Food Chemistry

Food Chemistry 100 (2007) 736741 www.elsevier.com/locate/foodchem

By-products from dierent citrus processes as a source of customized functional bres

n a,*, Cristina Soler-Rivas a, Obdulio Benavente-Garc a b, Francisco R. Mar b c A. Pe rez-Alvarez Julian Castillo , Jose
a c

noma de Madrid (UAM), Ctra de Colmenar, Km 15, 28049 Madrid, Spain Unit of Food Science and Technology, Universidad Auto Research and Development Department, Furfural Espan ol, S.A. Camino Viejo de Pliego s/n, Alcantarilla, E-80320 Murcia, Spain a Agroalimentaria, E.P.S.O. Universidad Miguel Herna ndez, Ctra de Beniel, Km 3.2, 03312 Orihuela (Alicante), Spain Dep Tecnolog
b

Received 18 April 2005; received in revised form 18 April 2005; accepted 18 April 2005

Abstract Dietary bre encompasses a huge variety of macromolecules, exhibiting a wide range of physicochemical properties. Citrus bre can be obtained from dierent industrial sources and dierent kinds of citrus. The chemical components of bre (pectin, lignin, cellulose and hemicellulose), together with other compounds, such as avonoids, were analysed in nine dierent industrial sources. Final bre composition was more dependent on the industrial process than on the type of citrus. The chemical changes gone by citrus bre showed losses of functional values; i.e. soluble dietary bre and ascorbic acid content decreased when waste products were transformed into bres. The water-holding and lipid-holding capacities of analysed citrus bres suggested a non-linear behaviour of these properties for the analysed citrus bres. 2005 Published by Elsevier Ltd.
Keywords: Citrus; Fibre; Flavonoids; Ascorbic acid; TAA; Technological properties

1. Introduction Citrus is the most abundant crop in the world. Its worldwide production is over 88 106 tons and one-third of the crop is processed. Oranges, lemons, grapefruits and mandarins represent approximately 98% of the entire industrialized crop, oranges being the most relevant with approximately 82% of the total. Citrus fruits are processed, mainly to obtain juice, but also, in the canning industry, to produce marmalade, segments of mandarin and by the chemical industry to extract avonoids and essential oils (Izquierdo & Sendra, 2003). Worldwide industrial citrus wastes may be estimated at more than 15 106 tons, as the amount of residue obtained from the fruits accounts for 50% of the original whole fruit

Corresponding author. Tel.: +34 914973776; fax: +34 914973579. n). E-mail address: francisco.marin@uam.es (F.R. Mar

mass (Cohn & Cohn, 1997). Citrus wastes constitute a severe environmental problem (Laufenberg, Kunz, & Nystroem, 2003; Montgomery, 2004). They consist of peels (albedo and avedo), which are almost one-fourth of the whole fruit mass, seeds and fruit pulp, remaining after juice and essential oil extraction (Braddock, 1999). On the other hand, the consumption of dietary bre plays an important role in the prevention of diseases, such as constipation, haemorrhoids, hypercholesterolemia and ndez-Lo pez et al., 2004; Guillon colorectal cancer (Ferna & Champ, 2000; Harris & Ferguson, 1999; Lipkin, Reddy, Newmark, & Lamprecht, 1999). Dietary bres are not only desirable for their nutritional value but also for their functional and technological properties (Thebaudin, Lefebvre, Harrington, & Bourgeois, 1997). The main advantage of dietary bre from citrus fruits, when compared to other alternative sources, such as cereals, is its higher proportion of soluble dietary bre (Gorinstein et al., 2001; Griguelmon-Bel, 1999; Prosky, Asp, Schweizer, De Miguel & Mart

0308-8146/$ - see front matter 2005 Published by Elsevier Ltd. doi:10.1016/j.foodchem.2005.04.040

