Fo o d C h emi s tr y 1 10 ( 2 00 8 ) 1 0 3 6 1 0 4 0

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Analytical Methods

Effect of gamma irradiation on the stability of anthocyanins and shelf-life of various pomegranate juices
H. Alighourchi, M. Barzegar *, S. Abbasi
Dep a rtm en t o f Fo od Sc i en c e an d Tec h n ol og y, Ta rb i at Mo d ar es Un i ver si t y, P. O. Bo x 1 41 1 5- 3 36 , T eh ra n , I r an

article info abstract

Food irradiation is a process which exposing food to ionizing radiations and it can improve the safety of
A rt ic l e h i s tor y: Rece i ve d 29 S ep t emb er 2 0 07

food. The pomegranate juice contained considerable anthocyanins and has become a new functional food
Rece i ve d i n re vi s e d for m 1 8 Fe b ru a ry 2 0 08

available for dieting and health. In the present study, the effects of gamma irradiation (010 kGy) on the
Acce p ted 4 M ar ch 2 00 8

stability of anthocyanins and inhibition of microbial growth in pomegranate juice during storage were investigated. Results indicated that the irradiation at all applied doses, significantly reduced total and individual anthocyanins. Moreover, irradiation with higher dosages (3.510 kGy) had undesirable effect
Keyw or ds :

on the total content of anthocyanins. However, irradiation at 2.0 kGy had effectively diminished the total
Fo od i r r ad i at i o n

bacteria and fungi count and retarding microbial growth during storage. Based on adverse effect of
Ion i z i n g r ad i a ti o n s

gamma irradiation on ACs content of studied juices, it is not recommended to irradiate pomegranate juice
Po me gr an a te j u i c e

with dosage higher than 2.0 kGy.

An th o cy an i n

2008 Elsevier Ltd. All rights reserved.

Fu n g i

1. Introduction Neves, & Martins, 2004). Pomegranate ACs are labile compounds and easily susceptible to degradation during storage and processPomegranate (Punica granatum L.) is one of the oldest known ing. Mart et al. (2000) found that the total ACs content was reedible fruits and is grown in Afghanistan, China, India, Iran, Japan, duced to 20% after storage of pomegranate juice at 5 C for 6 Mediterranean countries, Russia and the US (Shahidi & Naczk, months. In addition, Miguel et al. (2004) observed that ACs content 2004). In Iran, pomegranate is eaten as fresh as well as processed of Assaria variety of pomegranate decrease significantly during for jams, jellies, syrups and pomegranate juice products. This fruit 72 h storage at 4 C. is one of the most important commercial fruits in Iran and its to- In the recent years, the tendency of consumers to fresh fruit tal production in year 2005 was $670,000 tons (Anonymous, juices has been increased due to their better organoleptic proper2005). ties than pasteurized ones. Previously, it was generally assumed Anthocyanins (ACs) are glycosides of polyhydroxy and poly- that pathogenic microorganisms could not survive in high acid methoxy derivatives of avylium cation. Recent studies demon- foods. However, recent outbreaks of foodborne illnesses from strated that polyphenolic avonoids exhibit a wide range of unpasteurized fruit juices have indicated the necessity of pasteurbiological, pharmacological and chemoprotective properties as free ization for all fruit juices (Parish, 1997). Microorganisms, in particradical scavengers preventing oxidation and cancer initiation ular acid tolerant bacteria and fungi such as yeasts and molds, can (Kong, Chia, Goh, Chia, & Brouillard, 2003). easily spoil fruit juices (Tournas, Heeres, & Burgess, 2006). During Based on some research, pomegranate juice is a rich source of the last decade, with increasing demand for nutritious, fresh-like different phenolic compounds, with ACs to be one of the most food products with high organoleptical quality and an adequate important classes (Du, Wang, & Francis, 1975). The main ACs of shelf-life, non-thermal inactivation techniques have been a major pomegranate arils identified were as delphinidin 3,5-O-diglucoside research issue. Some of the investigated technologies are irradia(Dp 3,5-O-dG), 3-O-glucoside (Dp 3-O-G), cyanidin 3,5-O-digluco- tion, high hydrostation pressure (HHP), pulsed electrical fields, side (Cy 3,5-O-dG), 3-O-glycoside (Cy 3-O-G), pelargonidin 3,5-O- and UV decontamination (Devlieghere, Vermeiren, & Debevere, diglycoside (Pg 3, 5-O-dG), and 3-O-glycoside (Pg 3-O-G) (Mart, 2004). Preservation and shelf-life extension have been the historiPrez-Vicente, & Garca-Viguera, 2000; Miguel, Dundlen, Antunes, cal focus of research on irradiation of juices (Niemira & Deschnes, 2004, chap. 11). Food irradiation is a means of food preservation that has been in development since the early part of the 20th cen* C or r es p on d i n g au t h or . Te l .: + 9 8 2 1 4 4 19 6 5 2 2 ; fa x: +9 8 2 1 4 41 9 6 5 24 .

