Industrial orange juice debittering: Effect on volatile compounds and overall quality attributes

Original article Industrial orange juice debittering: effect on volatile compounds and overall quality attributes Roc ıo Fern andez-V azquez, 1 Carla M. Stinco, 1 Dolores Hernanz, 2 Francisco J. Heredia 1 & Isabel M. Vicario 1 * 1 Food Colour & Quality Laboratory, Department Nutrition & Food Science, Facultad de Farmacia, Universidad de Sevilla, 41012 Sevilla, Spain 2 Department Analytical Chemistry, Facultad de Farmacia, Universidad de Sevilla, 41012 Sevilla, Spain (Received 26 November 2012; Accepted in revised form 19 March 2013) Summary This study was aimed to analyse the impact of the debittering process (DP) in the overall sensory properties of orange juice (OJ) (Citrus sinensis L. Var. Salustiana). The fresh industrial squeezed orange juice (FOJ) and the corresponding OJ after the DP (DOJ) were taken. No significant differences were found in acidity, pH and total soluble solids. Colour was evaluated by image analysis (DigiEye System). Hue and lightness were lower (more reddish and darker) after debittering (P<0.001).Odour profile (limonene, a-pinene, ethyl butanoate, octanal, linalool, citral and terpineol) decreased significantly (from 16% to 61% on average) as well as total phenolic compounds measured by Folin–Ciocalteu (P<0.05).The influence of the debittering on the perceived colour, smell and taste was evaluated by paired comparison tests. The na€ ıve panellists found significant differences not only in taste but also in colour and aroma between FOJ and DOJ; however, preference was not clear. Keywords Aroma, colour, debittering, orange juice. Introduction Orange juice is one of the most popular fruit juices and the most consumed in the world (Melendez- Martinez et al., 2004) largely due to its pleasant sensory properties. One of the major problems in the juice industry worldwide is the formation of bitter compounds after extraction from the fruit (Fayoux et al., 2007). It is due to the formation of limonin from its precursor, limonoate A-ring lactone, when it is released in an acid medium after juice extraction from the membrane tissues (Maier & Beverly, 1968). High levels of bitterness in orange juices cause dislike among consumers, so debittering of orange juice (OJ) is necessary in early and mid-season oranges to improve the acceptability of OJ by consumers. The most commonly used method to reduce the bitterness in processed citrus juices is the use of polymeric adsorbents to selectively remove limonoids (Konno et al., 1982; Johnson & Candler, 1986). Other compounds rather than limonoids may be reduced by the debittering process (DP) such as vitamin C, hesperidin and naringin or total phenolic compounds (Kimball & Norman, 1990; Lee & Kim, 2003; Kranz et al., 2011). In addition to the sorption of bitter phenolic compounds, polymeric adsorbents also have the ability to adsorb volatile flavour compounds, as reported for other industrial processes (Ericson et al., 1992; Krings et al.,1993; Tseng et al.,1993; Selli et al.,2003). In this sense, in debittered orange juice, a ‘flavour scalping’ effect has been described (Fayoux et al., 2007). However, flavour adsorption during the DP may have a positive effect on the sensory quality of orange juices when off-flavours or their precursors are adsorbed (Ma & Lada, 2003). Off-flavours can be formed during heat treatment and storage of juices and have a negative sensory impact on the product. The major off-flavour compound in citrus juices is a-terpineol, which is formed from d-limonene and linalool (Haleva-Toledo et al., 1999). Other sensory properties, like colour, could be modified due to a possible absorption of pigments by the resins during the DP (Lee & Kim, 2003). The aim of this work was to explore the effect of the DP on some quality parameters of OJ (Citrus sinensis L. Var. Salustiana) related with the sensory perception (phenolic compounds, volatile composition and colour parameters) that could be also affected by the DP, beyond the bitter taste itself. Furthermore, the effect on the perception of OJ overall quality was also explored by a na€ ıve panel. *Correspondent: E-mail: [email protected] International Journal of Food