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Vries, & Furda, 1988). This is important, considering that the requirement for dietary bre intake must be balanced, i.e. the water-soluble fraction should represent between 30% and 50% of the total dietary bre (Eastwood, 1987; Spiller, 1986). Moreover, citrus fruits have better quality than other sources of dietary bre due to the presence of associated bioactive compounds (avonoids and vitamin C) with antioxidant properties, which may provide addiSample CIB-1 CIB-2 CIB-3 CIB-4 CIB-5 CIB-6 CIB-7 CIB-8 CIB-9 Citrus kind Sour orange Sour orange Satsuma Grapefruit Satsuma Lemon Lemon Sweet orange Sweet orange Ripening Immature Mature Mature Mature Mature Mature Mature Mature Mature

sinensis (L.) Osb.), as raw material for industrial processes, were obtained from commercial orchards located in SouthEast of Spain. 2.2. Samples The citrus industrial by-products obtained from dierent types of industrial process were the following: Process Extraction of avonoids Extraction of avonoids Extraction of avonoids Extraction of avonoids Canning industry Juice industry Juice industry Juice industry Juice industry Fruit part Whole Whole Peels Whole Peels Peels Pulp Peels Pulp

CIB: Citrus industrial by-product.

a, Castional health-promoting eects (Benavente-Garc n, Ortun o, 1997; Mar n, Frutos, tillo, Mar o, & Del R rez-Alvarez, Mart nez-Sa nchez, & Del R o, 2002). Pe Chemical and physical properties of citrus bres have been widely studied (Gorinstein et al., 2001; Griguelmon-Bel, 1999). Many authors have described Miguel & Mart citrus dietary bre content (Schieber, Stintzing, & Carle, 2001), while others have mainly focussed on their avonoid concentrations (Coll, Coll, Laencina, & Tomas-Barberan, 1998). Other chemical properties have been extensively ndez-Lo pez et al. (2004). However, no reviewed by Ferna exhaustive studies combining the above-mentioned analysis have been reported to study both the eect of the process to obtain bre and the eect of previous industrial processes. This work describes the chemical composition of citrus by-products from three of the most important bre sources: wastes from chemical companies (whose interest lies in the extraction of natural compounds, such as avonoids), wastes from canning companies (whose main activity is canning citrus segments) and wastes from citrus juice production. Moreover, the chemical changes produced due to the processing of citrus wastes into bre are studied, together with an evaluation of chemical properties, such as total antioxidant activity, and physical properties, such as LHC (lipid-holding capacity) and WHC (water-holding capacity). 2. Material and methods 2.1. Plant material Sour orange (Citrus aurantium L.), satsuma (Citrus unshiu (Mak) Marc., grapefruit (Citrus paradisi Macf), lemon (Citrus limon (L.) Burm f.), and sweet orange (Citrus

Samples CIB-1, 2, 3 and 4 were obtained as o-products from an industrial process of avonoid extraction, in which aqueous and hydroalcoholic solvents were used. CIB-5 corresponds to peels obtained from the chemical peeling during the satsuma segments canning process. CIB-6, 7, 8 and 9 were obtained from the citrus juices industry as residue from the extraction process performed by using the in line extraction system from FMC (Food Machinery Corporation, USA). 2.3. Analytical methods 2.3.1. Proximate analyses Ash was determined according to method 923.03 of AOAC (2000). Total carbohydrates were determined by the LuSchorl method, according to the reference of the Journal Ocial enes, n L. 155/32. des Comunantes Europe Fats were determined according to method 960.39 of AOAC (2000). Proteins were determined according to method 955.04 of AOAC (2000). 2.3.2. Compounds of functional value The content of ascorbic acid, expressed in mg/l, was obtained by means of method no. 43056 of the AOAC. For the quantication of avonoids, an ODS-C18 (250 4.6 mm i.d.) analytical column was used with an average particle size of 5 mm, using water:acetonitrile:methanol:acetic acid (15:2:2:1) as the eluent with a ow rate of 1 ml/min at 30 C. The absorbance change was monitored at 280 and 350 nm with a Hewlett Packard mod. HP 1100 UV/Vis diode array detector (Castillo, a, & Del R o, 1994). Benavente-Garc

