j.1365-2621.2010.02486.x

Original article

Anthocyanins: optimisation of extraction from Cabernet Sauvignon

grapes, microcapsulation and stability in soft drink

Vıvian 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 different carrier agents in an isotonic soft drink system

under different 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 first-order

kinetics in the situations evaluated. Maltodextrin, maltodextrin ⁄ c-cyclodextrin and maltodextrin ⁄ arabic gumwere 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.

Keywords Anthocyanins, grape, microencapsulation, spray drying.

Introduction

Colour is one of the most important attributes in foodand drinks, as it affects the acceptability of products byconsumers. The replacement of synthetic dyes withnatural food colourants has increased considerably. Toprovide the colour red, anthocyanins are an attractivealternative, as they provide high colourant power, lowtoxicity and water solubility, which permit their incor-poration inmany food systems (Ersus &Yurdagel, 2007).Anthocyanins are the largest group of water-soluble

pigments obtained from plants and they can be definedchemically as polyhydroxylated or polymethoxylatedglycosides or acylglycosides of anthocyanidins belong-ing to the flavonoids class. These compounds aredifferentiated by the number of hydroxyl groups andthe nature, number and position of sugars attached tothe molecule (Kong et al., 2003). The main anthocya-nins found in grapes are cyanidin, peonidin, delphinidin,petunidin and malvidin (Munoz-Espada et al., 2004),and their concentration is dependent on the variety andenvironmental factors.Studies carried out in vitro and in vivo have shown

that anthocyanins are responsible for important thera-

peutic properties such as: antioxidant, anticarcinogenicand anti-inflammatory activity, low density lipoprotein(LDL) oxidation inhibition and a decrease in the risk ofcardiovascular disease, and thus their application asnatural pigments in food is of great interest (Wang &Xu, 2007; Rosso et al., 2008). However, anthocyaninsare unstable pigments and can be converted to colour-less compounds. Their stability is affected by manyfactors including pH, temperature, light, oxygen and thefood matrix (Wang & Xu, 2007). Also, there arelimitations to the application of anthocyanins as pig-ments in food systems related to the process conditions,formulation and storage conditions. This has stimulateda search for new methods to enhance their stability andincrease the possibilities for the use these compounds(Giusti & Wrolstad, 2003).To determine the best conditions for the extraction of

natural pigments using organic solvents, researchershave applied the response surface methodology (RSM).This methodology represents a combination of mathe-matical and statistical techniques aimed at optimisingthe final response. The main advantage of this method isthe reduced number of experiments required to providesufficient information to obtain statistically acceptableresults (Cho et al., 2009). The RSM with central com-posite design provides a complete factorial investigationof the simultaneous, systematic and efficient variation of

*Correspondent: Fax: +55 48 3721 9943;

e-mail: [email protected]

International Journal of Food Science and Technology 2011, 46, 186–193186

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

important components, identifying the possible interac-tions, main effects and optimal conditions of operation(Roriz et al., 2009).A very useful method to enhance the stability of

natural pigments is microencapsulation by the spraydrying technique, which provides powders with lowhumidity and good quality that are easy to store. Thedegradation protection and consequent preservation ofcompounds is due to the inclusion of these pigments inmacromolecules (Cano-Chauca et al., 2005; Ngo &Zhao, 2009). The active compounds in dehydratedformulations and microcapsules destined for con-trolled-released are protected during processing andstorage. Knowledge of the carrier agent properties iscrucial in order to optimise the process and reduce thecost (Kurozawa et al., 2009). In particular, the coatingagents submitted to spray drying must show a goodcapacity for emulsification and film forming, along withhigh solubility, low viscosity and hygroscopicity(Loksuwan, 2007). The most notable of the commonlyused carrier agents are natural gums and dextrins(Kanakdande et al., 2007). In general, the coating agentalone does not offer all the properties required to ensurea good microencapsulation and thus a mix of one ormore components is frequently employed to enhance theencapsulation (Turchiuli et al., 2005).The aim of this study was to establish the optimal

conditions for anthocyanin extraction from CabernetSauvignon (Vitis vinifera L.) grapes, employing theRSM. The stability of these anthocyanins encapsulatedwith different carrier agents in a soft drink model systemwas then evaluated under different light and temperatureconditions.

