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Composition, content of bioactive compounds, and antioxidant activity of fruit pulps from the Brazilian Amazon biome

Composição, teor de compostos bioativos e atividade antioxidante de polpas de frutas do bioma Amazônia

Abstract:

The objective of this work was to evaluate the physicochemical composition, the bioactive compounds, and the total antioxidant activity of the fruit pulps of abiu (Pouteria caimito), achachairu (Garcinia humilis), araza (Eugenia stipitata), bilimbi (Averrhoa bilimbi), and yellow mangosteen (Garcinia xanthochymus) from the Brazilian Amazon biome. Total soluble solid content, total and reducing sugar contents, titratable acidity contents, pH, ascorbic acid content, moisture, protein, lipid, ash, and caloric value were determined. The contents of total anthocyanins, yellow flavonoids, and total carotenoids, besides total phenolic compounds and antioxidant activity, were also evaluated. The abiu pulp showed the highest contents of ash, total sugars, reducing sugars, total soluble solids, calorie value, and pH, among the analyzed fruits. The achachairu pulp showed the highest content of total phenolic compounds, with significant antioxidant activity, while the araza pulp had the highest total carotenoid content, and the yellow mangosteen, the highest yellow flavonoid content. The positive correlation between antioxidant activity and total phenolic compounds suggests that these compounds are the most important for determining antioxidant activity in the evaluated fruit pulps.

Index terms:
Amazon biome; exotic fruit; fruit physicochemical composition; macronutrients; micronutrients

Resumo:

O objetivo deste trabalho foi avaliar a composição físico-química, os compostos bioativos e a atividade antioxidante total das polpas das frutas de abiu (Pouteria caimito), achachairu (Garcinia humilis), araçá-boi (Eugenia stipitata), biri-biri (Averrhoa bilimbi) e mangostão-amarelo (Garcinia xanthochymus), procedentes do bioma Amazônia no Brasil. Foram determinados os teores de sólidos solúveis totais, açúcares redutores e totais, acidez total titulável, pH, ácido ascórbico, umidade, proteínas, lipídeos, cinzas e valor energético total. Os teores de antocianinas totais, flavonoides amarelos e carotenoides totais, além dos compostos fenólicos totais e atividade antioxidante, também foram avaliados. A polpa de abiu apresentou os maiores teores de cinzas, açúcares totais, açúcares redutores, sólidos solúveis totais, valor energético total e pH, entre as frutas avaliadas. A polpa de achachairu apresentou o maior teor de compostos fenólicos totais com significativa atividade antioxidante, enquanto a polpa de araçá-boi apresentou o maior teor de carotenoides totais, e a polpa de mangostão-amarelo apresentou o maior teor de flavonoides amarelos. A correlação positiva entre a atividade antioxidante total e o teor de compostos fenólicos totais mostra que estes são os compostos mais importantes para a determinação da atividade antioxidante das polpas das frutas analisadas.

Termos para indexação:
bioma Amazônia; frutas exóticas; parâmetros físico-químicos das frutas; macronutrientes; micronutrientes

Introduction

Fruit consumption has increased in both Brazilian and international markets as a result of the increased awareness of the nutritional value and health benefits of fruit, and also as a result of new consumer habits generated by increased concerns over the relationship between diet and health (Liu, 2013LIU, R.H. Health-promoting components of fruits and vegetables in the diet. Advances in Nutrition an International Review Journal, v.4, p.384S-392S, 2013. DOI: 10.3945/an.112.003517.
https://doi.org/10.3945/an.112.003517....
). Some bioactive constituents present in fruit, such as vitamins, carotenoids, and phenolic compounds, are associated with the prevention of a series of chronic pathologies including cancer, cardiovascular disease, type-2 diabetes, and Alzheimer’s disease (Carter et al., 2010CARTER, P.; GRAY, L.J.; TROUGHTON, J.; KHUNTI, K.; DAVIES, M.J. Fruit and vegetables intake and incidence on type 2 diabetes mellitus: systematic review and meta-analysis. British Medical Journal, v.341, p.c4229, 2010. DOI: 10.1136/bmj.c4229.
https://doi.org/10.1136/bmj.c4229....
; Wang et al., 2011WANG, S.; MELNYK, J.P.; TSAO, R.; MARCONE, M.F. How natural dietary antioxidants in fruits, vegetables and legumes promote vascular health. Food Research International, v. 44, p.14-22, 2011. DOI: 10.1016/j.foodres.2010.09.028.
https://doi.org/10.1016/j.foodres.2010.0...
).

Brazil’s territory extends over 8.5 million km2. Its temperate and tropical climates, and soil conditions make fruit production one of the country’s main agribusiness activities. The country is the world’s third largest fruit producer, and is also one of the most biologically diverse regions in the world. Brazil’s large quantities of native fruit species, which are considered exotic in flavor, attract consumers from around the globe (FAO, 2013FAO. Food and Agriculture Organization of the United Nations. Top fruit producers and their productivity. In: FAO. Food and Agriculture Organization of the United Nations. FAO Statistical Yearbook 2013. Rome: FAO, 2013.; Anuário…, 2015ANUÁRIO BRASILEIRO DA FRUTICULTURA 2014. Santa Cruz do Sul: Gazeta Santa Cruz, 2015. 104p.).

