Influence of extrusion process on the release of phenolic compounds from mango (Mangifera indica L.) bagasse-added confections and evaluation of their bioaccessibility, intestinal permeability, and antioxidant capacity.

Extruded polyphenol-rich by-products like mango bagasse (MB) could be used to manufacture functional confections. However, few reports have assessed the extrusion impact on MB polyphenols within a food matrix. This research aimed to evaluate the impact of extrusion on the bioaccessibility, intestinal permeability, and antioxidant capacity of phenolic compounds (PC) from non-extruded and extruded MB-added confections (EMBC and MBC, respectively). The inhibition of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl radicals and in silico approaches were used to evaluate the antioxidant capacity. MBC displayed the highest gastric bioaccessibility (%) of xanthones and flavonoids, whereas selective release of gallic acid, mangiferin, and quercetin glucoside was shown for EMBC. Lower PC' apparent permeability coefficients were found in EMBC compared to MB (0.11 to 0.44-fold change, p < 0.05). EMBC displayed the highest antioxidant capacity by the DPPH method for the non-digestible fraction, being mangiferin the highest in silico contributor (-4 kcal/mol). Our results showed that the extrusion process helps release selective phenolics from MBC, which increases their bioaccessibility and intestinal permeability.

Multi-response optimal hot pressurized liquid recovery of extractable polyphenols from leaves of maqui (Aristotelia chilensis [Mol.] Stuntz).

Multi-response optimization of hot pressurized liquid extraction (HPLE) was applied for the first time to obtain maqui (Aristotelia chilensis [Mol.] Stuntz) leaf extracts. The total polyphenol content (TPC), the antioxidant capacity (AC) as well as the total polyphenol purity of the maqui leaf extracts were accurately predicted (RSD < 8%) at the evaluated extraction scales. The optimum HPLE conditions that prioritized TPC and AC equally (OPT1) recovered ~3 times more TPC (205.14 mg GAE/g leaves) than maqui leaf extracts obtained by maceration, while the extract that prioritized purity over TPC and AC presented the highest purity (36.29%) and an EC50 ~3 times lower than currently reported values. It was found by multi-response optimization that maqui leaves and HPLE are among the best natural sources and extraction techniques, respectively, to recover protocatechuic acid, quercetin, and catechin.

Edible flowers from Theobroma speciosum: Aqueous extract rich in antioxidant compounds.

Theobroma speciosum, known as "cacauí" in Brazil, is considered a prominent unconventional food plant. The objective of this work was to evaluate the chemical profiles, antioxidant capacity and minerals of the aqueous extract and fractions from its flowers. The identified compounds were sugars, organic acids and phenolics compounds such as citric, malic and protocatechuic acids, quercetin, quercetin pentoside and quercetin-3-O-glucoside. The extract was rich in phenolic compounds (640 mg GAE g-1). Furthermore, fractions also presented phenolic compounds from 170.7 to 560.7 mg GAE g-1 (mainly protocatechuic acid, quercetin and derivatives), which influenced on the high antioxidant capacity in DPPH, ABTS, FRAP and co-oxidation ß-carotene/linolenic acid assays. Flowers presented potassium (115 ± 2 µg mL-1), magnesium (18.4 ± 0.2 µg mL-1), phosphorus (7.0 ± 0.0 µg mL-1) and calcium (3.1 ± 0.1 µg mL-1). Moreover, the flowers aqueous extract represents a new promising food source rich in antioxidant compounds.

Phenolic composition, antioxidant capacity and α-glucosidase inhibitory activity of raw and boiled Chilean Araucaria araucana kernels.

The Araucaria araucana kernels are a traditional food in southern Chile and Argentina. The aim of this work was to determine the composition of the phenolic-enriched extracts (PEEs) of the boiled kernels as well as their antioxidant capacity, inhibitory activity on metabolic syndrome-associated enzymes and effect on postprandial oxidative stress in a simulated gastric digestion model. The PEEs composition was assessed by HPLC-DAD-MS/MS. The main PEEs constituents were catechin and epicatechin in the unbound form, while hydroxybenzoic acids occurred mainly in the bound form. The unbound phenolics from boiled kernels showed significant correlations with DPPH, FRAP, TEAC (Pearson's r of 0.481, 0.331 and 0.417, respectively) and lipid peroxidation (r = 0.381) and were more active than the bound phenolics. The extracts were highly active against α-glucosidase (IC50: 0.33-3.15 µg/mL) and reduced lipoperoxidation. Traditional processing increases the flavan-3-ol content. Our results suggest that this traditional food has potential health promoting properties.