Materials and methods

Chemicals

The study was carried out with Cabernet Sauvignon(Vitis vinifera L.) grapes from the region of SaoJoaquim, Santa Catarina, Brazil. Maltodextrin (Mor-Rex 1920 DE 19, Plury Quimica, Diadema, Brazil),arabic gum (Colloides Naturels, Sao Paulo, Brazil) andc-cyclodextrin (Cerestar, Minneapolis, MN, USA) wereused for the encapsulation by spray drying. All otherreagents were of analytical grade.

Experimental design

The RSM was used to optimise the conditions for theextraction of anthocyanins from Cabernet Sauvignongrapes. The experimental design was carried out using acentral composite design with three independent vari-ables solvent (x1), pH (x2) and extraction time (x3) andthree levels coded as –1, 0 and 1, resulting in seventeenruns including three replicates at the central point to

estimate the experimental error. All runs were carriedout with 10 g of grape skins. The response chosen forthe design was the total monomeric anthocyaninscontent (mg ⁄100 g grape skin) quantified by the pH-differential method.A second-order polynomial model was constructed to

estimate the response (eqn 1):

y ¼ b0 þXk

i¼1bixi þ

Xk

i¼1biix

2i þ

Xk�1

i¼1

Xk

j>1

bijxixj þ e ð1Þ

where y is the estimated response (dependent variable),b0 the model constant, bi the linear effect coefficient, bii

the quadratic effect coefficient, bij the interaction coef-ficient for two factors, xi, xj the independent variables, ethe error, k the number of variables considered, i and jthe codified factors of the system.

Extraction and quantification of anthocyanins

According to the results obtained in the experimentalplanning, the anthocyanin extract was prepared throughthe maceration of 100 g of Cabernet Sauvignon grapeskins in ethanol:1.5 N HCl (85:15) solution (solid:liquidratio 1:4) for 29.4 h in the dark under refrigerationtemperature conditions (4.0 ± 1 �C). The extractobtained was filtered and kept at 4.0 ± 1 �C in anamber flask until analysis.The total monomeric anthocyanins content was

determined using the pH-differential method describedby Giusti & Wrolstad (2001), measuring the absorbancein a UV-Vis absorption spectrophotometer (HitachiU2010, Tokyo, Japan). The content of total monomericanthocyanins in the grapes was expressed as mg malvi-din-3-glycoside per 100 g of grape skin (MW= 529 gmol)1 and e = 28 000).

Preparation of microcapsules and spray drying process

The extract was concentrated in a low-pressure rotaryevaporator, approximately 20% of ethanol (QUIMIS,Q.344.2, SP, Brazil) at 35 ± 2 �C and the carrier agentswere then added in concentrations calculated onthe basis of the solids content of the crude extract.The samples were codified 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 carrieragents, the samples were stirred in a homogeniser (B.Brain Biotech International, CERTOMAT�MO, Mels-ungen, Germany) at 100 rpm for 2.5 h in the dark insealed flasks.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 airtemperature 150 �C and outlet temperature 50 �C,

Anthocyanin: microcapsulation and stability V. M. Burin et al. 187

� 2010 The Authors International Journal of Food Science and Technology 2011

International Journal of Food Science and Technology � 2010 Institute of Food Science and Technology

pump power 25% and maximum aspirator rate. Duringthe process, the temperature of the feed solution was25 �C. The powder obtained was stored in an amberflask under refrigeration temperature (4.0 ± 1 �C) inthe dark.