In the Amazon biome, the largest one in Brazil, the greatest extent of biodiversity in the world can be found (Costa et al., 2013COSTA, A.G.V.; GARCIA-DIAZ, D.F.; JIMENEZ, P.; SILVA, P.I. Bioactive compounds and health benefits of exotic tropical red-black berries. Journal of Functional Foods, v.5, p.539-549, 2013. DOI: 10.1016/j.jff.2013.01.029.
https://doi.org/10.1016/j.jff.2013.01.02...
). This biodiversity includes both flora and fauna, as well as fruit with unique sensory characteristics and increased nutrient levels (Dembitsky et al., 2011DEMBITSKY, V.M.; POOVARODOM, S.; LEONTOWICZ, H.; LEONTOWICZ, M.; VEARASILP, S.; TRAKHTENBERG, S.; GORINSTEIN, S. The multiple nutrition properties of some exotic fruits: biological activity and active metabolites. Food Research International, v.44, p.1671-1701, 2011. DOI: 10.1016/j.foodres.2011.03.003.
https://doi.org/10.1016/j.foodres.2011.0...
). Fruit species such as abiu (Pouteria caimito), achachairu (Garcinia humilis), araza (Eugenia stipitata), bilimbi (Averrhoa bilimbi), and yellow mangosteen (Garcinia xanthochymus) have widely appreciated flavors by Brazilian consumers and are fruits of moderate importance to the economy. They show potential for commercialization in both domestic and international markets (Rógez et al., 2004RÓGEZ, H.; BUXANT, R.; MIGNOLET, E.; SOUZA, J.N.S.; SILVA, E.M.; LARONDELLE, Y. Chemical composition of the pulp of three typical Amazonian fruits: araça-boi (Eugenia stipitata), bacuri (Platonia insignis) and cupuaçu (Theobroma grandiflorum). European Food Research and Technology, v.218, p.380-384, 2004. DOI: 10.1007/s00217-003-0853-6.
https://doi.org/10.1007/s00217-003-0853-...
; Cavalcante et al., 2006CAVALCANTE, I.H.L.; JESUS, N. de; MARTINS, A.B.G. Physical and chemical characterization of yellow mangosteen fruits. Revista Brasileira de Fruticultura, v.28, p.325-327, 2006. DOI: 10.1590/S0100-29452006000200039.
https://doi.org/10.1590/S0100-2945200600...
; Lorenzi et al., 2006LORENZI, H.; BACHER, L.; LACERDA, M.; SARTORI, S. Frutas brasileiras e exóticas cultivadas (de consumo in natura). São Paulo: Instituto Plantarum de Estudos da Flora, 2006.; Duarte, 2011DUARTE, O. Achachairú (Garcinia humilis (Vhal) C. D. Adam). Postharvest Biology and Technology of Tropical and Subtropical Fruits: Açai to Citrus, v.54e, p.48-53, 2011.; Souza et al., 2011SOUZA, P.A. de; SENHOR, R.F.; COSTA, F.B. da; FREITAS, R.V. da S.; SILVA, M.S. Caracterização físico-química de frutos de bilimbí (Averrhoa bilimbi L.) produzidos no estado do RN. Revista Verde, v.6, p.270-273, 2011.; Garzón et al., 2012GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f.). These fruits are eaten raw; however, they can be used also in juices, ice creams, jams, and other sweets. Despite their nutritional and economic potential, these fruits lack an established commercialized market. They have still to be marketed properly because they are almost always picked from the wild. An additional consequence is that technical and scientific data on them are scarce, or nonexistent. These factors limit the consumption of these fruits to their regions of production; consequently, other national and international consumers (most of whom have more purchasing capacity) have limited access to them. Also, these fruits are not included in the Brazilian export basket items, a factor which leads to a loss in the productive potential of the Amazon region (Ribeiro & Ferreira, 2008RIBEIRO, G.D.; FERREIRA, M. das G.R. Comportamento inicial de duas fruteiras amazônicas e duas exóticas tropicais, em Porto Velho, Rondônia: abiu gigante (Pouteria caimito (Ruiz & Pav.) Radlk), araçá-boi (Eugenia stipitata Mc Vaugh.), abricó (Mammea americana Jacq.) e rambutan (Nephelium lappaceum L.). Porto Velho: Embrapa Rondônia, 2008. (Embrapa Rondônia. Comunicado técnico, 355).).

There is currently a need for interventions to stimulate small rural growers in the Amazon region to enable them to commercialize their fruits in markets that require high-quality products. Productive alternatives should also be developed to promote simultaneous species conservation, increased product value, and economic growth among rural farmers (Nobre et al., 2016NOBRE, C.A.; SAMPAIO, G.; BORMA, L.S.; CASTILLA-RUBIO, J.C.; SILVA, J.S.; CARDOSO, M. Land-use and climate change risks in the Amazon and the need of a novel sustainable development paradigm. Proceedings of the National Academy of Sciences of the United States of America, v.113, p.10759-10768, 2016. DOI: 10.1073/pnas.1605516113.
https://doi.org/10.1073/pnas.1605516113....
).