Bioguided extraction of phenolic compounds and UHPLC-ESI-Q-TOF-MS/MS characterization of extracts of Moringa oleifera leaves collected in Brazil.

Moringa oleifera leaves are used in Brazilian folk medicine for their hypoglycemic and nutritional properties. In this context, the chemical and biological characteristics were determined. Conventional successive solid-liquid extraction with simultaneous bioguided purification using solvents with different polarities was performed with M. oleifera leaves, yielding six fractions and extracts. The fractions showed better results for antioxidant activity than the extracts. All of them were evaluated by scavenging of synthetic free radicals and reactive oxygen species, and Fr-Ace and Fr-EtOAc showed >100 mg GAE g-1 of phenolic content, while for FRAP and ORAC assays the values were higher than 1600 µmol Fe2+ g-1 and 3500 mmol TEAC g-1 respectively. The UPLC-ESI-QTOF-MS analysis of hydroalcoholic extract (HE) allowed identifying 24 compounds, with flavonoid derivatives being the most abundant group. Furthermore, the alkaloid trigonelline and sesquiterpenoid abscisic acid were identified for the first time in M. oleifera leaves. Finally, gallic acid, caffeic acid, rutin and quercetin were found in concentrations of 16.5, 2129, 477.4 and 127.5 µg g-1 respectively in HE, all of which were higher in fractions and extracts. These results suggest that bioguided extraction is an important technique, due to its ability to concentrate active compounds in a logical and rational way. In addition, M. oleifera leaves grown in Brazil are an important source of phenolic compounds with antioxidant activity that can be used in food, nutraceutical and pharmaceutical products.

Wild Brazilian species of Eugenia genera (Myrtaceae) as an innovation hotspot for food and pharmacological purposes.

The Myrtaceae is one of the most common plant families in Brazil, including >1000 species of native fruit spread from North to South, where around 50% of all species are endemic to the Atlantic Rain Forest Biome. Most Brazilian species belong to Eugenia, Campomanesia, Psidium and Myrciaria genera. In general, they are characterized by the presence of leaves opposite, simple, entire, with pellucid glands containing ethereal oils and produces berry-like, fleshy fruits with a wide diversity of beneficial characteristics. Several parts of these plants are widely used in regional folk medicine as an astringent, anti-inflammatory, antihypertensive, to treat gastrointestinal disorders, and so on. Some species have already been studied and most of their metabolic effects are attributed to the presence of polyphenols, carotenoids as well as sesquiterpenes and monoterpenes. In addition, the Myrtaceae family has great economic potential, because of the sensory characteristics which encourages their commercial exploitation plus the presence of phytochemicals which play an important role in managing several degenerative chronic diseases besides representing a hotspot of technological innovation for food, cosmetic and pharmaceutical purposes. In this context, this review discusses about the importance of some Eugenia genera representatives such as Eugenia uniflora, Eugenia stipitata, Eugenia brasiliensis, Eugenia pyriformis and Eugenia dysenterica, emphasizing its profile of bioactive compounds as well as the knowledge of the nutritional and functional fruit potential, providing information for further studies aiming to stimulate the cultivation of Brazilian wild berries, for consumption and new food ingredients.

Plum (Prunus salicina) peel and pulp microparticles as natural antioxidant additives in breast chicken patties.

