Adriana Zerlotti Mercadante

Evaluation of the genotoxic and antigenotoxic effects after acute and subacute treatments with açai pulp (Euterpe oleracea Mart.) on mice using the erythrocytes micronucleus test and the comet assay.

Açai, the fruit of a palm native to the Amazonian basin, is widely distributed in northern South America, where it has considerable economic importance. Whereas individual polyphenolics compounds in açai have been extensively evaluated, studies of the intact fruit and its biological properties are lacking. Therefore, the present study was undertaken to investigate the in vivo genotoxicity of açai and its possible antigenotoxicity on doxorubicin (DXR)-induced DNA damage. The açai pulp doses selected were 3.33, 10.0 and 16.67g/kg b.w. administered by gavage alone or prior to DXR (16mg/kg b.w.) administered by intraperitoneal injection. Swiss albino mice were distributed in eight groups for acute treatment with açai pulp (24h) and eight groups for subacute treatment (daily for 14 consecutive days) before euthanasia. The negative control groups were treated in a similar way. The results of chemical analysis suggested the presence of carotenoids, anthocyanins, phenolic, and flavonoids in açai pulp. The endpoints analyzed were micronucleus induction in bone marrow and peripheral blood cells polychromatic erythrocytes, and DNA damage in peripheral blood, liver and kidney cells assessed using the alkaline (pH >13) comet assay. There were no statistically significant differences (p>0.05) between the negative control and the groups treated with the three doses of açai pulp alone in all endpoints analyzed, demonstrating the absence of genotoxic effects. The protective effects of açai pulp were observed in both acute and subacute treatments, when administered prior to DXR. In general, subacute treatment provided greater efficiency in protecting against DXR-induced DNA damage in liver and kidney cells. These protective effects can be explained as the result of the phytochemicals present in açai pulp. These results will be applied to the developmental of food with functional characteristics, as well as to explore the characteristics of açai as a health promoter.

Carotenoids and ascorbic acid from cashew apple (Anacardium occidentale L.): variety and geographic effects

Abstract Cashew apple is the pseudofruit of the cashew tree, native to Brazil. Different varieties of cashew apple were collected in Brazil, three being from Piaui State (Northeast) and two from Sao Paulo State (Southeast). In all the fruits, β-carotene (16.6–67.9 μg/100 g), β-cryptoxanthin (7.7–64.4 μg/100 g), α-carotene (5.9–51.9 μg/100 g) and 9- cis - + 13- cis -β-carotene (3.3–15.6 μg/100 g) were found. In general, the levels of carotenoids were higher in the red than in the yellow cashew apples, from both regions; for example, the levels of α- and β-carotene were about 1.8 and 1.3 times higher in the red than in the yellow fruits from the Southeast and Northeast, respectively. In contrast, ascorbic acid (AA) values were slightly higher in the yellow variety. Elongated red and yellow fruits also presented slightly higher AA contents than the rounded red ones. The total carotenoid levels of the rounded red fruits were 1.5 and 1.7 times lower than those found in the yellow and red varieties, respectively, all being from the Northeast region. Yellow fruits from the Northeast presented 1.7 times higher provitamin A levels than those from the Southeast whereas, for the red variety, the values were similar. The yellow and red varieties from the Northeast showed non statistically higher AA levels than those from the Southeast.

Identification of bioactive compounds from jambolao (Syzygium cumini) and antioxidant capacity evaluation in different pH conditions

The composition of carotenoids and phenolic compounds from jambolão fruits (Syzygium cumini) was determined by HPLC-DAD-MS/MS. Two main carotenoids were found in the fruits, all-trans-lutein (43.7%) and all-trans-β-carotene (25.4%). The anthocyanin composition was characterised by the presence of 3,5-diglucosides of five out of six aglycones commonly found in foods. This pattern was also observed for the other flavonoids, since diglucosides of dihydromyricetin, methyl-dihydromyricetin and dimethyl-dihydromyricetin, along with myricetin glucoside and a galloyl-glucose ester were identified. Furthermore, the antioxidant capacity of a functional extract rich in anthocyanins was evaluated through the scavenging capacities of ABTS(+) and peroxyl radical (ORAC) and the protective effect against singlet oxygen ((1)O2). The TEAC values indicated that the hemiacetals/chalcones and quinonoidal bases species (pH⩾5) possess higher scavenging capacity as compared to the flavylium cation (pH<3). The functional extract also showed 60% of dimethylanthracene protection against (1)O2 and an ORAC value of 16.4μmolTrolox/gfruit.

