( T h is i s a sa mpl e co ve r im ag e fo r t h is i ss ue . T h e act u al co ver i s no t yet ava il ab l e at t hi s t i me.

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Food Research International

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Chemical, technological and in vitro antioxid ant properties of cocoa (Theobroma cacao L.) co-produ cts
R. Martnez a, P. Torres a, M.A. Meneses a, J.G. Figueroa a, J.A. Prez-lvarez b, M. Viuda-Martos b,
a U ni ver si d ad T c ni c a P ar ti cu l ar de L o ja . C ent ro de Tr an sf er en ci a de Tec n ol og a e I nv esti ga c i n A g roi n du s tri a l ( C ETT I A) , San Ca yeta no A lto , L o ja , Ec u ad or

b IP OA R esea rc h Gro up ( UM H- 1 a nd RE V I V- Gen era l it at V al en c ia na ) , A gr oFo od T ec hn ol og y Dep ar tmen t, Es cu el a P ol it c ni c a Su pe ri or de O ri hu el a, Mi gu el Her n n dez Un i vers i ty, C rt a. B en i el km . 3 ,2 . E- 0 33 12 Ori h ue la , A l ic a nte, Spa i n

article info
A r ti c le h i sto ry : R e c ei ved 9 M ay 2 0 12 Ac c e p ted 2 Aug u s t 20 1 2 Av ai l ab l e o nl i n e 1 9 A ug u st 20 1 2 K ey wor ds : C o c oa C o -p r od u c t An t i ox id a nt Fi b er

abstract
The aim of this work was to determine the chemical, technological and in vitro antioxidant properties of cocoa co-products such as cocoa pod husks, cocoa bean shell and cocoa mucilage to determine the potential used as a dietary ber source for food enrichment. The proximate composition and total (TDF), insoluble (IDF) and soluble dietary ber (SDF) content were determined. The water holding, oil holding and swelling capacities and total phenol content (TPC) were also determined. For the antioxidant activity, three different analytical assays were used (ABTS, DPPH and FRAP). The cocoa co-products dietary ber obtained in this study ranged between 16.86 and 55.59 g/100 g. The TPC of cocoa pod husk ranging between 206.67 and 365.33 mg gallic acid equivalent (GAE)/100 g sample, depending the locality and solvent system used while in as regards to cocoa bean shell and cocoa mucilage the TPC levels were signi cantly lower (80.17144.83 mg GAE/100 g and 102.00182.63 mg GAE/100 g respectively). All samples analyzed showed a good antioxidant capacity in the three different methods used with values ranging between from 2.48 to 22.93 MTrolox Equivalents (TEs)/g in ABTS assay; 1.5733.93 M TEs/g in DPPH assay and 0.67 and 4.69 M TEs/g sample in FRAP assay. The results of this study indicate that cocoa co-products may be considered a good sour ce of natural compounds with signi cant antiox idant activity. 2012 Elsevier Ltd. All rights reserved.

1. Introduction Cocoa-derived foods (cacao powders, chocolate, cocoa-related products) are phenolic-rich foods derived from the fermented, roasted and milled seeds of Theobroma cacao L. (Sterculiaceae) (Arlorio et al., 2005) and are now among the most widely consumed of processed foods. Evidence based on epidemiological studies suggests that consumption of cocoa-containing products may confer cardiovascular protection, reducing the risk of CVD mortality (Djouss et al., 2011). In addition, cocoa products increase plasma antioxidant capacity (Lotito & Frei, 2006), may reduce blood pressure (Visioli et al., 2009), inhibit the oxidation of LDL particles in humans ex vivo (Khan et al., in press) and reduce biomarkers of oxidation such as F2-isoprostanes and malondialdehyde (Wiswedel et al., 2004).

pod rot (Barazarte, Sangronis, & Unai, 2008) because they are not composted. However, their composition gives them the potential to be used for other end, for example to obtain bioactive compounds and dietary ber which could be used as ingredient in food processing. Recently, the valorization of agricultural co-products has received growing attention, because of the increasing shortage of natural resource and serious environmental problems. Many researchers have attempted to convert such co-products into food ingredients and for use in other value-added applications (Fernndez-Lpez et al., 2009; Viuda-Martos, Ruiz-Navajas et al., 2010; Viuda-Martos et al., 2011). Thus, the useof these co-products for further exploitation as food additives or supplements of high nutritional value has gained increasing interest becausethesearehigh-valueproducts and their recovery may be economically attractive (Murthy & Naidu, 2012).

Once the cocoa dry bean has been obtained, the co-products that remain are composed mainly by three fractions (i) cocoa pod husk, (ii) cocoa bean shells and (iii) cocoa mucilage. In most cases, these co-products are underexploited and considered an undesirable waste of the cocoa/chocolate industry. Normally, they are left to rot on the cacao plantation, which can cause environmental problems. Besides producing foul odors, they can propagate diseases, such as black

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The food industry is experiencing a constantly growing demand for new ingredients from natural sources. This demand has therefore drawn researchers to these ingredients obtained from agro-industrial co-products (Guerrero, Torres, & Nuez, 2008). Depending on the availability of an adequate technology, the co-products can be converted into commercial products either as raw materials for secondary processes (intermediate foods ingredients), as operating supplies, or as ingredients of new products (Snchez-Zapata et al., 2009). Although plant food co-products are interesting from a nutritional

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point of view, only a few of them have been successfully developed from the vast quantities of plant co-products produced by the food processing industry. Several potential uses can be considered for cocoa co-products; however very few studies have looked at how they can be used. The aim of this work therefore was to determine the chemical, technological and in vitro antioxidant properties of cocoa co-products, such as cocoa pod husks, cocoa bean shell and cocoa mucilage obtained from the industrialization of cocoa to determine the potential use as an antioxidant dietary ber source for food enrichment.

2.5. Total phenol content The total phenol content (TPC) was determined using the Folin-Ciocalteu's reagent (Singleton & Rossi, 1965). A volume of 0.3 mL of the ethanol or methanol:acetone solutions of the different samples (30 g/L) was introduced into test tubes followed by 2.5 mL of Folin Ciocalt eu's reagent (diluted 10 times with water) and 2 mL of sodium carbonate (7.5% w/v). The tubes were shaking; covered with para lm and incubated at 50 C for 5 min. Absorption at 760 nm was measured with a HP 8451 spectrophotometer (Hewlett Packard, Cambridge, UK) and compared to a gallic acid calibration curve. The results were expressed as mg gallic acid equivalents (GAE)/g dry mass, as mean of three replicates.

2. Materials and methods 2.1. Materials

2.6. Sample preparation to determine the antioxidant activity Cocoa (Theobroma cacao L cv. complejo nacional x trinitario) co-products from industrialization to obtain chocolate such as, cocoa pod husk, cocoa bean shell and cocoa mucilage from two different localizations (Cone and Taura; Ecuador) were used. 2.2. Obtaining ber concentrates Cocoa pod husk and cocoa bean shells were washed twice with warm water (30 C) in a proportion of 1:1 (v:w); then, after dried at 60 C for 12 h in an air tunnel drier, they were ground to a particle size of 220640 m. The material was washed under mild conditions to avoid or minimize loss of soluble ber components (such as pectin and pentoses) bioactive components (such as avonoids, phenolic acids and tannins). The washing process reduces the free sugar and ash contents, while drying at temperatures below 65 C avoids changes in the functional properties and in the bioactive compounds. The relative large particle size after grinding was intended not affect the hydration characteristics of the ber concentrates. To obtain extracts for antioxidant activity two different methodologies were used. The rst consisted of dynamic maceration in absolute ethanol for 24 h at room temperature with a solvent material ratio of 1:20. In the second procedure cocoa pod husk and cocoa bean shells (5 g) and cocoa mucilage (0.5 g) were extracted by shaking at room temperature for 60 min with 20 mL of methanol-water-HCl (50:50 v/v, pH 2) and by shaking at room temperature for 60 min with 20 mL of acetonewater (70:30 v/v). After centrifugation (15 min, 25 C, 3000 g) supernatants were combined and used to determine extractable antioxidant capacity.