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For the identication of avonoids, the procedure described by Castillo et al. (1994) was used. All compounds isolated were identied by their melting points (Gallenkamp, England), mass spectrum (EIMS) (Hewlett-Packard Co., USA) and the 1H NMR (200 MHz) and 13C NMR (50 MHz) spectra (Brucker, Germany) in hexadeuteroDMSO. 2.3.3. Chemical bres Pectin content was determined by alkaline hydrolysis and colorimetric determination of galactouronic acid derivatized with carbazole (Rangana, 1986). Cellulose was determined according to the Kurschner method, by aceticnitric acid digestion of citrus by-products, washed with alcohol, benzole and ether (Lees, 1982). Hemicellulose content was determined by the Arsenal Royal Woolwich procedure (Powell and Whittaker modication) by titration with thiosulphate (Lees, 1982). Lignin content was analysed by the Springer method (Lees, 1982), using sequential extractions in a Soxhlet extractor with ether, alcohol and water, and subsequent acid hydrolysis. Dietary bre composition (soluble, insoluble and total bre) was determined as previously described by Prosky et al. (1988). 2.4. Wastes processing The citrus industrial by-products (CIB-1 to CIB-9) were processed for bre extraction using the following procedure: samples were vacuum-packed as fresh raw material in 2 kg pouches and promptly frozen to 30 C until further use. After thawing (24 h/25 C) the material was scalded in a water bath to remove potential pathogenic microorganisms (vegetative cells). Afterwards, the bre was pressed using a helical press to remove excess liquid prior to drying. Drying was carried out in an oven at 50 5 C during 24 h to improve the bre shelf life without addition of any chemical preservative. A grinder mill and sieves were used to obtain a powder particle size of less than 0.2 mm. The nal product was a powder with 7% ndez-Lo pez et al. (2004). moisture, as described by Ferna

2.5. Technological properties Water-holding capacity (WHC) of citrus by-products was determined as previously described by Robertson and Eastwood (1981) by using a simple ltration vacuum suction system. Experimental conditions were as follows: particle diameter <0.2 mm; lter of 40 lm; vacuum pressure of 70 mm Hg; ltration time of 15 min, and 30 C. Lipid-holding capacity (LHC) of citrus by-products was measured by a method similar to that previously described for WHC, using maize oil. Experimental conditions were as follows: particle diameter <0.2 mm; lter of 40 lm, vacuum pressure of 70 mm Hg, ltration time of 15 min, and 30 C. Commercial maize oil was used. 2.6. Antioxidant activity The TAA (total antioxidant activity) in the bres and by-products was measured spectrophotometrically, using the Miller, Rice-Evans, Davies, Gopinathan, and Milner (1993) method, based on the relative capacity of dierent substances to stabilize the ABTS (2,2 0 -azinobis (3-ethylbenzothiazoline-6-sulfonic acid)) radical cation, compared to a standard antioxidant (trolox) by means of a responsedosage curve. 3. Results and discussion The chemical composition of the citrus industrial byproducts (Table 1), according to the analysed parameters, seems to be more dependent on the industrial source than on the citrus species used. By-products obtained from the chemical industries (CIB-1, 2, 3 and 4) showed lower contents of pectin and avonoids, and higher contents of lignin and ash, than by-products from food industries (CIB-5, 6, 7, 8 and 9). This may easily be explained by the fact that, in the chemical industry, hydroalcoholic solvents are used during the process, which are able to extract both, avonoids and pectins. On the other hand, by-products from the food industry showed 3- to 10-fold higher amounts of total avonoids than by-products from the chemical industry. By-