Fiber microparticles (MPCs) separately obtained from peel and pulp of Japanese plum residues contained co-extracted ß-carotene, lutein, and α- and γ-tocopherols, as well as polyphenols (cyanidins, quercetin derivatives, pentameric proanthocyanidins). Peel and pulp MPCs were then separately evaluated as natural antioxidant additives (2.0% w/w level) in raw breast chicken patty, susceptible to oxidation. Their effect on technological properties was also analyzed. MPCs reduced in 50% the formation of thiobarbituric acid reactive substances (TBARS) in raw patties during 10-days storage at 4.0°C. Ferric reducing power (FRAP) was 77-157% higher in MPCs-added patties, especially with peel MPCs, being then attributed to the antioxidants supplied by these MPCs. It can be also associated to the highest α- and γ-tocopherol levels found in the peel MPCs-added patties, which remained high after cooking as well. Also, higher pectin and low lignin contents of pulp MPCs determined greater hydration, stabilized the cyanidins and, hence, the red color transferred to raw patties, and increased springiness of cooked patties. Plum peel and pulp MPCs are efficient additives for chicken meat products.

Baccharis trimera protects against ethanol induced hepatotoxicity in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis trimera has been traditionally used in Brazil to treat liver diseases. AIM OF THE STUDY: To evaluate the protective effect of Baccharis trimera in an ethanol induced hepatotoxicity model. MATERIALS AND METHODS: The antioxidant capacity was evaluated in vitro by the ability to scavenged the DPPH radical, by the quantification of ROS, NO and the transcription factor Nrf2. Hepatotoxicity was induced in animals by administration of absolute ethanol for 2 days (acute) or with ethanol diluted for 28 days (chronic). The biochemical parameters of hepatic function (ALT and AST), renal function (urea and creatinine) and lipid profile (total cholesterol, triglycerides and HDL) were evaluated. In addition to antioxidant defense (SOD, catalase, glutathione), oxidative damage markers (TBARS and carbonylated protein), MMP-2 activity and liver histology. RESULTS: Baccharis trimera promoted a decrease in ROS and NO, and at low concentrations promoted increased transcription of Nrf2. In the acute experiment it promoted increase of HDL, in the activity of SOD and GPx, besides diminishing TBARS and microesteatosis. Already in the chronic experiment B. trimera improved the hepatic and renal profile, decreased triglycerides and MMP-2 activity, in addition to diminishing microesteatosis. CONCLUSION: We believe that B. trimera action is possibly more associated with direct neutralizing effects or inhibition of reactive species production pathways rather than the modulation of the antioxidant enzymes activity. Thus it is possible to infer that the biological effects triggered by adaptive responses are complex and multifactorial depending on the dose, the time and the compounds used.

The flavonoid quercetin inhibits titanium dioxide (TiO2)-induced chronic arthritis in mice.

Titanium dioxide (TiO2) is a common component of orthopedic prosthesis. However, prosthesis wear releases TiO2, which induces inflammation and osteolysis in peri-prosthetic tissues. Quercetin is a flavonoid widely present in human diet, which presents biological activities such as antinociceptive, anti-inflammatory and antioxidant effects. Therefore, the effect of intraperitoneal treatment with quercetin in TiO2-induced arthritis model was evaluated. In the first set of experiments, mice received injection of TiO2 (0.1-3 mg/knee joint) and articular mechanical hyperalgesia, edema and histopathology analysis were performed in a 30 days protocol. The dose of 3 mg of TiO2 showed the most harmful effect, and was chosen to the following experiments. Subsequently, mice received 3 mg of TiO2 followed by post-treatment with quercetin during 30 days. Quercetin (10-100 mg/kg) inhibited in a dose-dependent manner TiO2-induced knee joint mechanical hyperalgesia, edema and leukocyte recruitment and did not induce damage in major organs such as liver, kidney and stomach. The dose of 30 mg/kg was chosen for the subsequent analysis, and reduced histopathological changes such as leukocyte infiltration, vascular proliferation and synovial hyperplasia (pannus formation) on day 30 after TiO2 challenge. The protective analgesic and anti-inflammatory mechanisms of quercetin included the inhibition of TiO2-induced neutrophil and macrophage recruitment, proteoglycan degradation, oxidative stress, cytokine production (TNF-α, IL-1ß, IL-6, and IL-10), COX-2 mRNA expression, and bone resorption as well as activation of Nrf2/HO-1 signaling pathway. These results demonstrate the potential therapeutic applicability of the dietary flavonoid quercetin to reduce pain and inflammatory damages associated with prosthesis wear process-induced arthritis.

