186

International Journal of Food Science and Technology 2011, 46, 186193

Original article Anthocyanins: optimisation of extraction from Cabernet Sauvignon grapes, microcapsulation and stability in soft drink
Vvian M. Burin, Priscilla N. Rossa, Nayla E. Ferreira-Lima, Maria C. R. Hillmann & Marilde T. Boirdignon-Luiz*
Departamento de Ciencia e Tecnologia de Alimentos CAL CCA, Universidade Federal de Santa Catarina, Rodovia Admar Gonzaga, 1346, CEP: 88034-001, Itacorubi, Florianopolis, SC, Brazil (Received 28 July 2010; Accepted in revised form 27 September 2010)

Summary

The aim of this study was to establish the optimum conditions for the extraction of anthocyanins from Cabernet Sauvignon (Vitis vinifera L.) grapes using the response surface methodology and to evaluate the stability of these anthocyanins encapsulated with dierent carrier agents in an isotonic soft drink system under dierent light and temperature conditions. The extraction process was optimised with the response surface methodology to obtain the highest anthocyanin concentration (40 mL of ethanol:1.5 N HCL (85:15) as solvent, extraction time 29.4 h at pH 2.4). The degradation of the anthocyanins followed rst-order kinetics in the situations evaluated. Maltodextrin, maltodextrin c-cyclodextrin and maltodextrin arabic gum were tested as carrier agents and the combination of maltodextrin arabic gum presented the longest half-life time and lowest degradation constant for all the conditions evaluated. The formation of microcapsules was observed through scanning electron microscopy.
Anthocyanins, grape, microencapsulation, spray drying.

Keywords

Introduction

Colour is one of the most important attributes in food and drinks, as it aects the acceptability of products by consumers. The replacement of synthetic dyes with natural food colourants has increased considerably. To provide the colour red, anthocyanins are an attractive alternative, as they provide high colourant power, low toxicity and water solubility, which permit their incorporation in many food systems (Ersus & Yurdagel, 2007). Anthocyanins are the largest group of water-soluble pigments obtained from plants and they can be dened chemically as polyhydroxylated or polymethoxylated glycosides or acylglycosides of anthocyanidins belonging to the avonoids class. These compounds are dierentiated by the number of hydroxyl groups and the nature, number and position of sugars attached to the molecule (Kong et al., 2003). The main anthocyanins found in grapes are cyanidin, peonidin, delphinidin, petunidin and malvidin (Munoz-Espada et al., 2004), and their concentration is dependent on the variety and environmental factors. Studies carried out in vitro and in vivo have shown that anthocyanins are responsible for important thera*Correspondent: Fax: +55 48 3721 9943; e-mail: bordign@cca.ufsc.br

peutic properties such as: antioxidant, anticarcinogenic and anti-inammatory activity, low density lipoprotein (LDL) oxidation inhibition and a decrease in the risk of cardiovascular disease, and thus their application as natural pigments in food is of great interest (Wang & Xu, 2007; Rosso et al., 2008). However, anthocyanins are unstable pigments and can be converted to colourless compounds. Their stability is aected by many factors including pH, temperature, light, oxygen and the food matrix (Wang & Xu, 2007). Also, there are limitations to the application of anthocyanins as pigments in food systems related to the process conditions, formulation and storage conditions. This has stimulated a search for new methods to enhance their stability and increase the possibilities for the use these compounds (Giusti & Wrolstad, 2003). To determine the best conditions for the extraction of natural pigments using organic solvents, researchers have applied the response surface methodology (RSM). This methodology represents a combination of mathematical and statistical techniques aimed at optimising the nal response. The main advantage of this method is the reduced number of experiments required to provide sucient information to obtain statistically acceptable results (Cho et al., 2009). The RSM with central composite design provides a complete factorial investigation of the simultaneous, systematic and ecient variation of

doi:10.1111/j.1365-2621.2010.02486.x
2010 The Authors. International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

Anthocyanin: microcapsulation and stability V. M. Burin et al.

