Effects of starch hydrolysatesRevised

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International Journal of Modern Agriculture, Volume 7, No.1, 2018

International Journal of Modern Agriculture, Volume 7, No.1, 2018 ISSN: 2305-7246

EFFECTS OF STARCH HYDROLYSATES ON THE PHYSICAL, CHEMICAL AND SENSORY CHARACTERISTICS OF FILLED

BONBONS

Flavia Maria Vasques Farinazzi-Machado1*, Juliana Audi Giannoni1, Paulo Sérgio Marinelli1, Josué Ribeiro de Jesus1, Natalia Conceição dos Santos1, Tamira Miranda Cabral1

1Faculty of Food Technology of Marilia (FATEC). Marilia/SP – Brazil. CEP: 17506-000 – Marília – SP – Brasil. *Corresponding author (e-mail: [email protected])

Abstract The objective of this study was to use starch hydrolysates - Mylose and Glucomalt - in the fillings of dark and milk chocolate bonbons and check the influence on their physical, chemical and sensory characteristics. Soluble solids, water activity, titratable acidity, moisture and instrumental texture were measured after 0, 15 and 30 days of storage, besides the sensory analysis. The bonbons made with Glucomalt had the variations in the levels of water activity and soluble solids when the storage time was considered. The disruption forces were greater in bonbons with dark chocolate when compared to milk chocolate. The results of the sensory analysis showed that there was a greater variation in the appearance, color and texture attributes between the different chocolates made at time zero, with the dark chocolate samples having the highest scores. After 30 days, however, there was no statistically significant difference between the milk and dark chocolate bonbons. This study presents important results to researches in chocolate manufacturing areas and contribute with sensorial profile and physicochemical parameters collected during the shelf life of products. Key words: Chocolate; Dark; Stability; Texture. Introduction The consumption of chocolates has acquired a positive contribution in human nutrition, because despite the lipid and sugar content, cocoa has high concentrations of antioxidant compounds, mainly polyphenols, including flavonoids, such as epicatechins, catechins and, particularly, procyanidins (Efraim et al., 2011; Oracz et al., 2015; Nabavi et al., 2015). Due to the presence of these compounds, scientific studies have demonstrated that the consumption of chocolates with high cocoa content is associated with the control of risk factors for cardiovascular disease, such as reduced total cholesterol and blood pressure, decreased platelet aggregation, reduced damage to the vascular endothelium and increased plasma levels of high density lipoprotein (HDL-c), in addition to reductions in inflammatory markers (Hooper et al., 2012; Zhang et al., 2013; Colombo et al., 2015; Goya et al., 2016). The confectionery chocolate business has increased 11% between 2000 and 2008 in some countries, including Brazil, which corresponds to an annual growth rate of 1.3% (Sato and Pépece, 2013). According to the Associação Brasileira da Indústria de Chocolates, Cacau, Amendoim, Balas e Derivados (Brazilian Chocolate, Cocoa, Peanut, Candy and Derivatives Association), there has been an increase of 34% in production and of 43% in the

consumption of chocolates between 2009 and 2013 in the country (Brasil, 2016). According Nattress et al (2004) the shelf-life of chocolate depends upon a number of criteria including whether the chocolate is filled or contains inclusions, the addition of other ingredients including fats, the storage temperature and humidity, the initial temper, and the packaging materials used. Increasingly higher environmental temperatures, however, have compromised the technological and sensory quality of filled chocolates and bonbons as a result of the oscillations in temperatures in the market supply chain and the low thermal stability of some products. The ingredients called starch hydrolysates used in the preparation of bonbon fillings, which include glucose syrup, invert sugar and maltose syrup, among others, have various sensory and technological properties, which not only decrease the water activity of fillings, but also extend shelf life and improve such properties as texture and brightness (Jeffery, 1993; Richter and Lannes, 2007a). Technological anti-crystallization, sweetness reduction, solids increase, viscosity control, taste and color improvement, and moisture variation control properties have been attributed to glucose syrups (Farahnaky et al., 2010; Richter and Lannes, 2007b). When the fillings of bonbons are considered, the factor of greatest influence on the