Science and Technology 2013, 48, 1861–1867 doi:10.1111/ijfs.12163 ©2013 The Authors. International Journal of Food Science and Technology ©2013 Institute of Food Science and Technology 1861 Material and methods Chemicals The standards limonene, apinene, linalool, octanal, citral, ethyl butyrate, terpineol and gallic acid were purchased from Sigma-Aldrich (Poole, United Kingdom). Methanol (HPLC grade) was purchased from Panreac (Barcelona, Spain), and Na 2 CO 3 and Folin– Ciocalteu from Sigma-Aldrich (Poole, UK). Orange juices samples Orange juices C sinensis cv Salustiana were directly taken from commercial OJ production line at the firm ‘Zumos Pascual’ (Palma del Rio, Cordoba, Spain) at different times during 2011 season (from February to March). In the industry, the fresh OJ is mechanically extracted with an FMC © in line Premium Juice Extractor (FMC Food Tech Citrus System, Lakeland, USA). The extracted juice is then conveyed to two finishing operations to separate juice sacs from the juice. The fresh industrial squeezed OJ (FOJ) samples were taken at this stage. The first step of the DP of FOJ is a pretreatment of centrifugation (Centrifuge, GEA Westfalia Group, Munich, Germany) to remove the excess of pulp (that could cause clogging) until 1–3%. Subsequently, the juice is subjected to the DP, which consists in passing the juice through a column packed with a neutral resin with highly specific adsorption to selectively remove the bitter juice components. The debittering of FOJ was undertaken under own conditions set up at the industry (Zumos Pascual S.A.) The Lewatit VPOC 1064 MD PH (Bayer, Germany) polymeric resin was used as absorbent. This is a macroporous hydrophobic adsorbent resin without functional groups based on a cross-linked polystyrene in the form of porous beads of uniform size (0.44– 0.54 mm). The specific area and pore volume of the resin were 800 m 2 g 1 and 1.2 cm 3 g 1 , respectively. After passing thought the resin, the juice is reconstituted by adding pulp to the desired level. The debittered industrial OJ samples (DOJ) were taken at this stage. Twenty seven samples of FOJ and DOJ counterparts were taken at nine different dates (three samples per day) and immediately frozen until they were analysed. All analyses were done in triplicate. Physicochemical characterisation of orange juices Acidity, pH, total soluble solids were measured according to AOAC methods (AOAC, 1995). The ratio was calculated by dividing the total soluble solids by the acidity. Measures were made in triplicate. Total phenolic compounds Total phenolic compound contents were determined by the Folin–Ciocalteu method (Singleton & Rossi, 1965). Total phenol values are expressed in gallic acid (mg L 1 ), which is a common reference compound. All analyses were made in triplicate. GC analysis of volatile flavour compounds Aroma compounds were extracted according to the method described by Jella et al. (1998). Briefly, 4 mL of pentane–diethyl ether mixture (1:1) was added to 20 mL OJ and mixed for 12 h. 25 lL of propyl benzene (50 mg L 1 ) was added to the samples prior to extraction as an internal standard. The resulting emulsion was broken by centrifugation (5 min 95000 g). The upper layer was carefully removed, dried and finally concentrated to a final volume of 1000 lL. The extractions were carried out in triplicate, and each sample was injected into the GC system three times. 1lL was analysed by GC system. Isolated solvent was chromatographically separated on a capillary column DB-Wax (60 m 90.25 mm 90.25 lm) from Varian (Walnut Creek, CA, USA) using an Agilent Technology 6890N GC-FID system (Palo Alto, CA, USA). The following conditions were used: injector temperature 250 °C; detector temperature 250 °C; carrier gas flow (He) 1 mL min 1 . The injections were made in split mode (split ratio 1:25), and the oven temperature was maintained at 40 °C for 3 min, then increased from 40 to 200 °Cat6°C min 1 and eventually held for 6 min. The quantification of all the compounds was carried out by the internal standard method. The flavour components were identified by comparison of their retention times with those of standard compounds. Linearity of responses was r 2 >0.998 across a standard concentration of