/mL green tea extract, there were 15 µg/mg epigallocatechin, 95 µg/mg epigallocatechin gallate, 20.8 µg/mg epicatechin gallate, and 4.9 µg/mg gallocatechin gallate. Green tea administered concomitantly with a high-fat diet increased HSL, ABHD5, and perilipin in mesenteric adipose tissue, and this was associated with reduced body weight and adipose tissue gain. Further, we observed that green tea supplementation reduced inflammatory cytokine TNFα levels, as well as TLR4, MYD88, and TRAF6 proinflammatory signalling. Our results show that green tea increases the lipolytic pathway and reduces adipose tissue, and this may explain the attenuation of low-grade inflammation in obese mice.

Stimulation of acidic reduction of nitrite to nitric oxide by soybean phenolics: possible relevance to gastrointestinal host defense.

This study aimed to evaluate the potential of soybean-promoted acidic nitrite reduction and to correlate this activity with the content of phenolics and with the bactericidal activity against Escherichia coli O157:H7. Extracts of embrionary axes and cotyledons enriched in phenolics increased •NO formation at acidic pH at values that were 7.1 and 4.5 times higher, respectively, when compared to the reduction of the nonenriched extracts. Among the various phenolics accumulated in the soybean extracts, five stimulated nitrite reduction in the following decreasing order of potency: epicatechin gallate, chlorogenic acid, caffeic acid, galic acid and p-coumaric acid. Extracts of embrionary axes presented higher contents of epicatechin gallate and caffeic acid, compared to that of cotyledons, indicating a positive correlation between activity of the extracts and content of phenolics with regard to nitrite reducing activity. Soybean extracts enriched in phenolics interacted synergistically with acidified nitrite to prevent E. coli O157:H7 growth. The results suggest that soybean phenolics may interfere with the metabolism of •NO in an acidic environment by accelerating the reduction of nitrite, with a potential antimicrobial effect in the stomach.

Essential oil of Lithraea molleoides (Vell): chemical composition and antimicrobial activity

Lithraea molleoides (Vell.) (Anacardiaceae) is a tree found in Brazil, Paraguay, Bolivia, Uruguay, Argentina and Chile. It is popularly used in the form of alcoholic extracts, decoctions and infusions for the treatment of cough, bronchitis, arthritis, diseases of the digestive system and as diuretic, tranquilizer, haemostatic and tonic agent. The objectives of this study were the extraction of the essential oil of the mature fruit, leaves and aerial parts of the plant and quantification of the yield thereof; the identification and quantification of the principal components of the essential oil and the determination of its antimicrobial activity against bacteria and yeast. The essential oil yield of the mature fruits was 1.0 percent but essential oil was not detected in leaves and flowery branches. The gas chromatograph-mass spectrometer (GC-MS) analysis of the essential oil of the mature fruits detected the presence of limonene (89.89 percent), alpha-pinene (3.48 percent), beta-pinene (2.63 percent), alpha-terpineol (1.27 percent), myrcene (0.64 percent), sabinene hydrate (0.54 percent), 4-terpineol (0.28 percent), camphene (0.22 percent) and delta-3-carene (0.13 percent). The essential oil was active against just a few of the Gram-positive bacteria and yeast tested and did not present antimicrobial activity against Gram-negative bacteria.

Lithraea molleoides(Vell.) (Anacardiaceae) é uma árvore encontrada no Brasil, Paraguai, Bolívia, Uruguai, Argentina e Chile. É popularmente usada na forma de extrato alcoólico, decocção e infusão para o tratamento de tosse, bronquite, artrite, doenças do sistema digestivo, como diurético, tranqüilizante, hemostático e tônico. O objetivo do presente estudo foi a extração do óleo essencial dos frutos maduros, folhas e outras partes aéreas da planta e o rendimento do mesmo; a identificação e quantificação dos principais componentes e a determinação da atividade antimicrobiana. O rendimento do óleo essencial dos frutos maduros foi de 1 por cento, entretanto, não foi encontrado óleo essencial nas partes aéreas da planta. A análise do óleo essencial por cromatografia gasosa com espectrometria de massa, mostrou a presença de limoneno (89,89 por cento), alfa-pineno (3,48 por cento), beta-pineno (2,63 por cento), alfa-terpineol (1.27 por cento), mirceno (0,64 por cento), sabineno (0,54 por cento), 4-terpineol (0,28 por cento), canfeno (0,22 por cento) e delta-3-careno (0,13 por cento). O óleo essencial foi ativo contra algumas bactérias Gram positivas e leveduras testadas e não apresentou atividade contra bactérias Gram negativas.