Caffeoyl and coumaroyl derivatives from Acacia cochliacantha exhibit ovicidal activity against Haemonchus contortus.

ETHNOPHARMACOLOGY RELEVANCE: Acacia cochliacantha is a small tree whose foliage is traditionally used in Mexico for treatment of kidney pain, gastrointestinal illnesses and to kill intestinal parasites. In recent decades, the study of vegetal extracts has offered other possible alternatives for the control of Haemonchus contortus. Considering that this nematode affects dramatically the health and productivity of small ruminants, the aim of this study was to identify the anthelmintic compounds from A. cochliacantha hydro-alcoholic extract (HA-E) through an ovicidal test. MATERIAL AND METHODS: In vitro egg hatch assay was conducted to determinate the anthelmintic effects of a HA-E (60g). Liquid-liquid ethyl acetate/water extraction gave two fractions (EtOAc-F, 1.92g; Aq-F; 58.1g). The less polar compounds from ethyl acetate fraction were extracted by addition of dichloromethane offering a precipitate phase (Mt-F, 1.25g) and a soluble mixture (DCMt-F 1.15g). All fractions were evaluated for ovicidal activity obtaining the egg hatching inhibition (EHI, 0.07-25mg/mL). Ivermectin (0.5mg/mL) was used as a reference drug (positive control), and distilled water, 2.5% DMSO and 2% methanol were used as negative controls. The isolated compounds from the most active fractions were subjected to spectroscopic (1H NMR) Spectrometric (MS) and UV HPLC analysis in order to identify the bioactive compounds. RESULTS: The less polar treatments (AcOEt-F, DCMt-F, DCMt-P) showed the highest ovicidal activities (98-100% EHI; at 0.62-1.56mg/mL) and the major compounds found in these fractions were identified as caffeoyl and coumaroyl derivatives, including caffeic acid (1), p-coumaric acid (2), ferulic acid (3), methyl caffeate (4), methyl-p-coumarate (5), methyl ferulate (6) and quercetin. In case of the less active fractions (Aq-F, Mt-F) were constituted principally by glycosylated flavonoids. CONCLUSION: These results show that caffeoyl and coumaroyl derivatives from Acacia cochliacantha leaves had promising anthelmintic activity against Haemonchus contortus. This leguminous may offer an alternative source for the control of gastrointestinal nematodes of small ruminants.

Effect of the oral administration of nanoencapsulated quercetin on a mouse model of Alzheimer's disease.

Quercetin has been identified as a promising compound with a neuroprotective potential against age-related neurodegenerative diseases such as Alzheimer's disease (AD). Nevertheless, the clinical application of quercetin is hampered by its low oral bioavailability. The aim of this work was to evaluate the capability of nanoencapsulated quercetin in zein nanoparticles (NPQ), that significantly improves the oral absorption and bioavailability of the flavonoid, as potential oral treatment for AD. For this purpose, SAMP8 mice were orally treated for two months with either NPQ (25mg/kg every 48h) or a solution of quercetin (Q; 25mg/kg daily). NPQ displayed a size of 260nm and a payload of about 70µg/mg. For Q, no significant effects were observed in animals. On the contrary, the oral administration of NPQ improved the cognition and memory impairments characteristics of SAMP8 mice. These observations appeared to be related with a decreased expression of the hippocampal astrocyte marker GFAP. Furthermore, significant levels of quercetin were quantified in the brain of mice treated with nanoparticles. These findings highlight the potential of zein nanoparticles to promote the oral absorption of quercetin as well as the therapeutic potential of this flavonoid in AD pathogenesis.

Antinociceptive and anti-inflammatory activities of standardized extract of polymethoxyflavones from Ageratum conyzoides.