each compound. Digital image analyses: Instrumental colour measurement For colour specifications, digital images were used to obtain the total appearance of juice at depths observed by consumers. The OJs were placed in 75 mL capacity transparent plastic bottles. The samples were measured in triplicate against a grey surround and white background using image analyses. The DigiEye imaging system (Luo et al., 2001) was used to capture the digital images. The latter system includes a calibrated digital camera 10.2-megapixel Nikon D80 (Nikon Corporation, Tokyo, Japan) and an objective Nikkor 35-mm f/2D (Nikon Corporation), a colour sensor for display calibration, and an illumination box designed by VeriVide Ltd. (Leicester, UK). In these measurements, the samples were illuminated by a diffuse D65 ©2013 The Authors International Journal of Food Science and Technology ©2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2013 Overall quality of debittered orange juices R. Fern andez-V azquez et al.1862 13652621, 2013, 9, Downloaded from https://ifst.onlinelibrary.wiley.com/doi/10.1111/ijfs.12163 by Universidad De Sevilla, Wiley Online Library on [07/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License simulator. For obtaining CIELAB coordinates from RGB colour space, the DigiFood software (Heredia et al., 2006) was used. From the CIELAB uniform colour space, the psychophysical parameters chroma (C ab ) and hue (h ab ) are defined as: C ab ¼ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi a ðÞ 2þðbÞ2 q;hab ¼arctan b=a ðÞ Chroma (C ab ) is used to determine the degree of difference of a hue in comparison with a grey colour with the same lightness and is considered the quantitative attribute of colourfulness. Hue (h ab ) is the attribute according to which colours are usually defined as reddish, greenish, etc. and is used to define the difference of a colour with reference to a grey colour with the same lightness. This attribute is related to the differences in reflectance at different wavelengths and is considered the qualitative attribute of colour. Colour differences, which are very important to evaluate relationships between visual and numerical analyses (Melgosa et al., 1997), are calculated as the Euclidean distance between two points in the 3-D space defined by L*,a*and b*: DE ab ¼ffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi DL ðÞ 2þðDaÞ2þðDbÞ2 q Sensory analyses Evaluation of the perceived differences due to the DP in terms of colour, aroma and taste was completed by specific difference tests (Paired comparison, ISO 5495:2005). Untrained assessors were recruited from staff and students of University of Seville (n=12) to take part in the study. Panellists were not informed about the characteristics of the samples, so they did not know that samples were FOJ and DOJ. A paired comparison tests (FOJ versus DOJ, for each pair of industrial samples) were performed. Assessors were presented with two samples and asked to first look the sample and determine whether the samples were different in colour. In the next session, they were asked to smell the samples and determine whether there was any difference between each pair of samples, and finally, the last sessions were to ask them to taste the sample and determine whether the samples were different. In all the sessions, panellists could choose which sample they preferred and do some comments about the OJs. Samples (75 mL) were presented in bottles of transparent plastic, labelled with random three digit codes, in a randomised order across the panel. Mineral water (Evian, France) and unsalted crackers (Matzo, UK) were provided for palate cleansing, and all testing were performed in temperature-controlled individual test booths. Data were collected using Fizz software (Biosynthesis, France). Data analysis The statistical design consisted of one-way repeated measures analysis of variance (ANOVA). Two factors were considered: the process with two levels (fresh and debittered) and the outcome variables, including the volatile compounds, phenolic content and the physicochemical and the colorimetric variables. The twenty seven cases (OJ samples) were used as random factor, for the purpose of establishing differences between