Table 1 Chemical composition of dierent citrus by-products Ash (% of DW) CIB-1 CIB-2 CIB-3 CIB-4 CIB-5 CIB-6 CIB-7 CIB-8 CIB-9 13.22 0.26 11.03 0.29 10.03 0.15 8.09 0.41 5.05 0.22 2.52 0.15 2.54 0.05 2.56 0.10 2.55 0.09 Sugar (% of DW) 3.52 0.14 8.01 0.23 8.50 0.20 8.02 0.36 10.07 0.54 6.52 0.48 9.01 0.87 9.57 0.22 6.04 0.41 Fat (% of DW) 1.58 0.08 0.55 0.03 1.57 0.06 0.52 0.02 1.59 0.09 1.51 0.11 3.09 0.12 4.00 0.15 1.52 0.05 Protein (% of DW) 8.82 0.42 12.87 0.90 7.33 0.33 12.51 0.87 7.50 0.21 7.00 0.44 8.72 0.36 9.06 0.38 6.55 0.32 Flavonoid (% of DW) 2.70 0.08 1.51 0.02 3.06 0.06 3.04 0.03 5.09 0.05 12.54 0.62 4.52 0.10 4.50 0.15 11.00 0.54 Pectin (% of DW) 3.52 0.27 6.54 0.56 2.58 0.22 8.53 0.68 16.01 1.21 13.00 1.06 22.53 1.95 23.02 2.12 12.07 1.12 Lignin (% of DW) 12.52 1.11 14.73 1.32 13.54 1.26 11.56 0.98 8.59 0.76 7.56 0.54 7.55 0.66 7.52 0.59 7.51 0.62 Cellulose (% of DW) 39.00 3.12 20.74 1.92 30.53 2.35 26.57 2.01 22.55 2.22 23.06 2.11 36.22 3.24 37.08 3.10 24.52 2.00 Hemicellulose (% of DW) 8.32 0.79 6.57 0.56 11.04 0.98 5.59 0.42 6.01 0.56 8.09 0.81 11.05 1.09 11.04 1.05 7.57 0.66

The results are expressed as an average ES (n = 7).

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products from the canning industry (CIB-5) showed lower contents of avonoids than samples CIB-6, 7, 8 and 9 and higher than CIB-1, 2, 3 and 4. This might be because the canning industry uses hot chemical peeling with NaOH. Under these conditions, the avonoid skeleton open to the chalcone form, becoming more soluble and leaving a solid matrix. The pectin content of food industry by-products (CIB-5, 6, 7, 8 and 9) was 2- to 10-fold higher than those obtained from the chemical industry. In CIB-1, 2, 3 and 4, pectin contributed to the total amount of chemical bres in a percentages ranging from 4% to 16% while, in the food industry by-products (CIB-5, 6, 7, 8 and 9), the percentage ranged from 2530%. Lignin in by-products originating from the chemical industry, contributed up to 2030% of the total cell wall components of bre. Lignin and ash showed higher contents in these by-products compared to those originating from the food industry. These fractions are less soluble and, therefore, hydroalcoholic solvents should not extract them; this produces a concentration eect on the studied by-products. The highest content in the chemical components of bre corresponded to CIB-7 and 8, which are the pulp fraction obtained from lemon and orange juice production, respectively. CIB-7 and 8 showed a total amount of chemical bre constituents of approximately 77% DW (dry weight). On the other hand, the peel fraction (CIB-6 and 9), together with CIB-2, showed the lowest values of the four analysed polymers. In this case, two dierent waste-streams from the same fruit and same industrial process resulted in dierent compositions. This could be due to the fact that the in line extraction system (FMC, USA) performs an extraction by squeezing after incisions have been made in the polar areas of the fruit. Afterwards, the pulp and juice