187

important components, identifying the possible interactions, main eects and optimal conditions of operation (Roriz et al., 2009). A very useful method to enhance the stability of natural pigments is microencapsulation by the spray drying technique, which provides powders with low humidity and good quality that are easy to store. The degradation protection and consequent preservation of compounds is due to the inclusion of these pigments in macromolecules (Cano-Chauca et al., 2005; Ngo & Zhao, 2009). The active compounds in dehydrated formulations and microcapsules destined for controlled-released are protected during processing and storage. Knowledge of the carrier agent properties is crucial in order to optimise the process and reduce the cost (Kurozawa et al., 2009). In particular, the coating agents submitted to spray drying must show a good capacity for emulsication and lm forming, along with high solubility, low viscosity and hygroscopicity (Loksuwan, 2007). The most notable of the commonly used carrier agents are natural gums and dextrins (Kanakdande et al., 2007). In general, the coating agent alone does not oer all the properties required to ensure a good microencapsulation and thus a mix of one or more components is frequently employed to enhance the encapsulation (Turchiuli et al., 2005). The aim of this study was to establish the optimal conditions for anthocyanin extraction from Cabernet Sauvignon (Vitis vinifera L.) grapes, employing the RSM. The stability of these anthocyanins encapsulated with dierent carrier agents in a soft drink model system was then evaluated under dierent light and temperature conditions.
Materials and methods

estimate the experimental error. All runs were carried out with 10 g of grape skins. The response chosen for the design was the total monomeric anthocyanins content (mg 100 g grape skin) quantied by the pHdierential method. A second-order polynomial model was constructed to estimate the response (eqn 1): y b0
k X i1

bi xi

k X i1

bii x2 i

k1 k XX i1 j>1

bij xi xj e

where y is the estimated response (dependent variable), b0 the model constant, bi the linear eect coecient, bii the quadratic eect coecient, bij the interaction coefcient for two factors, xi, xj the independent variables, e the error, k the number of variables considered, i and j the codied factors of the system.
Extraction and quantication of anthocyanins

According to the results obtained in the experimental planning, the anthocyanin extract was prepared through the maceration of 100 g of Cabernet Sauvignon grape skins in ethanol:1.5 N HCl (85:15) solution (solid:liquid ratio 1:4) for 29.4 h in the dark under refrigeration temperature conditions (4.0 1 C). The extract obtained was ltered and kept at 4.0 1 C in an amber ask until analysis. The total monomeric anthocyanins content was determined using the pH-dierential method described by Giusti & Wrolstad (2001), measuring the absorbance in a UV-Vis absorption spectrophotometer (Hitachi U2010, Tokyo, Japan). The content of total monomeric anthocyanins in the grapes was expressed as mg malvidin-3-glycoside per 100 g of grape skin (MW= 529 g mol)1 and e = 28 000).
Preparation of microcapsules and spray drying process

Chemicals

The study was carried out with Cabernet Sauvignon (Vitis vinifera L.) grapes from the region of Sao Joaquim, Santa Catarina, Brazil. Maltodextrin (MorRex 1920 DE 19, Plury Quimica, Diadema, Brazil), arabic gum (Colloides Naturels, Sao Paulo, Brazil) and c-cyclodextrin (Cerestar, Minneapolis, MN, USA) were used for the encapsulation by spray drying. All other reagents were of analytical grade.
Experimental design

The RSM was used to optimise the conditions for the extraction of anthocyanins from Cabernet Sauvignon grapes. The experimental design was carried out using a central composite design with three independent variables solvent (x1), pH (x2) and extraction time (x3) and three levels coded as 1, 0 and 1, resulting in seventeen runs including three replicates at the central point to

The extract was concentrated in a low-pressure rotary evaporator, approximately 20% of ethanol (QUIMIS, Q.344.2, SP, Brazil) at 35 2 C and the carrier agents were then added in concentrations calculated on the basis of the solids content of the crude extract. The samples were codied as: extract maltodextrin (M) (1:1 w w), extract maltodextrin c-cyclodextrin (MC) (1:1:1.5 w w w) and extract maltodextrin arabic gum (MG) (1:1:1.5 w w w). After addition of the carrier agents, the samples were stirred in a homogeniser (B. Brain Biotech International, CERTOMATMO, Melsungen, Germany) at 100 rpm for 2.5 h in the dark in sealed asks. The drying process was carried out using a Buchi Mini Spray Dryer (BUCHI Mini Spray Dryer B-290, Flawil, Switzerland). The drying conditions were: inlet air temperature 150 C and outlet temperature 50 C,