tury. If applied properly, irradiation can be an effective way to reE- m ai l ad dr es s es: mb b @mo d ar es . ac. i r, mo h se n b b@ gma i l .co m, b a rz ega r m@

duce the incidence of foodborne diseases and also inactivates food

ya h oo . com (M. B ar ze ga r) . 03 0 8 -8 1 4 6 /$ - s e e fr o n t ma tt er 2 0 0 8 E l s ev i er Lt d . A l l r i gh ts re s er ve d. d oi : 10 . 1 01 6 /j . fo od ch e m.2 0 0 8. 0 3 .0 1 3

H. A l ig ho u rc h i et a l . / F ood C h emi s tr y 1 1 0 ( 20 0 8) 10 3 6 1 04 0 1 0 3 7

spoilage organisms, including bacteria, molds, and yeasts in our leathery skin, which encloses hundreds of eshy sacs, was food supply (Morehouse, 2002). The FAO/IAEA/WHO joint commit- removed. The arils were manually separated and pressed. The tee on the wholesomeness of irradiated food approved irradiation juices (50 mL) were centrifuged (2 min at 10,000 rpm at 4 C), technology in 1981. It was stated that, irradiation of food at doses and divided into small vials and kept frozen at 18 C for one up to 10 kGy introduced no special nutritional problem (Stevenson, day upon analysis. 1994). The key organisms in food irradiation were yeasts and molds, 2.4. Irradiation treatment
which have a higher D10 value than bacterial pathogens (Monk, Beuchat, & Doyle, 1994). In fruit juices, D1 0 values have been re-

The pomegranate juices were filled in sterilized amber Wheaported for yeasts and molds are in the range of 13 kGy (Narton-320 glass vials (8 mL, from Sigma, USA). Irradiation treatment vaiz, Lescano, & Kairiyama, 1992) and 0.30.7 kGy for was carried out at various doses (0, 0.5, 2.0, 3.5, 5.0 and 10.0 kilopathogenic bacteria (Buchanan, Edelson, Snipes, & Boyd, 1998; gray (kGy)) at a dose rate of 1.43 kGy/h using a Gamma cell-220 Niemira, Sommers, & Boyd, 2001). Buchanan et al. (1998) irradiator (Nordion, Canada). After gamma irradiation, the samples showed that, a dose of 1.8 kGy should be sufficient to achieve were immediately placed in refrigerator at 4 C and they were the 5D inactivation of Escherichia coli recommended by the Naimmediately analyzed by HPLC/UV. Three samples were prepared tional Advisory Committee for Microbiological Criteria for Foods for each treatment. irradiation. In addition, Chachin and Ogata (1969) investigated the effect of sterilizing (280 kGy) doses of gamma irradiation 2.5. Microbiological analysis in grape juice. Based on our knowledge, there is no information about the The enumeration of microorganisms was made using stananthocyanin contents of major varieties of Iranian pomegranates dard techniques (AOAC, 1984), and included plate counts, yeasts and their variations after gamma irradiation. Therefore, the objecand molds and coliform. Microbial analyses of treated and untives of the present study were as follows: (i) identifying main ACs treated pomegranate juices were determined during two weeks of some pomegranate varieties; (ii) subjecting pomegranate juices storage at 4 C (1, 2, 3, 7, and 14 days). Serial dilutions of orito various levels of gamma irradiating and investigating its effect ginal samples were prepared in sterile peptone water (0.1%) to on ACs content and microbiological properties or shelf-life; and reduce the microbial population sufficiently to obtain separate (iii) investigating the effect of pomegranate variety on ACs content colonies when plating. Total plate counts were determined of juices after gamma irradiation. using pour plate method on Plate Count Agar. One milliliter of several diluted samples were mixed with 15 mL liquid plate count agar that had been cooled to about 45 C, and poured 2. Materials and methods immediately into sterile 90 mm Petri dishes. After the agar had hardened, incubation was performed at 35 C for 4872 h. 2.1. Samples Total fungi (moulds and yeasts) were carried out in triplicate using pour plate method on plates of Potato Dextrose Agar Six various pomegranate varieties were obtained from mature (PDA) medium, and then were enumerated after incubation fruits growing in agricultural research center of Yazd in south west for 5 days at 25 C. For a coliform count, samples were plated of Iran. Commercially ripe fresh fruits were harvested during the directly onto MacConkey agar and incubated at 37 C for 24 September of 2006 from different mature trees and they were ran36 h. Each value represents the mean of three samples and redomly selected to represent the population of the plantation. Fruits sults were expressed as colony-forming units (CFU) per were transported by ventilated car to the laboratory, where pomemilliliter. granates with defects (sunburns, cracks, cuts and bruises in husk) were discarded. The different varieties selected for this study were as follows: Gorche Shahvar Yazdi (GSY), Malase Yazdi (MY), Vahshe 2.6. Anthocyanin analysis Kane Tehran (VKT), Mesri Torshe Kazeron (MTK), Jangali Pust Germeze Rodbare Torsh (JPGRT) and Torshe Mamoli Lasjer (TML). Approx- ACs content of juices were determined using a Waters HPLC imately, 2 kg (n = 10) of pomegranates at harvesting maturity was system equipped with an Empower software, a pump (Waters 600, USA), a Rheodyne 7125i six-way injector with 20 lL sample sampled for each variety. loop, and a UVVis detector (Waters model 2487). A column
lBondapackTM C1 8 (4.6 250 mm, dp 10 lm) from Waters (Ire-