2.7. Antioxidant activity determinations 2.7.1. ABTS assays For ABTS assay, the procedure followed the method by Arnao, Cano, and Acosta(2001) with some modi cations. The stock solutions included 7.4 mM ABTS + solution and 2.6 mM potassium persulfate solution. The working solution was then prepared by mixing the two stock solutions in equal quantities and allowing them to react for 12 h at room temperature in the dark. The solution was then diluted by mixing 1 mL ABTS+ solution with 60 mL methanol to obtain an absorbance of 1.10.02 units at 734 nm using the spectrophotometer. Fresh ABTS + solution was prepared for each assay. Cocoa co-products extracts (150 L) were allowed to react with 2850 LoftheABTS+ solution for 2 h in a dark condition and then the absorbance was measured at 734 nm. The standard curve was linear between 25 and 600 mM Trolox. Results are expressed in M Trolox equivalents (TE)/gdry mass, as mean of three replicates.

2.3. Chemical analysis Ash, moisture, protein and fat content were determined by AOAC methods (AOAC, 1997). Moisture (g water/100 g dry mater (d.m.)) was determined by drying a 3 g sample at 105 C to constant weight. Ash (g ash/100 g d.m.) was performed at 550 C for 2 h. Protein (g protein/100 g d.m.) was analyzed according to the Kjeldahl method. Fat (g fat/100 g d.m.) was calculated by weight loss after a 6-cycle extraction with petroleum ether in a Soxhlet apparatus. Total dietary ber (TDF) (g TDF/100 g d.m.) and insoluble dietary ber (IDF) were determined following 991.43 AOAC methods (AOAC, 1997). Soluble dietary ber (SDF) was calculated by subtracting the IDF proportion from the TDF. Carbohydrates were determined by difference from the total dietary ber, lipids, protein and ash contents. Each assay was carried out in triplicate.

2.4. Technological properties. The water-holding capacity (WHC) and oil holding capacity (OHC) were determined according to Robertson et al. (2000) with some modi cations. Twenty- ve milliliters of buffer phosphate (1 M, pH 6.3) or commercial olive oil were added to 250 mg of dry sample, stirred and left at room temperature for 1 h. After centrifugation, the residue was weighed. The WHC was expressed as g of water held per g of sample, while the OHC was expressed as g of oil held per g of sample. Swelling capacity was determined according to Robertson et al. (2000). An accurately weighed dry sample (0.2 g) was taken in a graduated test tube and then 10 mL of phosphate buffer (1 M, pH 6.3) was added and hydrated for 18 h. After 18 h, the nal volume attained by ber was measured. Each assay was carried out in triplicate.

2.7.2. DPPH assay The DPPH assay followed the method of Brand-Williams, Cuvelier, and Berset (1995) with some modi cations. The stock solution was prepared by dissolving 24 mg DPPH with 100 mL methanol and then stored at -20 C until needed. The working solution was obtained by mixing 10 mL stock solution with 45 mL methanol to obtain an absorbance of 1.10.02 units at 515 nm using the spectrophotometer. Cocoa co-products extracts (150 L) were allowed to react with 2850 L of the DPPH solution for 24 h in the dark. Then the absorbance was taken at 515 nm. The standard curve was linear between 25 and 800 mM Trolox. Results are expressed in M TE/g dry mass as mean of three replicates. 2.7.3. FRAP assay The FRAP assay was carried out according to Benzie and Strain (1996) with some modi cations. The stock solutions included 300 mM acetate bufferpH 3.6, 10 mM TPTZ (2,4,6-tripyridyl-s-triazine) solution in 40 mM HCl, and 20 mM FeCl3 6H2O solution. The fresh working solution was prepared by mixing 25 mL acetate buffer, 2.5 mL TPTZ solution, and 2.5 mL FeCl36H2O solution and then warmed at 37 C before using. Cocoa co-products (150 L) were

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allowed to react with 2850 L of the FRAP solution for 30 min in the dark. Readings of the colored product [ferrous tripyridyltriazine complex] were then taken at 593 nm. The standard curve was linear between 25 and 800 mM Trolox. Results are expressed in M TE/g dry mass as mean of three replicates.

2.8. Statistically assay Statistical analysis and comparisons among means were carried out using the statistical package SPSS 19.0 (SPSS Inc., Chicago, IL.). The data collected for chemical and technological properties, as well as for the antioxidant activity were analyzed by two-way analysis of variance to test the effects of three xed factors: ber source (cocoa pod husk, cocoa bean shell and cocoa mucilage), locality (Cone and Taura) and extracting agent (ethanol, methanol:acetone). The Tukey's post hoc test was applied for comparisonsofmeans,anddifferenceswere considered signi cant at p> 0.05. Correlations between antioxidant capacity and total phenolic content weredetermined using Pearson's Correlation Coef cient Test.

Table 2 shows the contents of total dietary ber (TDF), insoluble dietary ber (IDF), soluble dietary ber (SDF) and the ratio between IDF and SDF of cocoa co-products. As regards cocoa mucilage, the TDF content range between 16.75 and 16.86 g/100 g d.m. with no statistically differences (p> 0.05) between them. SDF (the main form) values ranged between 16.06 and 16.11 g/100 g d.m., while the small amount of IDF ranged between 0.69 and 0.78 g/100 g d.m. The IDF and SDF content of Taura was lower (pb 0.05) than that found for Cone. These values mean that the mucilage obtained from cocoa co-products can be regarded as an ingredient of great interest to be incorporated in several foods, due to the capacity of SDF to retain water and increase satisfaction after eating, as well as its ability to decrease the time of nutrient absorption. In addition, these co-products would act technologically as important thickening agents, gelling and stabilizers of foams and emulsions, and

3. Results and discussion 3.1. Chemical analysis The chemical properties of the different cocoa co-product fractions are shown in Table 1. The highest level of proteins (pb 0.05) were found in cocoa bean shells with no statistically differences (p> 0.05) between the localities of Cone and Taura. These values are very similar to those reported by Lecumberri et al. (2007) for cocoa hulls (16.71 g/100 g of sample) or Viuda-Martos et al. (2012) in pomegranate arils bagasse (12.60 g/100 g of sample). The protein content in cocoa mucilage and cocoa pod husk ranged (p b0.05) between 4.21 and 5.56 g/100 g d.m. with no statistically differences (p> 0.05) between the respective localities. These values agree with those found in other fruit co-products such as mango (4.28 g/100 g of sample) or apple (5.21 g/100 g of sample) (Vergara-Valencia et al., 2007; do Esprito Santo et al., 2012). The cocoa co-products had a lipid content with values ranging (p b0.05) between 1.91 and 2.34 g/100 g of d.m. which is lower than that reported, for example, in tomato peel ber (6.01 g/100 g dry sample) (Navarro-Gonzlez, Garca-Valverde, Garca-Alonso, & Periago, 2011) or tiger nuts co-products (8.85 g/100 g dry sample) (Snchez-Zapata et al., 2009). The cocoa co-products showed a very high ash content, with values (pb 0.05) between 6.76 g/100 g d.m. in cocoa bean shell and 8.42 g/100 g d.m. in cocoa pod husk, both of which are higher than the values found in other fruit co-products such as passion fruits seeds (1.34 g/100 g) or apple pomace (0.50 g/100 g) (Chau & Huang, 2004; Sudha, Baskaran, & Leelavathi, 2007). The high ash content could be a problem in the potential application of these co-products in food, since the amounts of metal ions would increase considerably and might facilitate the oxidation of the product in which they are incorporated.

lm-forming and fat-mimetic agents. They may also have a potential use in encapsulation. TDF content of cocoa pod husks was 55.99 and 56.10 g/100 g d.m. for Taura and Cone respectively, with no statistically signi cant differences (p>0.05) between them. The IDF content of cocoa pod husk from Taura was higher (p b0.05) than that obtained from Cone and, consequently, the SDF content of cocoa pod husk from Cone was higher (p b0.05) than that obtained from Taura. As regards cocoa bean shell, the TDF content ranged between 52.23 and 56.98 g/100 g d.m. with statistically signi cant differences (pb 0.05) between them. As in the case of cocoa pod husk, the IDF content of the bean shells from Taura was higher (p b0.05) than that from Cone, while the SDF content of Cone was higher (p b0.05) than that from Taura. It should be noted that cocoa pod husks had an IDF/SDF ratio ranging (pb 0.05) between 12.60 and 18.44, while the same ratio in cocoa bean shell ranged (p b0.05) between 2.19 and 2.90. It is very important for a ber source to show a good balanced between IDF and SDF since, in general terms, SDF has a high hydration capacity and swells to form viscous solutions. It also adsorbs and retains other substances like minerals, non-polar molecules, and glucose, while IDF can also adsorb and retain water within its brous matrix and adsorb other components such as SDF but without forming viscous solutions (Al-Sheraji et al., 2011).