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

International Journal of Food Science and Technology 2011

188

Anthocyanin: microcapsulation and stability V. M. Burin et al.

pump power 25% and maximum aspirator rate. During the process, the temperature of the feed solution was 25 C. The powder obtained was stored in an amber ask under refrigeration temperature (4.0 1 C) in the dark.
Colour characterisation of encapsulated anthocyanins

the SEM samples were attached to stubs using two-sided carbon adhesive tape. The surface was sputter-coated with a thin layer of gold and then examined at 35, 500 and 1000 magnications. An acceleration potential of 10 kV was used to obtain the micrographs.
Statistical analysis

The colour parameters of the encapsulated pigments were determined using the CIELab system (colourimeter Chroma Meter CR-400, Konica Minolta, Osaka, Japan) with the coordinates a* (chromatic index red green), b* (chromatic index yellow blue) and L* (lightness).
Application and evaluation of encapsulated anthocyanin stability

All analyses were carried out in triplicate with two repetitions. The statistica software (v. 8.0, StatSoft, Inc., Tulsa, OK, USA) was used to verify the central composite design, to calculate the coecients, mean values and standard deviations, and to carry out the analysis of variance (anova) and Tukey test (P < 0.05).
Results and discussion

The encapsulated anthocyanin extracts were added to an isotonic soft drink system (sucrose 4%, citric acid 0.38%, glucose 0.15%, sodium chloride 0.12%, sodium citrate 0.11% and potassium monophosphate 0.022%), considering a commercial isotonic soft drink colour as the reference. To evaluate the colourant stability, 5 mL samples of the isotonic soft drink system were transferred to screw-top test tubes and kept under the following three conditions: under a 40 W uorescence lamp at 25 1 C; in the absence of light at 25 1 C; and in the absence of light at 4 1 C. Samples were withdrawn to quantify the content of anthocyanins at regular intervals for 40 days. Based on the plots of log of total anthocyanins concentration versus time for the dierent conditions analysed, the rst-order kinetics constants (k) and halflife times (t) of the pigments in the isotonic soft drink system were calculated using eqs 2 and 3, respectively. Slope of the straight line k=2:303 t1=2 ln0:5 k1 2 3

Experimental design for anthocyanin extraction

The RSM evaluated the eects and interactions of the solvent volume, extraction time and pH of the solution in terms of anthocyanin extraction from Cabernet Sauvignon grapes. Table 1 shows the values for the factors and the total monomeric anthocyanins content obtained for each extraction condition. The signicance of the factors was conrmed by anova, where it was possible to observe that the quadratic eect of solvent volume and the linear eects of pH and extraction time were signicant. The pH was shown to have the weakest inuence on the responses, and interactions between the variables were not observed (Table 2). The predicted

Table 1 Full factorial central composite design matrix of three variables in coded and natural units along with the observed responses Coded units of variable Run 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 x1 )1 )1 )1 )1 1 1 1 1 )1,68 1,68 0 0 0 0 0 0 0 x2 )1 )1 1 1 )1 )1 1 1 0 0 )1,68 1,68 0 0 0 0 0 x3 )1 1 )1 1 )1 1 )1 1 0 0 0 0 )1,68 1,68 0 0 0

Natural units of variable Solvent (mL) 20.00 20.00 20.00 20.00 60.00 60.00 60.00 60.00 6.40 73.60 40.00 40.00 40.00 40.00 40.00 40.00 40.00 Time (h) 8.00 8.00 24.00 24.00 8.00 8.00 24.00 24.00 16.00 16.00 2.56 29.40 16.00 16.00 16.00 16.00 16.00 pH 1.20 3.60 1.20 3.60 1.20 3.60 1.20 3.60 2.40 2.40 2.40 2.40 0.40 4.40 2.40 2.40 2.40 Response 465.96 356.32 580.84 407.52 500.88 407.88 592.01 463.06 153.38 519.36 270.11 611.25 550.02 370.11 593.33 590.08 575.72

where k is the rst-order kinetics constant. The percentage of colour retention was calculated as described by Katsaboxakis et al. (1998) using eqn 4. %R At =A0 100 4

where At and A0 are the absorbance at time t and zero respectively.