2.2. Chemicals land)) was used for the separation. Clarified juice (20 lL) was injected onto the HPLC. The eluDelphinidin 3,5-O-diglucoside (Dp 3,5-O-dG), delphinidin 3-Otion was carried out at room temperature using 5% formic acid glucoside (Dp 3-O-G), cyanidin 3,5-O-diglucoside (Cy 3,5-O-dG), aqueous solution (A) and methanol (B) in a linear gradient cyanidin 3-O-glucoside (Cy 3-O-G), pelargonidin 3,5-O-diglucofrom 15% to 35% B at 15 min, followed by isocratic run until side (Pg 3,5-O-dG), and pelargonidin 3-O-glucoside (Pg 3-O-G) 20 min. Flow rate was 1 mL/min with UVVis detector at standards were purchased from Apin Chemicals Co. Ltd. (Oxford510 nm (Miguel et al., 2004). Calculation of the concentrations shire, UK). Methanol (HPLC grade) was purchased from Caledon was based on the external standard method and ACs were laboratories (Ontario, Canada). Formic acid and all bacteriological identified by comparison of their retention times with those media were obtained from Merck (Darmstadt, Germany). The ulof pure standards (Table 1). For each sampling point, there tra pure water was prepared with the Purise system (Seoul, South were three replicates. Korea). 2.7. Statistical analysis 2.3. Preparation of raw pomegranate juice One-way analysis of variance was used to analyze the data. A p Fruit from each pomegranate variety was washed in cold tap value of 0.05 or less was considered as significant (using SAS water and drained. They were manually cut-up and the outer software).

1 03 8 H. A l i gh ou rc h i et al . / Foo d C h em is tr y 11 0 (2 00 8) 1 03 6 1 0 40 Ta b le 1

3. Results and discussion

Li n ear cal i b r ati o n eq u ati o n s of i nd i v id u al a n th oc yan i n s ta n da rd s

3.1. Effect of irradiation on ACs content

C omp o u n d t R (mi n ) Li n ea r ra n ge (mg /L) Li n e ar eq u at i o n r Dp 3 , 5 -O -d G 9 .9 1 1 0 0 A = 1 8 71 8 C 4 1 45 5 a 0 .9 9 9 5

Based on our findings, some of 6 different ACs, including

C y 3 , 5 -O -d G 1 0 .6 1 5 0 0 A = 1 8 72 0 C 5 7 35 1 0 .9 9 9 8 Pg 3 , 5 - O- d G 1 1 .2 0 .5 5 0 A = 2 3 44 4 C + 43 1 1 .0 0

delphinidin 3,5-O-diglucoside, 3-O-glucoside, cyanidin 3,5-ODp 3 - O- G 1 1 .9 1 2 5 0 A = 2 2 93 3 C 4 7 82 1 0 .9 9 9 9

diglucoside, 3-O-glucoside, pelargonidin 3,5-O-diglucoside, and

C y 3 - O- G 1 2 .7 1 2 5 0 A = 7 6 95 0 C 4 3 96 1 5 0 .9 9 8 9

3-O-glucoside were identified in freshly prepared juices by comPg 3 - O- G 1 3 .5 0 .0 2 50 A = 6 6 46 8 C 1 7 32 0 0 .9 9 9 5

paring their retention times with those of authentic standards

a A : p ea k a r ea; C : co n cen t ra ti o n (mg /L ).