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shelf life of these products is the gain or loss of water, which can be influenced by the glucose syrups used as ingredients, among other factors, such as packaging and storage (Soler et al., 1996; Hartel, 2013). The objective of this study was to use starch hydrolysates - Mylose and Glucomalt - in the fillings of dark and milk chocolate bonbons and check the influence on the physical, chemical and sensory characteristics of the bonbons. Material and methods Preparation of the products The chocolates used for the preparation of the products were granted by the company Barry Callebaut, head quartered in the city of São

Paulo/SP, and are described as milk chocolate 823lb (33.6 % cocoa) and dark chocolate 811lb (54.5 % cocoa). The starch hydrolysates were donated by the company Tereos, located in the city of Palmital/SP, and they included Glucomalt 570 (high maltose content) and Mylose 475, with dextrose equivalents (DE) of 44% to 46% and 38% to 40%, respectively (Tereos, 2013a; Tereos, 2013b). The other ingredients, passion fruit and butter, were obtained from a local market in the city of Marília/SP. The bonbons were produced in the Food Processing Laboratory of the Fatec Marília/SP in an air-conditioned room with a temperature of 21 ºC ± 2 ºC. Four formulations were developed, which are described in Table 1.

Table 1. Ingredients used in the manufacture of milk and dark chocolates filled with the Mylose and Glucomalt starch hydrolysates fillings, described as percentages Ingredients BMGL* BMMY BDGL BDMY Milk Chocolate 75 75 - - Dark Chocolate - - 73 73 Passion Fruit Pulp 10 10 11 11 Butter 9 9 10 10 Glucomalt 6 - 6 - Mylose - 6 - 6 *BMGL: Bonbons filled with milk chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDGL: Bonbons filled with dark chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Mylose For the preparation of the fillings, the passion fruit pulp, butter and glucose syrups (starch hydrolysates) were mixed and put in a microwave until they reached the temperature of 80 ºC, measured with a digital thermometer, and then mixed with the chocolates. After reducing the temperature to 32 ºC, the fillings were placed in rectangular stainless steel container, covered with a polyvinyl polyvinylchloride (PVC) plastic film and put under refrigeration for 24 hours (8° C). The samples were cut manually in standard pieces of 3 cm by 3 cm and 15g in weight, after which they were coated with the respective chocolates of the filling. After the tempering, the pieces were once again cooled to the temperature of 8 ºC, and packaged in laminate. Physical and chemical analysis The samples were analyzed at the Physical and Chemical Analysis Laboratory of the Fatec Marília/SP, Brazil, regarding their water activity, soluble solids (º Brix) and titratable acidity (g 100-

1g of citric acid) at the times of 0, 15 and 30 days, according to the methodology of the Institute Adolfo Lutz (2008). The analyses were performed in triplicate. Texture

The texture analysis were performed in the Physical and Chemical Analysis Laboratory of the Fatec Marília/SP. The analyses were performed in triplicate according to the Institute Adolfo Lutz (2008), for three storage times (0, 15, 30 days), at which times the filled bonbons were removed from their original packaging immediately before the test. The device used was the Stable Micro Systems TA. XT plus, with the Probe HDP/BS, using the compression test with two ruptures. The device was calibrated with a weight of 10 Kg, using a return distance of 17 mm, a speed of 2.0 mm/s, a pre-test speed of 1.5 mm/s and a post-test speed of 10mm/s. In the first two tests, the breaking sensitivity was 1000N, and in the last test it was 3000N. Sensory analysis The sensory analysis of the filled bonbons was performed at the company Tereos, Palmital-SP, Brazil, with 60 untrained tasters at the times of 0 and 30 days. The acceptability test employing the affective method was used, using a nine-point hedonic scale with the extremes 1 - disliked a lot, and 9 - liked a lot, evaluating the attributes appearance, color, aroma, texture, taste and overall acceptance for the four prepared samples, which were offered in monadic form to the tasters (Teixeira et al., 1987; Dutcosky, 1996).