The TDF content of all the samples analyzed was similar to that found in other fruits co-products such as banana ber (51.19 % dry sample) (do Esprito Santo et al., 2012), or in ber from chia (51.98 g/100 g dry sample) (Capitani, Spotorno, Nolasco, & Toms, 2012) and lower than that reported in dietary ber powder obtained from lime co-products (70.76 g/100 g) (Peerajit, Chiewchan, & Devahastin, 2012), date ber concentrates 88.0 g/100 g dry sample) (Elleuch et al., 2008) or orange co-product (71.62 g/100 g dry sample) (Fernndez-Lpez et al., 2009). The IDF fraction was higher than the SDF fraction in both cocoa pod husks and cocoa bean shells and would be a good indicator of the considerable amount of celluloses and hemicelluloses present in them.

Dietary ber intake in industrialized countries is currently estimated to be less than 25 g per person per day. However, nutritionists recommend intakes of 35 g per person per day (Vuksan et al., 2008).

T ab le 1 C h em i c al c o mp o s it i on o f th e d i ffer en t c o c oa c o - pr o du c t s ( c o c oa p o d hu s k s, c oc o a b ean s h el l , c o c oa m u c i l ag e) (M e an SD ) . C o -p r od u c t Loc a li za ti on Pr ot ei n As h Fa t C ar bo h yd r ate s (g /1 00 g d. m .) ( g /1 00 g d . m .) ( g/ 10 0 g d .m . ) (g /1 0 0 g d. m .)

C o c oa p o d h u s ks Co n e 4 .2 1 0 .1 6a , 8. 42 0 .1 0 a 2 .3 4 0 .0 8 a 2 9. 0 4 0 . 18 a 6 .5 3 0. 13 a Tau r a 4 .2 2 0 .0 7a 8. 32 0 .0 7 a 2 .2 4 0 .1 0 a 2 8. 7 3 0 . 11 a 6 .7 2 0. 17 a C o c oa b ea n s h el l Co n e 15 .8 5 0 .1 7b 7. 35 0 .1 5 b 2 .0 2 0 .0 3 b 17 .8 0 . 09 b 7 .7 1 0. 12 b Tau r a 15 .7 9 0 .0 4b 6. 76 0 .0 9 c 2 .0 5 0 .0 6 b 2 3. 1 7 0 . 12 b 7 .8 0 0. 17 b C o c oa m u c i l age Co n e 5 .4 7 0 .1 2c 7. 51 0 .1 4 b 1 .9 2 0 .0 6 c 6 8. 3 5 0 . 16 c 9 .6 4 0. 13 c Tau r a 5 .5 6 0 .1 0c 7. 68 0 .1 8 b 1 .9 1 0 .0 4 c 6 7. 9 9 0 . 14 c 9 .2 7 0. 17 c

M oi s tu r e ( g/1 0 0 g)

Val u es fol l o wed b y th e sa me l ett er w i th i n th e s am e c o l um n ar e n o t s ig n i ca n tl y d i ffer en t ( p > 0 .0 5) ac c o r di n g to T u key 's M u l ti p l e R an ge Tes t. D ry m att er: d. m .

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Chel-Guerrero, & Betancur-Ancona, 2009). This is an important propTa ble 2 To tal d i eta ry b r e ( TD F) , i n s ol u b le d ie tar y b re ( ID F) , s o l ub l e d i eta ry br e ( SD F) oferty o f of dietary ber from both physiological and technological points th e d i ffer en t c o c oa c o- p ro d u c ts ( c o c oa p od h u s ks , c o c oa be an s h el l , c oc o a m uc of i l ag e) view. (M e an SD ) .
C o -p r od u c t Lo c al i zat i on T DF ( g/1 00 g d .m .) SDF (g /1 0 0 g d. m .) IDF ( g /1 00 g d . m. ) R at io I D F/SD F

C o c oa T au ra 1 6. 75 0 . 07 a 16 .0 6 0 .2 0 0 .6 9 0. 1 5 0. 0 4 0. 35 m u c i l agC e o n e 1 6. 89 0 . 18 a 16 .1 1 0 .2 5 0 .7 8 0. 1 5 0. 0 4 0. 19 C o c oa T au ra 5 5. 99 0 . 22 a 2 .8 8 0 .0 3 5 3 .1 1 0. 8 4 1 8. 4 4 0. 04 C o n e 5 6. 10 0 . 13 a 4 .1 2 0 .0 7 5 1 .9 8 0. 4 8 1 2. 6 0 0. 12 pod h u s ks C o c oa b ea n s h el l
T au ra 5 6. 70 0 . 11 a 14 .5 3 0 .1 0 4 2 .1 7 0. 1 3 2. 9 0 0. 11 C o n e 5 1. 88 0 . 14 b 16 .2 4 0 .0 4 3 5 .6 4 0. 2 0 2. 1 9 0. 09

The SWC (Table 3) of cocoa pod husk was 5.70-5.81 mL/g sample, with no statistically signi cant differences (p >0.05) between the localities of Taura and Cone. This was lower than the SWC of ber-rich cocoa product and dietary ber extracted from sweet potato varieties (6.51, 8.11-10.56 mL/g, respectively) (Lecumberri et al., 2007; Mei, Mu, & Han, 2010). The cocoa bean shell showed a SWC ranging between 3.68 and 3.87 mL/g sample, with no statistically signi cant differences (p >0.05) between Taura and Cone. As occurred with WHC, cocoa pod husks showed a higher (p b0.05) SWC than cocoa bean shell. It is known that the structural characteristics and the chemical composition of ber (including the water af nity of its components) play an important role in the kinetics of water uptake. According to

Val u es fo ll o we d b y t h e s am e l et ter wi th i n th e s a me c ol u m n are n ot s i gn i c an t ly di ffe ren t (p >0 . 05 ) ac c or d i ng to Tu k ey' s Mu l ti p l e R an g e Te s t. Dr y m att er : d .m .

One way to increase the amount of ber in the diet without radically changing eating habits would be to consume foods enriched with ber (Viuda-Martos, Ruiz-Navajas et al., 2010). Thus, a high proportion of IDF in the cocoaco-products could be considered an advantage because of its potential application as a functional ingredient in confectionery or in the preparation of low-fat, high- ber dietetic products. DF exerts a buffering effect and binds excess hydrochloric acid in the stomach, increases the fecal bulk and stimulates intestinal peristaltic, as well as provides a favorable environment for the growth of the desired intestinal ora (Viuda-Martos, Lpez-Marcos et al., 2010).

Lpez et al. (1996) water may be held in capillary structures within the ber because of its high surface tension; in addition, water is capable of interacting with various molecular components of ber through hydrogen bonding or dipole formation. As regards OHC, cocoa pod husk samples were characterized by a low values (Table 3) which ranged between 1.18 and 1.20 g oil/g sample with no statistically signi cant differences (p> 0.05) between the localities, while cocoa been shells exhibited an OHC (p >0.05) that was 1.30 and 1.24 times its own weight. No statistically differences (p >0.05) were found between cocoa pod husk and cocoa bean shells. These results obtained were lower than those for some agricultural co-products and DF concentrates, such as native banana pseudo-stem our and boiled tender core of the banana pseudo-stem our (5.48 and 3.88 g oil/g sample) (Aziz et al., 2011), M. pajang ber (4.65 g oil/g sample) (Al-Sheraji et al., 2011) or date ber concentrates (9.6 g oil/g sample) (Elleuch et al., 2008). The hydration properties of DFs determine their optimal usage levels in foods as they provide desirable texture properties. These properties may be portrayed by measuring water absorption, water holding capacity and swelling. These hydration properties of DFs are related to the chemical structure of the component polysaccharide components, and other factors such as porosity, particle size, ionic form, pH, temperature, ionic strength, type of ions in solution and stresses upon bers (Elleuch et al., 2011). Thus, the use of speci c bers in food products is largely determined by their functionality, which depends on physicochemical properties, and by food processing. However their impact on sensory characteristics of food needs to be taken into consideration (Tosh & Yada, 2010).