process stages (fresh and debittered). Statistically significant differences (P<0.05) were determined using the Tukey’s multiple comparison procedure. Pattern recognition techniques, such as stepwise linear discriminant analysis (SLDA), were applied on experiment standardised data to distinguish between different types of orange juices. All the statistical analyses were performed with Statistica v.8.0 software (StatSoft, 2007). Sensory data of pair comparisons tests were analysed using ISO 5495:2005 to determine whether the panellists were able to discriminate between samples (P<0.05). Results and discussion Physicochemical characterisation Physicochemical characterisation of each of the samples included in the study is shown in Table 1. The DP did not result in significant variations in the physicochemical parameters after the application of Table 1 Physicochemical characteristics and colour coordinates in FOJ and DOJ (Mean SD) Parameters FOJ DOJ pH 3.9 0.1 4.0 0.2 Acidity † 0.7 0.1 0.7 0.1 Total soluble solids ‡ 11.7 0.9 11.2 0.5 Ratio § 15.8 1.4 15.2 1.2 L* 63.03 1.29 61.08 1.32*** a* 7.08 1.93 8.94 1.72*** b* 59.77 1.49 5951 1.22 C* ab 60.24 1.35 60.22 1.20 h ab 83.23 1.94 81.45 1.66*** † Gram of citric acid per 100 mL of orange juice. ‡ Expressed as ºBrix. § Total soluble solids/acidity. Different superscripts within the same row indicate statistically significant differences. *P<0.05, **P<0.01 and ** * P<0.001. ©2013 The Authors International Journal of Food Science and Technology ©2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2013 Overall quality of debittered orange juices R. Fern andez-V azquez et al.1863 13652621, 2013, 9, Downloaded from https://ifst.onlinelibrary.wiley.com/doi/10.1111/ijfs.12163 by Universidad De Sevilla, Wiley Online Library on [07/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License repeated measures ANOVA analysis. FOJ showed a pH ranging from 3.7 to 4.1. This result is in agreement with Del Caro et al. (2004) who reported a pH in this variety of 3.9. In the case of DOJ, pH ranged from 3.8 to 4.3. Acidity in FOJ and DOJ ranged from 0.6 to 0.8 g of citric acid per 100 mL. Other studies reported values a bit higher (0.9 and 1.1 g of citric acid per 100 mL) in fresh orange juices (Del Caro et al., 2004; G omez-L opez et al., 2010). Kola et al. (2010) neither found a significant decrease in the acidity in samples after debittering, reporting 1.4 g citric acid per 100 mL in FOJ and from 1.3 to 1.4 in DOJ. Total soluble solids ranged from 10.1 to 12.5 ºBrix (average 11.7 ºBrix) in FOJ, this average value is close to those reported for other OJ (11.9–12.8 ºBrix) (Del Caro et al., 2004; Tribess et al., 2009). In accordance with Kola et al. (2010), DOJ total soluble solids were 11.2 ºBrix, without a significant decrease in relation to FOJ. Total phenolic compounds Based on the absorbance values of extract solutions that reacted with Folin–Ciocalteu reagent, using gallic acid as standard, total phenolic content of juices varied between FOJ and DOJ. The total phenolic content in FOJ was 716 mg gallic acid per L. This result agree with Rapisarda et al. (2008) who reported the presence of moderate phenol amounts (524–696 mg of gallic acid per L) in five orange genotypes (Tarocco Messina, Tarocco Meli, Moro, Ovale and Valencia) from Italy. Tounsi et al. (2011) also studied total phenolic compounds in orange juices, reporting 255 mg of gallic acid per L in blood orange juice. In contrast, DOJ suffered a significant (P<0.05) decrease in total phenolic compounds due to the DP (24%), so the final concentration in DOJ was 547 mg of gallic acid per L. Digital image analyses: instrumental colour measurement The application of digital image (DigiEye analysis) to evaluate the colour offers a more reliable measurement of the food colour than traditional methods. A better correlation with sensory analysis can be achieved (Fern andez-V azquez et al., 2011). Table 1 shows the colour parameters obtained in fresh and debittered OJ. The values of the coordinate L*ranged from 60.02 to 64.30 and from 59.29 to 63.19, in FOJ and DOJ, respectively, showing a decrease in average values of 12.36% in DOJ. Hue ranged from 77.94 (the most reddish OJ) to 84.86 (the most yellow) before