Colour characterisation of encapsulated anthocyanins

The colour parameters of the encapsulated pigmentswere determined using the CIELab system (colourimeterChroma 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 anthocyaninstability

The encapsulated anthocyanin extracts were added to anisotonic soft drink system (sucrose 4%, citric acid0.38%, glucose 0.15%, sodium chloride 0.12%, sodiumcitrate 0.11% and potassium monophosphate 0.022%),considering a commercial isotonic soft drink colour asthe reference. To evaluate the colourant stability, 5 mLsamples of the isotonic soft drink system were trans-ferred to screw-top test tubes and kept under thefollowing three conditions: under a 40 W fluorescencelamp at 25 ± 1 �C; in the absence of light at 25 ± 1 �C;and in the absence of light at 4 ± 1 �C. Samples werewithdrawn to quantify the content of anthocyanins atregular intervals for 40 days.Based on the plots of log of total anthocyanins

concentration versus time for the different conditionsanalysed, the first-order kinetics constants (k) and half-life times (t½) of the pigments in the isotonic soft drinksystem were calculated using eqs 2 and 3, respectively.

Slope of the straight line ¼ �k=2:303 ð2Þ

t1=2 ¼ �ln0:5� k�1 ð3Þ

where k is the first-order kinetics constant.The percentage of colour retention was calculated as

described by Katsaboxakis et al. (1998) using eqn 4.

%R ¼ ðAt=A0Þ � 100 ð4Þ

where At and A0 are the absorbance at time t and zerorespectively.

Scanning electron microscopy

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

the SEM samples were attached to stubs using two-sidedcarbon adhesive tape. The surface was sputter-coatedwith a thin layer of gold and then examined at ·35, ·500and ·1000 magnifications. An acceleration potential of10 kV was used to obtain the micrographs.

Statistical analysis

All analyses were carried out in triplicate with tworepetitions. The statistica� software (v. 8.0, StatSoft,Inc., Tulsa, OK, USA) was used to verify the centralcomposite design, to calculate the coefficients, meanvalues and standard deviations, and to carry out theanalysis of variance (anova) and Tukey test (P < 0.05).

Results and discussion

Experimental design for anthocyanin extraction

The RSM evaluated the effects and interactions of thesolvent volume, extraction time and pH of the solutionin terms of anthocyanin extraction from CabernetSauvignon grapes. Table 1 shows the values for thefactors and the total monomeric anthocyanins contentobtained for each extraction condition. The significanceof the factors was confirmed by anova, where it waspossible to observe that the quadratic effect of solventvolume and the linear effects of pH and extraction timewere significant. The pH was shown to have the weakestinfluence on the responses, and interactions between thevariables were not observed (Table 2). The predicted

Table 1 Full factorial central composite design matrix of three vari-

ables in coded and natural units along with the observed responses

Run

Coded units of

variable Natural units of variable

Responsex1 x2 x3 Solvent (mL) Time (h) pH

1 )1 )1 )1 20.00 8.00 1.20 465.96

2 )1 )1 1 20.00 8.00 3.60 356.32

3 )1 1 )1 20.00 24.00 1.20 580.84

4 )1 1 1 20.00 24.00 3.60 407.52

5 1 )1 )1 60.00 8.00 1.20 500.88

6 1 )1 1 60.00 8.00 3.60 407.88

7 1 1 )1 60.00 24.00 1.20 592.01

8 1 1 1 60.00 24.00 3.60 463.06

9 )1,68 0 0 6.40 16.00 2.40 153.38

10 1,68 0 0 73.60 16.00 2.40 519.36

11 0 )1,68 0 40.00 2.56 2.40 270.11

12 0 1,68 0 40.00 29.40 2.40 611.25

13 0 0 )1,68 40.00 16.00 0.40 550.02

14 0 0 1,68 40.00 16.00 4.40 370.11

15 0 0 0 40.00 16.00 2.40 593.33

16 0 0 0 40.00 16.00 2.40 590.08

17 0 0 0 40.00 16.00 2.40 575.72

Anthocyanin: microcapsulation and stability V. M. Burin et al.188

International Journal of Food Science and Technology 2011 � 2010 The Authors


values for total monomeric anthocyanins are close to theexperimental values demonstrating that the model isapplicable, which showed determination coefficient (R2)of 0.80. The mathematical relationship between theindependent and response variables was evaluatedthrough the quadratic model constructed in the regres-sion analysis (eqn 5).

