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IllEuropean Drying ConferencePalma de Mallorea (Spain), Octobcr 26-28.2011

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European Drying Conference - EuroDrying'2011Palma. Balearic Island, Spain, 26-28 October 20 II

MICROENCAPSULATION OF BIOACTIVE COMPOUNDS FROMPOMEGRANATE (PUNICA GRANATUM L) JUICE USING SPRAY DRYING

Regina I. Nogueira I, Manuela Cristina P. de A. Santiago', Flávia dos Santos Gomes',Sérgio Macedo Pontes', Paulo Roberto Coelho Lopes", Suely Pereira Freitas3

IEmbrapa Food Tecnology, Av. Das Américas, 29501, Rio de JaneirolRJ, 23020-470,Brazil Te/.:+55 2/362296//, E-mail: nogueira@ctaa.embrapa.br

2 Embrapa Tropical Semi-Arid, BR 428, Km 152, Zona Rural - Caixa Postal 23 Petro/ina/PE

3Universidade Federal do Rio de Janeiro - Escola de Química. Av. Horácio Macedo,2030, Bloco E sala E-2//, Ilha do Fundão, Rio de Janeiro/Ri, 2/94/-909, Brazil Te/.:

+5521256277J7,jreitasp@eq.ujrj.br

Abstract: The aim of this work was to evaluate the effect of operational conditions on anthocyaninsconcentration and antioxidant capacity of the microcapsules from pomegranate juice obtained by spray drying.The effect of air conditions (temperature and air flow rate) on the anthocyanins concentration of pomegranatemicrocapsules was investigated. The experimental results showed that the identified bioactive compounds werepreserved for ali conditions evaluated.

Keywords: anthocyanins, functional compounds, spray drying

The pomegranate (Punica granatum L.) is a fruitfrom the Middle East and grows in arid regions. It isa fruit with a long medical history, widely used bymany people, especially Asians. The fruit has seedscovered with a reddish pulp attended by phenoliccompounds, especially anthocyanins. This is animportant commercial fruit widely cultivated in Asia,North Africa, the Mediterranean and Middle East(Sarkhosh et al. 2006), and it has a good consumerpreference for its attractive juice, sweet acidic andrefreshing arils. There are great interests of scientistswho engage themselves in pharmaceutical,nutriological and pharmacological research, and newdrug development, due to its distinctive multipleofficinal parts and multiple bioactive suchhypolipidemic, antioxidant, antiviral, anti-neoplastic,antibacterial, anti-diabetic, antidiarrheal andhelminthic effects (Wang et ai, 2010). Humanconsumption of anthocyanins is increasing becauseofthe rising awareness and interests in their potentialhealth benefits and pomegranate is one of the majorsources of polyphenolic such as the anthocyanins.

There are a lot of products made from pomegranate,like jellies juices and dry fruit. However, during theprocessing, may be occur the degradation of theirconstituents. The transformation process shouldpreserve the nutritional and functional characteristicsof the raw material from which it originates. Jaiswal

et aI (20 IO) evaluated the effect of dryingpomegranate arils anthocyanins and observedanthocyanins losses of 60 to 80% by vacuum and sundrying, respectively. They concluded thatanthocyanins are stable at high temperatures, but theenzymatic actions contributed to color changes in theproduct.

ln conventional drying processes, high anthocyaninslosses are observed, but the use ofmicroencapsulation by spray drying may allow thepreservation of bioactive compounds present inpomegranate juice. Microencapsulation is a processof packaging of solid, liquid or gas in extremelysmall capsules which can release content in acontrolled manner and under specific conditions,increasing the product stability (Favaro-Trindade etai, 2008).

The aim of this work was to evaluate the effect ofoperational conditions (inlet air temperature and airflow rate) on anthocyanins concentration andantioxidant capacity of the microcapsules frompomegranate juice obtained by spray drying.

MATERIALS AND METHODS

Fresh pomegranates (Punica granatum L.) weresupplied by Special Fruit farm, located in Braziliansemiarid. They were selected, washed and manuallyprepared to separate the arils from the shell.