Scanning electron microscopy

The particle structures of the microcapsules containing maltodextrin (M), maltodextrin c-cyclodextrin (MC) and maltodextrin arabic gum (MG) were evaluated with a scanning electron microscope (SEM) (JEOL, model JSM-6390LV, Tokyo, Japan). For visualisation

International Journal of Food Science and Technology 2011

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

Anthocyanin: microcapsulation and stability V. M. Burin et al.

189

Table 2 Regression coefcients and anova results for total monomeric anthocyanins extracted from Cabernet Sauvignon grapes (1) Solvent (L) Solvent (Q) (2) Time (L) Time (Q) (3) pH (L) pH (Q) 1L 2L 1L 3L 2L 3L Error Total SS

Regression coefcients 56.2880 )72.6772 64.8549 )35.7908 )59.3673 )29.0830 )2.4720 7.6265 )12.4545

Sum of squares (SS) 43231.4 59530.0 57321.1 14365.1 47776.3 9281.2 48.9 465.3 1240.9 56078.9 268636.1

DF 1 1 1 1 1 1 1 1 1 7 16

Average square 43231.4 59530.0 57321.1 14365.1 47776.3 9281.2 48.89 465.31 1240.92 8011.27

F value 5.396327 7.430784 7.155060 1.793111 5.963640 1.158518 0.006102 0.058082 0.154896

P value 0.053159 0.029511 0.031773 0.222395 0.044624 0.317464 0.939921 0.816461 0.705612

DF, degrees of freedom; L, linear effect, Q; quadratic effect.

values for total monomeric anthocyanins are close to the experimental values demonstrating that the model is applicable, which showed determination coecient (R2) of 0.80. The mathematical relationship between the independent and response variables was evaluated through the quadratic model constructed in the regression analysis (eqn 5). Y 581:2832 64:8549t 59:3673pH 72:6772S2 5 where Y is the total monomeric anthocyanins content (mg 100 g grape skin); t the extraction time (hours); and S the solvent volume (mL).
(a)
Anthocyanin content (mg 100 g1)
700 600 500 400 300 200 100 0 100 200

The response surface plots indicated the eects of two variables on the anthocyanins content, with a third independent variable (central point) xed to the central experimental level (Fig. 1). The anthocyanin content signicantly increased with extraction time and solvent volume, reaching the maximum response (611.25 mg 100 g) at 40 mL ethanol:1.5 N HCl (85:15) solution and 29.4 h of extraction time (Fig. 1a). A study carried out by Chen et al. (2007) with dierent solid liquid ratios for anthocyanin extraction also demonstrated that a ratio of 1:4 (solid:liquid) gave the highest content of extracted pigments, which is consistent with the results reported herein. The same authors arm that during solid liquid extraction an excessive increase in

(b)
Anthocyanin content (mg 100 g1)
800 600 400 200 0 200

15 10 5

Ti m

(h )

Eth

o an

l (m

L)

0 5. .5 4 .0 4 .5 3 .0 3 .5 2 0 pH 2. 1.5 0 1. .5 0 .0 0

35

80 70 60 50 40 30 20 10

30

80 70 60 50 40 30

15

n tha

ol

(m

L )

20

Figure 1 Response surface for total monomeric anthocyanins content considering: (a) extraction time and solvent volume with a constant pH of 2.4; (b) solution pH and solvent volume with an extraction time of 16 h; and (c) solution pH and extraction time at a constant solvent volume of 40 mL.