Y ¼ 581:2832þ 64:8549t� 59:3673pH� 72:6772S2

ð5Þ

where Y is the total monomeric anthocyanins content(mg ⁄100 g grape skin); t the extraction time (hours); andS the solvent volume (mL).

The response surface plots indicated the effects of twovariables on the anthocyanins content, with a thirdindependent variable (central point) fixed to the centralexperimental level (Fig. 1). The anthocyanin contentsignificantly increased with extraction time and solventvolume, reaching the maximum response (611.25mg ⁄100 g) at 40 mL ethanol:1.5 N HCl (85:15) solutionand 29.4 h of extraction time (Fig. 1a). A study carriedout by Chen et al. (2007) with different solid ⁄ liquidratios for anthocyanin extraction also demonstratedthat a ratio of 1:4 (solid:liquid) gave the highest contentof extracted pigments, which is consistent with theresults reported herein. The same authors affirm thatduring solid ⁄ liquid extraction an excessive increase in

Table 2 Regression coefficients and anova

results for total monomeric anthocyanins

extracted from Cabernet Sauvignon grapes

Regression

coefficients

Sum of

squares (SS) DF

Average

square F value P value

(1) Solvent (L) 56.2880 43231.4 1 43231.4 5.396327 0.053159

Solvent (Q) )72.6772 59530.0 1 59530.0 7.430784 0.029511

(2) Time (L) 64.8549 57321.1 1 57321.1 7.155060 0.031773

Time (Q) )35.7908 14365.1 1 14365.1 1.793111 0.222395

(3) pH (L) )59.3673 47776.3 1 47776.3 5.963640 0.044624

pH (Q) )29.0830 9281.2 1 9281.2 1.158518 0.317464

1L ⁄ 2L )2.4720 48.9 1 48.89 0.006102 0.939921

1L ⁄ 3L 7.6265 465.3 1 465.31 0.058082 0.816461

2L ⁄ 3L )12.4545 1240.9 1 1240.92 0.154896 0.705612

Error 56078.9 7 8011.27

Total SS 268636.1 16

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

510

1520

1530

35

Time (h)

(a)(b)

(c)

700600500400300200100

0

0.00.5

0

10 5

1520

1530

35

1.01.52.02.53.03.54.04.55.0

Ant

hocy

anin

con

tent

(mg

100

g–1 )

pH

Time (h)

–200–200–100

0100200300400500600700

0

200

400

600

800

0.00.5

1020

3040

5060

7080

1.01.52.02.53.03.54.04.55.0Ant

hocy

anin

con

tent

(mg

100

g–1 )

Ant

hocy

anin

con

tent

(mg

100

g–1 )

pHEthanol (m

L )

1020

3040

5060

7080

Ethanol (mL)

Figure 1 Response surface for total mono-

meric 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.




solvent volume can result in losses through evaporationor saturate the extractor medium. This may be thereason for the decrease in the anthocyanins content atethanol volumes higher than 60 mL in this study.The reduction in pH and concomitant increase in

solvent volume also caused an increase of the anthocy-anin content, with an optimum extraction region ataround pH 2 (Fig. 1b). Montes et al. (2005) alsoreported an ethanol solution acidified with hydrochloricacid and a pH of around 2 as the best conditions foranthocyanin extraction. Torskangerpoll & Andersen(2005) affirmed that the colour stability of anthocyaninsis mainly dependent on the pH and the structure of themolecule, since in acid solutions anthocyanins remainpreferentially in the flavylium cation form, which giveshigher wavelengths in UV–Vis absorption spectra. Kircaet al. (2007) observed a considerable decrease in antho-cyanin stability for pH values up to 5.0.On increasing the extraction time it was observed that