3.2. Technological properties The results for the water-holding capacity (WHC) oil-holding capacity (OHC) and swelling capacity (SWC) of cocoa pod husks and cocoa bean shell are presented in Table 3. The cocoa pod husks showed a WHC ranged between 5.81 and 5.86 g water/g sample with no statistically signi cant differences (p >0.05) between the localities of Taura and Cone. In the same way, no statistically differences (p> 0.05) were found between cocoa bean shells obtained from Taura and Cone with values ranging between 4.63 and 4.72 g water/g sample. Cocoa pod husks showed a higher (pb 0.05) WHC than cocoa bean shell. This WHC of cocoa co-products suggests that this material could be used as a technological tool for food product development. However, the cocoa co-products showed a lower WHC than fruit ber concentrates, such as that obtained from Mexican lime peel (12.84 g water/g ber) (Ubando-Rivera, Navarro-Ocaa, & Valdivia-Lpez, 2005), Mangifera pajang peel (11.6 g water/g product) (Hassan, Ismail, Hamid, Azlan, & Al-sheraji, 2011) or ber powders from asparagus co-products (20.3 g water/g ber) (Fuentes-Alventosa et al., 2009). The WHC is the ability of a moist material to retain water when subjected to an external centrifugal gravity force or compression. It consists of the sum of bound water, hydrodynamic water and, mainly, physically trapped water (Vzquez-Ovando, Rosado-Rubio,

3.3. Total phenol content The total phenol content (TPC) of cocoapod husk, cocoa bean shell and cocoa mucilage obtained from two localities (Taura and Cone) and extracted with two solvents (ethanol and methanol:acetone) are shown in Fig. 1. The TPC of cocoa pod husk, expressed as gallic acid equivalent, ranged (p b0.05) from 352.67 to 365.33 mg/100 g sample, in the extraction made with methanol:acetone while in the extraction made with ethanol, the TPC values were lower (p b0.05), with values ranging (p b0.05) from 206.67 to 227.00 mg/100 g sample. This is higher than the values reported in other bers; for example, tomato ber (158.1 mg GAE/100 g) (Navarro-Gonzlez et al., 2011), cherimoya peel (Annona cherimolia) (323 mg GAE/100 g) or Ta ble 3 (Fragaria ananasa) (238 mg GAE/100 g) (Vasco, Ruales, Tec h n o lo g ic a l p ro p er ti es of th e di ffe re n t c o c o a c o -p r od u c ts ( co c o a p o d h u s ks strawberry , c oc o a bea n sh el l , c oc o a m u c il a ge) ( m ean S D) . & Kamal-Eldin, 2008). However, TPC levels were considerably lower than in pomegranate arils bagasse powder (1005 mg GAE/100 g) C o -p r od u c t Lo c al i zat i on W HC SW C OH C (Viuda-Martos et al., 2011) or fresh carrot peels (979 mg GAE/ ( m L/g) (g /g ) ( g/ g) 100 g) (Zhang & Hamauzu, 2004). As regards to cocoa bean shell, the TPC levels were signi cantly greater (p b0.05) in the extraction C o c oa po d h us k s T au r a 5 .8 1 0. 23 a 5. 7 0 0 . 23 a 1. 18 0 .0 7 a made with methanol:acetone (154.43 and 144.83 mg GAE/100 g for C o n e 5 .8 6 0. 19 a 5. 8 1 0 . 09 a 1. 20 0 .0 9 a Cone and Taura, respectively) than in the extraction made with ethaC o c oa be an sh e ll T au r a 4 .6 3 0. 20 b 3. 8 7 0 . 14 b 1. 30 0 .1 5 a nol (80.17 and 82.37 mg GAE/100 g for Coneand Taura, respectively).
C o n e 4 .7 2 0. 16 b 3. 6 8 0 . 35 b 1. 24 0 .0 6 a

WH C : W ate r h ol d i n g c a pa c it y; S WC : s we l li n g c ap ac i ty; OH C : oi l h ol d i n g c ap ac i t y.

Val u es fo ll o we d by th e s am e l e tte r w i th i n th e sa m e c o l um n a re n ot s ig n i ca n tl y di ffe ren t (p >0 . 05 ) ac c or d i ng to Tu k ey' s Mu l ti p l e R an g e Te s t. 42

Author's personal copy

R. Ma rt n ez et al . / F ood Res ea rc h I n ter na ti on al 49 ( 20 12 ) 39 4 5 43

inhibiting lipid peroxidation or chelating metal ions (Alothman, Bhat, & Karim, 2009). In this study, three different methods have been used to evaluate the antioxidant capacity of the cocoa co-products extracts: the ABTS assay, the ferric reducing/antioxidant power assay (FRAP assay) and the DPPH free radical-scavenging assay. The results of these assays, expressed as M Trolox Equivalents (TEs)/g sample, are summarized in Table 4. In ABTS assay the antioxidant activity of the methanol:acetone cocoa pod husk extracts tested varied (p b0.05) from 37.97 to 10 0 22.93 M of TEs/g; while in the ethanolic extracts the antioxidant ca15 0 pacities were lower (pb 0.05) with values ranged between 23.03 and 20 0 50 25 0 24.13 M of TEs/g. Cocoa bean shell showed a lower antioxidant ac30 0 0 tivity (p b0.05) than cocoa pod husk, with values for the ethanolic ex35 0 C oco a po d C ocoa pod Co coa b ean Coco a be an C oco a Co coa traction (p b 0.05) of 2.56 and 2.89 M of TEs/g, and for the methanol: 40 0 h usks C one h usks T au ra sh el l Con e sh el l Ta ura muci la ge muci lag e acetone extraction (pb 0.05) of 4.45 and 4.56 M of TEs/g. In the case Con e Ta ura of cocoa mucilage, the ethanolic extracts showed lower (p b 0.05) antioxidant activity than methanol:acetone extracts, with values rangEt ha nol Met ha nol :Ace to ne ing from 2.48 and 2.56 M of TEs/g compared with 4.10-4.17 M of respectively. F ig. 1. To tal p h en ol i c c o nt en t o f t h e di ffe re nt c oc o a c o- p ro d u c ts ( c o c oa p o d h u s TEs/g, ks ,
c o c oa b ean s he l l, c o c o a m u c i l age ) ex p res s ed as m g ga l li c ac i d eq u i val e nt /1 00 g s am p l e.