Anthocyanin content (mg 100 g1)

20 10

(c)
700 600 500 400 300 200 100 0 0 5. .5 4 .0 4 .5 3 3.0 .5 pH 2 2.0 .5 1 .0 1 .5 0 .0 0

Tim

e(

35 30

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

International Journal of Food Science and Technology 2011

15 20 15

h)

10 5

190

Anthocyanin: microcapsulation and stability V. M. Burin et al.

solvent volume can result in losses through evaporation or saturate the extractor medium. This may be the reason for the decrease in the anthocyanins content at ethanol volumes higher than 60 mL in this study. The reduction in pH and concomitant increase in solvent volume also caused an increase of the anthocyanin content, with an optimum extraction region at around pH 2 (Fig. 1b). Montes et al. (2005) also reported an ethanol solution acidied with hydrochloric acid and a pH of around 2 as the best conditions for anthocyanin extraction. Torskangerpoll & Andersen (2005) armed that the colour stability of anthocyanins is mainly dependent on the pH and the structure of the molecule, since in acid solutions anthocyanins remain preferentially in the avylium cation form, which gives higher wavelengths in UVVis absorption spectra. Kirca et al. (2007) observed a considerable decrease in anthocyanin stability for pH values up to 5.0. On increasing the extraction time it was observed that the anthocyanin content progressively increased. Corrales et al. (2009) established that the contact of grape skins with the solvent for a longer time provides a higher diusion of compounds such as anthocyanins. Revilla et al. (1998) evaluated the extraction of phenolic compounds from Cabernet Sauvignon grapes under dierent conditions and observed that a longer extraction time provided a higher amount of extracted anthocyanins. These results together show that the optimum combination of independent variables for anthocyanin extraction was 40 mL ethanol:1.5 N HCl (85:15) solution and an extraction time of 29.4 h at pH 2.4. These parameters were therefore chosen to carry out the extraction of anthocyanins for later encapsulation.
Colour parameters of encapsulated pigments

Log of anthocyanin content (mg L1)

1.8

(a)

1.4

0.6 0 10 20 30 40

Time (days) Log of anthocyanin content (mg L1)

1.8

(b)

1.4

0.6 0 10 20 30 40

Time (days) Log of anthocyanin content (mg L1)

1.8

(c)

1.4

0.6 0 10 20 30 40

Time (days)
Figure 2 Degradation kinetics for anthocyanins encapsulated with maltodextrin ( ), maltodextrin c-cyclodextrin ( ) and maltodextrin arabic gum ( ) in an isotonic soft drink system submitted to dierent conditions: 25 C in the presence of light (a); 25 C in the absence light (b) and 4 C in the absence of light (c).

The colour parameters obtained for samples with dierent carrier agents were analysed by the CIELab method. It could be observed that the encapsulated extract with maltodextrin (M) presented the lowest values for lightness (L* = 21.49), while maltodextrin ccyclodextrin (MC) showed the highest value (L* = 42.68). All formulations presented positives values for a*, 47.75, 50.43 and 40.58 for M, MC and MG respectively, indicating the predominance of a red hue for the extracts.
Evaluation of encapsulated anthocyanin stability

The stability of the anthocyanins (encapsulated with the carrier agents M, MC and MG) added to the isotonic soft drink system was studied under dierent temperature and light conditions. The degradation curves are shown in Fig. 2. The degradation of the anthocyanins tted a rstorder reaction model under all conditions evaluated,

which is evidenced by the linear relationship in the log of total anthocyanins concentration versus time plots (Fig. 2). The carrier agent inuenced the reaction constants, but not the reaction order. Kirca & Cemeroglu (2003) and Wang & Xu (2007) also reported rst order kinetics for the degradation of anthocyanins from dierent sources. Table 3 shows the percentage of colour retention and the kinetics parameters calculated for the samples. The analysis time (40 days) did not allow the calculation of the half-life and rst-order kinetics constant for the samples stored at 4 C in the absence of light, considering the lower degradation of the pigment in these conditions. The rst-order kinetics constant (k) and the half-life time (t) of the anthocyanins were aected by the carrier agent and by the ambient conditions (light and temperature) (Table 3). The samples stored at 4 C showed

International Journal of Food Science and Technology 2011

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

Anthocyanin: microcapsulation and stability V. M. Burin et al.