the anthocyanin content progressively increased. Cor-rales et al. (2009) established that the contact of grapeskins with the solvent for a longer time provides a higherdiffusion of compounds such as anthocyanins. Revillaet al. (1998) evaluated the extraction of phenolic com-pounds from Cabernet Sauvignon grapes under differentconditions and observed that a longer extraction timeprovided a higher amount of extracted anthocyanins.These results together show that the optimum com-

bination of independent variables for anthocyaninextraction was 40 mL ethanol:1.5 N HCl (85:15) solu-tion and an extraction time of 29.4 h at pH 2.4. Theseparameters were therefore chosen to carry out theextraction of anthocyanins for later encapsulation.

Colour parameters of encapsulated pigments

The colour parameters obtained for samples withdifferent carrier agents were analysed by the CIELabmethod. It could be observed that the encapsulatedextract with maltodextrin (M) presented the lowestvalues for lightness (L* = 21.49), while maltodextrin ⁄ c-cyclodextrin (MC) showed the highest value (L* =42.68). All formulations presented positives values fora*, 47.75, 50.43 and 40.58 for M, MC and MGrespectively, indicating the predominance of a red huefor the extracts.

Evaluation of encapsulated anthocyanin stability

The stability of the anthocyanins (encapsulated with thecarrier agents M, MC and MG) added to the isotonicsoft drink system was studied under different tempera-ture and light conditions. The degradation curves areshown in Fig. 2.The degradation of the anthocyanins fitted a first-

order reaction model under all conditions evaluated,

which is evidenced by the linear relationship in the log oftotal anthocyanins concentration versus time plots(Fig. 2). The carrier agent influenced the reactionconstants, but not the reaction order. Kirca & Cemer-oglu (2003) and Wang & Xu (2007) also reported first-order kinetics for the degradation of anthocyanins fromdifferent sources.Table 3 shows the percentage of colour retention and

the kinetics parameters calculated for the samples. Theanalysis time (40 days) did not allow the calculation ofthe half-life and first-order kinetics constant for thesamples stored at 4 �C in the absence of light, consid-ering the lower degradation of the pigment in theseconditions.The first-order kinetics constant (k) and the half-life

time (t½) of the anthocyanins were affected by the carrieragent and by the ambient conditions (light and temper-ature) (Table 3). The samples stored at 4 �C showed

0.6

1

1.4

1.8

0 10 20 30 40

Time (days)

Log

of a

ntho

cyan

in c

onte

nt

(mg

L–1)

0.6

1

1.4

1.8

0 10 20 30 40

Time (days)

Log

of a

ntho

cyan

in c

onte

nt(m

g L–1

)

0.6

1

1.4

1.8

0 10 20 30 40

Time (days)

Log

of a

ntho

cyan

in c

onte

nt

(mg

L–1)

(a)

(b)

(c)

Figure 2 Degradation kinetics for anthocyanins encapsulated with

maltodextrin ( ), maltodextrin ⁄ c-cyclodextrin ( ) and maltodextrin ⁄arabic gum ( ) in an isotonic soft drink system submitted to different

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).




lower degradation than the samples stored at 25 �C forall the tested carrier agents. These results are consistentwith research by Kirca et al. (2007), who observed a half-life of 4.1 weeks for black carrot juice anthocyaninsamples stored at 37 �C, but when refrigerated at 4 �Cthe half-life was 71.8 weeks, showing that the storagetemperature had a strong influence on the pigmentdegradation. Ersus & Yurdagel (2007) found thatencapsulated black carrot anthocyanins stored underrefrigeration (4 �C) showed a half-life three times higherthan those stored at 25 �C. Research conducted byAmendola et al. (2010) with grapes concluded that anincrease in the storage temperature from 4 to 25 �C led toa change in the characteristic colour index (bright red) ofanthocyanins, and a darkening to brick red was noted.On evaluating the degradation of anthocyanins sub-

