Rapid extraction method followed by a d-SPE clean-up step for determination of phenolic composition and antioxidant and antiproliferative activities from berry fruits.

Despite the numerous studies that have shown a wide range of biological activities to berry fruits, scientific data focusing on modern, rapid and simple extraction methods followed by a clean-up step is still lacking. Therefore, the present work was aimed at investigating the use of a fast one-step solid-liquid extraction procedure followed by a dispersive solid-phase extraction (d-SPE) clean-up step to evaluate the phenolic composition, antioxidant and antiproliferative activities from three of the principal berries found in Brazil, pomegranate (Punica granatum L.), blackberry (Rubus ulmifolius Schott.), and strawberry (Fragaria × ananassa Duch.). Under the optimized extraction conditions, sixteen phenolic compounds were determined by UHPLC-MS/MS analysis and all berry extracts showed antioxidant activity and antiproliferative effects on cervical (HeLa) and colon (HT-29) cancer cells. Overall, these results highlight the importance of the clean-up step for more reliable data in studies of health-promoting proprieties from berry fruits.

Chlorogenic and caffeic acids in 64 fruits consumed in Brazil.

This study presents unpublished data on the chlorogenic acid and caffeic acid content in fruits. Sixty-four fruits consumed in Brazil, most of which were produced domestically, were evaluated based on their levels of 3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and caffeic acid. The study investigated 15 fruits that have not been reported in the literature in relation to these compounds, including several native species. The highest concentration of mono-caffeoylquinic acid was observed in strawberry, cherry, bilberry, quince and mulberry, while the dicaffeoylquinic acid was present with highest concentration in kumquat, passion fruit and sweet granadilla. Regarding caffeic acid, the highest content was found in bilberry and yellow pitaya. Considering the sum of the concentrations of these compounds, quince, cherry, bilberry, mulberry and sweet granadilla were exceptional, with concentrations between 200.0 and 569.7 mg kg-1.

UHPLC-MS Metabolome Fingerprinting: The Isolation of Main Compounds and Antioxidant Activity of the Andean Species Tetraglochin ameghinoi (Speg.) Speg.

The seriated extracts of petroleum ether (PE-E), dichloromethane (DCM-E) and methanol extracts (MeOH-E) from the aerial parts of the native South American plant Tetraglochin ameghinoi (Rosaceae), were evaluated regarding their antioxidant and antibacterial activities. The antioxidant properties were evaluated by free radical scavenging methods (DPPH and TEAC), ferric-reducing antioxidant power (FRAP) and lipoperoxidation in erythrocytes (LP), while the antibacterial activity was performed against Gram-positive and Gram-negative bacteria according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. The chemical and biological analyses of this plant are very important since this bush is currently used in traditional medicine as a cholagogue and digestive. The polar MeOH-E showed the highest antioxidant activities (17.70 µg/mL in the DPPH assay, 381.43 ± 22.38 mM TE/g extract in the FRAP assay, 387.76 ± 91.93 mg TE/g extract in the TEAC assay and 93.23 + 6.77% in the LP assay) and it was selected for chromatographic isolation of its components. These components were found to be four acetophenones, including the new phloracetophenone glucoside: 4',6',-dihydroxy-2'-O-(6″-acetyl)-ß-d-glucopyranosylacetophenone or IUPAC name: (6-(2-acetyl-3,5-dihydroxyphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)methyl acetate, whose structure was elucidated by NMR and MS methods. In addition, twenty-six compounds, including five of these acetophenone derivatives, two sugars, six flavonoids, eleven phenolic acids and two triterpenes, were identified based on UHPLC-OT-MS and PDA analysis on the MeOH-E. The results support the medicinal use of the plant.

Evolutionary diversification of galactinol synthases in Rosaceae: adaptive roles of galactinol and raffinose during apple bud dormancy.

Galactinol synthase (GolS) is a key enzyme in the biosynthetic pathway of raffinose family oligosaccharides (RFOs), which play roles in carbon storage, signal transduction, and osmoprotection. The present work assessed the evolutionary history of GolS genes across the Rosaceae using several bioinformatic tools. Apple (Malus × domestica) GolS genes were transcriptionally characterized during bud dormancy, in parallel with galactinol and raffinose measurements. Additionally, MdGolS2, a candidate to regulate seasonal galactinol and RFO content during apple bud dormancy, was functionally characterized in Arabidopsis. Evolutionary analyses revealed that whole genome duplications have driven GolS gene evolution and diversification in Rosaceae speciation. The strong purifying selection identified in duplicated GolS genes suggests that differential gene expression might define gene function better than protein structure. Interestingly, MdGolS2 was differentially expressed during bud dormancy, concomitantly with the highest galactinol and raffinose levels. One of the intrinsic adaptive features of bud dormancy is limited availability of free water; therefore, we generated transgenic Arabidopsis plants expressing MdGolS2. They showed higher galactinol and raffinose contents and increased tolerance to water deficit. Our results suggest that MdGolS2 is the major GolS responsible for RFO accumulation during apple dormancy, and these carbohydrates help to protect dormant buds against limited water supply.