191

Table 3 First-order kinetics constant (k), halflife (t), and percentage of colour retention (R%) for anthocyanins encapsulated with dierent carrier agents in isotonic soft drink system stored in the presence and absence of light at 25 C, and in absence of light at 4 C

Carrier agent M

Conditions Light Light Light Light Light Light Light Light Light absent (25 C) present (25 C) absent (4 C) absent (25 C) present (25 C) absent (4 C) absent (25 C) present (25 C) absent (4 C)

k 10)3 (h)1) 1.245 1.825 1.170 1.804 1.075 1.316 0.01 (0.9396) 0.012 (0.9484) 0.014 (0.9624) 0.011 (0.9698) 0.013 (0.9165) 0.017 (0.9548)

t (h) 556.2 379.7 591.8 384.1 644.6 526.3 0.2 0.3 0.4 0.1 0.6 0.3

R% 35.59 31.38 94.01 39.89 37.78 94.57 42.76 39.22 94.66 0.14 0.81 0.94 0.57 0.44 0.53 0.29 0.74 0.88

MC

MG

M, maltodextrin, MC; maltodextrin c-cyclodextrin; MG, maltodextrin arabic gum.

lower degradation than the samples stored at 25 C for all the tested carrier agents. These results are consistent with research by Kirca et al. (2007), who observed a halflife of 4.1 weeks for black carrot juice anthocyanin samples stored at 37 C, but when refrigerated at 4 C the half-life was 71.8 weeks, showing that the storage temperature had a strong inuence on the pigment degradation. Ersus & Yurdagel (2007) found that encapsulated black carrot anthocyanins stored under refrigeration (4 C) showed a half-life three times higher than those stored at 25 C. Research conducted by Amendola et al. (2010) with grapes concluded that an increase in the storage temperature from 4 to 25 C led to a change in the characteristic colour index (bright red) of anthocyanins, and a darkening to brick red was noted. On evaluating the degradation of anthocyanins submitted to a temperature of 25 C in the presence and absence of light, it was observed that samples exposed to light have higher k values, and consequently shorter half-life times (Table 3). Falcao et al. (2004) evaluated Cabernet Sauvignon anthocyanin stability in relation to temperature, light and pH and also showed that light is an important accelerating factor in the degradation of anthocyanins. In studies conducted by Janna et al. (2007) it was noted that when exposed to light at 25 C anthocyanins showed a signicant decrease in the pigment content, with a reduction of more than 50% on the third day of exposure. The samples encapsulated with maltodextrin (M) showed higher k values and a lower percentage of colour retention (R%) than samples encapsulated with a combination of agents (MC and MG), indicating that these combinations were more ecient in terms of anthocyanin protection. The low lm-forming ability of the maltodextrin may have aected the organisation of the microcapsule structure when used alone. According to Gharsallaoui et al. (2007) a single carrier agent will not possess all of the material properties required for good encapsulation and thus the use of carrier mixtures is a viable and eective way to protect the compounds. Under the conditions evaluated, samples encapsulated with the maltodextrin arabic gum mixture had the lowest k values (Table 3), suggesting higher

encapsulation eciency and thus greater anthocyanin protection. The higher eciency of the carrier agent arabic gum may be related to its structure. Arabic gum is a highly branched heteropolymer of sugars, containing a small amount of protein covalently linked to the carbohydrate chain, acting as an excellent lm-forming agent and thus better trapping the encapsulated molecule. This makes the avylium cation less vulnerable to nucleophilic attack by water molecules, increasing the stability of the anthocyanins (Dangles & Brouillard, 1992). In experiments performed by Krishnan et al. (2005) with dierent carrier agents, the mixture of maltodextrin and arabic gum also showed higher eciency in the encapsulation of food avouring, providing a greater trapping of the constituents, and the higher the proportion of arabic gum, the longer the half-life time and the better the protection of the compound.
Structural evaluation of microcapsules

The surface characteristics, size and shape of microcapsules containing maltodextrin (M), maltodextrin ccyclodextrin (MC) and maltodextrin arabic gum (MG) observed by SEM are shown in Fig. 3. The MC and MG samples showed microcapsules with similar size and more spherical shape (Figs 3b,c, respectively) than those with maltodextrin only (Fig. 3a). These results were in agreement with the anthocyanin stability study, in which the MG sample presented the longest half-life time, followed by the MC sample. According to a study carried out by Cano-Chauca et al. (2005), the addition of arabic gum allows particles with better distribution and uniformity to be obtained. In oleoresin encapsulation studies, arabic gum was also revealed to be the best carrier agent (Kanakdande et al., 2007; Kaushik & Roos, 2007).
Conclusions

The optimisation of anthocyanin extraction from Cabernet Sauvignon grapes was achieved using the RSM, reaching a maximum content of total monomeric anthocyanins when 40 mL ethanol:1.5 N HCl (85:15)

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

International Journal of Food Science and Technology 2011

192

Anthocyanin: microcapsulation and stability V. M. Burin et al.