ETHNOPHARMACOLOGICAL RELEVANCE: Ageratum conyzoides L. is a plant widely used in traditional medicine in tropical and subtropical regions of the world due to its anti-inflammatory, antinociceptive and antibacterial properties. AIM OF THE STUDY: To characterize the standardized extract of polymethoxyflavones (SEPAc) from the plant and evaluate its antinociceptive and anti-inflammatory effects. MATERIALS AND METHODS: The SEPAc purified from the ethanol extract of the plant leaves was characterized by high resolution mass spectrometry and the methoxyflavones were quantified by a validated UPLC-PDA method. The antinociceptive and anti-inflammatory activities of the SEPAc were evaluated after oral administration on the acute nocifensive behavior of mice induced by formalin, prostaglandin E2 (PGE2) and proinflammatory cytokines (interleukin-1beta (IL-1ß)) and tumor necrosis factor-alpha (TNF-α) in mice. RESULTS: Qualitative analyses revealed the presence of seven methoxyflavones in the SEPAc, also a simple UPLC-PDA method was developed and validated for the quantification of 5,6,7,3',4',5'-hexametoxyflavone; nobiletin; 5'-methoxynobiletin and eupalestin, major compounds in the extract. The SEPAc exhibited antinociceptive and anti-inflammatory activities in both formalin phases, with significant inhibition of the paw edema formation and significant reduction of the nocifensive response induced by an intraplantar injection of PGE2 and intrathecal injection of interleukin-1ß. CONCLUSIONS: The SEPAc exhibited significant antinociceptive and anti-inflammatory effects. These results provided scientific suggestion of its potential as a source of new medicines to treat inflammatory diseases, such rheumatoid arthritis.

Characterization of Coffea arabica monofloral honey from Espírito Santo, Brazil.

In this study, samples of coffee honey produced in Espírito Santo State, Brazil, were characterized based on their melissopalynology, physicochemical and nutritional properties, and mineral and caffeine contents. The caffeine content in the nectar from coffee flowers was measured by high performance liquid chromatography (HPLC). Blends of honey were obtained from three Coffea arabica crops, each with 10 colonies of Africanized Apis mellifera. All honey samples contained monofloral (75-78%) pollen belonging to C. arabica. Physicochemical parameters (total acidity, pH, moisture, dry matter, ash, and qualitative hydroxymethylfurfural) were within the approved limits established by EU legislation. Coffee honey contains high levels of ascorbic acid (294.68 mg kg(-1)) and low amounts of total flavonoids (3.51 ± 0.18 mg QE kg(-1)). The most abundant minerals were potassium and calcium (962.59 ± 154.3 and 343.75 ± 25.56 mg kg(-1), respectively). The caffeine content in coffee nectar (1.64 mg kg(-1)) was approximately 8-fold lower than that in honey (12.02 ± 0.81 mg kg(-1)).

Medicinal plants used in the traditional management of diabetes and its sequelae in Central America: A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Globally 387 million people currently have diabetes and it is projected that this condition will be the 7th leading cause of death worldwide by 2030. As of 2012, its total prevalence in Central America (8.5%) was greater than the prevalence in most Latin American countries and the population of this region widely use herbal medicine. The aim of this study is to review the medicinal plants used to treat diabetes and its sequelae in seven Central American countries: Belize, Costa Rica, El Salvador, Guatemala, Honduras, Nicaragua and Panama. MATERIALS AND METHODS: We conducted a literature review and extracted from primary sources the plant use reports in traditional remedies that matched one of the following disease categories: diabetes mellitus, kidney disease, urinary problems, skin diseases and infections, cardiovascular disease, sexual dysfunctions, visual loss, and nerve damage. Use reports were entered in a database and data were analysed in terms of the highest number of use reports for diabetes management and for the different sequelae. We also examined the scientific evidence that might support the local uses of the most reported species. RESULTS: Out of 535 identified species used to manage diabetes and its sequelae, 104 species are used to manage diabetes and we found in vitro and in vivo preclinical experimental evidence of hypoglycaemic effect for 16 of the 20 species reported by at least two sources. However, only seven of these species are reported in more than 3 studies: Momordica charantia L., Neurolaena lobata (L.) R. Br. ex Cass., Tecoma stans (L.) Juss. ex Kunth, Persea americana Mill., Psidium guajava L., Anacardium occidentale L. and Hamelia patens Jacq. Several of the species that are used to manage diabetes in Central America are also used to treat conditions that may arise as its consequence such as kidney disease, urinary problems and skin conditions. CONCLUSION: This review provides an overview of the medicinal plants used to manage diabetes and its sequelae in Central America and of the current scientific knowledge that might explain their traditional use. In Central America a large number of medicinal plants are used to treat this condition and its sequelae, although relatively few species are widely used across the region. For the species used to manage diabetes, there is variation in the availability and quality of pharmacological, chemical and clinical studies to explain traditional use.