Few scientific studies have characterized abiu, achachairu, araza, bilimbi, and yellow mangosteen fruits. The physicochemical characteristics of abiu have been determined (Canuto et al., 2010CANUTO, G.A.B.; XAVIER, A.A.O.; NEVES, L.C.; BENASSI, M. de T. Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre. Revista Brasileira de Fruticultura, v.32, p.1196-1205, 2010. DOI: 10.1590/S0100-29452010005000122.
https://doi.org/10.1590/S0100-2945201000...
), as have those of achachairu (Pimentel, 2012PIMENTEL, M.R. da F. Caracterização qualitativa de frutos de achachairu (Garcinia humilis (Vahl) C. D. Adam) cultivados em Moreno-PE. 2012. 76p. Dissertação (Mestrado) - Universidade Federal Rural de Pernambuco, Recife. Available at: ˂Available at: ˂http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5091 ˃. Accessed on: Apr. 7 2017.
http://www.tede2.ufrpe.br:8080/tede2/han...
), araza (Rógez et al., 2004RÓGEZ, H.; BUXANT, R.; MIGNOLET, E.; SOUZA, J.N.S.; SILVA, E.M.; LARONDELLE, Y. Chemical composition of the pulp of three typical Amazonian fruits: araça-boi (Eugenia stipitata), bacuri (Platonia insignis) and cupuaçu (Theobroma grandiflorum). European Food Research and Technology, v.218, p.380-384, 2004. DOI: 10.1007/s00217-003-0853-6.
https://doi.org/10.1007/s00217-003-0853-...
; Canuto et al., 2010CANUTO, G.A.B.; XAVIER, A.A.O.; NEVES, L.C.; BENASSI, M. de T. Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre. Revista Brasileira de Fruticultura, v.32, p.1196-1205, 2010. DOI: 10.1590/S0100-29452010005000122.
https://doi.org/10.1590/S0100-2945201000...
), and bilimbi (Lima et al., 2001LIMA, V.L.A.G. de; MÉLO, E. de A.; LIMA, L. dos S. Physicochemical characteristics of bilimbi (Averrhoa bilimbi L). Revista Brasileira de Fruticultura, v.23, p.421-423, 2001. DOI: 10.1590/S0100-29452001000200045.
https://doi.org/10.1590/S0100-2945200100...
; Araújo et al., 2009ARAÚJO, E.R.; ALVES, L.I.F.; RÊGO, E.R. do; RÊGO, M.M. do; CASTRO, J.P. de; SAPUCAY, M.J.L. da C. Caracterização físico-química de frutos de biri-biri (Averrhoa bilimbi L.). Biotemas, v.22, p.225-230, 2009. DOI: 10.5007/2175-7925.2009v22n4p225.
https://doi.org/10.5007/2175-7925.2009v2...
; Souza et al., 2011SOUZA, P.A. de; SENHOR, R.F.; COSTA, F.B. da; FREITAS, R.V. da S.; SILVA, M.S. Caracterização físico-química de frutos de bilimbí (Averrhoa bilimbi L.) produzidos no estado do RN. Revista Verde, v.6, p.270-273, 2011.). Meanwhile, the bioactive compounds and total antioxidant activity have been determined in abiu (Canuto et al., 2010CANUTO, G.A.B.; XAVIER, A.A.O.; NEVES, L.C.; BENASSI, M. de T. Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre. Revista Brasileira de Fruticultura, v.32, p.1196-1205, 2010. DOI: 10.1590/S0100-29452010005000122.
https://doi.org/10.1590/S0100-2945201000...
), achachairu (Pimentel, 2012PIMENTEL, M.R. da F. Caracterização qualitativa de frutos de achachairu (Garcinia humilis (Vahl) C. D. Adam) cultivados em Moreno-PE. 2012. 76p. Dissertação (Mestrado) - Universidade Federal Rural de Pernambuco, Recife. Available at: ˂Available at: ˂http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5091 ˃. Accessed on: Apr. 7 2017.
http://www.tede2.ufrpe.br:8080/tede2/han...
), araza (Canuto et al., 2010CANUTO, G.A.B.; XAVIER, A.A.O.; NEVES, L.C.; BENASSI, M. de T. Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre. Revista Brasileira de Fruticultura, v.32, p.1196-1205, 2010. DOI: 10.1590/S0100-29452010005000122.
https://doi.org/10.1590/S0100-2945201000...
; Garzón et al., 2012GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f.), and yellow mangosteen (Cavalcante et al., 2006CAVALCANTE, I.H.L.; JESUS, N. de; MARTINS, A.B.G. Physical and chemical characterization of yellow mangosteen fruits. Revista Brasileira de Fruticultura, v.28, p.325-327, 2006. DOI: 10.1590/S0100-29452006000200039.
https://doi.org/10.1590/S0100-2945200600...
). No studies in the literature were found to provide a combined analysis of the macronutrients and micronutrients of these fruits from the Amazon biome.

The objective of this work was to evaluate the physicochemical composition, the bioactive compounds, and the total antioxidant activity of the fruit pulps of abiu, achachairu, araza, bilimbi, and yellow mangosteen from the Brazilian Amazon biome.

Materials and Methods

Abiu, achachairu, araza, bilimbi, and yellow mangosteen fruits, from the 2012/2013 harvest (November 2012-February 2013, during the summer), were acquired directly from the growers in the city of Cacoal, Rondônia state, Brazil (11°26'19"S, 61°26'50"W, at 200 m altitude). According to the Köppen-Geiger´s classification system for climate, the state of Rondônia has a climate that corresponds to the Aw type (tropical wet climate), with average temperatures in the coldest month above 18ºC, and a clearly defined dry season during the winter. Average annual rainfall ranges from 1,750 to 2,750 mm. Average annual temperatures vary between 24ºC and 26ºC, and relative humidity values stay between 80 and 85% (Climate-Data.Org, 2017CLIMATE-DATA.ORG. Clima: Cacoal. Available at: ˂Available at: ˂https://pt.climate-data.org/location/31797/ ˃. Accessed on: Apr. 7 2017.
https://pt.climate-data.org/location/317...
).

All fruits are cultivated in the Brazilian Amazon biome (Table 1). The fruits were harvested in a sufficiently ripe state for consumption, and they all came from the same rural property. The fruits were washed, sanitized, and dried. Those which exhibited mechanical damages and visible signs of contamination on the skins were discarded. The edible portion of each fruit was mashed and homogenized to produce a pulpy mass. The pulp from the fruits (approximately 5 kg) were packed in labeled plastic bags, and stored at -18ºC for analyses.

Table 1.
Common name, scientific name, family, and analyzed edible and nonedible portions of fruits from the Brazilian Amazon biome.

