Annona Crassiflora Mart.) As a Potential Source of Natural Antioxidant Compounds
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International Food Research Journal 25(5): 2005-2012 (October 2018) Journal homepage: http://www.ifrj.upm.edu.my Brazilian Cerrado fruit araticum (Annona crassiflora Mart.) as a potential source of natural antioxidant compounds *Arruda, H. S., Pereira, G. A. and Pastore, G. M. Bioflavors and Bioactive Compounds Laboratory, Department of Food Science, School of Food Engineering, University of Campinas, Monteiro Lobato Street, 80, Campinas P.O. Box 13083-862, São Paulo, Brazil Article history Abstract Received: 30 May 2017 Araticum is an underutilized Brazilian Cerrado fruit with few reports in the literature about Received in revised form: its bioactive compounds and functional properties. This study aimed to determine the amount 15 July 2017 of total phenolic compounds, total flavonoids and antioxidant activity of araticum pulp. Total Accepted: 25 July 2017 phenolic compounds and total flavonoids were determined spectrophotometrically using the Folin-Ciocalteu and aluminium chloride colorimetric assays, respectively. In addition, antioxidant activity from araticum fruit was measured by DPPH, TEAC and ORACFL assays. Keywords The araticum pulp showed high amount of total phenolic compounds (2.62 g GAE/100 g dw) and total flavonoids (1.79 g CE/100 g dw). Flavonoids were the predominant class of phenolic Antioxidant activity compounds, representing about 68% of the phenolic compounds content. Furthermore, the fruit Bioactive compounds showed high antioxidant activity for all assays performed (93.76 µg/mL, 231.79 µmol TE/g Flavonoids and 902.27 µmol TE/g for DPPH IC50, TEAC and T-ORACFL, respectively). Quercetin showed Phenolic compounds •+ Phytochemicals the most potent antioxidant activity in DPPH• and ABTS radical scavenging capacities, whereas the kaempferol was the compound with the highest scavenging capacity peroxyl radicals. Araticum fruit showed phenolic compounds content and antioxidant activity hardly found in common fruits. These results indicate that araticum is a potential source of bioactive compounds. © All Rights Reserved Introduction their safety and potential nutritional and therapeutic effects (Souza et al., 2012). Antioxidants are defined Reactive species are highly reactive molecules as one heterogeneous family of molecules, which containing one or more unpaired electrons in atomic can prevent or reduce the extension of the oxidative or molecular orbitals which can be generated from damage to lipids, proteins and nucleic acids as environmental factors (smoking, certain drugs, they scavenge reactive species. The most abundant pollution, poor diet, sedentary lifestyle and stress- antioxidants in fruits are the phenolic compounds inducing agents) and during normal cellular (mostly flavonoids), carotenoids and vitamins A, B, metabolism (respiration, phagocytosis, intoxication C and E (Lim et al., 2007). and fatty acid metabolism) (Thatoi et al., 2013; Brazil boasts one of the richest biodiversity in the Martins et al., 2015). When there is an imbalance world, which includes a large number of fruit species. between the production of reactive species and However, many native and exotic fruits species in antioxidants these species can join with cellular Brazil are underexploited or unknown (Souza et components (DNA, lipids and proteins) and destroy al., 2012). In the last years, the Cerrado fruits have them. This condition known as oxidative stress is attracted the attention of consumers and researchers involved in the generation or aggravation several of because to their nutritional and functional properties pathological conditions (Schaffer et al., 2016), such combined with the potential to add value and preserve as metabolic, neurodegenerative, cardiovascular the biodiversity of this biome (Malta et al., 2013). and mitochondrial diseases, aging process and even The Cerrado is the second largest biome in Brazil cancer (Martins et al., 2016). occupying approximately 25% of the country and it is Fruits are notable sources of antioxidants that considered one of the 25 sites of greatest biodiversity can provide protection against disorders promoted in the world. The Cerrado species covers about 30% by oxidative stress (Malta et al., 2012). Natural of all existing species in Brazil, it is estimated the antioxidants present in food have attracted the presence of approximately 11,000 plant species, 837 interest of various sectors of society because of bird species and 199 mammal species (Arruda and *Corresponding author. Email: [email protected], [email protected] 2006 Arruda et al/IFRJ 25(5): 2005-2012 Almeida, 2015). Brazil). All others chemical and solvents were of Cerrado flora holds several fruit species with analytical grade. great