mitted to a temperature of 25 �C in the presence andabsence of light, it was observed that samples exposed tolight have higher k values, and consequently shorterhalf-life times (Table 3). Falcao et al. (2004) evaluatedCabernet Sauvignon anthocyanin stability in relation totemperature, light and pH and also showed that light isan important accelerating factor in the degradation ofanthocyanins. In studies conducted by Janna et al.(2007) it was noted that when exposed to light at25 �C anthocyanins showed a significant decrease in thepigment content, with a reduction of more than 50% onthe third day of exposure.The samples encapsulated with maltodextrin (M)

showed higher k values and a lower percentage ofcolour retention (R%) than samples encapsulated with acombination of agents (MC and MG), indicating thatthese combinations were more efficient in terms ofanthocyanin protection. The low film-forming abilityof the maltodextrin may have affected the organisationof the microcapsule structure when used alone. Accord-ing to Gharsallaoui et al. (2007) a single carrier agentwill not possess all of the material properties requiredfor good encapsulation and thus the use of carriermixtures is a viable and effective way to protect thecompounds. Under the conditions evaluated, samplesencapsulated with the maltodextrin ⁄arabic gum mixturehad the lowest k values (Table 3), suggesting higher

encapsulation efficiency and thus greater anthocyaninprotection. The higher efficiency of the carrier agentarabic gum may be related to its structure. Arabic gumis a highly branched heteropolymer of sugars, contain-ing a small amount of protein covalently linked to thecarbohydrate chain, acting as an excellent film-formingagent and thus better trapping the encapsulated mole-cule. This makes the flavylium cation less vulnerable tonucleophilic attack by water molecules, increasing thestability of the anthocyanins (Dangles & Brouillard,1992). In experiments performed by Krishnan et al.(2005) with different carrier agents, the mixture ofmaltodextrin and arabic gum also showed higherefficiency in the encapsulation of food flavouring,providing a greater trapping of the constituents, andthe higher the proportion of arabic gum, the longer thehalf-life time and the better the protection of thecompound.

Structural evaluation of microcapsules

The surface characteristics, size and shape of microcap-sules containing maltodextrin (M), maltodextrin ⁄ c-cyclodextrin (MC) and maltodextrin ⁄arabic gum (MG)observed by SEM are shown in Fig. 3. The MC and MGsamples showed microcapsules with similar size andmore spherical shape (Figs 3b,c, respectively) than thosewith maltodextrin only (Fig. 3a). These results were inagreement with the anthocyanin stability study, in whichthe MG sample presented the longest half-life time,followed by theMC sample. According to a study carriedout by Cano-Chauca et al. (2005), the addition of arabicgum allows particles with better distribution and unifor-mity 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 Cab-ernet Sauvignon grapes was achieved using the RSM,reaching a maximum content of total monomericanthocyanins when 40 mL ethanol:1.5 N HCl (85:15)

Table 3 First-order kinetics constant (k), half-

life (t½), and percentage of colour retention

(R%) for anthocyanins encapsulated with

different 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 Conditions k · 10)3 (h)1) t½ (h) R %