The fruit pulps were evaluated for their contents of moisture, protein, lipid, and ash, and for total soluble solid, titratable acidity, total and reducing sugars, and pH, using the method described by Horwitz (2005)HORWITZ, W. (Ed.). Official Methods of Analysis of the AOAC International. 18th ed. Gaithersburg: Association of Official Analytical Chemists, 2005. 771p.. All analyses were performed in triplicate. Total caloric value was calculated as the sum of the calories (kcal) provided by carbohydrates, lipids, and protein, whose values were multiplied in grams by the respective conversion factors of 4 kcal g-1, 9 kcal g-1, and 4 kcal g-1 (Merril & Watt, 1973MERRIL, A.L.; WATT, B.K. Energy value of foods: … basis and derivation. Washington: Usda, 1973. (Usda. Agriculture Handbook, 74).). Total carbohydrate content was defined by subtracting the protein, lipid, ash, and moisture content from 100%.

Ascorbic acid content of the pulps was determined (mg ascorbic acid 100 g-1 pulp) using the method described by Horwitz (2005)HORWITZ, W. (Ed.). Official Methods of Analysis of the AOAC International. 18th ed. Gaithersburg: Association of Official Analytical Chemists, 2005. 771p., and modified by Benassi & Antunes (1988)BENASSI, M. de T.; ANTUNES, A.J. Comparison of metaphosphoric and oxalic acids as extractants solutions for the determination of vitamin C in selected vegetables. Arquivos de Biologia e Tecnologia, v.31, p.507-513, 1988. , and the analyses were performed in triplicate.

Anthocyanin and yellow flavonoid contents of the fruit pulps were determined following the method described by Francis (1982)FRANCIS, F.J. Analysis of anthocyanins in foods. In: MARKAKIS, P. (Ed.). Anthocyanins as food colors. New York: Academic Press, 1982. p.181-207. DOI: 10.1016/B978-0-12-472550-8.50011-1.
https://doi.org/10.1016/B978-0-12-472550...
. Using the Beckman DU-640 spectrophotometer (Fullerton, CA, USA), absorbance was measured at 535 nm for total anthocyanin content, and at 374 nm for yellow flavonoid content. Yellow flavonoids and total anthocyanin contents were calculated (mg total anthocyanins or yellow flavonoids 100 g-1 of pulp) using the absorption coefficients (ε1cm1%) of 982 and 766, respectively (Silva et al., 2014SILVA, L.M.R. da; FIGUEIREDO, E.A.T. de; RICARDO, N.M.P.S.; VIEIRA, I.G.P.; FIGUEIREDO, R.W. de; BRASIL, I.M.; GOMES, C.L. Quantification of bioactive compounds in pulps and by-products of tropical fruits from Brazil. Food Chemistry, v.143, p.398-404, 2014. DOI: 10.1016/j.foodchem.2013.08.001.
https://doi.org/10.1016/j.foodchem.2013....
). The analyses were performed in triplicate.

Total carotenoid content of the fruit pulps was determined using the method described by Rodriguez-Amaya & Kimura (2004)RODRIGUEZ-AMAYA, D.B.; KIMURA, M. HarvestPlus Handbook of Carotenoid Analysis. Washington: IFPRI; Cali: CIAT, 2004.. Absorbance was measured at 450 nm in the Beckman DU-640 spectrophotometer (Fullerton, CA, USA). Total carotenoid content was calculated (μg of β-carotene 100 g-1 of pulp) using the absorption coefficients of β-carotene (ε1cm1%= 2592) in petroleum ether. The analyses were performed in triplicate.

The extracts and analyses of total phenolic compounds and antioxidant activity were obtained using the methodology described by Macoris et al. (2012)MACORIS, M.S.; DE MARCHI, R.; JANZANTTI, N.S.; MONTEIRO, M. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp. Journal of the Science of Food Agriculture, v.92, p.1886-1891, 2012. DOI: 10.1002/jsfa.5556.
https://doi.org/10.1002/jsfa.5556....
. Fruit pulps were blended with a 70% acetone extraction solution. Three extracts were obtained from each type of pulp. Total phenolic compounds of fruit pulps were determined using the Folin and Ciocalteu method (Waterhouse, 2002WATERHOUSE, A.L. Folin-Ciocalteau Micro Method for Total Phenol in Wine. 2002. Available at: ˂Available at: ˂http://waterhouse.ucdavis.edu/faqs/folin-ciocalteau-micro-method-for-total-phenol-in-wine ˃. Accessed on: Oct. 30 2014.
http://waterhouse.ucdavis.edu/faqs/folin...
; Macoris et al., 2012MACORIS, M.S.; DE MARCHI, R.; JANZANTTI, N.S.; MONTEIRO, M. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp. Journal of the Science of Food Agriculture, v.92, p.1886-1891, 2012. DOI: 10.1002/jsfa.5556.
https://doi.org/10.1002/jsfa.5556....
). Absorbance was measured at 720 nm in the Beckman DU-640 spectrophotometer (Fullerton, CA, USA). The results are expressed as mg gallic acid equivalent (GAE) 100 g-1 of pulp. The analyses were performed in triplicate for each type of pulp extract. Antioxidant activity of pulps was determined by capturing the ABTS•+ (2,2’-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) diammoninum salt) free radical (Rufino et al., 2010RUFINO, M. do S.M.; ALVES, R.E.; BRITO, E.S. de; PÉREZ-GIMÉNEZ, J.; SAURA-CALIXTO, F.; MANCINI-FILHO, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry, v.121, p.996-1022, 2010. DOI: 10.1016/j.foodchem.2010.01.037.
https://doi.org/10.1016/j.foodchem.2010....
; Macoris et al., 2012MACORIS, M.S.; DE MARCHI, R.; JANZANTTI, N.S.; MONTEIRO, M. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp. Journal of the Science of Food Agriculture, v.92, p.1886-1891, 2012. DOI: 10.1002/jsfa.5556.
https://doi.org/10.1002/jsfa.5556....
), as well as by capturing the DPPH• (2,2-diphenyl-1-picryl-hydrazyl) free radical (Rufino et al., 2010RUFINO, M. do S.M.; ALVES, R.E.; BRITO, E.S. de; PÉREZ-GIMÉNEZ, J.; SAURA-CALIXTO, F.; MANCINI-FILHO, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry, v.121, p.996-1022, 2010. DOI: 10.1016/j.foodchem.2010.01.037.
https://doi.org/10.1016/j.foodchem.2010....
), and by measuring iron-reducing activity using the ferric reducing antioxidant power (FRAP) assay (TPTZ reagent: 2,4,6-Tris (2-pyridyl)-s-triazine) (Rufino et al., 2010RUFINO, M. do S.M.; ALVES, R.E.; BRITO, E.S. de; PÉREZ-GIMÉNEZ, J.; SAURA-CALIXTO, F.; MANCINI-FILHO, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry, v.121, p.996-1022, 2010. DOI: 10.1016/j.foodchem.2010.01.037.
https://doi.org/10.1016/j.foodchem.2010....
). Absorbance was measured in the Beckman DU-640 spectrophotometer (Fullerton, CA, USA) at 734 nm for ABTS method, at 515 nm for DPPH method, and at 595 nm for FRAP method. The analyses were performed in triplicate for each fruit-type pulp extract, and the results were expressed as μmol of trolox 100 g-1 of pulp.