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Statistics The results were presented in the form of average and standard deviation. The design used was complete randomized design (CRD) complemented with Tukey test (Ayres, 2007; Bussab, 2011). Results and Discussion Physical and chemical analysis

The moisture values of the milk and dark chocolate bonbons did not differ at the time zero. However, during the storage period, there was a reduction in the moisture of the bonbons and a statistically significant difference between the samples at time zero and after 30 days of storage (Table 2), with the exception of the dark chocolate bonbons with Mylose (BDMY), whose moisture values were numerically smaller, but without a statistically significant difference.

Table 2. Moisture content in a vacuum (60ºC) of milk and dark chocolate with starch hydrolysates during storage Sample Moments

0 15 30 BMGL* 7.243 ± 1.506 B¹a² 5.867 ± 0.984 ABa 3.190 ± 0.445 Aa BDGL 7.643 ± 0.484 Ba 4.920 ± 0.139 Aa 4.053 ± 0.352 Aab BMMY 7.113 ± 0.683 Ba 5.673 ± 0.414 Aa 5.410 ± 0.252 Abc BDMY 7.113 ± 0.683 Aa 5.293 ± 0.710 Aa 6.250 ± 0.924 Ac *BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose. (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test. Among other properties, the glucose syrups used in sugar products control moisture variation, hygroscopicity and the crystallization of the sugars in the products. These properties are dependent on the dextrose equivalent (DE) of the different syrups, which varies according to the degree of hydrolysis of the starches used. Glucose syrups with low DE are more effective in controlling these properties, since their components have a high molecular weight (Oetterer et al., 2006). The syrups used in this study had a DE that is considered low, between 44 to 46 % for Glucomalt 570, and 38 to 40 % for Mylose 475. As can be seen in Table 2, the moisture contents of the bonbons made with the Mylose syrup had less variation numerically when compared to the other samples. The moisture values for the filled bonbons were smaller than those described in crunchy (12.47 to 16.38 g 100g-1) and strawberry bonbons (21.74 to 25.29 g 100g-1) in a study conducted by Reis

(2011), and also smaller than the moisture values observed in fruit-based truffles (10.2 to 12.35 %) in a study by Miquelim (2006). The results of the water activity analyses performed on the chocolates during the three storage times (0, 15 and 30 days) are described in Table 3. Regarding the samples at time zero, the chocolates with Glucomalt had higher water activity values, which was statistically different between the milk chocolate bonbon with Glucomalt and the Mylose samples. Considering the storage time, there was a significant difference for the BMMY, BMGL and BDGL samples, with a significant difference being observed for the last two samples at 15 and 30 of storage. These findings are in line with the moisture values, since in the chocolates made with Mylose syrup (BMMY and BDMY), the variations in water activity were lower and not significant between 15 and 30 days of storage.

Table 3. Water activity of milk and dark chocolate with starch hydrolysates during storage Sample Moments

0 15 30 BMGL* 0.744 ± 0.023 B1c2 0.731 ± 0.015 Bb 0.650 ± 0.048 Aa BDGL 0.686 ± 0.022 ABbc 0.751 ± 0.007 Bb 0.663 ± 0.047 Aa BMMY 0.602 ± 0.046 Aa 0.682 ± 0.030 Bb 0.706 ± 0.008 Ba BDMY 0.651 ± 0.005 Aab 0.587 ± 0.054 Aa 0.634 ± 0.004 Aa *BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose. (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test.

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Some factors interfere directly with the water activity variations of foods, such as residual moisture, packaging, storage conditions, including ambient temperature and humidity, availability of nutrients and pH (Vasconcelos, 2001). In addition, the fact that the bonbons were produced manually, with a non-industrial technique, is considered important, since it wasn't possible to follow an accurate uniformity regarding the size and layer thickness of the chocolates. And similarly, the samples were packaged by hand. The variations in

water activity values, therefore, may be the result of these interfering factors. When the soluble solids content is considered, there was no statistically significant difference between the samples of dark chocolate with Mylose (BAMY) and the other samples at time 0 (Table 4). However, during the storage period, the greatest variation in the soluble solids content was observed between the samples prepared with the syrup with a high content of maltose (Glucomalt), with a statistically significant difference between the milk chocolates with this starch hydrolysate.