Carotenoid Biosynthesis in Intraerythrocytic Stages of Plasmodium falciparum

Carotenoids are widespread lipophilic pigments synthesized by all photosynthetic organisms and some nonphotosynthetic fungi and bacteria. All carotenoids are derived from the C40 isoprenoid precursor geranylgeranyl pyrophosphate, and their chemical and physical properties are associated with light absorption, free radical scavenging, and antioxidant activity. Carotenoids are generally synthesized in well defined subcellular organelles, the plastids, which are also present in the phylum Apicomplexa, which comprises a number of important human parasites, such as Plasmodium and Toxoplasma. Recently, it was demonstrated that Toxoplasma gondii synthesizes abscisic acid. We therefore asked if Plasmodium falciparum is also capable of synthesizing carotenoids. Herein, biochemical findings demonstrated the presence of carotenoid biosynthesis in the intraerythrocytic stages of the apicomplexan parasite P. falciparum. Using metabolic labeling with radioisotopes, in vitro inhibition tests with norflurazon, a specific inhibitor of plant carotenoid biosynthesis, the results showed that intraerythrocytic stages of P. falciparum synthesize carotenoid compounds. A plasmodial enzyme that presented phytoene synthase activity was also identified and characterized. These findings not only contribute to the current understanding of P. falciparum evolution but shed light on a pathway that could serve as a chemotherapeutic target.

In vitro scavenging capacity of annatto seed extracts against reactive oxygen and nitrogen species.

Bixa orellana L. (annatto), from Bixaceae family, is a native plant of tropical America, which accumulates several carotenoids (including bixin and norbixin), terpenoids, tocotrienols and flavonoids with potential antioxidant activity. In the present study, the in vitro scavenging capacity of annatto seed extracts against reactive oxygen species (ROS) and reactive nitrogen species (RNS) was evaluated and compared to the bixin standard. Annatto extracts were obtained using solvents with different polarities and their phenolic compounds and bixin levels were determined by high performance liquid chromatography coupled to diode array detector. All annatto extracts were able to scavenge all the reactive species tested at the low μg/mL range, with the exception of superoxide radical. The ethanol:ethyl acetate and ethyl acetate extracts of annatto seeds, which presented the highest levels of hypolaetin and bixin, respectively, were the extracts with the highest antioxidant capacity, although bixin standard presented the lowest IC(50) values.

Bioactive compounds from several tropical fruits and correlation by multivariate analysis to free radical scavenger activity

O alto consumo de frutas tem sido associado a baixa incidencia de doencas cronicodegenerativas, provavelmente devido a presenca de compostos bioativos, como os antioxidantes, nestes alimentos. Os teores de compostos bioativos -acido ascorbico, fenolicos totais, flavonoides e carotenoides totais - foram determinados em polpas obtidas a partir de 18 frutas tropicais adquiridas no Brasil. A atividade anti-radical livre foi avaliada pelo metodo ABTS. As frutas que apresentaram maior teor de compostos bioativos foram buriti, caja-manga, canistel, murici, physalis, piquia e tucuma. Considerando a composicao dos compostos bioativos analisados e as atividades anti-radical livre, as frutas foram divididas em 5 grupos, segundo Analise Hierarquica de Agrupamentos. Aplicando a Analise de Componentes Principais, a atividade anti-radical livre mostrou alta correlacao com compostos fenolicos totais (r = 0,99) e flavonoides (r = 0,86); porem, a correlacao encontrada foi muito pequena para acido ascorbico (r = 0,02) e carotenoides totais (r = 0,08). High ingestion of fruits has been associated with low incidence of chronic-degenerative diseases, probably due to the presence of bioactive compounds in these foods, such as antioxidants. The levels of bioactive compounds - ascorbic acid, total phenolics, flavonoids and carotenoids - were determined in 18 pulps obtained from tropical fruits acquired in Brazil. The free radical scavenger activity was evaluated by the ABTS assay. The fruits that showed higher levels of bioactive compounds were buriti, otaheite apple, egg-fruit, golden spoon, physalis, piquia and star nut palm. Considering the composition of bioactive compounds and free radical scavenger capacities the fruits were divided into five groups, according to Hierarchical Cluster Analysis. Applying Principal Component Analysis, free radical scavenger showed high correlation with total phenolic compounds (r = 0.99) and flavonoids (r = 0.86); however, the correlation was found to be very poor with ascorbic acid (r = 0.02) and with total carotenoids levels (r = 0.08).

Identification and quantification, by HPLC-DAD-MS/MS, of carotenoids and phenolic compounds from the Amazonian fruit Caryocar villosum.

