Food Chemistry 127 (2011) 641644

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Food Chemistry
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Quality attributes of starfruit (Averrhoa carambola L.) juice treated with ultraviolet radiation
Rajeev Bhat a,, Suhaida Binti Ameran a, Han Ching Voon a, A.A. Karim a, Liong Min Tze b
a b

Food Technology Division, School of Industrial Technology, Universiti Sains Malaysia, Penang 11800, Malaysia Bioprocess Technology Division, School of Industrial Technology, Universiti Sains Malaysia, Penang 11800, Malaysia

a r t i c l e

i n f o

a b s t r a c t
Starfruit juice were exposed to ultraviolet (UV-C) light for 0, 30 and 60 min at room temperature (25 1 C). On exposure, the titratable acidity signicantly decreased, while the decrease in Brix and pH were not signicant. With regard to colorimetric parameters, L value increased signicantly with a subsequent decrease in a and b values corresponding to UV treatment time. Except for the ascorbic acid, other antioxidants measured (% DPPH inhibition, total phenols, avonols, avonoids and antioxidant capacity) showed enhancement on expsoure to UV (signicant at 60 min). Microbial studies showed reduction in APC, yeasts and mould counts by 2-log cycle on UV treatments. These results supports the application of UV as a measure of non-thermal and physical food preservation technique for starfruit juice that can be explored commercially to benet both the producers and consumers. 2011 Elsevier Ltd. All rights reserved.

Article history: Received 27 September 2010 Received in revised form 29 November 2010 Accepted 11 January 2011 Available online 18 January 2011 Keywords: Ultraviolet treatment Non-thermal processing Starfruit juice Antioxidants Microorganisms Safety

1. Introduction Epidemiological studies have indicated that regular consumption of fruits and vegetables can reduce the risk of cardiovascular disease, ageing, cancer, etc. (Alothman, Bhat, & Karim, 2009a). Presently, consumers demand for safer and high quality foods with extendable shelife and with minimal quantity of chemical residues has increased tremendously. Application of short wave length ultraviolet light rays (UV-C) for decontamination and shelife improvement of fruits or their products is one of the emerging novel technologies the food industry can rely on. The lethal germicidal effect of UV-C on bacteria, yeasts and moulds have been used as an effective means for microbial inactivation and for preservation of overall quality of fruits or their products (Alothman, Bhat, & Karim, 2009b). Starfruit is a popular tropical fruit, which is consumed either fresh or made into juice to be served as a cooling beverage. The mature fruit is star-shaped, sweet and juicy and golden-yellow in appearance. Starfruits are used for preparing fruit salads or made into jellies and preserves (Macleod & Ames, 1990). In Malaysia, starfruits are mixed with apples and stewed with sugar and cloves or is cooked along with seafood or meat. In Ayurvedic medicine, starfruits or their juices are recommended to relieve pain from indigestion and bleeding haemorrhoids, and to reduce fever (Khare, 2004).
Corresponding author. Tel.: +60 4653 5212; fax: +60 4657 3678.
E-mail address: rajeevbhat@usm.my (R. Bhat). 0308-8146/$ - see front matter 2011 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodchem.2011.01.042

To our knowledge, ultraviolet (UV-C) radiation has not been applied for starfruits or their juices. Hence, the purpose of this study was to evaluate the effectiveness of UV on the physico-chemical parameters, antioxidant activities and on the microbiological quality of starfruit juices. It is envisaged that the baseline information generated will be useful for the successful implementation of UV technology on a pilot scale, to benet health conscious consumers as well as being useful in implementing the hazard analysis and critical control point (HACCP) approach for preservation and shelf life improvement of starfruit and their juice. 2. Material and methods 2.1. Plant material Locally grown, fresh, matured Arkin cultivar of starfruits (Averrhoa carambola L. Family: Oxalidaceae) (average weight, 168.0 0.1 g; 1112 mm length and 7375 mm in cross section) were purchased from the local supermarket (Jusco, Penang, Malaysia). After transporting to the laboratory under sterile conditions, the fruits were washed in distilled water and selected for healthy fruits with uniformity in size, shape and peel colour. 2.2. Juice preparation and UV treatment Starfruits cut into identical cubes were blended in a kitchen mixer (3 min) followed by ltration (Whatman lter paper No. 1) to obtain clear juice. This juice was poured into sterile disposable