Table 4. Soluble solids of milk and dark chocolate with starch hydrolysates during storage Sample Moments

0 15 30 BMGL* 80.767 ± 0.473 C¹b² 75.133 ± 0.833 Aa 78.767 ± 0.351 Bab BDGL 82.200 ± 0.458 Bb 78.400 ± 0.361 Ab 80.367 ± 1.332 ABb BMMY 82.333 ± 0.764 Bb 76.133 ± 0.666 Aab 75.833 ± 2.248 Aa BDMY 76.833 ± 1.604 Aa 79.867 ± 1.686 ABb 80.933 ± 0.907 Bb *BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose. (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test. According to Soler et al. (1996), solids content and water activity are important parameters when formulating a filling, which must be controlled because of their influence on the bonbons’ shelf-life. In studies conducted by Torrey (1974), a solids concentration above 65 °Brix enables the preservation of products at ambient temperature, preventing their deterioration. The results obtained for the soluble solids content of the samples had higher values, ensuring, by means of this parameter, the durability of the bonbons.

Medeiros et al. (2012) found mean values of 66 °Brix in truffles filled with a sweet cashew nut pastry and passion fruit albedo. The titratable acidity values were higher for the bonbons made with dark chocolate, being statistically significant, probably due to the greater acidity of the cocoa solids in this type of chocolate (54.5 % cocoa) in relation to milk chocolate (Table 5).

Table 5. Titratable acidity values of milk and dark chocolate with starch hydrolysates during storage

Sample Moments 0 15 30

BMGL* 4.753 ± 0.395 Aab 6.770 ± 0.581 Bb 5.443 ± 0.388 Ab BDGL 5.650 ± 0.200 B¹b² 5.697 ± 0.142 Bab 2.280 ± 0.615 Aa BMMY 3.740 ± 0.623 Aa 6.640 ± 0.605 Cb 5.193 ± 0.314 Bb BDMY 4.367 ± 0.378 Ba 4.973 ± 0.159 Ba 3.103 ± 0.445 Aa *BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose. (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test.

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Figure 1 – Breaking strength of milk and dark chocolate with starch hydrolysates during storage

BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose; * p < 0.05, by Tukey Test. The adhesiveness of the samples did not differ statistically at time zero or during the storage time of products, with the exception of the sample BDMY, whose values were higher in the 15th day of storage (Figure 2). According to Anzaldua-Morales (1994), adhesiveness is the effort required to overcome the forces of attraction between the surface of the food and the surface of other materials with which the food comes into contact

Storage, packaging and handling are considered variable conditions under which the sample is tested. It is important to identify the storage conditions and keep them constant. According Afoakwa et al (2007), differences in the texture characters of chocolate can be attributed to use of different cocoa types, variations in ingredient proportions, use of milk crumb instead of milk powder, blending techniques and processing methods.

Figure 2 - Adhesiveness of milk and dark chocolate with starch hydrolysates during storage

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BMGL: Bonbons filled with milk chocolate and Glucomalt; BDGL: Bonbons filled with dark chocolate and Glucomalt; BMMY: Bonbons filled with milk chocolate and Mylose; BDMY: Bonbons filled with dark chocolate and Mylose. Numerically, we can observe in figure 2 that the bonbons made with Mylose syrup had a higher adhesiveness, probably because of the higher viscosity of the syrup, between 12,000 to 17,000cp, in relation to the viscosity of Glucomalt (between 5,000 to 9.000cp) (Tereos, 2013c). Sensory analysis

The mean age of the tasters was 29.6 ± 8.3 for women and 33.4 ± 12.4 for men at time zero and 31.2 ± 12.0 for women and 28.4 ± 8.2 for men after 30 days. Table 7 presents the results of the sensory analysis performed with the bonbons at time zero. The results showed that there was a significant difference between the milk and dark chocolates with Glucomalt for the appearance, color, texture and flavor attributes, with the highest scores awarded to the dark chocolate bonbons. Likewise, the Mylose-based bonbons prepared with dark chocolate had statistically higher scores for the attributes appearance, color and texture.