(a)

(b)

(c)

Figure 3 SEM micrographs of microcapsules of anthocyanin pigments encapsulated with maltodextrin (a), maltodextrin c-cyclodextrin (b) and maltodextrin arabic gum (c).

solution (solid:liquid ratio 1:4) and 29.4 h of extraction time at pH 2.4 were employed. It could be observed that the anthocyanins added to an isotonic soft drink system showed rst-order reaction kinetics of degradation in all situations evaluated. The kinetics variables were inuenced by the carrier agent used. Of the carrier agents tested in this study, the combination of maltodextrin arabic gum led to the longest anthocyanin half-life time and lowest degradation constant under all conditions evaluated, and thus provided better protection of the anthocyanin pigments. This also was conrmed through SEM, which showed the combination of maltodextrin arabic gum presented uniform microcapsules and with spherical surface.
References
Amendola, D., Faveri, D.M. & Spigno, G. (2010). Grape marc phenolics: extraction kinetics, quality and stability of extracts. Journal of Food Engineering, 97, 384392. Cano-Chauca, M., Stringheta, P.C., Ramos, A.M. & Cal-Vidal, J. (2005). Eect of the carriers on the microstructure of mango powder obtained by spray drying and its functional characterization. Innovative Food Science and Emerging Technologies, 6, 420428. Chen, F., Sun, Y., Zhao, G. et al. (2007). Optimization of ultrasoundassisted extraction of anthocyanins in red raspberries and identication of anthocyanins in extract using high-performance liquid chromatographymass spectrometry. Ultrasonics Sonochemistry, 14, 767778. Cho, S.Y., Lee, Y.N. & Park, H.J. (2009). Optimization of ethanol extraction and further purication of isoavones from soybean sprout cotyledon. Food Chemistry, 117, 312317. Corrales, M., Garc a, A.F., Butz, P. & Tauscher, B. (2009). Extraction of anthocyanins from grape skins assisted by high hydrostatic pressure. Journal of Food Engineering, 90, 415421.

Dangles, O. & Brouillard, R. (1992). A spectrophotometric method based on the anthocyanin copigmentation interaction and applied to the quantitative study of molecular complexes. Journal Chemical Society Perkin Trans, 2, 247257. Ersus, S. & Yurdagel, U. (2007). Microencapsulation of anthocyanin pigments of black carrot (Daucus carota L.) by spray drier. Journal of Food Engineering, 80, 805812. Falcao, L.D., Gauche, C., Barros, D.M. et al. (2004). Stability of anthocyanins from grape (Vitis vinifera L.) skins with tannic acid in a model system. Italian Journal of Food Science, 16, 323332. Gharsallaoui, A., Roudaut, G., Chambin, O., Voilley, A. & Saurel, R. (2007). Applications of spray-drying in microencapsulation of food ingredients: an overview. Food Research International, 40, 1107 1121. Giusti, M.M. & Wrolstad, R.E. (2001). Characterization and measurement of anthocyanins by UV-visible spectroscopy. Unit F1.2. In: Current Protocols in Food Analytical Chemistry (edited by R.E. Wrolstad & S.J. Schwartz). Pp. F1.2.1F1.2.13. New York, NY: John Wiley & Sons, Inc. Giusti, M.M. & Wrolstad, R.E. (2003). Acylated anthocyanins from edible sources and their applications in food systems. Biochemical Engineering Journal, 14, 217225. Janna, O.A., Khairul, A.K. & Maziah, M. (2007). Anthocyanin stability studies in Tibouchina semidecandra L. Food Chemistry, 101, 16401646. Kanakdande, D., Bhosale, R. & Singhal, R.S. (2007). Stability of cumin oleoresin microencapsulated in dierent combination of gum arabic, maltodextrin and modied starch. Carbohydrate Polymers, 67, 536541. Katsaboxakis, K., Papanicolaou, D. & Melanitou, M. (1998). Stability of pigmented orange anthocyanins in model and real food system. Italian Journal Food Science, 10(1), 1725. Kaushik, V. & Roos, Y.H. (2007). Limonene encapsulation in freezedrying of gum Arabicsucrosegelatin systems. LWT Food Science and Technology, 40, 13811391. Kirca, A. & Cemeroglu, B. (2003). Degradation kinetics of anthocy anins in blood orange juice and concentrate. Food Chemistry, 81, 583587.