The Amazonian region from Brazil has a wide variety of native and wild noncommercially cultivated fruits. This article reports for the first time the composition of carotenoids and phenolic compounds from Caryocar villosum fruit pulp, and, in addition, its proximate composition and antioxidant capacity (ORAC assay) were determined. According to the nutritional composition, water (52%) and lipids (25%) were the major components found in the pulp, and the total energetic value was 291 kcal/100 g. The major phenolic compounds identified by HPLC-DAD-ESI-MS/MS were gallic acid (182.4 μg/g pulp), followed by ellagic acid rhamnoside (107 μg/g pulp) and ellagic acid (104 μg/g pulp). The main carotenoids identified by HPLC-DAD-APCI-MS/MS were all-trans-antheraxanthin (3.4 μg/g pulp), all-trans-zeaxanthin (2.9 μg/g pulp), and a lutein-like carotenoid (2.8 μg/g pulp). The antioxidant capacity of the pulp (3.7 mMol Trolox/100 g pulp) indicates that it can be considered a good peroxyl radical scavenger.

Development of a novel micro-assay for evaluation of peroxyl radical scavenger capacity: Application to carotenoids and structure–activity relationship

A micro-assay was developed and validated, using a microplate reader in 96-well format, C(11)-BODIPY(581/591) as fluorescent probe and AIBN as ROO() generator. The structure-activity relationship was established for 15 carotenoid standards, indicating that the opening of the β-ionone ring and the increase of chromophore extension in the carotenoid structure were the major factors leading to the increase of ROO() scavenging capacity. The values for ROO() scavenging capacity were calculated using α-tocopherol as reference compound. Among the studied carotenoids, all-trans-lycopene was the most efficient ROO() scavenger (8.67±0.74) followed by all-trans-astaxanthin (6.50±0.62). All the carotenoids showed to be more effective ROO() scavengers than α-tocopherol and some hydrophilic compounds. Finally, the method was successfully applied to assay the ROO() scavenging capacity of carotenoid extracts from two Amazonian fruits, peach palm (7.83±0.21) and mamey (6.90±0.44).

Phenolic Compounds and Carotenoids from Four Fruits Native from the Brazilian Atlantic Forest

Fruits from the Atlantic Forest have received increasing interest because they contain high levels of bioactive compounds with notable functional properties. The composition of carotenoids and phenolic compounds from fruits found in the Atlantic Forest (jussara, uvaia, araça, and grumixama) was determined by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors. Uvaia showed the highest levels of carotenoids (1306.6 μg/100 g fresh matter (f.m.)). Gallic acid was the major phenolic compound in araça (12.2 mg GAE/100 g f.m.) and uvaia (27.5 mg GAE/100 g f.m.). In grumixama, eight quercetin derivatives were found; the main carotenoids included all-trans-β-cryptoxanthin (286.7 μg/100 g f.m.) and all-trans-lutein (55.5 μg/100 g f.m.). Uvaia and grumixama contain high amounts of carotenoids, while jussara showed greater levels of phenolic compounds (415 mg GAE/100 g f.m.), particularly anthocyanins (cyanidin 3-rutinoside: 179.60 mg/100 g f.m.; cyanidin 3-glucoside: 47.93 mg/100 g f.m.).

Carotenoids and Phenolic Compounds from Solanum sessiliflorum, an Unexploited Amazonian Fruit, and Their Scavenging Capacities against Reactive Oxygen and Nitrogen Species

The composition of carotenoids and phenolic compounds from mana-cubiu (Solanum sessiliflorum), a fruit native to Amazonia, was determined by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-DAD-MS(n)). The antioxidant capacities of the hydrophilic and carotenoid extracts against some reactive oxygen (ROO(•), H(2)O(2), HOCl, and HO(•)) and nitrogen (ONOO(-)) species were also determined. Seventeen carotenoids and three phenolic compounds were found in mana-cubiu. The major carotenoids were (all-E)-β-carotene (7.15 μg/g of dry weight) and (all-E)-lutein (2.41 μg/g of dry weight). The 5-caffeoylquinic acid (1351 μg/g of dry weight) was the major phenolic compound, representing more than 78% (w/w) of the total phenolic compounds. Moreover, two dihydrocaffeoyl spermidines were found in the hydrophilic extract. Both mana-cubiu extracts were able to scavenge all the tested reactive species. The carotenoid extract was shown to be a potent scavenger of peroxyl radical, while the hydrophilic extract was a potent hydrogen peroxide and hypochlorous acid scavenger.