Quercetin derivative induces cell death in glioma cells by modulating NF-κB nuclear translocation and caspase-3 activation.

Treated glioblastoma multiforme (GBM) patients only survive 6 to 14months after diagnosis; therefore, the development of novel therapeutic strategies to treat gliomas remains critically necessary. Considering that phenolic compounds, like quercetin, have the potential to be used in the chemotreatment of gliomas and that some flavonoids exhibit the ability to cross the BBB, in the present study, we investigated the antitumor effect of flavonoids (including chalcones, flavones, flavanones and flavonols). Initially their activities were tested in C6 glioma cells screened using the MTT method, resulting in the selection of chalcone 2 whose feasibility was confirmed by a Trypan Blue exclusion assay in the low µM range on C6 glioma cells. Cell cycle and apoptotic death analyses on C6 glioma cells were also performed, and chalcone 2 increased the apoptosis of the cells but did not alter the cell cycle progression. In addition, treatments with these two compounds were not cytotoxic to hippocampal organotypic cultures, a model of healthy neural cells. Furthermore, the results indicated that 2 induced apoptosis by inhibition of NF-κB and activation of active caspase-3 in glioma cells, suggesting that it is a potential prototype to develop new treatments for GBM in the future.

Effect of drying method on volatile compounds, phenolic profile and antioxidant capacity of guava powders.

We studied the chemical composition of oven and freeze dried guava powders for future use as antioxidant-rich flavour enhancers. Among thirty-one volatiles in guava powders, terpenes were predominant, even after both drying processes. In contrast, esters and aldehydes, volatiles characteristic of fresh guava fruit, appeared to have been decreased by drying. Insoluble phenolics were predominant and among the sixteen compounds identified, quercetin-3-O-rutinoside and naringenin corresponded to 56% of total phenolics. Drying processes decreased total phenolics contents by up to 44%. Oven drying promoted the release of insoluble flavonoids, generating mainly quercetin. Antioxidant capacity also decreased due to both drying processes, but guava powders still presented similar antioxidant capacity in comparison to other tropical fruit powders. Our results suggest that oven drying is a viable option for the production of a functional ingredient that would improve the phenolic content of cereal foods while adding desirable guava flavour.

Toxicological evaluation of the natural products and some semisynthetic derivatives of Heterotheca inuloides Cass (Asteraceae).