Cocoa mucilage showed higher TPC values (p b0.05) than cocoa bean shell but lower (pb 0.05) than that observed in cocoa pod husk. Again, the extraction made with methanol:acetone showed higher TPC values (173.67 and 182.63 mg GAE/100 g for Cone and Taura, respectively) than the extraction made with ethanol (102.00 and 109.00 mg GAE/100 g for Cone and Taura, respectively). The results obtained agree with Naczk and Shahidi (2006), who mentioned that the recovery of polyphenols from plant materials is in uenced by the solubility of the phenolic compounds in the solvent used for the extraction process. Furthermore, solvent polarity will play a key role in increasing phenolic solubility. 3.4. Antioxidant properties It is very dif cult to assess the antioxidant activity of a product on the basis of a single method. A single method will provide basic information about antioxidant properties, but a combination of methods will describe the antioxidant properties of the sample in more detail ( et al., 2010). Antioxidant activity assessment may require acombination of different methods because there are substantial differences in sample preparation, extraction of antioxidants, the selection of end-points and the way in which the results are expressed, even for the same method, so that any comparison between the values reported by different laboratories may be quite dif cult (Viuda-Martos, Ruiz-Navajas, Snchez-Zapata et al., 2010). Usually, the antioxidant methods measure the ability of antioxidants, in a particular plant material, to scavenge speci c radicals, by

As regards the DPPH radical scavenging assay, the antioxidant capacities of the ethanolic cocoa pod husk extracts tested varied (p b0.05) from 18.40 to 21.37 M of TEs/g for Cone and Taura, respectively; while in the methanol:acetone extracts the antioxidant capacities were higher (p b0.05) with values ranging between 33.07 and 33.93 M of TEs/g for the same localities. Cocoa bean shell showed lower antioxidant activity (p b0.05) than cocoa pod husk with values for the ethanolic extraction (p b0.05) of 1.57 and 1.71 M of TEs/g for Cone and Taura, respectively. The methanol:acetone extracts provided higher (p b0.05) antioxidant activity values than the ethanol extracts, with values of 3.81 and 4.05 M of TEs/g for Cone and Taura, respectively with no statistically differences (p b0.05) between them. In the case of cocoa mucilage, the antioxidant capacities were very similar to the cocoa bean shells values. As in the other extracts, ethanolic extracts showed lower (pb 0.05) antioxidant activity than methanol:acetone extracts, with values ranging from 1.77 and 2.25 M of TEs/g for Cone and Taura, respectively, compared with the 3.45 - 3.86 M of TEs/g obtained for the methanol:acetone extracts(Cone and Taura, respectively).

In the FRAP assay the analyzed cocoa co-product showed a wide range of antioxidant capacities, with values ranging from 0.67 M TE/g for cocoa bean shell and 2.00 M TE/g for cocoa pod husks in the extracction made with ethanol with statistically signi cant differences (p b0.05) between samples. In the extraction made with methanol:acetone the antioxidant activity values ranged between 1.51 M TE/g for cocoa bean shell and 4.69 M TE/g for cocoa pod husks with statistically signi cant differences (p b0.05) between samples. As in the case of ABTS and DPPH, the extraction made with

T ab le 4 An t i ox id a nt ac t iv i ty o f th e di ffe re n t c o c oa c o -p r od u c ts ( c o c oa p od h u s ks , c oc o a bea n s h el l , c o c o a m u c i la ge ) m eas u re d by th r ee d i ffer en t t es t sy s tem s ( AB T S, DP PH a nd FR AP) . ( M ea n SD ). C o -p r od u c t S ol ve n t Lo c al i zat io n AB T S D PP H FR AP ( M T E/ g) ( M T E/ g) ( M T E/g ) C o c oa p o d h u s ks

E th an o l C o ne 2 4. 1 3 0. 15 aA 1 8 .4 0 0. 26 a A 2. 00 0 .0 1 aA T au ra 2 3. 0 3 0. 15 b B 2 1 .3 7 0. 31 b B 1. 98 0 .0 2 aB M e th an ol : Ac eto n e C o ne 3 7. 9 7 0. 25 aC 3 3 .0 7 0. 16 a C 4. 69 0 .0 5 aC T au ra 4 2. 9 3 0. 15 b D 3 3 .9 3 0. 55 b D 4. 49 0 .0 4 bD E th an o l C o ne 2. 5 6 0. 01 aA 1 .5 7 0. 18 a A 0. 67 0 .0 1 aA T au ra 2. 8 9 0. 01 b B 1 .7 1 0. 04 b B 0. 74 0 .1 7 aB M e th an ol : Ac eto n e C o ne 4. 5 6 0. 03 aC 3 .8 1 0. 03 a C 1. 51 0 .0 7 aC T au ra 4. 4 5 0. 02 b D 4 .0 5 0. 01 b D 1. 78 0 .0 1 bD

C o c oa b ea n s h el l

C o c oa m u c i l age

E th an o l C o ne 2. 4 8 0. 01 aA 1 .7 7 0. 03 a A 0. 88 0 .0 2 aA T au ra 2. 5 6 0. 01 b B 2 .2 5 0. 02 b B 0. 88 0 .0 7 aA M e th an ol : Ac eto n e C o ne 4. 1 0 0. 23 aC 3 .4 5 0. 15 a C 1. 74 0 .0 9 aB T au ra 4. 1 7 0. 09 aC 3 .8 6 0. 06 b D 1. 97 0 .0 7 bC

Fo r a s am e m eth o d a s am e s ol v en t, val u es fo ll o we d b y t h e s am e l ow er c as e l ett er ar e n ot s i g ni c an t ly d i ffer en t ( p > 0. 0 5) ac c o rd i n g t o T uk ey 's M u lt i pl e R a n ge T es t. Fo r a s am e m eth o d a s am e c o- p ro d u c t, v al u es fol l o wed b y t he sa m e u pp er c as e l et ter ar e n ot s ig n i c a n tl y d i ffer en t ( p > 0 .0 5) ac c o r di n g to T u ke y's M u l ti p l e R an ge Tes t .

Author's personal copy

44 R .R. M Ma ar t rtnez n ez et et a l. al/ .Food / F ood R esea Res ea rc h rcIn h ter Inn ter ati na on ti al on4al 9 49 ( 20 ( 20 12 12 ) 39 ) 39 4 4 5 5 45

L p ez, G ., R o s, lower G ., R iantioxidant n c n , F. , Per i ag o, M . J ., M ar t n ez , than M . Cex. , & O rt u o , J . ( 19References 9 6) To. sh R el , S. at M ., & Y a da, S. ( 2 01 0) . Di eta ry br es i n p u ls e se ed s an d fra ct io n s: C ha rac te ri zati o ethanol, provided activity values (pb 0.05) io n n, f un c ti on al att ri bu te s, an d ap p l ic at io n s. Food Res earc h In ter nati on al , 4 3, 4 5 0 46 0. s hmade i p b etw eenmethanol:acetone. p h y si c al an d h yd ra ti on pr o pe rt ie s o f s ol u b le an d i n s ol u b le b er of traction with