are separated by means of a prenisher tube, determining dierent tissue compositions of wastes. During the processing of citrus by-products into bre, the raw material undergoes two critical steps: scalding (which includes washing) and drying. CIB-1 and CIB-7 were selected to evaluate the eect of the described process. CIB-1 is the most abundant by-product from the chemical industry, and CIB-7 has a pale yellow colour that may be more convenient for food formulations. In general terms, contents of ash, sugar, protein, avonoid and pectin decreased, while the contents of fat, lignin, cellulose and hemicellulose increased after this treatment (Table 2). The most relevant eect due to processing is observed on pectin and lignin contents. Pectin levels decreased by 58% and 35% compared to the original amount in CIB-1 and CIB-7, respectively. On the other hand, lignin levels increased up to 89% and 46% with respect the initial values found for CB-1 and 7, respectively. The processing of by-products into bre also aects those parameters known to determine the nutritional functional value of a product. In the studied samples, losses in soluble dietary bre, avonoids, ascorbic acid and total antioxidant activity (TAA) were observed while insoluble dietary bre levels increased (Table 3). Losses in soluble dietary bre may be explained by pectin lost as a consequence of the washing eect of scalding. On the other hand, the increase in insoluble dietary bre may be explained by a concentration eect, due to the former scalding step. Other valuable compounds, such as avonoids, drop slightly while ascorbic acid practically disappears from the processed material. Values of ascorbic acid in CIB-1 and CIB-7 were extremely low when compared with that reported by other authors (Gorinstein et al., 2001). However, these studies were done on pulp and peel fractions obtained by peeling fresh fruits. In the present study,

Table 2 Eect of the transforming process on citrus bre composition Ash (% of DW) CIB-1 CIB-1 Fibre CIB-7 CIB-7 Fibre 13.22 0.26 6.57 0.10 2.54 0.05 2.54 0.09 Sugar (% of DW) 3.52 0.14 2.54 0.10 9.01 0.87 2.50 0.08 Fat (% of DW) 1.58 0.08 1.86 0.07 3.09 0.12 6.01 0.23 Protein (% of DW) 8.82 0.42 7.55 0.33 8.72 0.36 5.50 0.12 Flavonoid (% of DW) 2.70 0.08 2.47 0.06 4.52 0.10 4.22 0.02 Pectin (% of DW) 3.52 0.27 1.59 1.02 22.53 1.95 15.04 1.12 Lignin (% of DW) 12.52 1.11 23.79 1.85 7.55 0.66 11.04 0.88 Cellulose (% of DW) 39.00 3.12 40.51 3.64 36.22 3.24 39.02 3.55 Hemicellulose (% of DW) 8.32 0.79 10.07 0.99 11.05 1.09 12.01 1.11

The results are expressed as an average ES (n = 7).

Table 3 Eect of transforming process on citrus bre functional compounds and properties Soluble Dietary Fibre (% of DW) CIB-1 CIB-1 Fibre CIB-7 CIB-7 Fibre 6.75 0.50 3.10 0.22 23.31 1.76 17.15 1.43 Insoluble Dietary Fibre (% of DW) 56.00 5.24 74.62 6.99 54.03 4.15 64.56 6.32 Flavonoid mg/g 27.01 0.84 24.20 0.63 45.19 1.03 40.22 0.2 Ascorbic Ac mg/g 0.21 0.01 ND 5.64 0.11 0.42 0.01 TAA mg/g 20.32 0.40 18.15 0.12 60.07 1.03 47.34 0.66

The values are expressed as an average value ES (n = 3). ND: Not detected.