(and by spiking), which confirm previously reports on the other pomegranate varieties (Mart et al., 2000; Miguel et al., 2004).

20 80 MTK JPGRT 16 TML 60 MY VKT 12 GSY 40 8 20 4 00 0 0.5 2 3.5 5 10 0 0.5 2 3.5 5 10

Radiation dose (kGy) Radiation dose (kGy)

35 180 30 150 25 120 20 90 15 60 10 30 5 0 0 0 0.5 2 3.5 5 10 0 0.5 2 3.5 5 10

Radiation dose (kGy) Radiation dose (kGy)

10
1.5

8
1.2

6
0.9

4
0.6

2
0.3

0
0 0 0.5 2 3.5 5 10

0 0.5 2 3.5 5 10

Radiation dose (kGy) Radiation dose (kGy)

F ig . 1 . E vo l u ti o n o f t h e i n d i vi d u a l an t h ocy an i n c on ce n tr at i on s o f co n t ro l (0 kG y ) a n d i r r ad i at ed ( 0 .5 1 0 k Gy ) p o meg r an at e j u i ce s .

H. A l ig ho u rc h i et a l . / F ood C h emi s tr y 1 1 0 ( 20 0 8) 10 3 6 1 04 0 1 0 3 9

Irradiation was carried out at several doses to find the optimal the irradiation (Song et al., 2006). In contrast, Ayed, Yu, and Lacroix dose for getting maximum microbial reduction and minimum deg- (1999) investigated the effect of irradiation (09 kGy) of grape radation of ACs as the color of pomegranate juice is directly de- pomace and they observed that AC contents increased at all doses pended on its content and composition. The current study of irradiation, especially at 6 kGy. appears, is one of the only ones that examined the effect of irradi- In our study, irradiation at 5 and 10 kGy did not show signifiation by different doses on the AC contents and microbiological cant difference (p P 0.01) between the amounts of total ACs of behavior of pomegranate juices. Although there are many studies MY, VKT and GSY varieties. The average loss of the total AC conon the effect of irradiation on microbiological and organoleptic tents in all juices, after gamma irradiation (010 kGy), were beattributes of other fruit and vegetable juices (Buchanan et al., tween 22.0% and 90.0%. In fact, pomegranate juices that 1998; Chachin & Ogata, 1969; Kim, Song, Lim, Yun, & Chung, irradiated at 0.5 and 10 kGy retained approximately 80% and 10% 2007; Song et al., 2006). of the AC content compared with the control, respectively. These Fig. 1 shows that gamma irradiation of studied juices cause a results indicated that the stability of ACs against the irradiation significant decrease on the individual ACs concentration in com- was related to juice composition. The differences among juices parison with the initial AC contents (dose 0 kGy) due to degrada- originated from fruits variety, and the relative stability of an AC detion of ACs (p < 0.05). As it can be seen, the stability of pends on its matrix, structural features, and the processing condidiglycosides ACs to irradiation were higher than monoglycosids tions (Es-Safi, Cheynier, & Moutounet, 2002; Talcott, Brenes, Pires, at lower doses of gamma irradiation (0.5 and 2 kGy). For example, & Del Pozo-Insfran, 2003; Torskangerpoll & Andersen, 2005). Howcyanidin 3,5-O-diglucoside was more stable (up to 2 kGy) than the ever, there are few pieces of research in the literature about the efother ACs. However, decreasing trends of diglycosides were similar fect of the fruit variety on the radiation sensitivity of the resulting to monoglycosides in the higher level of gamma irradiation. In juices. addition, higher stability of diglycosides ACs at different conditions were previously reported by other researchers (Holcroft, Gil, & Kader, 1998; Timberlake & Bridle, 1977). 3.2. Effect of irradiation on microbial ora Table 2 shows total AC contents of all studied pomegranates before and after irradiation by gamma ray (010 kGy). It can be seen The predominant organisms found in the various types of juices that the different applied doses had significant in uence on the to- were yeasts (Hatcher, Parish, Weih, Splittstoesser, & Woodward, tal ACs contents of all juices after irradiation (p < 0.05). Chachin 2000; Tournas et al., 2006). These organisms could grow during and Ogata (1969) have previously investigated the effect of steril- refrigeration (4 C) and cause spoilage of samples. In food irradiaizing (280 kGy) doses of gamma irradiation in grape, apple, and
tion, the D10 values of yeasts and molds were higher than bacterial

orange juices. They reported that gamma irradiation, at pathogens. Thus, most research efforts related to irradiation of P10 6 80 kGy, obviously caused losing of grape juice ACs. In the juices have targeted spoilage organisms such as yeasts and molds irradiated kale juice (0, 3, and 5 kGy), the amount of total phenolic rather than bacterial pathogens (Grinbaum, Ashkenazi, Treister, content was significantly lower than the control immediately after Goldschmied-Requven, & Block, 1994; Monk et al., 1994).