a rti c h o ke . J ou rn al of A gr i c ul tu ra l a nd Foo d C hem i str y, 44 , 2 77 3 2 7 78 . Ub an d o- R i ver a, J . , N ava rr o- O ca a, A. , & Va ld i vi a -L p ez, M . A. ( 20 0 5) . M e xi c an l i me h lma at,ra Rti ., ve &s Kar im , A. 0 09 Ai nt i ox i da an d co n- There numerous studies in. which the Lo ti t o, are S.B ., & Fre i, B. ( 20 0 6) C o ns u m pantioxidant ti on o f a vo activity n oi d -r i of c hthe f oo ds an d i nc r eaAl se ot dp pas e n, el - M : C.,oB mhpa tu d y on c A. on (2 te nt s o) f. d eta ry b re nt an c d ap as ac so ic ty i ate dp anhe ti n o ol xi idcan t ac ten t iovi f se ec te trhoemi pi cs atry l fr u 9, it s m1M ty .l Fo oddC ,8 5 fr 7o 6 . al ay s i a, ex tr ac te d w i th di ffe re nt so l ve nt s. a an ti o xi d an t c ap ac i t y i n h u et m al., an s 2010; : C au sBubonja-Sonje, e, c o n s eq ue n c e, o r ep i p h en om en o n ? tFree cocoa m has been determined (Schinella Fooc d CC hem is a trle y,s, 11 5,& 78 5ma 7 88 . l d in , A. ( 2 0 08 ) . T ot al ph e n ol i c c om p o un d s an d an ti o xi R ad i c al Bi ol o gy & M edi c i ne, 41 , 1 72 7 1 7 46 . Vas o, ., Ru J ., Ka l -E Giacometti, & Abram, 2011). However, few studies show the antioxiAl -S era j i, . H. , ti Is ma , Aj ., an ,M .Y ., ua M do u srtafa ,S s of, R ., M an -ph M e i, X. , M u , T. H. , & H an , J . J . ( 2 0 10 ) . C o mp o s it i on a nd p h ys i c o c he m ic a l p ro ert d a i nt cS ap ac i es o filma orM fru i ap ts fr om Ec . Foo d ., CY hu emi st ry 11 .1, ,& 8H 16as s 82 3., F. A. ( 2 dant activity of thecocoa co-products. Thus, Lecumberri et al. (2007) an01 1 ). Fu n c ti on al p r op e rti e s an d c ha ra c ter i zati o n of di e tar y b er fr om M an gi f era pa j an g es ofd i et ary b er ex tr ac ted f ro m re s id u es o f 1 0 var i eti e s of sw eet p ot ato b y a s i evi n g Vzq u ez- Ov an d o, A., R os ad o -R u b i o, G. , C h el - Gu er re ro , L ., & B eta n c ur -A n co n a, D. alyzedm the antioxidant capacity of ber-rich cocoa product and found ko rt fr 09 u it lp Jc oo ur l oi fc Ag ul tu r al emi try ,5 39 (39 5.ia h i sp an i et h od . J o ur na l of A gri c u lt ur al an d Fo od Ch emi s try , 5 8, 73 0 5 73 1 0. ( 2. 0 ) pu . Ph y .si c na h em al ri pc r op e rti e an s odf Fo a bod ro C uh s fr acsti on fr9, om c 80 hi a Sa8lv AOA C ( 19 9 7) . Of c ia l M eth ods of A n al ys i s of A O AC I n ter na ti on al ( 16 th ed .) . W as h in g to n , FRAP and ABTS values of 72.32 and 7.73 MTE/gofsamplerespectively. M e zad ri , T ., Vi l l a o, D ., Fe rn n d ez- P ac h n , M . S. , G ar c a-P ar r il l a, M . C ., & Tr on c ocsao, A. M DC : As s oc i a ti on o f Of c i al An al y ti c al C h em i st s . These values with in d this On the .) a. Fo od Sc an d an T ech n ol ogy 4E 2, ., 16 8 1 7 -A 3. c eve do , E. , T ov ar , J ., R u al es , J . , & . ( 2 00 8 )contrast . An t io x id anthose t c omobtained p o un d s an an tstudy. io x id an t ac ti other v i ty in a c er ol a (M al p ig Ver hi a L gar Val en c ien i a, c Ne ., Gr ad os - P re, z, Aga ma lo r io , M ., C o ss o n , J . D., Tr ava gl i a, F. , Var sa l di , F. , M i gl i o, G ., Lo m b ard i , G ., et al . ( 2 hand, numerous studies have the antioxidant activity ofa nd A n al ys i s, 2 1,Ar em ar gi na ta D C. ) fr u it sdetermined an d de ri v ati v es . Foo d Co mp os i ti on 28 2 2 90 e ll. o -P r ez, L. A. ( 2 00 7 ). Fi br e c o nc e n tra te fr om m an go fru i t: C h ar ac ter i zat io n , as 00 5 An ). B ti o xi d an t an d bi o lo g ic a l a c ti vi ty of p h en ol i c po i gm fr er om eobr om cg ac a oi en t . L fruit or vegetable co-products. example, oo c na i ate d an ti o xi d an t c ap ac i t y an d ap p l i c ati n asen a ts bak y Th p ro d uc t ia n red Mu r th y, P . S ., & N ai d u , M . For M.( 2 01 2 ). RNavarro-Gonzlez ec ov er y o f p h en o et l i al. c a nt i ox i da n ts a n d fu n cs ti h ul l s e xt rac t ed w i th s u p er c ri ti c al C O2 . J . D ., . Foo d Res ear c h I nt ern ati o na l, 3 8, WTl (2011) the tomato ber c reported o m po u nthat ds fro m antioxidant c o ffee in d u activity s tr y b y-of p ro d u c tspeel . Fo od anmead B io pr oc es s Tec hn ol og 10 y, 0 1 Sc 01 i4enc . e an d Tec hn ol og y, 4 0 , 7 22 72 9 . F 9 ood sured with assay 5 ( 3) ABTS , 89 7 9 03 . was 3.90 MTE/gforhydrophilicextract,while Vis i ol i , F. , B ern aer t, H . , Co r ti, R. , Fer ri , C . , He pt in s tal l , S. , M ol i n ari , E ., e t al . ( 20 09 ) . Ar na ho o, c Mo. B . , C an o, A., & A c os ta , M . ( 20 0 1) . T h e h yd ro p h i li c an d li p o ph i l i c c on tr i b ut N ac zk , M ., & Sh ah i d i, F. ( 2 00 6) . P h en o l ic s i n c e re al s, fru i ts an d v eg eta bl es : O c c C ur r l en atce, e,l i fes tyl e an d h eal th . C ri ti ca l R evi ews i n Foo d Sc i enc e an d Nu tr iti on , 4 9, 2 99 31 2. io n to tot al an ti o xi d an t a ct i vi ty . Fo od C h emi s try , 7 3, 2 3 9 2 44 . e xtr ac t io n a n d an al ys i s . J ou rn al of P ha rm ac eu ti ca l an d B io med i c al A n al ys i s, Vi 4 1, u d a- M ar to s , M ., L p ez -M a rc o s , M . C ., F er n n d ez -L p ez , J ., S en d r a, E ., L p ez -V ar Azigas z, N. A., H o, L. H. , Azah ar i , B . , B h at, R ., C h en g, L. H ., & I b ra hi m , M . N . M . ( 2 01 1 5 23 15 42 . , .A. H ., & P re z- l va re z, J . A . ( 2 01 0 a) . R o l e o f b e r i n car d io v as cu l ar di s e as es : A r eCapitani et al. (2012) reported that the antioxidant activity of the 1 ). Ch J e mi c a l an d fun c t io n al p ro pe rt i es o f th e n ati v e b an an a ( M us a ac u mi n ata x i ew N av ar ro -G on z le z, I . , G ar c a -Va lv er d e, V ., Gar c a- Al on s o, J ., & P eri a go , M . J . ( 20 1v 1) . . C om p reh en s i ve R evi ews i n Fo od S c i enc e a nd Foo d Sa f ety , 9 ,240 2 58 . co-products of chia measured with ABTS assay ranged between 226.6 ba lb i si an a C ol l a c v . Aw ak) p s eu d o- s tem an d p s eu do - st em te nd er c o re o u rs . Food C h em i c al p r o l e, fu nc t i on al a nd a nt i ox i da nt p ro p er ti es o f to m ato p eel b er . Fo od Vi ud a- M ., 4 R8 u i75 z- 3 Na Ch emi s trar y,to 1 s, 28M ,7 . vaj as , Y. , Fe rn n d ez- L p ez, J . , & P r ez- l va rez , J . A. ( 2 0 10 and 557.2 MTE/gsample. R es ear ch I n tern at io na l, 44 , 1 5 28 15 3 5. b) . E ffe ct o f ad d ed c i t ru s b re an d s p i c e e ss en t ia l oi l s o n q u al i ty c h ara c ter i s ti c s an d B ara zar te, H ., S an gr on i s , E ., & a i,Sc E .ien ( 20 0 8) . La ar hn el f- l i fe o f mor tad el l a. Un M eat c e, 8 5, 56 c8as 5c 7 6.a d e c a c ao ( Th eob ro ma c ac ao L .) : P ee raj i t, P. , Ch i ew c h an , N. , & De vah as t in , S . (2 0 12 ) . Effe c ts o f pr et re atm en t m et ho d s o The antioxidant activity of cocoa co-products could be attributed to a apo s ar ib to l es, fM ue ., ntR eu c ioz-N m er c aj ia as l d, e ti nnas Aezrc h iv os ati a no s das eN ic io h ea l th -r el at ed fu n c ti o na l p ro p er ti es o f h i gh d i et ary b re p ow d er fro