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Petri dishes (15 100 mm; 10 mm depth) and exposed under the UV lamp (pre-calibrated for 1 h, germicidal uorescent lamp with a peak emission of 254 nm, ESCO Airstream laminar ow cabinet, Selangor, Malaysia) for 30 and 60 min separately, maintained at a distance of 30 cm. On an average, each of the exposed samples received a UV radiation dose of 2.158 J/m2 (digital radiometer). A corresponding control was maintained under same experimental conditions (25 1 C) (n = 3). 2.3. Physico-chemical analysis (pH, titratable acidity and total soluble solids content) The pH of the fruit juice was determined using a pH Meter (HORIBA F-21, Japan). For titratable acidity, 1 g of sample was titrated with 0.1 N sodium hydroxide (NaOH) to the phenolphthalein end-point (1%, pH 8.2 0.1). The total acidity was calculated using the following equation, with reference to citric acid:

2.7. Total phenolics content, antioxidant capacity, avonoids, avonols and ascorbic acid The total phenolics content was determined using the FolinCiocalteu (FC) reagent method (Singleton & Rossi, 1965) and the results expressed as milligrammes of gallic acid equivalents per gramme of sample. The method described by Prieto, Pineda, and Aguilar (1999) was adapted to measure the antioxidant capacity and the results were expressed relative to that of ascorbic acid (mg/ml). To determine total avonoids, the method described by Sakanaka, Tachibana, and Okada (2005) was employed and the results were expressed as mg of (+)-catechin equivalent per gramme of extract. The total avonols content was estimated by adapting the method described by Miliauskas, Venskutonis, and Van Beek (2004) with slight modications and the content was expressed as mg quercetin equivalent per gramme of extract. Ascorbic acid (Vitamin C) content were determined based on the 2,6-dichloroindophenol titrimetric method (AOAC, 1995). Results were expressed as milligrammes of ascorbic acid per 100 ml sample and the results calculated as:

TA%

V 0:1 N NaOH 0:067 100 m

where, V is titre volume of NaOH and m is the weight of starfruit juice (g). The total soluble solids content was determined according to the method described by Matsumoto, Obara, and Luh (1983). One drop of the juice was placed inside the refractometer ATAGO HSR-500 (Japan) and the results were expressed in Brix. 2.4. Colour analysis The colour of the samples was measured using Minolta Spectrophotometer (Model: CM-3500D). Colour appearance was taken as L, a and b values. L was a measure of lightness and varied from 100 for perfect white to zero for black, while a measured the redness when positive and greenness when negative, and b was a measure of yellowness when positive and blueness for negative. The total colour difference (TCD) or DE was measured by following equation:

Ascorbic acid mg=100 g or ml Titre dye factor concentration 100 =Extract aliquot used for estimation volume of sample use for estimation:
2.8. Microbial analysis (aerobic plate count, yeast and mould counts) UV treated and non-treated juice were analysed for the status of aerobic plate count (APC), yeast and mould counts. A series of decimal dilutions (101105) was prepared with 0.1% (w/v) peptone water. Further, one millilitre of decimal dilution of the sample was pipetted into Petri dish and the total plate counts were enumerated using the pour plate method. The plates were incubated at 37 1 C for 48 h. The total yeast and moulds were enumerated by pour plate method using potato dextrose agar (PDA) media. To inhibit the growth of others microbes, 10% tartaric acid was added into PDA agar (nal pH 3.53.7). The plates were incubated at 25 C for 5 7 days in the incubator. The results were expressed as log colony-forming units (cfus) per millilitre of juice (n = 5). 2.9. Solvents and reagents All the solvents and reagents used in the present study were procured from Sigma Aldrich (Laborchemikalien GmbH, 30926, Seelze, Germany) and R&M Chemicals (Essex, UK). Peptone water (buffer), plate count agar (PCA) and potato dextrose agar (PDA) were procured from MERCK (Darmstadt, Germany). 2.10. Statistical analysis The results obtained in the present study are represented as mean values of three individual replicates standard deviation (SD). Analysis of variance (one way-ANOVA) was performed and the signicant differences between mean values were determined by Tukeys pair-wise comparison test at a signicance level of p < 0.05. Statistical analyses were conducted by using SPSS 12.01 software (SPSS Inc., Chicago, IL, USA). 3. Results and discussion 3.1. pH, total soluble solids and titratable acidity The results on pH, total soluble solids and titratable acidity are shown in Table 1. The pH and acidity are important criteria during