ETHNOPHARMACOLOGICAL RELEVANCE: Heterotheca ineuloides Cass (Asteraceae), popularly known as árnica mexicana, is widely used in Mexican traditional medicine to treat bruises, dermatological problems, rheumatic pains, and other disorders as cancer. The major constituents in H. inuloides are cadinane type sesquiterpenes, flavonoids and phytosterols. Compounds with a cadinane skeleton have been proved to possess cytotoxic activity against human-tumor cell lines and brine shrimp, and display toxic effects in different animal species. Although this plant has been widely used, there is little available information on the safety and toxicity especially of pure compounds. AIM OF THIS STUDY: Evaluate the potential toxicity of the natural products isolated from H. inuloides and some semisynthetic derivatives. MATERIALS AND METHODS: The toxic aspects of the following natural products isolated from dried flowers of H. inuloides: 7-hydroxy-3,4-dihydrocadalene (1), 7-hydroxycadalene (2), 3,7-dihydroxy-3(4H)-isocadalen-4-one (3), (1R,4R)-1-hydroxy-4H-1,2,3,4- tetrahydrocadalen-15-oic acid (4), D-chiro-inositol (5), quercetin (6), quercetin-3,7,3'-trimethyl ether (7), quercetin-3,7,3',4'-tetramethyl ether (8), eriodictyol-7,4'-dimethyl ether (9), α-spinasterol (10), caryolan-1,9ß-diol (11) and 7-(3,3-dimethylallyloxy)-coumarin (12) as well as the toxic aspects of the semisynthetic compounds 7-acetoxy-3,4-dihydrocadalene (13), 7-benzoxy-3,4-dihydrocadalene (14), 7-acetoxycadalene (15), 7-benzoxycadalene (16), quercetin pentaacetate (17), 7-hydroxycalamenene (18), 3,8-dimethyl-5-(1-methylethyl)-1,2-naphthoquinone (19), and 4-isopropyl-1,6-dimethylbenzo[c]oxepine-7,9-dione (20). Toxic activities of compounds were determined by sulforhodamine B (SRB) assay, Artemia salina assay, RAW264.7 macrophage cells. Additionally, the acute toxicity in mouse of compound 1, the major natural sesquiterpene isolated from the acetone extract, was evaluated. RESULTS: The best cytotoxicity activity was observed for mansonone C (19) on K562 cell line with IC50 1.45 ± 0.14 µM, for 7-hydroxycadalene (2) on HCT-15 cell line with IC50 18.89 ± 1.2 µM, and for quercetin pentaacetate (17) on MCF-7 cell line with IC50 22.57 ± 2.4 µM. Sesquiterpenes mansonone C (19) and 7-hydroxy-3,4-dihydrocadalene (1) caused the strongest deleterious effects against A. salina with IC50 39.4 ± 1.07, and 45.47 ± 1.74 µM, respectively. The number of viable RAW 264.7 cells was reduced with sesquiterpenes 1 and 2 by more than 90%. In addition, the acute study of 1 revealed no lethal effects at 300 mg/kg body weight, however, a reduction in the body weight of mice, morphological changes in the tissues of the liver and kidney and toxic signs were observed at very high doses (2000 mg/kg). CONCLUSION: The results provided evidence for the cytotoxicity of Mexican arnica (H. inuloides) metabolites and may be correlated with one of the popular uses of this plant, in traditional Mexican medicine, as anticancer remedy. Among the active compounds contained in the acetone extract, the cytotoxic activity is mainly ascribable to cadinene type sesquiterpenes. In addition, evidence of acute toxicity suggests that 7-hydroxy-3,4-dihydrocadalene (1) may lead to toxicity at very high doses.

Biochemical and hematological effects of acute and sub-acute administration to ethyl acetate fraction from the stem bark Scutia buxifolia Reissek in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutia buxifolia is a native tree of Southern Brazil, Uruguay, and Argentina, which is popularly known as "coronilha" and it is used as a cardiotonic, antihypertensive and diuretic substance. The aim of this study was to assess the acute and sub-acute toxicity of the ethyl acetate fraction from the stem bark Scutia buxifolia in male and female mice. MATERIALS AND METHODS: The toxicity studies were based on the guidelines of the Organization for Economic Cooperation and Development (OECD-guidelines 423 and 407). In an acute study, a single dose of 2000 mg/kg of Scutia buxifolia was administered orally to male and female mice. Mortality, behavioral changes, and biochemical and hematological parameters were evaluated. In the sub-acute study, Scutia buxifolia was administered orally to male and female mice at doses of 100, 200, and 400mg/kg/day for 28 days. Behavioral changes and biochemical, hematological, and histological analysis were evaluated. RESULTS: The acute administration of Scutia buxifolia did not cause changes in behavior or mortality. Male and female mice presented decreased levels of platelets. Female mice presented decreased levels of leukocytes. On the other hand, in a sub-acute toxicity study, we observed no behavioral changes in male or female mice. Our results demonstrated a reduction in glucose levels in male mice treated to 200 and 400mg/kg of Scutia buxifolia. Aspartate aminotransferase (ASAT) activity was increased by Scutia buxifolia at 400mg/kg in male mice. In relation to the hematological parameters, male mice presented a reduction in hemoglobin (HGB) and hematocrit (HCT) when treated to 400mg/kg of plant fraction. Female mice showed no change in these parameters. Histopathological examination of liver tissue showed slight abnormalities that were consistent with the biochemical variations observed. CONCLUSION: Scutia buxifolia, after acute administration, may be classified as safe (category 5), according to the OECD guide. However, the alterations observed, after sub-acute administration with high doses of ethyl acetate fraction from the stem bark Scutia buxifolia, suggest that repeated administration of this fraction plant can cause adverse hepatic, renal, and hematological effects.