M Light absent (25 �C) 1.245 ± 0.01 (0.9396) 556.2 ± 0.2 35.59 ± 0.14

Light present (25 �C) 1.825 ± 0.012 (0.9484) 379.7 ± 0.3 31.38 ± 0.81

Light absent (4 �C) – – 94.01 ± 0.94

MC Light absent (25 �C) 1.170 ± 0.014 (0.9624) 591.8 ± 0.4 39.89 ± 0.57

Light present (25 �C) 1.804 ± 0.011 (0.9698) 384.1 ± 0.1 37.78 ± 0.44

Light absent (4 �C) – – 94.57 ± 0.53

MG Light absent (25 �C) 1.075 ± 0.013 (0.9165) 644.6 ± 0.6 42.76 ± 0.29

Light present (25 �C) 1.316 ± 0.017 (0.9548) 526.3 ± 0.3 39.22 ± 0.74

Light absent (4 �C) – – 94.66 ± 0.88

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




solution (solid:liquid ratio 1:4) and 29.4 h of extractiontime at pH 2.4 were employed. It could be observed thatthe anthocyanins added to an isotonic soft drink systemshowed first-order reaction kinetics of degradation in allsituations evaluated. The kinetics variables were influ-enced by the carrier agent used. Of the carrier agentstested in this study, the combination of maltodex-trin ⁄arabic gum led to the longest anthocyanin half-lifetime and lowest degradation constant under all condi-tions evaluated, and thus provided better protection ofthe anthocyanin pigments. This also was confirmedthrough SEM, which showed the combination ofmaltodextrin ⁄arabic gum presented uniform microcap-sules and with spherical surface.

References

Amendola, D., Faveri, D.M. & Spigno, G. (2010). Grape marcphenolics: extraction kinetics, quality and stability of extracts.Journal of Food Engineering, 97, 384–392.

Cano-Chauca, M., Stringheta, P.C., Ramos, A.M. & Cal-Vidal, J.(2005). Effect of the carriers on the microstructure of mango powderobtained by spray drying and its functional characterization.Innovative Food Science and Emerging Technologies, 6, 420–428.

Chen, F., Sun, Y., Zhao, G. et al. (2007). Optimization of ultrasound-assisted extraction of anthocyanins in red raspberries and identifi-cation of anthocyanins in extract using high-performance liquidchromatography–mass spectrometry. Ultrasonics Sonochemistry, 14,767–778.

Cho, S.Y., Lee, Y.N. & Park, H.J. (2009). Optimization of ethanolextraction and further purification of isoflavones from soybeansprout cotyledon. Food Chemistry, 117, 312–317.

Corrales, M., Garcıa, A.F., Butz, P. & Tauscher, B. (2009). Extractionof anthocyanins from grape skins assisted by high hydrostaticpressure. Journal of Food Engineering, 90, 415–421.

Dangles, O. & Brouillard, R. (1992). A spectrophotometric methodbased on the anthocyanin copigmentation interaction and applied tothe quantitative study of molecular complexes. Journal ChemicalSociety Perkin Trans, 2, 247–257.

Ersus, S. & Yurdagel, U. (2007). Microencapsulation of anthocyaninpigments of black carrot (Daucus carota L.) by spray drier. Journalof Food Engineering, 80, 805–812.

Falcao, L.D., Gauche, C., Barros, D.M. et al. (2004). Stability ofanthocyanins from grape (Vitis vinifera L.) skins with tannic acid ina model system. Italian Journal of Food Science, 16, 323–332.

Gharsallaoui, A., Roudaut, G., Chambin, O., Voilley, A. & Saurel, R.(2007). Applications of spray-drying in microencapsulation of foodingredients: an overview. Food Research International, 40, 1107–1121.

Giusti, M.M. & Wrolstad, R.E. (2001). Characterization and mea-surement 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. F1.2.1–F1.2.13. New York, NY:John Wiley & Sons, Inc.

Giusti, M.M. & Wrolstad, R.E. (2003). Acylated anthocyanins fromedible sources and their applications in food systems. BiochemicalEngineering Journal, 14, 217–225.

Janna, O.A., Khairul, A.K. & Maziah, M. (2007). Anthocyaninstability studies in Tibouchina semidecandra L. Food Chemistry, 101,1640–1646.

Kanakdande, D., Bhosale, R. & Singhal, R.S. (2007). Stability ofcumin oleoresin microencapsulated in different combination of gumarabic, maltodextrin and modified starch. Carbohydrate Polymers,67, 536–541.