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by-products that underwent usual milling and extraction with hydroalcoholic solvents were used. For CIB-7, the pulp is squeezed through a sieve, the prenisher tube of the in line system. Later it is collected in batches from a tank or washed again. Whatever the case, pulp is easily broken up and ascorbic acid is either washed or degraded by air oxidation. Dietary bre values obtained in this work are in accordance with those obtained by Lario et al. (2004), and more than 2530% of those obtained by Braddock and Graumlich (1981); dierences may be due to the dierent analytical method and or raw material used in the work. As expected, total antioxidant activity also fell with the processing, being higher for CIB-7 and its transformed product, than for CIB-1 and its counterpart. In light of the ascorbic acid values, most of the contribution to TAA might be due to the avonoid antioxidant activity. The lower avonoid contents of CIB-1 and CIB-1 bres, resulting from the industrial process of avonoid extraction, were not the only reason for this lower TAA. The avonoids present in CIB-1, CIB-7, and their counterparts, correspond to those previously described for sour orange and lemon (Vandercook, Tisserat, & Berhow, 1990). In consequence, neoeriocitrin, naringin, neohesperidin and neodiosmin were found at concentrations of approximately 25%, 40%, 25% and 0.7% DW, respectively in CIB-1 (sour orange), and eriocitrin, hesperidin, diosmin and luteolin-7O-rutinoside, at concentrations of approximately 37%, 48%, 4% and 10%, respectively in CIB-7 (lemon). Sour orange avonoids have lower relative antioxidant potenn et al., 2002; tials than have lemon avonoids (Mar n, Martinez, Uribesalgo, Castillo, & Frutos, 2002; Mar Rice-Evans, Miller, & Paganda, 1997). For instance, naringin has a TEAC (trolox equivalent antioxidant capacity) of about 0.05 while neoeriocitrin and neohesperidin have a TEAC of approximately 1, while all the lemon avonoids have individual antioxidant potentials, expressed as TEAC, over 1. Furthermore, higher pharmacological value has been reported for lemon avonoids than for sour orange, specially regarding the presence of diosmin, that constitutes n et al., 2002; Mar n & Mara quarter of all of them (Mar tinez et al., 2002). This fact, together with the higher content of soluble dietary bre, suggests that lemon citrus bre may be of better nutritional value than sour orange bre. Besides, conditioning of citrus industrial by-products leads to a decrease in the functional value of the bre, which could be avoided using milder conditions. Finally, water-holding capacity (WHC) and lipid-holding capacity (LHC) were measured for each one of the by-products. Attempts have been made to correlate WHC and LHC with each one of the measured chemical constituents of bres and soluble and insoluble dietary bre, to propose a main contributor, in citrus bres, to these technological properties. The best t for WHC was found for soluble dietary bre content (Fig. 1) with correlation of r2 = 0.998 for a cubic polynomial expression (y = 0+ax +bx2+cx3). On the other hand, the best t for LHC was

WHC (g of water/g of product)

10

15

20

Soluble Fibre (%)

Fig. 1. Relationship between WHC and soluble dietary bre content of dierent citrus by-products and bres. The gures express an average content in water in g of water/g of product, the standard errors (SE) being shown by vertical bars (n = 3), when these are greater than the symbols.

0,35

LHC (g of oil/g of product)

0,30

0,25

0,20

0,15

0,10 8 10 12 14 16

Lignin (%)

Fig. 2. Relationship between LHC and lignin content of dierent citrus by-products and bres. The gures express an average content in water in g of oil/g of product, the standard errors (SE) being shown by vertical bars (n = 3), when these are greater than the symbols.

found for lignin content (Fig. 2) with a poorer correlation of r2 = 0.878 for an exponential expression (y = a [1ebx]). These results show that WHC and LBC, in citrus bres, do not behave linearly. On the contrary, maximum WHC for citrus bres seems to be above the soluble dietary bre values found in the analysed samples. However, LBC seems not to increase further with contents of lignin over 14%. These results may help to design better foods in the future by adjusting the contents of these constituents. 4. Conclusions Industrial citrus by-products have been traditionally used as raw material for pectin production (Laufenberg et al., 2003). On the other hand, many works have reported

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citrus by-products as putative sources of dietary bre. However, there are few publications regarding direct application on food. Recently, some researchers have published ndez-Lo pez the use of citrus bres for meat products (Ferna et al., 2004; Lario et al., 2004). Their organoleptic characteristics may mask the typical avour of citrus. Furthermore, the disparate collection of citrus bres may help for the design of new meat and other types of products. The composition of citrus industrial wastes shows large variations in their composition. Lignin, cellulose, hemicellulose and protein contents can vary 2-fold their contents while, in others, such as fat and avonoids, variations of 8-fold can be found. Processing of industrial wastes also carries some variation in citrus bre composition and its technological properties. For instance, scalding gives a maximum performance of LHC, but produces a sensitive decrease in soluble dietary bre, reducing the WHC of the nal product. Similarly, ascorbic acid and antioxidant activity of citrus bre are also reduced. Although more research is needed on processing of citrus waste into bres to optimise their properties, the broad composition of the dierent citrus industrial wastes, together with the changes produced during processing and an appropriate blending, might allow the design of specic bres for dierent purposes. References
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