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Chemical and Biological Characterization from Condalia Microphylla Fruits, a Native Species of Patagonia Argentina

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Chemical and Biological Characterization from Condalia Microphylla Fruits, a Native Species of Patagonia Argentina View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by CONICET Digital Journal of Agricultural Science and Technology B 7 (2017) 395-405 doi: 10.17265/2161-6264/2017.06.004 D DAVID PUBLISHING Chemical and Biological Characterization from Condalia microphylla Fruits, a Native Species of Patagonia Argentina Patricia Boeri1, Lucrecia Piñuel1, 2, Sandra Sharry1, 3, Andrea Tombari1 and Daniel Barrio1, 2 1. Departamento de Ciencias Exactas, Naturales y de Ingeniería, Sede Atlántica, Universidad Nacional de Río Negro, Don Bosco y Leloir, Viedma 8500, Río Negro, Argentina 2. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Sede Atlántica, Universidad Nacional de Río Negro (UNRN), Av. Don Bosco, Viedma 8500, Río Negro, Argentina 3. Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de La Plata, Calle 60 s/n. La Plata, Buenos Aires 1900, Argentina Abstract: Condalia microphylla Cav. (Rhamnaceae), popularly known as “piquillin”, is widely distributed in Patagonia. The drupes are consumed as fresh fruits by Argentine communities. The aim of this work was to quantify the nutritional value of C. microphylla fruit and the phenolic compounds present and to determine the functional antioxidant properties in vitro and in vivo. The nutritional value was determined according to the Association of Official Analytical Chemists (AOAC) methodology, and phenolic compounds were quantified by diode-array detection (HPLC-DAD). Antioxidant activity in vitro and in vivo was analyzed through the use of the radical species 2,2-diphenyl-1-picrylhydrazyl (DPPH) and zebrafish model, respectively. Quercetin-3-O-rutinoside (rutin) was the single principal phenolic compound. The extracts contained in vitro antioxidant activities and total phenolic contents (TPCs) between 1,143 ± 112 µg and 4,633 ± 174 µg gallic-acid equivalents (GAEs) per 100 g dry weight (DW), though no relationship was found between the latter parameter and the antioxidant activity of the extracts. When zebrafish larvae were exposed to oxidative stress (2.4% v/v H2O2), a concentration as low as 1.44 µg of GAEs/mL of piquillin-derived polyphenols inhibited lipid oxidation by up to 40%. Thus, in view of these advantageous functional food properties and the opportunity to exploit this Patagonian natural resource, piquillin consumption should be promoted worldwide. Key words: Piquillin, antioxidant activity, zebrafish, in vivo activity. 1. Introduction pulp, and similar to those of Ziziphus, another genus in the Rhamnaceae family, has been extensively Piquillin (Condalia microphylla Cav.), a thorny studied [2, 3]. Phytochemical investigations have evergreen rhamnaceous plant, is a dominant shrub of revealed that Ziziphus fruits are rich sources in the Patagonian Monte semidesert. This native species bioactive compounds with biologic properties provides wood, food and various active principles [1]. potentially beneficial to human health, such as The fresh fruit of piquillin is widely consumed by antiarrhythmic [4], antioxidant [5], antiseizure [6] and many communities of Patagonia, Argentina, and for a antitumor activities [7]. Many degenerative diseases long time has been used in many preparations of are associated with an imbalance between prooxidants traditional medicine [1]. The fruits are a and antioxidants within the body, a phenomenon reddish-brown spherical drupe with a sticky, sweet referred to as oxidative stress [8]. In recent decades, phytochemicals, such as flavonoids, catechins, tannins Corresponding author: Patricia Boeri, Ph.D., research fields: plant biotechnology and bioprospecting. and anthocyanins, among other polyphenols, have 396 Chemical and Biological Characterization from Condalia microphylla Fruits, a Native Species of Patagonia Argentina attracted a growing attention, because their redox diverse substances in addition to other types of study properties can prevent or delay the development of [11]. The present work employed an in vivo model degenerative diseases [9]. These secondary based on the zebrafish (Danio rerio), in which the metabolites, however, are often present in low biochemical parameters of embryos were analyzed to quantities in plant material; and their extraction, determine the antioxidant effects in vivo of extracts purification and characterization still remain a major from the fruit of C. microphylla. challenge in drug discovery and development. In environments, such as the Patagonia, Argentina Therefore, the most appropriate extraction where nutrient availability is very limited, the plants methodology and the most suitable analytical have developed a wide