Characterization of Rubus fruticosus mitochondria and salicylic acid inhibition of reactive oxygen species generation at Complex III/Q cycle: potential implications for hypersensitive response in plants.

In addition to adenosine triphosphate (ATP) production, mitochondria have been implicated in the regulation of several physiological responses in plants, such as programmed cell death (PCD) activation. Salicylic acid (SA) and reactive oxygen species (ROS) are essential signaling molecules involved in such physiological responses; however, the mechanisms by which they act remain unknown. In non-photosynthesizing tissues, mitochondria appear to serve as the main source of ROS generation. Evidence suggests that SA and ROS could regulate plant PCD through a synergistic mechanism that involves mitochondria. Herein, we isolate and characterize the mitochondria from non-photosynthesizing cell suspension cultures of Rubus fruticosus. Furthermore, we assess the primary site of ROS generation and the effects of SA on isolated organelles. Mitochondrial Complex III was found to be the major source of ROS generation in this model. In addition, we discovered that SA inhibits the electron transport chain by inactivating the semiquinone radical during the Q cycle. Computational analyses confirmed the experimental data, and a mechanism for this action is proposed.

Fermentative behavior of Saccharomyces strains during microvinification of raspberry juice (Rubus idaeus L.).

Sixteen different strains of Saccharomyces cerevisiae and Saccharomyces bayanus were evaluated in the production of raspberry fruit wine. Raspberry juice sugar concentrations were adjusted to 16° Brix with a sucrose solution, and batch fermentations were performed at 22 °C. Various kinetic parameters, such as the conversion factors of the substrates into ethanol (Y(p/s)), biomass (Y(x/s)), glycerol (Y(g/s)) and acetic acid (Y(ac/s)), the volumetric productivity of ethanol (Q(p)), the biomass productivity (P(x)), and the fermentation efficiency (E(f)) were calculated. Volatile compounds (alcohols, ethyl esters, acetates of higher alcohols and volatile fatty acids) were determined by gas chromatography (GC-FID). The highest values for the E(f), Y(p/s), Y(g/s), and Y(x/s) parameters were obtained when strains commonly used in the fuel ethanol industry (S. cerevisiae PE-2, BG, SA, CAT-1, and VR-1) were used to ferment raspberry juice. S. cerevisiae strain UFLA FW 15, isolated from fruit, displayed similar results. Twenty-one volatile compounds were identified in raspberry wines. The highest concentrations of total volatile compounds were found in wines produced with S. cerevisiae strains UFLA FW 15 (87,435 µg/L), CAT-1 (80,317.01 µg/L), VR-1 (67,573.99 µg/L) and S. bayanus CBS 1505 (71,660.32 µg/L). The highest concentrations of ethyl esters were 454.33 µg/L, 440.33 µg/L and 438 µg/L for S. cerevisiae strains UFLA FW 15, VR-1 and BG, respectively. Similar to concentrations of ethyl esters, the highest concentrations of acetates (1927.67 µg/L) and higher alcohols (83,996.33 µg/L) were produced in raspberry wine from S. cerevisiae UFLA FW 15. The maximum concentration of volatile fatty acids was found in raspberry wine produced by S. cerevisiae strain VR-1. We conclude that S. cerevisiae strain UFLA FW 15 fermented raspberry juice and produced a fruit wine with low concentrations of acids and high concentrations of acetates, higher alcohols and ethyl esters.

Tribromophenol and pentachlorophenol uptake from sawdust to horticultural products.

This paper presents a study of the uptake of 2,4,6-tribromophenol (TBP), pentachlorophenol (PCP), and its metabolite pentachloroanisole (PCA) from contaminated sawdust from the forest industry in horticultural products such as apples, raspberries, and fodder maize for cattle feed. The samples were obtained from Bio-Bio Province in South Chile between 2002 and 2006. The analytical parameters of the methodology applied to the different matrices are presented and discussed. The chromatographic method was applied to determine the residues in 413 horticultural product samples. Eleven per cent of fodder maize samples showed detectable or quantifiable levels of PCP, TBP or PCA, 3% of samples presented quantifiable levels, although the concentrations surpassed the maximum allowed concentrations for vegetables (>10 microg kg(-1)) in only two samples. Traces of TBP were detected in eight samples, PCA was detected in 15, and PCP in 14 samples. Based on these results, a risk analysis was performed, indicating a low probability, 0.4% for PCA, 1.6% for TBP and 1.9% for PCP, to find concentrations higher than the allowed maximum. For apples and raspberries, no residues of these compounds were detected. These results indicate that those cultivars directly exposed to sawdust, like fodder maize, could contain detectable residues in several samples. To confirm this observation, a field assay was performed on fodder maize cultivated in the presence of sawdust artificially contaminated with 30 mg of TBP and/or PCP under controlled conditions. The results showed that under the experimental conditions used in the study, TBP can be transferred from sawdust to the plant, with an uptake rate of 0.04% from the TBP applied initially with sawdust but not to the corn ear. Also, the degradation of PCP to PCA was observed in the soil.