Ta b le 2 To tal a n th oc yan i n co n ten t s of i r r ad i ate d an d no n -i r r ad i ate d po m eg ra na te j ui ce s (m g/ L)

Irr ad i a ti o n d os e (k G y) P ome gr an a te va ri e ti e s a

MT K JPG RT T ML MY VKT GS Y 0 2 5 1. 7 0. 1 au 24 9 . 2 0 . 6b u 2 5 2 .2 1 .0 a u 31 . 7 0 . 6 eu 4 9. 4 0. 1 cu 37 . 2 0 . 0d u 0. 5 2 0 2. 7 1. 8 b v 19 7 . 3 0 . 0cv 2 0 6 .7 0 .3 a v 30 . 2 0 . 5 ev 3 9. 4 0. 4 d v 19 . 4 0 . 0f v 2 1 4 2. 1 2. 8 b w 14 1 . 2 2 . 3b w 1 5 2 .6 0 .4 a w 27 . 2 0 . 3 cw 2 1. 4 0. 3 d w 16 . 3 0 . 1 ew 3. 5 7 2. 2 0. 3 cx 7 8 . 2 0 . 4b x 9 8 .0 3 .0 a x 12 . 6 0 . 5 dx 9. 5 0. 0 ex 9 . 0 0 . 1 ex 5 6 9. 5 1. 0 b x 50 . 5 0 . 5c y 7 3 .1 1 .8 a y 8 . 4 0 . 1 dy 8. 6 0. 1 d y 7 . 0 0 . 1 dy 10 1 2. 9 0. 2 b y 10 . 7 0 . 1c z 2 0 .0 0 .4 a z 6 . 0 0 . 0d z 6. 2 0. 1 d z 6 . 1 0 . 2 dz

a V al u e s wi t h d i ffer en t l et te rs ( a d ) w i th i n a s i mi l ar r ow a n d (w z ) w i th i n a s i mi l ar co l u mn ar e s i g n i fican t l y d i f fer en t (p < 0. 0 5) .

Ta b le 3
To tal p l at e co u n ts (T P C) an d to ta l fu n gal co u n ts ( TFC ) of ir r ad i ate d an d n on - i rr ad i ate d M Y an d TM L p om eg ra n ate j u ic es , s t or ed at 4 C for 1 4 da ys a

Ju ic e Mi cr ob i a l co u n ts (c fu /mL) Irr ad i a ti o n d os e (k Gy ) S to r age ( da ys ) 1 23 7 14 MY T PC 0 4 7 0 0 1 4 5 74 7 0 85 0 1 9, 8 5 0 4 5 0 8 9 ,8 0 0 8 0 0 5 3 6 ,0 0 0 9 2 0 0 0. 5 8 4 0 30 12 5 0 14 0 2 40 0 3 80 1 9 ,8 0 0 1 0 5 0 4 9 ,2 5 0 25 0 0 2 0 0 <1 0 2 7 2 2 4 0 2 5 T FC 0 3 4 0 0 8 0 69 0 0 45 0 1 0, 1 5 0 1 1 5 0 1 8 5 ,5 0 0 7 2 0 0 7 1 5 ,0 0 0 3 5 4 0 0. 5 1 0 5 0 2 3 0 28 2 0 19 0 4 24 0 3 8 0 2 5 ,1 8 0 1 3 5 0 8 7 ,2 0 0 8 5 4 0 2 0 <1 0 0 1 4 5 4 3 12 TML T PC 0 2 0 6 0 4 5 0 42 0 0 43 0 8 25 0 6 9 0 3 8 ,6 0 0 4 8 0 0 3 0 3 ,8 0 0 7 8 0 0 0. 5 3 9 8 45 67 5 9 0 2 85 0 1 5 0 9 6 5 0 1 7 2 0 3 0 ,6 4 0 2 6 5 0 2 0 0 2 0 5 0 7 0 1 7 T FC 0 1 0 2 0 4 4 0 34 6 0 56 0 7 82 0 1 8 40 4 5 ,8 0 0 5 8 0 0 3 9 ,6 4 5 0 5 2 5 0 0. 5 5 9 0 80 10 8 0 45 2 74 0 2 4 0 1 1 ,8 5 0 1 5 0 0 4 2 ,8 9 0 33 0 0 2 0 0 0 3 8 12 8 5 3 2

a A t > 3 .5 kG y , t h er e w er e n o mi cr o or g an i s ms i n MY a n d TM L j u i ces .