International Journal of Food Science and Technology 2011

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

Anthocyanin: microcapsulation and stability V. M. Burin et al.

193

Kirca, A., Ozkan, M. & Cemeroglu, B. (2007). Eects of temperature, solid content and pH on the stability of black carrot anthocyanins. Food Chemistry, 101, 212218. Kong, J.M., Chia, L.S., Goh, N.K., Chia, T.F. & Brouillard, R. (2003). Analysis and biological activities of anthocyanins. Phytochemistry, 64, 923933. Krishnan, S., Bhosale, R. & Singhal, R.S. (2005). Microencapsulation of cardamom oleoresin: evaluation of blends of gum arabic, maltodextrin and a modied starch as wall materials. Carbohydrate Polymers, 61, 95102. Kurozawa, L.E., Park, K.J. & Hubinger, M.D. (2009). Eect of carrier agents on the physicochemical properties of a spray dried chicken meat protein hydrolysate. Journal of Food Engineering, 94, 326333. Loksuwan, J. (2007). Characteristics of microencapsulated b-carotene formed by spray drying with modied tapioca starch, native tapioca starch and maltodextrin. Food Hydrocolloids, 21, 928935. Montes, C., Vicario, I.M., Raymundo, M., Fett, R. & Heredia, F.J. (2005). Application of tristimulus colorimetry to optimize the extraction of anthocyanins from Jaboticaba (Myricia Jaboticaba Berg.). Food Research International, 38, 983988. Munoz-Espada, A.C., Wood, K.V., Bordelon, B. & Watkins, B.A. (2004). Anthocyanin quantication and radical scavenging capacity of Concord, Norton, and Marechal Foch Grapes and wines. Journal of Agricultural and Food Chemistry, 52, 67796786.

Ngo, T. & Zhao, Y. (2009). Stabilization of anthocyanins on thermally processed red DAnjou pears through complexation and polymerization. LWT Food Science and Technology, 42, 11441152. Revilla, E., Ryan, J.M. & Mart n-Ortega, G. (1998). Comparison of several procedures used for the extraction of anthocyanins from red grapes. Journal of Agriculture and Food Chemistry, 46, 45924597. Roriz, M.S., Osma, J.F., Teixeira, J.A. & Couto, S.R. (2009). Application of response surface methodological approach to optimise Reactive Black 5 decolouration by crude laccase from Trametes pubescens. Journal of Hazardous Materials, 169, 691696. Rosso, V.V., Hillebrand, S., Montilla, E.C., Bobbio, F.O., Winterhalter, P. & Mercadante, A.Z. (2008). Determination of anthocyanins from acerola (Malpighia emarginata DC.) and acai (Euterpe oleracea Mart.) by HPLCPDAMS MS. Journal of Food Composition and Analysis, 21, 291299. Torskangerpoll, K. & Andersen, O.M. (2005). Colour stability of anthocyanins in aqueous solutions at various pH values. Food Chemistry, 89, 427440. Turchiuli, C., Fuchs, M., Bohin, M. et al. (2005). Oil encapsulation by spray drying and uidised bed agglomeration. Innovative Food Science and Emerging Technologies, 6, 2935. Wang, W.D. & Xu, S.Y. (2007). Degradation kinetics of anthocyanins in blackberry juice and concentrate. Journal of Food Engineering, 82, 271275.

2010 The Authors International Journal of Food Science and Technology 2010 Institute of Food Science and Technology

International Journal of Food Science and Technology 2011