variety of metabolites as a technique must be sought in order to achieve the best result of both selective pressure and resource extraction, purification and final isolation of the availability [12]. In view of these considerations desired secondary metabolite. involving the ongoing quest to find new edible plants Numerous studies have confirmed that polyphenols with functional properties and to promote the exert a protective action on human health and are key reproduction and preservation of those taxa, the components of a healthy, balanced diet. experiments were undertaken to be described here on Epidemiological studies suggest that a high dietary the chemical and biologic properties of the fruit of C. intake of these compounds is associated with a reduced microphylla—an area as yet unexplored. The aim of incidence of chronic ailments, such as cardiovascular these investigations was to determine the nutritional disease, diabetes and cancer. In particular, a group of composition of the fruit C. microphylla grown in the polyphenols known as flavonoids have been shown to arid and semi-arid region from Patagonia, Argentina exert a variety of anticarcinogenic effects, including an with an aim at characterizing the different bioactive ability to induce apoptosis in tumor cells, inhibit cancer phytocompounds having potential antioxidant cell proliferation and prevent angiogenesis and tumor properties. cell invasion [10]. An interest in the investigation of the biologic activities and bioactive compounds of 2. Materials and Methods fruits and vegetables has increased significantly in 2.1 Chemical Reagents recent years. To the knowledge of the authors, no review has yet been published on the investigation of Folin-Ciocalteu reagent, 2,2-diphenyl-1- bioactive compounds in piquillin fruits and the picrylhydrazyl (DPPH), α-tocopherol, gallic acid, potential benefits that might be attributed to C. dimethyl sulfoxide (DMSO), D-glucose, thiobarbituric microphylla, if extracts of the fruit were to be used for acid (TBA) and Tween 20 were purchased from nutraceutical purposes. Sigma Chemical Co. (St Louis, MO, USA). The other The evaluation of the behaviour of a single chemicals used were of analytical grade. compound or chemical mixture can be approached 2.2 Samples though the use of biological models in vitro or the in vivo activity in whole animals. The zebrafish model 2.2.1 Plant Materials has been used as an alternative paradigm in ambits, The fruits from at least 20 individuals of C. such as the food and pharmaceutical industries in microphylla were collected during February in 2012, order to investigate the teratogenic, embryotoxic and 2013 and 2014 in Northern Patagonia (41°55′296″ S, genotoxic properties, as well as the beneficial 63°05′693″ W). embryologic and neurobiologic activities of very Ripe fruits were washed with distilled water and Chemical and Biological Characterization from Condalia microphylla Fruits, 397 a Native Species of Patagonia Argentina quickly processed to facilitate the separation of 2.4.1 Determination of Polyphenols manually removed endocarps. The pulp thus obtained The determination of polyphenol composition was was stored at -20 °C in the dark until use for analysis. performed at the Center for Research and Technical 2.2.2 Sample Processing Assistance to Industry (CIATI-AC), Villa Regina, Río Three types of pulp extracts obtained with the Negro. The total ascorbic acid, ferulic acid, gallic acid, following three solvents were evaluated: (1) methanol; rutin (quercetin-3-O-rutinoside), p-coumaric acid, (2) 70% v/v aqueous acetone; (3) 50% v/v aqueous catechin and epicatechin were determined by high DMSO. The pulp-to-solvent ratio was 1:10. In all performance liquid chromatography with diode-array preparations, the extraction was performed overnight detection (HPLC-DAD) (Shimadzu). at room temperature with shaking (500 rpm), followed 2.4.2 Quantification of Anthocyanins by centrifugation at 7,000 rpm for 15 min. For Total anthocyanin content—expressed as mg biologic activity determination, the acetone was cyanidin-3-glucoside equivalents (C3GE) per 100 g removed from the extract in a vacuum oven at 40 °C DW—was evaluated by the differential-pH method and the residue resuspended in 5% v/v aqueous [17]. DMSO. 2.5 Antioxidant Activity 2.3 Determination of Chemical Composition 2.5.1 Antioxidant Activity in Vitro The measurement of ash, dietary fiber, protein and The pulp extracts from ripe fruit of C. microphylla lipids was performed according to the methodology were evaluated for antioxidant activity. The ability to specified by the Association of Official Analytical capture free radicals by antioxidants was analyzed Chemists (AOAC) [13]. through the use of the radical species DPPH [18] by Sugar determination: total carbohydrates were following the decrease in that compound’s absorbance determined according
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