1 04 0 H. A l i gh ou rc h i et al . / Foo d C h em is tr y 11 0 (2 00 8) 1 03 6 1 0 40

The changes of microbial populations of MY and TML (pH 4.2

Ay ed , N . , Yu , H . L ., & La cro i x , M. ( 1 9 99 ) . Imp r ov eme n t o f an th o cy an i n yi e l d an d s h el f- l i fe ex t en s i on o f gr ap e po ma ce b y g amma i r r ad i at i o n. Foo d R es ea rc h

and 4.1, respectively) pomegranate juices are shown in Table 3.

I nt ern a ti on al , 3 2, 5 39 5 4 3 .

The microbial counts (bacteria and fungi) were measured by the

Azi z , N . H ., & Mo u s s a, L. A. A. (2 0 0 2) . In u en ce of ga mma- r ad i at i o n on myco t ox i n

plant counts and total fungi in control and irradiated pomegranate

p ro d u ci n g m ou l d s an d m yco to x i n s i n fr u i ts . Foo d Co nt ro l , 1 3, 2 8 1 2 8 8 . B u ch an an , R. L ., Ed e l so n , S. G . , Sn i p e s , K. , & B oy d , G . ( 1 99 8 ). In ac ti v ati o n o f

juices. The initial mean populations of the total bacterial and fungi
Es c he ri c h ia c ol i O 1 5 7: H 7 i n ap p l e j u i ce b y i r r ad i at i on . A pp l ie d an d E n vi ro n men ta l

counts for fresh juices of MY and TML (without treatment) were

Mi c r ob i ol og y, 64 (1 1 ), 4 5 3 3 4 5 3 5. C h ach i n , K ., & Og at a, K . (1 9 6 9 ). C h a n ge s o f ch em i cal co n st i tu e n ts an d q u al i ty i n $4.7 103 , $3.4 103 and $2.0 103 , $1.0 103 cfu/mL, respecs ome j u i ce i rr ad i a ted w i th th e s te r il i z i n g d os e l e ve l o f g amma r ay s . Fo od

tively. After irradiation and during storage, no coliform was deI rr ad i at io n , 4 (1 ), 8 5 90 .

tected. The total counts of the bacteria in the non-irradiated

De vl i e gh e re , F. , Ver me i re n , L. , & De b ev er e, J. ( 2 00 4 ). N e w p re s er va ti o n t ech n o l o gi e s:

juices of MY and TML, were rapidly increased to more than

Po s s i b i l i ti es a n d l i mi ta ti o n s . I n te rn at i on al D ai r y J o ur n al , 14 , 2 7 3 2 85 . Du , C . T ., W an g , P. L. , & Fr an ci s , F. J. (1 9 7 5 ). An t h ocy a ni n s of p ome gr an a te , P u n ic a 5.3 105 and 3.0 105 cfu/mL, respectively, during storage at gr an at um . J ou rn a l of Fo od Sc i en c e, 40 , 41 7 4 18 .

4 C. Gamma irradiation has prevented the microbial growth and

Es - Sa fi, N . , C h ey n i er , V . R. , & Mo u to u n et , M. (2 0 0 2) . Ro l e o f al d eh y d i c d er i va ti v es i n

by increasing the irradiation dosage a decreasing trend was obth e co n d en s a ti o n of p h e n ol i c co mp o u n ds w i th e mp ha s i s on t h e s en s o ri a l p ro p er ti e s o f fr u i t- d er i v ed fo o d s . J o u rn al of t he Sc i en c e o f Fo od a nd A g ri c u lt u re,

served (in studied varieties). Irradiation at 0.5 and 2 kGy reduced

50 , 55 7 1 55 8 5 .

the growth rate of bacteria and fungi of the selected pomegranate

Gr i n b au m, A., As h k en az i , I. , T re i s te r, G. , G o ld s ch mi e d -Re q u ve n, A. , & B l ock , C . S.

juices during the first 3 days of storage at 4 C. The microbial pop(1 9 9 4) . E x pl o d i n g b o tt l es : e ye i n j u ry d u e t o y ea s t fe rme n ta ti o n of a n

ulation reduced to below the detection limits at P3.5 kGy, in all

u n car b on a ted so ft d ri n k . Br i ti s h J o u rn al o f Op h th al m ol og y, 78 (1 1 ), 8 8 3. H atc h er , W . S ., Pa ri s h , M. E ., W ei h , J. L. , Sp l i t ts t oe s se r , D . F. , & W oo d w ar d, B . B .