Processing

The arils were processed in horizontal device marcltametal, model Bonina 0,25df, in order to separatethe juice from the seeds. The juice was frozen andmaintained at -20° C until the processing.

Formulation

For encapsulation purposes, a mixture of 50%modified starch (Capsulf.National Starch) and 50%maltodextrin (MD14P, National Starch), washydrated with pomegranate juice (15°Brix), to beused as the wall material. The ratio between the juicesolid content and the wall material was 1:I.

Spray drying

The formulated material was fed into a spray drying8uchii model B 190, and atomized with a nozzle. Thewater was evaporated by the hot air contacting theatomized material. The microcapsules were collectedafter they fali to the bottom of the dryer and kindpacked in stand up pouch, vacuum sealed and storedat 25° C until the analytical determinations. The yieldwas ca\culated as the ratio of the mass of product(dry basis) and mass ofsolids in the feed.

Analytical determinations

Anthocyanins analysis: about I g of sample wasweighed and the extraction was made with a solutionof methanol and formic acid in the ultrasonic bathwith subsequent centrifugation until discoloration ofthe solution. Then, an aliquot ofthe extract was driedunder compressed air being re-suspended inmethanol and formic acid for chromatographicanalysis. Chromatography was performed on aWaters® Alliance 2695 system, with a Waters®2996 photodiode array detector, with a Thermo®Scientific C18 8DS (100mm x 4.6mm; 2AJlm)column, flow IA mL / min., column temperature of40° C, injection volume of Züul. and gradient elutionmethod with acetonitrile and formic acid.Characterization of anthocyanins present inpomegranate was performed by comparing thechromatographic profile with the literature and bycomparing the retention times obtained withchromatograms of other fruits previouslycharacterized by the laboratory in similar conditionsanalysis (Santiago et ai, 20(0). The quantification ofanthocyanins was performed by external standardcurve and the results have been reported by theequivalence cyanidin-3-0-glycoside, which was theexternal standard used to plot the calibration curve.Anthocyanins determination was conducted induplicate analysis.

Antioxidant Activity: measured by the TroloxEquivalent Antioxidant Capacity (TEAC), alsoknown as ABTS cationic radical scavenging activity,according to the method proposed by Re et aI. (1999)and Rufino et aI. (2010). Antioxidant activityanalysis was performed in triplicate.

Experimental design

The response surface method was applied to optimizethe process parameters: inlet air temperature (XI) andair flow rate (X2) were the independent variablesstudied to optimize the process yield, anthocyaninsconcentration and antioxidant activity. The levels ofthe independent parameters were based onpreliminary experimental results.

Table 1. Uncoded and coded levels of independentvariables used in the experimental designo

Uncoded- Air inlet Air flowvariables Temperature eC) (m3/h)

-1 180 500O 190 6001 200 700

RESULTS

The overall yield (product mass in dry basis/solids inthe feed) reached was about 57%. The yield of spraydrying on a laboratory scale observed by Li et aI.(2010) was between 50 to 70%.

Characterization of phenolic compounds

The anthocyanins profiles and content of theformulated pomegranate juices and microcapsuleswere similar in a dry basis. So, the identifiedbioactive compounds were preserved during spraydrying processing for ali operational conditions(Table 1). The results showed that the averageantioxidant activity of microcapsules, expressed asumol Trolox eq/g in dry basis, was 121.77 ± 9.93representing about 21% losses as compared withformulatedjuice, 154.33 ± 5.75

TableI. Anthocyanins concentration (mg/IOO g db) inthe formulated juice and microcapsules

Anthocyanin Average concentration

Feed* Powder**

Delphinidin 3,5 -diglucoside 14.99 ± 14.38 ± 2.36Cyanidin 3,5 -diglucoside 33.00 ± 28.91 ±4.64Delphinidin 3 -glucoside 2.63 ± 2.06 ±0.33Pelargonidin 3,5 -diglucoside 2.07± 1.39 ±0.19Cyanidin 3-glucoside 5.87± 4.66±0.67Pelargonidin 3 -glucoside 1.29± 0.66±0.05

* Values expressed as means of two deterrninations ±standard error; ** values expressed as means of sevenexperiments ± standarderror.