The results of each parameter of fruit pulps were subjected to the analysis of variance, followed by the Tukey’s test, in order to compare the means. The differences were considered significant at 5% probability. A linear correlation analysis was also performed, which considered the contents of ascorbic acid, total carotenoids, anthocyanin, yellow flavonoids, total phenolic compounds, and the antioxidant activity. The physicochemical means were fixed in columns (variables), and the different pulps of fruits were fixed in rows (cases). The data were standardized before analyses. The principal component analysis (PCA) was performed with a correlation matrix, without the factor rotation. Percentage variations greater than 70%, explained by the two first principal components, indicate a strong correlation among variables, and they also indicate that PCA is an appropriate multivariate analysis to be applied to the data. Cluster analysis was performed with Euclidean distances as the distance measurements, and Ward’s hierarchy was applied as the amalgamation rule to the individual data from each species pulp. The midpoint of the largest increment was used to separate the groups. Multidimensional scaling analysis was then applied to the resulting correlation matrix, which considered the chemical makeup, bioactive components, and antioxidant activity of each fruit pulp. Multidimensional scaling analysis is a multivariate technique based on proximities between objects (pulps), which are used to produce a spatial representation of these items.

The Statistica 7.0 software (StatSoft, Tulsa, OK, USA) was employed for the analyses.

Results and Discussion

The physicochemical composition of the fruit pulps from the Brazilian Amazon biome showed significant differences among the evaluated parameters (Table 2).

Table 2.
Physicochemical composition of fruit pulps from the Brazilian Amazon biome(1).

Abiu showed the highest values for total soluble solids, reducing sugars, total sugars, pH, protein, lipid, ash, and total calories, and the lowest values for titratable acidity, and moisture. Achachairu had the highest value for titratable acidity, and the lowest value for lipid content. Araza showed the lowest values for total soluble solids, reducing sugars, total sugars, pH, and ash. Bilimbi showed the highest values for moisture, and the lowest values for total calories. Yellow mangosteen showed the lowest-protein content. The analyzed fruit pulps do not have identity and quality standards in the current Brazilian laws for comparison.

Pimentel (2012)PIMENTEL, M.R. da F. Caracterização qualitativa de frutos de achachairu (Garcinia humilis (Vahl) C. D. Adam) cultivados em Moreno-PE. 2012. 76p. Dissertação (Mestrado) - Universidade Federal Rural de Pernambuco, Recife. Available at: ˂Available at: ˂http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5091 ˃. Accessed on: Apr. 7 2017.
http://www.tede2.ufrpe.br:8080/tede2/han...
analyzed the achachairu pulp from the Brazilian state of Pernambuco. The pulp was found to have 78.95% moisture, 0.24% ash content, 0.48% protein, 0.21% lipids, and 78.69 kcal 100g-1 total calories; these amounts differed from those of the present study. When analyzed by Canuto et al. (2010)CANUTO, G.A.B.; XAVIER, A.A.O.; NEVES, L.C.; BENASSI, M. de T. Caracterização físico-química de polpas de frutos da Amazônia e sua correlação com a atividade anti-radical livre. Revista Brasileira de Fruticultura, v.32, p.1196-1205, 2010. DOI: 10.1590/S0100-29452010005000122.
https://doi.org/10.1590/S0100-2945201000...
, the abiu pulp from the Amazon biome was found to have 3.8 °Brix total soluble solids, a value which was lower than that found in the present study. Meanwhile, their titratable acidity content of 5.9 mg of citric acid 100 g-1 was higher than that of the present study. The lipid content (0.1 g 100g-1) and pH (5.0) were similar to those found in the present study. In a study on araza (Garzón et al., 2012GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f.), data on soluble solids (4.6 °Brix) and on pH (2.6) were similar to those of the present study. A study on bilimbi by Araújo et al. (2009ARAÚJO, E.R.; ALVES, L.I.F.; RÊGO, E.R. do; RÊGO, M.M. do; CASTRO, J.P. de; SAPUCAY, M.J.L. da C. Caracterização físico-química de frutos de biri-biri (Averrhoa bilimbi L.). Biotemas, v.22, p.225-230, 2009. DOI: 10.5007/2175-7925.2009v22n4p225.
https://doi.org/10.5007/2175-7925.2009v2...
) found total soluble solids (3.23 °Brix) and pH (2.49), similar to those of the present study. Souza et al. (2011)SOUZA, P.A. de; SENHOR, R.F.; COSTA, F.B. da; FREITAS, R.V. da S.; SILVA, M.S. Caracterização físico-química de frutos de bilimbí (Averrhoa bilimbi L.) produzidos no estado do RN. Revista Verde, v.6, p.270-273, 2011. studied the bilimbi, and reported 4.95% soluble solids, and 5.10% oxalic acid in total titratable acidity. Yellow mangosteen pulp (Cavalcante et al., 2006CAVALCANTE, I.H.L.; JESUS, N. de; MARTINS, A.B.G. Physical and chemical characterization of yellow mangosteen fruits. Revista Brasileira de Fruticultura, v.28, p.325-327, 2006. DOI: 10.1590/S0100-29452006000200039.
https://doi.org/10.1590/S0100-2945200600...
) from Jaboticabal, SP, Brazil, was found to have 11.73 °Brix total soluble solids, and 4.19% titratable acidity contents, which are higher than those found in the current study. The physicochemical characteristics of the fruits may vary as a result of many factors, including weather conditions, location, use of pesticides, state of ripeness, processing, and storage (Souza et al., 2012SOUZA, V.R. de; PEREIRA, P.A.P.; QUEIROZ, F.; BORGES, S.V.; CARNEIRO, J. de D.S. Determination of bioactive compounds, antioxidant activity and chemical composition of Cerrado Brazilian fruits. Food Chemistry, v.134, p.381-386, 2012. DOI: 10.1016/j.foodchem.2012.02.191.
https://doi.org/10.1016/j.foodchem.2012....
).