Table 7. Sensory analysis performed with the bonbons at time zero

Glucomalt Mylose Milk Dark Milk Dark

Appearance 5.73 ± 1.99 A1a² 6.75 ± 1.62 Ba 5.92 ± 2.06 Aa 7.12 ± 1.51 Ba Color 6.13 ± 1.99 Aa 6.98 ± 1.47 Ba 6.48 ± 1.86 Aa 7.22 ± 1.32 Ba Aroma 6.22 ± 2.11 Aa 6.68 ± 1.59 Aa 6.42 ± 1.97 Aa 6.82 ± 1.60 Aa Texture 5.80 ± 2.14 Aa 6.43 ± 1.91 Ba 5.67 ± 2.26 Aa 6.58 ± 1.87 Ba Flavor 6.15 ± 2.32 Aa 6.65 ± 1.89 Ba 6.13 ± 2.49 Aa 6.28 ± 1.90 Aa Global acceptance 6.07 ± 2.02 Aa 6.60 ± 1.75 Aa 6.25 ± 2.06 Aa 6.35 ± 1.59 Aa (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test. According to Misnawi et al. (2004) and Heath (2002), flavor is one of the most important characteristics in chocolate products, followed by the perception of the product's texture by the mouth. As such, the best scores for these attributes were given to the dark chocolate bonbons. For the overall acceptance, however, the scores did not differ statistically between the different bonbons. In a study conducted by Miquelim (2006), the chocolates made with passion fruit fondant had mean scores of 6.7 for the attribute overall acceptance. According to Lannes (1997), the texture of chocolates is the main parameter to evaluate the manufacturing process, influenced especially by the tempering step and the quality of the raw materials used. The snap, characterized by the "breaking" of

the chocolate, is therefore the result of a combination of these factors, and this attribute is the one about which the consumers of these products are most demanding. The data of the sensory analysis, in which the texture of the dark chocolate bonbons received significantly higher scores, corroborates the data of the instrumental texture analysis, in which the breaking strength was greater in these samples, indicating a greater hardness on the surface. Table 8 presents the results of the sensory analysis performed with the bonbons at time 30. The results showed that there was no statistically significant difference between the milk and dark chocolate bonbons, regardless of the glucose syrup used in the formulation of the products.

Table 8. Sensory analysis performed with the bonbons at time 30 during storage

Glucomalt Mylose Milk Dark Milk Dark

Appearance 7.15 ± 1.36 A¹a² 7.17 ± 1.08 Aa 7.27 ± 1.35 Aa 7.50 ± 1.14 Aa Color 7.18 ± 1.28 Aa 7.22 ± 1.09 Aa 7.28 ± 1.32 Aa 7.55 ± 1.06 Aa Aroma 7.30 ± 1.28 Aa 7.10 ± 1.31 Aa 7.20 ± 1.41 Aa 7.10 ± 1.26 Aa Texture 6.97 ± 1.52 Aa 7.02 ± 1.23 Aa 7.02 ± 1.36 Aa 7.38 ± 1.03 Aa Flavor 7.35 ± 1.38 Aa 7.00 ± 1.38 Aa 7.15 ± 1.66 Aa 6.77 ± 1.49 Aa Global acceptance 7.35 ± 1.29 Aa 7.35 ± 1.29 Aa 7.28 ± 1.39 Aa 7.02 ± 1.17 Aa (1) Averages followed by at least one same capital letter do not differ from each other, the sample being fixed. (2) Averages followed by at least one lowercase letter do not differ from each other, setting the time and the analysis, by Tukey Test.

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The scores were greater than 7.0, with the exception of the texture of the milk chocolate bonbon with Glucomalt (6.97) and the flavor for the dark chocolate bonbon with Mylose (6.77). Conclusion Considering the storage duration, there was greater variation in the water activity and soluble solids levels of the chocolates made with Glucomalt, which have a higher DE. Numerically, the breaking strengths were greater in bonbons with dark chocolate compared to milk chocolate, influenced by the composition of chocolate, but the bonbons made with Mylose syrup had more adhesiveness due to the high viscosity of the syrup (smaller DE). The starch hydrolysate did not interfere in the sensory analysis of the samples. References Afoakwa, E.O., Paterson, A., Fowler, M. 2007.

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Effects of starch hydrolysatesRevised

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