debittering and from 77.85 to 82.89 after debittering, decreasing 2.13% in DOJ. Ranges for the coordinate C ab were 58.20–62.16 and 58.59–62.50 for OJs and debittered OJs, respectively. Our findings agree with other studies in FOJ of different varieties measured by image analyses which showed values of L*ranged from 56.09 to 61.34, h ab ranged from 66.43 to 81.99, and finally range of chroma was 54.03–60.29 (Fern andez-V azquez et al., 2012). Coordinate h ab , the qualitative component of colour, was significantly different (P<0.001) being slightly lower after debittering. This means that debittering makes the samples a bit more reddish. On the other hand, L*coordinate was significantly higher before than after debittering, which means that samples were darker after the treatment. Finally, the quantitative component of colour (C ab) did not show a clear pattern comparing fresh OJs and debittered OJs, and there were not significant differences. Lee and Coates (2003) reported a lightness increase and a chroma decrease after debittering in red grapefruit juice concentrate. CIELAB colour differences (DE ab) were calculated between FOJ and DOJ. The average was 2.72, which is close to the threshold for human colour discrimination (Mart ınez et al., 2001). The CIELAB colour difference DE ab was split in the three components, called lightness difference (DL*), chroma difference (DC ab) and hue difference (DH ab). Figure. 1 shows the contribution (as percentage) of each component to the total colour differences detected in comparison with each date (average data of three samples). It can be observed that the main contribution to the whole colour difference was related to the qualitative component of colour (DH ab) and the lightness (DL*), while chroma difference had a minor contribution to the total difference in most of the cases. DH% DL% DC*ab% 123456789 Pairs of OJs 0 20 40 60 80 100 Figure 1 Contribution of DC ab,DL*and Dh ab to colour differences between FOJ and DOJ of each date (average data of three samples). ©2013 The Authors International Journal of Food Science and Technology ©2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2013 Overall quality of debittered orange juices R. Fern andez-V azquez et al.1864 13652621, 2013, 9, Downloaded from https://ifst.onlinelibrary.wiley.com/doi/10.1111/ijfs.12163 by Universidad De Sevilla, Wiley Online Library on [07/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License GC analysis of volatile flavour compounds Limonene, a-pinene, ethyl butanoate, octanal, linalool and citral were chosen because they have been identified as the most contributing compounds to orange flavour and can be even used in OJ authentication (Robards & Antolovich, 1995). Moreover, terpineol is a well-known off-flavour compound present in citrus products formed from limonene or linalool. This compound exceeds the taste threshold level under practical processing and storage conditions (Naim et al., 1998). Table 2 shows the concentrations for aroma compounds in FOJ and DOJ. All the aroma compounds decreased significantly in DOJ. Percentages of reductions were from 16% to 61%. The main compound in both groups was limonene with 1667 and 793 mg L 1 in FOJ and DOJ, respectively. Limonene has been reported as the major terpene in OJ (Jordan et al., 2001; Brat et al., 2003; Moufida & Marzouk, 2003; Rega et al., 2004; Fan et al., 2009; Kelebek & Selli, 2011); nevertheless, limonene is not necessarily the most important or overriding compound when evaluating flavour quality (Jia et al., 1998). The next odour compounds in concentration were the ethyl butyrate (108 mg L 1 in FOJ) followed by pinene (21 mg L 1 in FOJ). Recent studies reported concentration of ethyl butyrate of 102 mg L 1 analysed in C sinensis L.Osbeck (Kelebek & Selli, 2011) and of pinene of <3mgL 1 in a mixture of C sinensis Berna, Valencia Late and Salustiana (Jordan et al., 2003).These results are in accordance to those reported by Kranz et al. (2011) in grapefruit juice, who reported that the DP has a strong impact on volatile flavour compounds. Moreover, they also reported that the off-flavour compound a-terpineol had the higher adsorption affinity for the adsorption resin. However, our findings point out