Fruits are part of one of the food groups that are richest in bioactive compounds. They provide a series of health benefits, particularly because they act as antioxidants (Liu, 2013LIU, R.H. Health-promoting components of fruits and vegetables in the diet. Advances in Nutrition an International Review Journal, v.4, p.384S-392S, 2013. DOI: 10.3945/an.112.003517.
https://doi.org/10.3945/an.112.003517....
). Ascorbic acid content of the analyzed fruit pulps had the lowest values in abiu, and the highest ones in yellow mangosteen (Table 3). The daily recommended vitamin C intake (or daily recommended allowance) for adults is 90 mg for men, and 75 mg for women, and the maximum tolerable intake limit (or tolerable upper intake level) is 2 g per day (United States, 2000UNITED STATES. Institute of Medicine. Dietary reference intakes for vitamin C, vitamin E, selenium and carotenoids. Washington: National Academic Press, 2000.). According to the current Brazilian laws ((Anvisa, 2012ANVISA. Agência Nacional de Vigilância Sanitária. Resolução - RDC nº 54, de 12 de novembro de 2012. Dispõe sobre o Regulamento Técnico sobre Informação Nutricional Complementar. Diário Oficial da União, 13 nov. 2012. Seção 1, p.122-126.), for a food to be considered a “source” of vitamins, or a product with a “high content” of vitamins, 100 g or 100 mL of the food should contain 15 or 30% of the daily recommended allowance, respectively. From the analyzed fruits, yellow mangosteen can be considered a “source” of vitamin C, and could be marketed to female consumers, since 100 g of pulp contains 15.36% of their daily recommended allowance. Pimentel (2012)PIMENTEL, M.R. da F. Caracterização qualitativa de frutos de achachairu (Garcinia humilis (Vahl) C. D. Adam) cultivados em Moreno-PE. 2012. 76p. Dissertação (Mestrado) - Universidade Federal Rural de Pernambuco, Recife. Available at: ˂Available at: ˂http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5091 ˃. Accessed on: Apr. 7 2017.
http://www.tede2.ufrpe.br:8080/tede2/han...
observed 0.90 mg 100 g-1 ascorbic acid content, in a study on achachairu, a value much lower than the obtained one in the present study. Cavalcante et al. (2006)CAVALCANTE, I.H.L.; JESUS, N. de; MARTINS, A.B.G. Physical and chemical characterization of yellow mangosteen fruits. Revista Brasileira de Fruticultura, v.28, p.325-327, 2006. DOI: 10.1590/S0100-29452006000200039.
https://doi.org/10.1590/S0100-2945200600...
found 40.32 mg 100 g-1 ascorbic acid content, a much higher content than the ones reported here for yellow mangosteen, although their study used very different methods. Araújo et al. (2009)ARAÚJO, E.R.; ALVES, L.I.F.; RÊGO, E.R. do; RÊGO, M.M. do; CASTRO, J.P. de; SAPUCAY, M.J.L. da C. Caracterização físico-química de frutos de biri-biri (Averrhoa bilimbi L.). Biotemas, v.22, p.225-230, 2009. DOI: 10.5007/2175-7925.2009v22n4p225.
https://doi.org/10.5007/2175-7925.2009v2...
reported higher-ascorbic acid contents for bilimbi, at different stages of ripeness (50.82 mg 100 g-1 in unripe fruit, 41.69 mg 100 g-1 in semiripe fruit, and 32.01 mg 100 g-1 in ripe fruit).

Table 3.
Bioactive compounds and antioxidant activity in the pulp of fruits from the Brazilian Amazon biome(1).

Yellow flavonoid content had the lowest value in the abiu pulp, and the highest one in the yellow mangosteen pulp (Table 3). The content of yellow flavonoids in the yellow mangosteen pulp, found in the present study, was higher than the value reported by Rufino et al. (2010)RUFINO, M. do S.M.; ALVES, R.E.; BRITO, E.S. de; PÉREZ-GIMÉNEZ, J.; SAURA-CALIXTO, F.; MANCINI-FILHO, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry, v.121, p.996-1022, 2010. DOI: 10.1016/j.foodchem.2010.01.037.
https://doi.org/10.1016/j.foodchem.2010....
. The fruit pulps from the Amazon biome that were analyzed herein were found to contain low-anthocyanin contents (lower than the limit of quantification of 1 mg of anthocyanins 100 g-1 of pulp).