Figures 1, 2 and 3 are the Pareto chart showing theeffect of air inlet temperature and air flow on theprocess yield, antioxidant activity and cianidin-3-glucoside content, respectively. No significant effectof the studied independent variables was observed onprocess yield and anthocyanins content, at 95% ofconfidence. However, a negative effect onantioxidant activities occurred as air temperature andair flow increased simultaneously (Figure 2).

1bY-1! ·1.73208

(1JTemperaturer ·1,13087

(2)Airflowf ,6155342

Standardized Effect Estimate(Absotute Vatue)

FigureI. Pareto chart: Effect of air temperature andair flow on total mass product.

(1)Temperaturel

II

(2)Airflow'

,3728313

',146561

p=,05

Standardized Effect Estimate(Absolute Value)

Figure 2: Pareto chart: Effect of air temperature andair flow on Antioxidant activity.

I

(2)AirfIOwf ,582433

',477495

(1)Temperaturel

I',324879

Standardized Eff ect Estimate(Absolute Value)

Figure 3: Pareto chart: Effect of air temperature andair flow on Cianidina-3-5-diglicosídeo

CONCLUSIONS

This study investigated the effects of operational airconditions on anthocyanin content of pomegranatemicrocapsules prepared by spray drying fromformulated juice, using modified starch andmaltodextrin as wall materiaIs. No statistical

difference, at 95% of confidence, was observed forprocess yield and anthocyanins content, at theselected operational conditions. Very satisfactoryresults were observed for ali anthocyaninsnsretention in pomegranate microcapsules (about100%). So, the experimental resuIts showed that theidentified bioactive compounds were preserved forali conditions evaluated.

ACKNOWLEDGEMENTS

The authors (UF.) grateful\y acknowledge CNPq forfinancial support.

REFERENCES

Favaro-Trindade, C. S.; Pinho, S. c., Rocha, G. A.(2008). Revisão: Microencapsulação deingredientes alimentícios. Brazilian Joumal ofFood Tecnology, 11,2, 103-112.

laiswal,V.; Dermarderosian,A.; Porter, J.R.. (2010).Anthocyanins and poliphenol oxidase from driedarils of pomegranate (Punica granatum L.). FoodChemistry 118, 11-16.

Re, R.; Pellegrini, N.; Proteggente, A.; Pannala, A.;Yang, M.; Riceevans, C.. (1999). Antioxidantactivity applying na improved abts radical cátiondecolorization assay. Free Radic. Biol, Med., 26,9110, 1231 -1237.

Rufino, M.S.M., Alves, R.E., Brito, E.S., liménez,1.P., Calixto, F.S., Filho, J.M.( 2010). Bioactivecompounds and oxidant capacities of 18 non-traditional tropical fruits from Brazil. Foodchemistry, 121,996-1002.

Li, X.; Anton, N.; Aspargaus, c.. belleteix, F.;Thierry, F.V. (2010). Nanopartic\es by spraydrying using innovative new technology: TheBuchi Nano Spray Dryer B-90. Journal ofControlled Release, 147,304-310.

Sarkhosh, A., Zamani, Z., Fatahi, R., & Ebadi, A.(2006). RAPD markers reveal polymorphismamong some lranian pomegranate (Punicagranatum L). Scientia Horticulturae, 11,24-29.

Santiago, M. C. P. A. ; Gouvêa, A.C.M.S. ; Godoy,Ronoel Luiz de Oliveira ; Oiano-Neto, J. ;PACHECO, S. ; ROSA, 1. S. Adaptação de ummétodo por cromatografia líquida de altaeficiência para análise de antocianinas em suco deaçaí (Euterpe oleraceae Mart.). Rio de Janeiro:Embrapa Agroindústria de Alimentos, 2010(Comunicado técnico, 162. Biblioteca: CTAA(FL CTE 0162 UMT».

Wang, R.; Ding, Y.; Liu, R.; Xiang, L.; Du, L.Pomegranate: Constituents, Bioactivities andPharmacokinetics (2010). Fruit, Vegetable, andCereal Science and Biotechnology, 4 (Speciallssue 2), 77-87.