m l i m e r es Vi i Un - ud M av Yp. ec , Fer n. d Lp ez L ,J ., n S oa en mer d ra ic , E. , Say -Butr a rb er ,n, E phytochemical they contain especially, the polypheno6 4 7 0. d u es . Fo compounds od Ch emi s try , 1 32 , 1 89 1 and, 1 8 98 . .,58 &, P r ez- l v are z, J . A. ( 20 1 1) . An t io x id a nt p r op er ti es of p om e gr an ate ( Pu n ic a B en zi e, I . F . F. , & S tr ai n, J . J . (1 9 96 ) . Th e ferr i c r ed u c in g ab i l i ty of pl as m a (F R AP) a licR compounds mainly avan-3-ol compounds such as monomers cateo be rt so n , J . A., de Mo n re d on , F . D. , D ys s el er , P ., Gu i l lo n , F. , Am ad , R . , & T h ib augl ra t, na tu m L.) b ag as s es o b tai n ed as c o -p r od u c t i n th e j u i c e e xt rac t i on . Foo d R esea rc h s a m eas u re of a nt i ox i da nt p ow er : Th e FR AP as s ay. A na l yti c al B io c hem i str y, 23 9, chin and epicatechin, and the procyanidin J . F. ( 2 00 0 ) . Hy dr at ati o n p dimer ro p er ti es o f di e tar B2 y br(Lamuelae an d r es i st an t st ar c h: A Eu r op ea n I n ter na ti on al , 4 4, 12 1 7 1 22 3 . 70 76 . Ravents, Romero-Prez, Andrs-Lacueva, & Tornero, 2005). Recent c o l l abo r ati v e s tu d y. L eb en sm it tel -Wi s sen s c ha ft un d T ech n ol ogi e, 33 , 7 2 7 9. Vi ud a- M ar to s, M ., R u i z-N av aj as , Y ., Ma rt i n- S nc h e z, A. , Sn c h ez- Z ap ata , E. , Fer n n d - Wi l l iJ. am s ,n W Cu er ., (M .E .,& rs et C ,. p (1 )o .U of fr ee ra didcfu al n mc et top r ez-d L studies have demonstrated the activity of J phenoS n ch ez -Za pa ta, E ., Fue nt es individual -Z arag oza ,antioxidant E ., Fe rn n d ez-L p ez, ., Sen d ra, E. , Sa yas ,B E ran ., p ez, , Se dr.,a, Ev ., el e ti al 20 1 2) . CBheem i c, al h 99 ys 5 ic -c se h em ic al an ti h o od n al eva l ua te an ti o xi d an t ac ti v it y. L eb ens mi tt el- Wi s sen s ch af t u nd T ech n ol ogi e, 2 8, 2 5 3 0. N ava rr o, in C ., et al . (systems 2 00 9) . Pr ep the ara ti on o f di et ary b re p ow de r fro m ti ge r n ut ( Cy peru s op lic compounds model and interaction between the m that nj ai -So nf jp e, o M , Gian acat oe m(P et u tini ,J amm ,M ( 2gas 01 1 ). An ti ox an p t an a nt ti l is eri ao lf ac e rn rt es o m. egr c ., a& gr Abr an atu L.). ba s es po wd ei rd c oro d uc .J out rn al es c or ul entu s) m il kthe (h or ch ata ) by pr od u c ts a is, nd it s psynergistic h ys ic o c he or mi c al p ro per ti B esub . Jo ou al increases decreases antioxidant activity, that the ti vi F tood y of En ol gi i ve neoeri i ln ,c g,oc 1 10 o a, an 2 20 d ros 22 em 4 .ar y ex tr ac t p ol y ph e no l s . Foo d Ch emi s tr y, 12 7, o f A gr ic u ltu ra l a nd Food C h emi str y, 5 7, 7 71 9 7 72 5. Vi ud a- M ar to s, M ., R u iz -N ava j as , Y . , S n c he z-Z ap at a, E . , Fe rn n d ez- L p ez, J . , & antagonistic effect of the same compounds (Hidalgo, Snchez-Moreno, 18 2 1 1 82 7 . S c hi n el l a, G., M os c a, S ., C i en fu eg os - Jo v el l an These os , E .,compounds P as am ar , M . A. , M u gu er za, B . ,P Rr am ezn Al , v are z, J . A. ( 20 1 0c ) . An ti o xi d an t ac ti v i ty of es s en ti al oi l s o f v e sp i c e p l an &Pascual-Teresa,2010;Terpinc&Abramovic,2010). api it ts D ., et a l . ( 20 constituents 1 0) . An ti o xi d an t p due r op er es o fantioxidant p ol y ph en oactivity l -r i c h c oc o a p r od u C c ts n an du is ,-M . I ., Sp o to rn o , V., No l as c o, S . M ., & T om s , M . C . ( 2 01 2) . P h ys i c oc h e mi c a l are very important of fruit totitheir an d fu nc t i on al ar teM za ti ,er on o ne fki b an ypr o t. s, fr m cvi hi a gr (S al aJo h i sp ca ee f an w i d el y ed a ed it Fc la r nd Fra ce ur n an al i,ip 2 5,L.) 13 19 . o t ri al l y p r oc e ss e d. Foo d R esea rc h I nte rn ati on a l, 43 , 1 61 4 1 6 23 . Vuk s an , V. ,uc Jsh en kiac inns , ri A. L. J ra en n sd , ie D .du A. Jt.vou Ro oa go k, A. an L. vi , Si ev en p er, J .s L. , ds & J ov which involves chelating redox-active metal ions, inactivating lipid free en ti n a. L WT- Foo d Sc i en ce a nd Tec h no lo gy, 45 , 9 4 1 0 2. oArg vs E ki S in gl et on , V. L., & Ro s si , J . A . ( 1 96 5) . C ol or i me try o f to tal p h en ol i c s wi th p h o sp h om ol . ,(2 y bd 0 08 i c ) . U s i ng c er eal t o i n c r eas e d i et ar y b er i nt ak e to th e rec o m m en d ed l ev el radical chains and preventing hydroperoxide conversion into reactive C ha u, C . F. , & H u an g, Y . L. ( 20 0 4) . C h ar ac te ri za ti on o f p as si o n fru i t s ee d b r es a p op h os p ho tu n gs ti c a c id re agen ts . A mer i ca n J ou rn al o f E nol og y and V iti c ul tu re, 1 6, a n d t he effec t of b er on bo w el fun c t io n i n h e al th y p er s on s c on s u m i ng No r th oxyradicals. The ten A t ia b rceasnou r et c e. od Ch emi 8 5, 9 1 m leri di s. Fo A mer ic a n Jsotry ur ,na l o18 f Cl i ni9c4. al N utr i ti on , 8 8( 5 ) , 1 25 6 1 2 62 . 1 44 15 8. TPC showed a strong correlation with antioxidant activ , M ., o v , H . , D en ev , P ., Kra tc h an ov a, M ., Sl avo v , A. , & Loj e k, A. ( 2 01 0 ) . D iff ere n t ity. signi correlation was observed TPC and S Thus, ud h a, a M. L., B cant as k ar an , V. , & Lee l ava th i , K. between ( 2 00 7 ). Ap pl e p o the m ac e as a so u rc e of d iWi etar s we y de l , I ., H i rs c h , D. , Kr op f, S. , Gr u en i n g, M . , P s te r, E ., S c he we , T. , et al . ( 2 00 m. eth ds fo or co nthro om s oer ns of e an ti F(2 o xi)d is anot p p ro r op e rti eo s of et ati ab o lens s .i n DPPH radical-scavenging activity, R2 =0.977, 4) b er an d p ol y p he n ol s an d i ts effe c t on thof e rcocoa h eo lo co-products g ic a l c h ar ac te ri s ti c s an d c a ke F lo avan l -r ic cl o an c odacdr i npkari l ow p th l as ma s tan ec n cveg en tr Foo 4) , 51 8 l5 3. ogy & M ed ic i n e, 3 7( 3 ) , 4 11 42 1 . extracted with the correlation between TPC hd uC mon an tro s . l, Fr21 ee(R ad ic a B2 i ol m ak i n g.methanol:acetone, F ood Ch emi s tr y, 1 while 04 , 6 8 6 6 92 . C on tr er as -C al d er n, J . , C a ld e r n -J ai m es , L. , G u er ra- H er n n de z, E. , & G ar c a -Vi l l an o and radical-scavenging samples extracted ethanol T the er p DPPH in c , P. , & Ab r am ov i c , H . of (2 0 10 ) . A ki n et i c ap with p ro ac h fo r e val u at i on o f th e an Zh ti oan xi g - , D ., & H am au zu , Y . ( 20 0 4) . P he n ol i c c o m p ou n d s an d t h ei r an ti o xi d an t p ro p ert i va, B. ( 20 1 1) . An ti o xi d an t c ap ac i t y, ph e no l i c c on te n t an d vi t am i n C i n p u lp , p eel an d es i n d i ffer en t t is s u es of c a rr ot s . Foo d A gr i cu l tu re a nd En vi ro nm en t, 2 , 9 5 1 00 . d an t a c ti vi tythe of ssame e le c ted p ha en o li c s ac i d s . Fo od Cobtained h emi st ry , 12 1, 36 6 3 7 1. was R2 = 0.977. In way, high correlation was se ed fr o m 24 e xo ti c f ru i ts fr o m C ol o mb i a. Fo od R esea rc h I n tern at io na l , 4 4, between ABTS assay and TPC of cocoa co-products extracted with 20 4 7 2 05 3 .