studied juices. Similar results were observed in other pomegranate

(2 0 00 ) . Fr u i t be ve ra ge s . In F . P . Do wn e s & K . It o ( E ds . ), M eth od s f or th e

varieties (GSY, VKT, MTK, JPGRT, data not shown). Based on the reMi c r ob i ol og i ca l Ex am in a ti on o f Foo ds ( pp . 5 6 5 5 6 8 ). Wa s h i n gt on , D C : Ame ri ca n

sults of Aziz & Moussa, 2002 and Kim et al. (2007), the 35 kGy
Pu b l i c H e al th As s oc ia ti o n . H ol cr o ft, D. M., Gi l , M. I., & Kad e r, A. A . (1 9 9 8 ). E ffe ct o f car b on d i o xi d e o n

radiation doses may prevent microbial growth in the kale juice

an th o cy an i n s , p h en y l al a n i n e ammo n i a l ya s e an d g l u co sy l t ra n sf er as e i n t h e ar i l s

during storage period. This result showed that, the inactivation of

of s t or e d p o meg r an at es . J ou rn a l o f th e A mer i c an So c ie ty f o r H or ti c u lt u ra l S ci en c e,

microorganisms in different juices depended on their composi12 3 , 1 3 6 1 4 0 . Ki m, D ., S o n g, H . , Li m, S ., Yu n , H . , & C h u n g , J. (2 0 0 7 ). E ffe cts of g amma i r r ad i at i o n o n

tions. Buchanan et al. (1998) reported that differences among the

th e r ad i a ti o n -r es i s t an t b ac ter i a a n d p o l yp h e n ol o xi d as e act i vi ty i n f re s h k al e

juices may be affecting the radiation resistance of microorganism.

j u i ce. R ad i ati o n Ph ys i c s a n d C h em is tr y, 7 6, 1 21 3 1 2 17 .

Their results showed that low-dose gamma radiation treatment

Ko n g, J. M. , C h i a, L. S ., G o h , N . K ., C h i a, T . F. , & B ro u i l l ar d , R. (2 0 0 3 ). An a l ys i s a n d bi o l o gi ca l ac ti vi t i es o f a n th o cy an i n s . P h yto c he mi s tr y, 64 , 92 3 9 33 .

potentially could be eliminating E. coli without any detectable

Mar t , N ., P r ez- Vi ce n te , A. , & G ar c a- Vi g u er a, C . ( 2 00 0 ). In u en ce of s to ra ge

effect on the organoleptic quality of the product. In our study, we

tem pe r atu r e a n d a s co rb i c ac i d a d d i ti o n o n p o meg ra n ate j u i ce. J ou rn a l o f th e

found that irradiation doses upper than 2 kGy were sufficient to

Sc i enc e of Fo od a nd A gr i c u lt ur e, 82 (2 ) , 2 1 7 22 1 . Mi gu e l , G. , D u n dl e n , S. , An t u ne s , D ., N ev es , A ., & Ma r ti n s , D . (2 0 04 ) . Th e e ffe ct of tw o

completely inactivate the studied microorganisms.

met h od s o f p o meg ra n at e j u i ce ex tr ac ti o n on qu a l i ty d u ri n g s to r ag e a t 4 C . J ou rn a l of B i om ed ic i n e a nd B i ot ec hn o lo gy , 5 , 3 3 2 3 37 .

4. Conclusions
Mo nk , J. D ., B e u ch at , L. R. , & Do y l e, M. P . ( 19 9 4 ). Ir r ad i at i on i n act i va ti o n o f fo od bo r n e mi cr o o rg an i s ms . J ou rn a l of Fo od P ro tec ti o n, 5 8( 2 ), 19 7 2 08 . Mo re h ou s e , K. M. (2 0 0 2) . Fo o d i r r ad i at i o n US re gu l a to ry co n s id e ra ti o n s . R ad i at i on

Gamma irradiation can be applied to produce fruit juices with

Ph ys i c s an d C h em i str y, 6 3, 2 8 1 2 8 4 .

improved shelf-life. The different applied gamma doses signifiN ar va i z, P . , L es can o , G. , & Ka i ri y am a, E. (1 9 9 2 ). Irr a di a ti o n o f al mo n d s an d ca s h ew n u ts . Lebe ns m it tel - Wi s sen s c ha f t u n d- Te c hn ol o gi e, 25 , 23 2 2 35 .