potential for food exploration, since they contain high levels of sugars, proteins, vitamins, Plant material and sample preparation minerals and fibers, besides attractive sensory Araticum fruits (Annona crassiflora Mart.) with characteristics such as peculiar and intense color, full physiological maturity were collected during flavor and aroma (Arruda et al., 2014; Arruda et harvest season (from January to March) in the rural al., 2015). Among the Cerrado fruit species, the areas of the municipality of Carmo do Paranaíba, araticum (Annona crassiflora Mart.) stands out for its Minas Gerais State, Brazil. The fruits were washed strong presence in this region and for its excellent with tap water to remove surface dirt. The fruit pulp sensory characteristics, much appreciated by the was manually separated from the seeds and peel, local population (Silva et al., 2013), and it is among freeze-dried (LIOTOP, model L101, São Carlos, the 20 most commonly used species in regional food Brazil), and ground using a knife grinder (Marconi, preparation (Arruda et al., 2017). Araticum belongs model MA340, Piracicaba, Brazil). The powder to the Annonaceae family and receives various other obtained was stored at -80°C until analysis. popular names such as “araticum-do-Cerrado”, “ariticum”, “articum”, “marolo”, “bruto”, “cabeça- Extraction procedure de-negro” and “pasmada” (Villela et al., 2013). The extraction conditions were defined according Several parts of the plant are used in traditional to the analysis protocols of total phenolic compounds, medicine to treat wounds, sexually transmitted total flavonoids and antioxidant activity (Leite- diseases (STDs), ophidian bites, degenerative Legatti et al., 2012; Pavan et al., 2014) with slight diseases, as well as antimicrobial, antidiarrheal, modifications. Briefly, 10 mg of freeze-dried fruit pulp antitumor and antirheumatic agents and in the combat were transferred to a microtube and mixed with 1.0 of lice (Dragano et al., 2010). The fruit pulp is very mL of corresponding solvent to each analysis (total popular and rich in fibers, carotenoids, vitamins phenolic compounds and DPPH assays: methanol; A and E, ascorbic acid and folates (Cardoso et al., total flavonoids and TEAC assays: ultrapure water; 2013). Recently, studies have showed that the fruit H-ORACFL assay: potassium phosphate buffer (75 presents biological properties such as antimicrobial mM, pH 7.4); and L-ORACFL assay: RMCD 7% (w/v; (Silva et al., 2014) and hepatoprotective activities acetone:water 1:1). The mixtures were ultrasonicated (Roesler, 2011). (UNIQUE, model UCS-2850, 25 kHz, 120W, Brazil) Despite the great potential presented by this fruit, for 30 minutes at 10°C and then centrifuged (Hettich there are few reports in the literature about its amount Zentrifugen, model Rotanta 460R, Tuttlingen, of bioactive compounds and antioxidant activity. Germany) at 10,000 rpm for 11 minutes at 5°C. The In this context, this study aimed to determine the supernatants were used for the analysis of phenolic amount of total phenolic compounds, total flavonoids compounds and antioxidant activity. and antioxidant activity of araticum pulp in order to provide relevant information to support its application Total phenolic compounds (TPC) as functional food and/or ingredient. TPC were determined using the Folin-Ciocalteu method according to Cicco et al. (2009) with Material and Methods modifications. Briefly, 100 µL of extract were mixed with 100 µL of Folin-Ciocalteu reagent (50% v/v) Chemicals and 800 µL of sodium carbonate (5% w/v). The Gallic acid, (+)-catechin, (-)-epicatechin, mixtures were incubated at 40°C for 20 minutes. (-)-gallocatechin, (-)-epigallocatechin, myricetin, The absorbance was measured at 760 nm against quercetin, kaempferol, 6-hydroxy-2,5,7,8- a blank on a spectrophotometer (Beckman, model tetramethylchromane-2-carboxylic acid (Trolox), DU600, CA, USA). The results were expressed as g 2,2-diphenyl-1-picrylhydrazil (DPPH), 2,2’-azobis(2- of gallic acid equivalent per 100 g of dried weight (g methylamidinopropane)-dihydrochloride (AAPH), GAE/100g dw). 2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)-diammonium salt (ABTS) and randomly Total flavonoids content (TFC) methylated beta-cyclodextrin (RMBC) were TFC were measured using a colorimetric assay purchased from Sigma-Aldrich (St. Louis, MO, USA). developed by Zhishen et al. (1999). Aliquots of Sodium carbonate, methanol and Folin-Ciocalteu 28 µL of extract and 110 µL of ultrapure water reagent were provided by Dinâmica (Diadema, SP, were homogenized and 8 µL of NaNO2 (5% w/v) Arruda et al/IFRJ 25(5): 2005-2012 2007 was added. After 2 minutes, 8 µL of AlCl3 (10% used as solvent. The lipophilic ORACFL (L-ORACFL) w/v) was added and the mixture was incubated for assay was conducted as follows: 20 μL of extract,