Carotenoids are natural pigments with many biological functions (Liu, 2013LIU, R.H. Health-promoting components of fruits and vegetables in the diet. Advances in Nutrition an International Review Journal, v.4, p.384S-392S, 2013. DOI: 10.3945/an.112.003517.
https://doi.org/10.3945/an.112.003517....
). β-carotene is commonly found in foods, and it is known for its provitamin A activity (Rodriguez-Amaya & Kimura, 2004RODRIGUEZ-AMAYA, D.B.; KIMURA, M. HarvestPlus Handbook of Carotenoid Analysis. Washington: IFPRI; Cali: CIAT, 2004.). Abiu showed the lowest value of total carotenoid content, and araza showed the highest value for this parameter. In their analysis of araza, Garzón et al. (2012)GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f. reported much higher-total carotenoid values than those found in the present study (806 μg β-carotene 100 g-1 pulp).

When it came to total phenolic compounds, the bilimbi pulp showed the lowest values, and the achachairu pulp showed the highest values. The total phenolic compound content (19.3 mg GAE 100 g-1) of araza from the Colombian Amazon (Garzón et al., 2012GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f.) was lower than the value found in the present study. The achachairu pulp analyzed by Pimentel (2012)PIMENTEL, M.R. da F. Caracterização qualitativa de frutos de achachairu (Garcinia humilis (Vahl) C. D. Adam) cultivados em Moreno-PE. 2012. 76p. Dissertação (Mestrado) - Universidade Federal Rural de Pernambuco, Recife. Available at: ˂Available at: ˂http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5091 ˃. Accessed on: Apr. 7 2017.
http://www.tede2.ufrpe.br:8080/tede2/han...
was found to have lower-total phenolic compound content (104.58 and 264.43 mg GAE 100 g-1 in hydromethanolic extracts and hydroacetone extracts, respectively) than the pulp analyzed in the present study. Vasco et al. (2008)VASCO, C.; RUALES, J.; KAMAL-ELDIN, A. Total phenolic compounds and antioxidant capacities of major fruits from Ecuador. Food Chemistry, v.111, p.816-823, 2008. DOI: 10.1016/j.foodchem.2008.04.054.
https://doi.org/10.1016/j.foodchem.2008....
classified the fruits as for polyphenol content and placed them into three categories: low (<100 mg 100 g-1 of GAE), moderate (100-500 mg 100 g-1 of GAE), and high (>500 mg 100 g-1 of GAE) content. This classification has been used in other studies (Rufino et al., 2010RUFINO, M. do S.M.; ALVES, R.E.; BRITO, E.S. de; PÉREZ-GIMÉNEZ, J.; SAURA-CALIXTO, F.; MANCINI-FILHO, J. Bioactive compounds and antioxidant capacities of 18 non-traditional tropical fruits from Brazil. Food Chemistry, v.121, p.996-1022, 2010. DOI: 10.1016/j.foodchem.2010.01.037.
https://doi.org/10.1016/j.foodchem.2010....
; Souza et al., 2012SOUZA, V.R. de; PEREIRA, P.A.P.; QUEIROZ, F.; BORGES, S.V.; CARNEIRO, J. de D.S. Determination of bioactive compounds, antioxidant activity and chemical composition of Cerrado Brazilian fruits. Food Chemistry, v.134, p.381-386, 2012. DOI: 10.1016/j.foodchem.2012.02.191.
https://doi.org/10.1016/j.foodchem.2012....
). Thus, bilimbi pulp can be classified as having low-polyphenol content, while abiu, achachairu, araza, and yellow mangosteen pulps can be classified as having moderate quantities of polyphenol content. The Folin-Ciocalteu reagent is frequently used in colorimetric in vitro assays because it is simple, and exhibits strong reproducibility. This method is employed to quantify all classes of polyhydroxy phenolic compounds. However, other nonphenolic compounds, such as ascorbic acid, certain sugars, and amino acids may also react with the Folin-Ciocalteu reagent (Rocha et al., 2011ROCHA, W.S.; LOPES, R.M.; SILVA, D.B. da; VIEIRA, R.F.; SILVA, J.P. da; AGOSTINI-COSTA, T. da S. Compostos fenólicos totais e taninos condensados em frutas nativas do Cerrado. Revista Brasileira de Fruticultura, v.33, p.1215-1221, 2011. DOI: 10.1590/S0100-29452011000400021.
https://doi.org/10.1590/S0100-2945201100...
; Wootton-Beard & Ryan, 2011WOOTTON-BEARD, P.C.; RYAN, L. Improving public health?: The role of antioxidant-rich fruit and vegetable beverages. Food Research International, v.44, p.3135-3148, 2011. DOI: 10.1016/j.foodres.2011.09.015.
https://doi.org/10.1016/j.foodres.2011.0...
; Macoris et al., 2012MACORIS, M.S.; DE MARCHI, R.; JANZANTTI, N.S.; MONTEIRO, M. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp. Journal of the Science of Food Agriculture, v.92, p.1886-1891, 2012. DOI: 10.1002/jsfa.5556.
https://doi.org/10.1002/jsfa.5556....
). Due to the low concentration of ascorbic acid and sugars in the analyzed fruits in the present study, an extraction step was not necessary to remove these interferents.