cantly affected the total and individual ACs content of all juices
N i emi r a, B . A. , & D es ch n es , L. (2 0 04 ) . Ion i z i n g ra d i ati o n p ro ces s i n g o f fr u i ts an d

and therefore pomegranate juice color. In fact, pomegranate juices

fru i t p r od u ct s . In D . M. B a rr e tt, L. So mo gy i , & H . Rama s wa my ( Ed s . ), Pr oc es s i ng

that irradiated at 0.5 and 10 kGy retained approximately 80% and

f ru i ts (2 n d e d. ). B o ca Ra to n , FL: C RC p re s s. N i emi r a, B . A ., S omme rs , C. H ., & B o y d , G. ( 2 00 1 ). Irr ad i a ti o n i n a cti v at i on o f fo u r

10% of the initial AC contents. Irradiation at doses upper than

Sal mo n e l la s p e ci es i n o ra n ge j u i ce s w i th va ry i n g tu r b i d i ty . J ou rn a l of Fo od

2 kGy can completely inactivate studied microorganisms as well

Pr ot ec ti on , 64 (5 ) , 6 1 4 6 1 7 .

as retarding microbial growth during storage. However, at higher

Pa ri s h , M . E. (1 9 97 ) . Pu b l i c h ea l th an d n o n p as t eu r i zed fr u i t j u i ces . C r it i c al R ev i ews i n

doses (>2 kGy), a considerable decrease of total AC content was obMi c r ob i ol og y, 23 (2 ) , 1 0 9 1 1 9 . Sh ah i d i , F ., & N a czk , M. ( 20 0 4 ). P h en o l i c c omp o u n d s i n fr u it s a n d v eg et ab l es .

served. Finally, it is not recommended to irradiate pomegranate

In Ph en ol i c in f oo d a n d n u tr ac eu ti c al s (p p . 1 3 1 2 39 ) . B o ca Ra to n , FL : C RC

juice with dosage higher than 2.0 kGy.

Pr es s . So n g, H . P ., Ki m, D . H ., Jo , C . , Lee , C . H. , Ki m, K. S. , & B yu n , M. W . (2 0 06 ) . Eff ect o f ga mma i r ra di a ti o n on t h e mi cro b i o l og i cal q u al i t y an d an t i o xi d an t act i vi t y o f

Acknowledgments
fre s h ve ge ta bl e j u i ce. Fo od M i c ro bi o lo gy , 2 3, 3 7 2 3 7 8 . St ev en s on , M. H . ( 19 9 4 ). N u tr i ti o n al a n d o t h er i mp l i cat io n s o f i r ra d i ati n g mea t.

The authors would like to acknowledge Research Council of

Pr oc eed i n gs of t he Nu tr i ti on So c i ety, 5 3, 3 1 7 3 2 5 . Ta l co tt , S. T . , B re n es , C . H . , P i re s , D . M. , & D el P ozo - In s fr an , D . ( 20 0 3 ). P h y to ch emi ca l

Tarbiat Modares University for its financial support and Agriculs tab i l i t y an d co l or re te n ti o n o f co p ig me nt ed an d p ro ce s se d m us c ad i n e gr ap e

tural Research Center of Yazd for providing fresh pomegranates.

j u i ce. J ou r na l of A g ri c u lt ur a l a n d F oo d C h em is tr y, 5 1, 9 5 7 9 6 3 . Ti mb e rl a ke , C . F., & B r i d l e, P . (1 9 7 7) . An th o cy an i n s : col o u r a ug men t at i on wi t h ate ch i n a n d ace tal d e h yd e . J o u rn al o f th e S c ie nc e of Fo od a n d A g ri c u l tu re, 2 8,

References
53 9 5 4 4. To u r n as , V . H ., H eer e s, J., & B u r ges s , L. ( 2 00 6 ). Mou l d s an d y eas t s i n fr u i t s a la d s an d An o n ymo u s (2 0 0 5) . Ir an S ta ti s ti c al Yea r B o o k 2 0 0 5. < ht tp : //e ama r. s ci .o r g. i r/ fru i t j u i ces . Fo od M i c ro bi ol o gy, 2 3, 6 8 4 6 8 8 . i n de x _e. as p x >. To r s ka n ge rp o l l , K. , & A n d er s en , Q . M. (2 0 05 ) . C o l ou r s tab i l i t y o f an th o cy an i n s i n AO AC (1 9 84 ) . B ac ter i ol o gi c al An a ly ti c al M an u al ( 6 th ed .) . W as h i n g to n , D C: aq u eo u s s o l ut i o n s at va ri o u s p H va l u es . Foo d Ch em i st ry , 8 9, 4 2 7 4 4 0 . As s o ci ati o n o f O ffici a l An al y t i cal C h em is t s .