methanol:acetone and ethanol with values of R 2 = 0.965 and R2 = 0.951, respectively. There was also a high correlation between the FRAP assay and the TPC of the of cocoa co-product samples extracted with methanol:acetone and ethanol: R 2 = 0.988 and R 2 = 0.984, respectively. The three methods involve single electron transfer mechanism, and a positive and signi cant correlation has also been obtained by other authors (Mezadri, Villao, Fernndez-Pachn, Garca-Parrilla, &Troncoso,2008;Alothmanetal.,2009;Contreras-Caldern, CaldernJaimes, Guerra-Hernndez, & Garca-Villanova, 2011).

Dj o u ss , L ., H op k in s , P . N. , No r th , K. E. , Pa n ko w, J . S. , Arn e tt, D . K., & E l l is o n , R. C . ( 2 01 1 Ch ). o c ol a te c on s u m pt i on i s i n ver s el y as s o c ia ted wi th pr ev al en t c o ro n ar y he ar t di s ea se : Th e N at i on al H ear t, Lu ng , an d B l oo d I n s ti t ut e Fam i l y H ea rt S tu d y. C l i ni c al Nu tr iti o n, 30 ( 2) , 1 8 2 1 87 . d o Es p ri to San to , A. P ., C ar to la n o, N . S., S i l va, T . F., S oar es , F. A . S . M ., G io i el l i , L. A., Pe re go , P ., et al . ( 2 01 2 ) . Fi b er s fro m fr ui t by - pr od u c ts e n ha nc e p ro b io ti c v i ab il i ty an d fat ty ac i d p ro l e a n d i nc r ea se C LA c o n te nt i n y og h ur ts . I nt ern ati o na l J ou rn al of Foo d M i cr ob i ol ogy , 1 54 , 1 35 14 4 . El l eu c h , M . , B ed i gi a n, D. , R o i se u x, O. , B es b es , S. , B l ec k er , C ., & A tti a, H. ( 20 1 1) . Di et ar ybr e a n d b r e- ri c h b y- p ro d uc t s of foo d p ro c es s i ng : C h ar ac te ri s at io n , tec h n o lo gi c al , f un c ti o n al i ty an d c om m e rc i al a pp l i c ati o n s : A r ev i ew. Fo od Ch emi s tr y, 12 4( 2 ), 41 1 4 2 1. El l eu c h , M. , Be sb es , S. , R o is e ux , O ., B l ec k er, C ., De ro an n e, C. , Dr i ra, N . E ., et al . ( 2 00 8 ). Dat e es h : C h em i c al c o mp o s it i on an d c h ar ac te ri s ti c s of th e d i eta ry br e. Food Ch emi s tr y, 1 11 , 6 7 6 68 2 . Fer n n d ez- Lp ez , J ., S en d ra -N ad al , E ., N av ar ro , C ., S aya s, E. , Vi u d a-M a rto s , M ., & P re z-Al v ar ez, J . A. ( 2 00 9 ). S to ra ge s tab i l it y of a h ig h d i et ary b er po wd er fro m o ran ge b y- p ro d uc t s. I n ter na ti on al Jo ur n al o f Foo d Sc i enc e an d Tec h no lo gy, 4 4, 74 8 7 5 6. Fu en te s- Al ve n tos a , J . M . , R o dr gu ez -G ut i r rez , G ., J a ram i l l o- C ar m on a, S., E sp ej o - Ca lv o, J . A. , R od r g u ez- Arc o s , R. , Fer n n d ez- Bo l a o s, J ., et al . ( 20 0 9) . Ef fec t o f e xtr ac t io n m eth o d o n c he m ic a l c o m p os i ti o n a nd fu n c ti on a l c h ar ac t eri s ti c s o f h i g h d i et ary br e po wd e rs o b tai n ed fr o m as p ar agu s b y- p ro d u c ts . Food C h emi st ry , 1 1 3, 6 65 67 1. Gu er re ro , M . S ., T or re s , J . S. , & N u e z, M . J . ( 20 0 8) . E x tra c ti on o f p o l yp h en ol s fro m wh i te d i st i ll e d gr ap e p om ac e : O p ti m i zati o n an d mo d el l i n g. B io res ou r ce Tec h no l ogy, 99 , 1 31 1 1 3 18 . Ha ss a n, F. A. , I s m ai l , A. , H am i d, A. A. , Azl an , A., & Al -s h er aj i , S . H . ( 20 1 1) . C h ar ac t eri s a- ti on o f br e- r ic h p o wd er an d a n ti ox i d an t c ap ac i ty of M an gi f er a pa j an g K. fr u i t pe el s . Fo od C h emi s try , 1 26 (1 ) , 2 8 3 2 88 . Hi d al g o, M ., Sn c h ez -M o re no , C. , & P as c ua l- T ere s a, S . ( 2 01 0 ) . Fl av on o i d - av on o i d in ter ac ti o n an d i ts effe c t on th ei r an ti ox i d an t ac ti v i ty. Food Ch emi s tr y, 1 21 , 6 91 69 6. Kh an , N. , M on ag as , M ., An d r es -L ac u eva , C. , C as as , R ., Ur p -S ard , M. , Lam u el a- R av en t s, R . M . , E s tr uc h , R . ( i n p re s s) . R egu l ar c o n su m p ti o n of c o c o a p ow d er wi th m i l k in c re as es H DL c h ol e st er ol a n d re d uc e s ox i d iz ed LD L l e vel s i n su b j ec ts at hi g h -r i sk of c ar d i ov as c ul a r d i se as e. N u tr it i on , M e tab ol i s m an d C ar d io va s c ul a r Di s ea se s, do i : h tt p: //d x. d oi . or g/ 10 .1 0 16 /j . n um e c d. 20 1 1. 0 2. 00 1 . Lam u el a- R ave n ts , R . M . , R om er o -P r ez, A . I ., An d r s -La c ue va, C. , & To r ne ro , A. ( 2 00 5 ). R evi e w: He al th effec t s o f c o c oa av on o i ds . Fo od Sc i en ce an d Tec h no l ogy I nt ern ati on al , 1 1, 15 9 1 7 6. Lec u m be rr i , E. , M ateo s , R ., I zq u ie rd o -P u l id o , M ., R u p re z, P. , Go ya, L. , & Br av o, L. ( 2 00 7 ). Di et ar y b r e c o m p os i ti o n , an ti ox i d an t c ap ac i ty an d p h ys i c o -c h e mi c al p ro p er ti es o f a b re -r i c h pr o du c t fro m c oc o a ( Th eob ro ma c ac ao L.) . Foo d C hem i str y, 10 4, 94 8 9 5 4.

4. Conclusions The co-products obtained from the industrial processing of cocoa constitute an important potential source of dietary ber for application in food industry, since they also contain polyphenol compounds in appreciable amounts. These ber-rich cocoa products could be used as intermediate foods ingredients (IFI) in functional food development for their interesting antioxidant activities and the good insoluble DF/soluble DF ratio. The geographical origin of the cocoa co-products does not affect its technological or compositional characteristics.

Acknowledgment The authors would like to thank Transmar Commodity Group for providing the cocoa co-products. The project CYTED-IBEROFUN Codec: 110 AC0386 and CajaMurcia for supporting the post-doctoral grant of one of the authors (M.V.M.).