Katsaboxakis, K., Papanicolaou, D. & Melanitou, M. (1998). Stabilityof pigmented orange anthocyanins in model and real food system.Italian Journal Food Science, 10(1), 17–25.

Kaushik, V. & Roos, Y.H. (2007). Limonene encapsulation in freeze-drying of gum Arabic–sucrose–gelatin systems. LWT – Food Scienceand Technology, 40, 1381–1391.

Kirca, A. & Cemeroglu, B. (2003). Degradation kinetics of anthocy-anins in blood orange juice and concentrate. Food Chemistry, 81,583–587.

(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).




Kirca, A., Ozkan, M. & Cemeroglu, B. (2007). Effects of temperature,solid content and pH on the stability of black carrot anthocyanins.Food Chemistry, 101, 212–218.

Kong, J.M., Chia, L.S., Goh, N.K., Chia, T.F. & Brouillard, R.(2003). Analysis and biological activities of anthocyanins. Phyto-chemistry, 64, 923–933.

Krishnan, S., Bhosale, R. & Singhal, R.S. (2005). Microencapsulationof cardamom oleoresin: evaluation of blends of gum arabic,maltodextrin and a modified starch as wall materials. CarbohydratePolymers, 61, 95–102.

Kurozawa, L.E., Park, K.J. & Hubinger, M.D. (2009). Effect of carrieragents on the physicochemical properties of a spray dried chickenmeat protein hydrolysate. Journal of Food Engineering, 94, 326–333.

Loksuwan, J. (2007). Characteristics of microencapsulated b-caroteneformed by spray drying with modified tapioca starch, native tapiocastarch and maltodextrin. Food Hydrocolloids, 21, 928–935.

Montes, C., Vicario, I.M., Raymundo, M., Fett, R. & Heredia, F.J.(2005). Application of tristimulus colorimetry to optimize theextraction of anthocyanins from Jaboticaba (Myricia JaboticabaBerg.). Food Research International, 38, 983–988.

Munoz-Espada, A.C., Wood, K.V., Bordelon, B. & Watkins, B.A.(2004). Anthocyanin quantification and radical scavenging capacityof Concord, Norton, and Marechal Foch Grapes and wines. Journalof Agricultural and Food Chemistry, 52, 6779–6786.

Ngo, T. & Zhao, Y. (2009). Stabilization of anthocyanins on thermallyprocessed red D’Anjou pears through complexation and polymer-ization. LWT – Food Science and Technology, 42, 1144–1152.

Revilla, E., Ryan, J.M. & Martın-Ortega, G. (1998). Comparison ofseveral procedures used for the extraction of anthocyanins from redgrapes. Journal of Agriculture and Food Chemistry, 46, 4592–4597.

Roriz, M.S., Osma, J.F., Teixeira, J.A. & Couto, S.R. (2009).Application of response surface methodological approach to opti-mise Reactive Black 5 decolouration by crude laccase from Trametespubescens. Journal of Hazardous Materials, 169, 691–696.

Rosso, V.V., Hillebrand, S., Montilla, E.C., Bobbio, F.O., Winterhal-ter, P. & Mercadante, A.Z. (2008). Determination of anthocyaninsfrom acerola (Malpighia emarginata DC.) and acai (Euterpe oleraceaMart.) by HPLC–PDA–MS ⁄MS. Journal of Food Composition andAnalysis, 21, 291–299.

Torskangerpoll, K. & Andersen, O.M. (2005). Colour stability ofanthocyanins in aqueous solutions at various pH values. FoodChemistry, 89, 427–440.

Turchiuli, C., Fuchs, M., Bohin, M. et al. (2005). Oil encapsulation byspray drying and fluidised bed agglomeration. Innovative FoodScience and Emerging Technologies, 6, 29–35.

Wang, W.D. & Xu, S.Y. (2007). Degradation kinetics of anthocyaninsin blackberry juice and concentrate. Journal of Food Engineering, 82,271–275.




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