The ABTS, DPPH, FRAP, and orygen radical absorbance capacity (ORAC) methods used to determine the antioxidant activity are most commonly used in fruits. Pérez-Jiménez et al. (2008)PÉREZ-JIMÉNEZ, J.; ARRANZ, S.; TABERNERO, M.; DÍAZ-RUBIO, M.E.; SERRANO, J.; GOÑI, I.; SAURA-CALIXTO, F. Updated methodology to determine antioxidant capacity in plant foods, oils and beverages: extraction, measurement and expression of results. Food Research International, v.41, p.274-285, 2008. DOI: 10.1016/j.foodres.2007.12.004.
https://doi.org/10.1016/j.foodres.2007.1...
recommend using more than one method (and preferably all of them), in order to acquire more complete information on the antioxidant capacity of foods. This variety of testing allows researchers to consider the advantages or disadvantages of each method, as well as each method’s acceptability. In all the applied assays herein, the achachairu pulp was found to have the highest antioxidant activity, and the bilimbi pulp was found to have the lowest one (Table 3). Garzón et al. (2012)GARZÓN, G.A.; NARVÁEZ-CUENCA, C.-E.; KOPEC, R.E.; BARRY, A.M.; RIEDL, K.M.; SCHWARTZ, S.J. Determination of carotenoids, total phenolic content, and antioxidant activity of Arazá (Eugenia stipitata McVaugh), an Amazonian fruit. Journal of Agricultural and Food Chemistry, v.60, p.4709-4717, 2012. DOI: 10.1021/jf205347f. studied araza, and reported a total antioxidant activity of 1.2 μmol g-1 of trolox when using the ABTS, 0.8 μmol g-1 of trolox when using the DPPH, and 3.5 μmol g-1 of trolox when using the FRAP method; all these values are lower than those found in the present study. The ABTS, DPPH, and FRAP methods showed a correlation only between antioxidant activity and total phenolic compounds (r=0.784, p≤0.05; r=0.829, p≤0.05; and r=0.936, p≤0.05, respectively). This correlation suggests that phenolic compound contents are the most important factor for determining the antioxidant activity in the analyzed fruit pulps. A positive linear correlation between antioxidant activity and total phenolic compounds has also been reported in other studies on fruits (Thaipong et al., 2006THAIPONG, K.; BOONPRAKOB, U.; CROSBY, K.; CISNEROS-ZEVALLOS, L.; BYRNE, D.H. Comparison of ABTS, DPPH, FRAP and ORAC assays for estimating antioxidant activity from guava fruit extracts. Journal of Food Composition and Analysis, v.19, p.669-675, 2006. DOI: 10.1016/j.jfca.2006.01.003.
https://doi.org/10.1016/j.jfca.2006.01.0...
; Macoris et al., 2012MACORIS, M.S.; DE MARCHI, R.; JANZANTTI, N.S.; MONTEIRO, M. The influence of ripening stage and cultivation system on the total antioxidant activity and total phenolic compounds of yellow passion fruit pulp. Journal of the Science of Food Agriculture, v.92, p.1886-1891, 2012. DOI: 10.1002/jsfa.5556.
https://doi.org/10.1002/jsfa.5556....
).

A principal component analysis was used to better visualize the physicochemical parameter, bioactive compounds, and antioxidant activity of the pulps of fruits (2012-2013 harvest) from the Amazon biome (Figure 1 A). The two first main components accounted for 76.54% of the variance. The first main component was positively associated with the moisture content and total carotenoid parameters, and it was negatively associated with ash content, total sugars, reducing sugars, total soluble solids, carbohydrates, total calories, and pH. The second main component was negatively associated with titratable acidity, total phenolic compounds, and antioxidant activity values acquired using the ABTS, DPPH, and FRAP assays. The fruit pulps were placed in different quadrants because their characteristics are all very different (Figure 1 B). The achachairu pulp was characterized to have the highest titratable acidity content, the largest phenolic compound content, and the highest antioxidant activity in the ABTS, DPPH, and FRAP assays. The yellow mangosteen pulp was characterized to have the highest yellow flavonoid content, while the abiu pulp was characterized to have the highest contents for ash, protein, lipid, total sugars, reducing sugars, and total soluble solids, besides total calories, and pH. The araza pulp was characterized to contain the highest carotenoid content and moisture content. Meanwhile, the bilimbi pulp was characterized to have the highest ascorbic acid content.

Figure 1.
Principal component analysis (A) and dendrogram (B) of the physicochemical parameters, bioactive compounds, and antioxidant activity of pulps of the fruits abiu, achachairu, araza, bilimbi, and yellow mangosteen, from the Brazilian Amazon biome. TSS, total soluble solids; RS, reducing sugars; TS, total sugars; TA, titratable acidity; AA, ascorbic acid; YF, yellow flavonoids; TC, total carotenoids; TPC, total phenolic compounds; M, moisture; P, protein; L, lipid; A, ash; C, carbohydrates; TCV, total caloric value.

These results can help the small-scale rural farmers in the Brazilian Amazon region to get their fruits marketed as high-quality products, and can also provide information that can contribute to the development of new products with higher concentration of nutrients and, therefore, of greater value. Abiu, achachairu, araza, bilimbi, and the yellow mangosteen fruits from other harvests in the Brazilian Amazon biome should be evaluated in further researches, in order to confirm the results obtained in the present study on their physicochemical parameters and bioactive compounds.

Conclusions

  1. The fruits abiu (Pouteria caimito), achachairu (Garcinia humilis), araza (Eugenia stipitata), bilimbi (Averrhoa bilimbi), and yellow mangosteen (Garcinia xanthochymus) show moisture contents above 80% and low-lipid contents; and araza and bilimbi are considered low-calorie fruits.

  2. As to bioactive components, the achachairu pulp has the highest-total phenolic compound content, while the araza pulp has the highest-carotenoid content, and the yellow mangosteen pulp has the highest-yellow flavonoid content.

  3. The antioxidant activity in these fruits is highly correlated with total-phenolic compound content.

Acknowledgments

To Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes), for the scholarship granted; and to Universidade Estadual Paulista (Unesp/Prope), for financial support.

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Publication Dates

  • Publication in this collection
    Oct 2017

History

  • Received
    05 July 2016
  • Accepted
    06 Mar 2017
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