that octanal is the compound which decreases in a higher proportion, from 13 to 5 mg L 1 (61% of decrease). SLDA analysis Total phenolic compounds, colour parameters and aroma content were included in a discriminant analysis to enhance the discrimination of the samples using debittering as criterion for comparison. Of these variables, seven were significant (P<0.05) and, in descending order of discriminating power, were total phenolic compounds, a-pinene, linalool, a-terpineol, a*, ethyl butyrate and b*. As only two sets were taken into consideration for the model, only one classification function was obtained, which yielded a good separation (100% correct classification) between the samples (Fig. 2). The colorimetric variables a*and b* had positive sign, while linalool, a-pinene, total phenolic compounds, a-terpineol and ethyl butyrate had negative sign. Sensory analyses To find out sensory differences in colour, aroma and taste, these attributes were evaluated separately. Results from paired comparison test of the visual evaluation provided sufficient evidence to conclude that a significant difference (P<0.05) existed between the colour of FOJ and DOJ. DE*of each pair of samples can explain these results, as it was mentioned before an average difference in colour of 2.72 was calculated. In terms of preference, there was not a clear pattern in the panellists. 37% of the responses did not show any preference to none of the sample, 48% preferred the colour after debittering and only 15% preferred the OJs without debittering treatment. On the other hand, aroma evaluation by panellists showed significant differences (P<0.05) comparing Table 2 Volatile components in FOJ and DOG (mg L 1 ) (Mean SD) Components FOJ DOJ Limonene 1667.63 330.39 792.94 395.15** Octanal 13.03 5.05 5.06 2.01*** Linalool 21.09 6.65 17.73 2.69* Ethyl Butanoate 107.80 23.04 85.96 21.23*** Terpineol 13.88 4.44 9.76 3.27* Citral 12.40 2.23 8.52 2.35*** Pinene 20.47 2.49 13.67 3.56*** Different superscripts within the same row indicate statistically significant differences. *P<0.05, **P<0.01 and ***P<0.001. Root 1 Samples –6 –4 –2 0 2 468 –10 0 10 20 30 40 50 60 FOJ DOJ Figure 2 Scatterplot of the orange juice samples in the plane defined by the canonical function when debittering is considered for discrimination. ©2013 The Authors International Journal of Food Science and Technology ©2013 Institute of Food Science and Technology International Journal of Food Science and Technology 2013 Overall quality of debittered orange juices R. Fern andez-V azquez et al.1865 13652621, 2013, 9, Downloaded from https://ifst.onlinelibrary.wiley.com/doi/10.1111/ijfs.12163 by Universidad De Sevilla, Wiley Online Library on [07/03/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License FOJ and DOJ. Moreover, 71% showed a preference for FOJ pointing out that the DP could have a negative effect in the OJs. This could be due to the aroma reduction in important flavour compounds in DOJ, as reported previously. For example, octanal, limonene and a-pinene, which were the compounds with the major reduction, are also responsible for the positive sensory properties of OJ like citrusy, lemon and mint, and pine, respectively (Arena et al., 2000, 2006; Rega et al., 2003). The off-flavour a-terpineol also had a significant reduction of 30%, but this effect was not enough to overcome the negative effect of this compound in the flavour. Results from paired comparison test for taste evaluation provided sufficient evidence to conclude that a significant difference (P<0.05) existed between FOJ and DOJ. However, it was unclear what the panellists’ preference was, as 53% of the answers showed preference for OJs without debittering, 35% preferred OJs after debittering and 13% did not show preference to any of the samples compared. This may be related to the fact that after debittering, OJs were less bitter, but also too smooth and artificial, as reported by some panellists. Conclusions In conclusion, total phenolic compounds and the entire aroma compounds analysed decreased significantly due to DP. Colour parameters also changed significantly in DOJ, being these orange juices more reddish and darker. 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