L Lo 2 a ao 2 b bo 2 1=2
where, Lo, ao and bo are colour values of untreated juice. 2.5. Antioxidant assays 2.5.1. Sample preparation Fruit samples (control and UV treated) cut into equal sized cubes (2 4 cm) were blended in a kitchen mixer (3 min) followed by extraction with the solvent (methanol, 1:10) with continuous magnetic stirring on a hot plate stirrer (JENWAY 1000 hot plate and stirrer, JENWAY Ltd., Staffordshire, UK) at 1100 rpm for 3 h (25 1 C). The extract was ltered through Whatman lter paper (No. 1), centrifuged (3000g, 15 min) (SIGMA Laborzentrifugen 6-10, Deutschland, Germany) and the supernatant obtained was concentrated (50 C) using a rotary evaporator (Buchi Rotavapor R-215 Postfach Flawil, Switzerland). This was freeze dried (LABCONCO, Free Zone 6 Liter, Kansas City, Missouri, USA) and stored at 4 C, until further analysis. 2.6. DPPH radical scavenging activity The free radical scavenging activity of the juice was measured by DPPH (1,1-diphenyl-2-picrylhydrazyl) assay based on the method described by Blois (1958) and the radical scavenging activity was calculated as:

%DPPH inhibition Ao A1 =Ao 100

where, Ao is the absorbance of the control and A1 is the absorbance of the extracts.

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fruit juice processing as they can prolong the shelife of the product and can be used as one of the most reliable indicator to evaluate the overall qualities. Whereas, the Brix can be used for measuring the amount of sugars present in fruits and their juices. In the present study, no signicant changes were observed in the pH and Brix on UV treatments. The pH ranged from 4.39 to 4.37 and the Brix between 9.13 and 8.87 in control and UV treated samples, respectively. UV treatments showed signicant decrease in the acidity, which ranged between 6.73%, 6.54% and 6.24% in control and UV treated samples (30 and 60 min), respectively. 3.2. Colour analysis Signicant differences in the colour parameters were recorded in starfruit juice after UV treatments (Table 2). An increase in the lightness (L) value was recorded corresponding to increased UV treatment time. This increase might be attributed to the degradation of the coloured compounds formed previously due to UV treatment, making the juice more transparent. However, parameters a and b showed a decreasing trend corresponding to UV time of exposure. Previously, Ibarz, Pagan, Panades, and Garza (2005) have made similar observations in fruit juices (apple, peach, lemon juice) wherein decrease in a and b was observed, which was attributed to the destruction of the coloured polymeric compounds and to the enhancement in the browning degree of the fruit juice. Additionally, DE representing the total colour difference (magnitude of colour change) showed a signicant increase after UV treatments.

3.3. Antioxidant assays The results on the effect of UV on antioxidants in starfruits extracted with methanol are shown in Table 3. Application of UV can either enhance or decrease the antioxidants, which are entirely dependent on the exposure time, delivered dose, solvents used and the basic raw material (Alothman et al., 2009a). In this study, methanol was used as an extracting solvent as it has been shown to exhibit potentially high antioxidant activities (Khattak, Simpson, & Ihasnullah, 2008; Ndhlala et al., 2008). A non-signicant increase in the percent inhibition of DPPH was recorded after UV treatments. The DPPH assay involves reaction of specic antioxidant with a stable free radical 2,2-diphenyl-1-picrylhydrazyl wherein a reduction in the DPPH concentration caused by the antioxidant decreases the optical absorbance of DPPH. The increase recorded was from 85.74% up to 87.27% and 88.08% in control and UV treated samples for 30 and 60 min, respectively. With the increase in UV time, a corresponding increase in the levels of total phenols (0.650.69 mg GAE/g at 0 and 60 min, respectively), avonoids (0.230.27 mg CE/g at 0 and 60 min, respectively), avonols (2.322.47 mg QE/100 g at 0 and 60 min, respectively) and antioxidant capacity (31.9333.01 mg/ml AA at 0 and 60 min, respectively) occurred. Overall, considering 0 time as 100%, the percentage change in total phenols after UV exposure was 3.1 and 6.2 at 30 and 60 min, while for avonoids it was 17.4 at 30 and 60 min, respectively. With regard to avonols, the percent change recorded was 1.7 and 6.5 at 30 and 60 min, while for antioxidant capacity it was 1.9 and 3.4 at 30 and 60 min of UV exposure time, respectively. Our results are on par with previous reports on fruits exposed to UV radiation treatments (Alothman et al., 2009b; Wang, Chen, & Wang, 2009). The increase might be attributed to the accumulation of phenolic compounds or their products as a means of defence against UV treatments. Additionally, the increase can be attributed to the enhanced phenylalanine ammonia-lyase and the polyphenol oxidase enzyme activities. Increase in the total phenolics, avonoids and avonols might be advantageous for health conscious consumers as these compounds are known to be potential antioxidants and active free radical scavengers (Alothman et al., 2009a; Balasundram, Sundram, & Samman, 2006). With regard to ascorbic acid, a signicant decrease was recorded which ranged between 27.31 up to 24.57 and 21.77 mg/
Table 4 Effect of UV exposure time on the survival of microorganisms in freshly prepared star fruit juices*. Treatment time (min) APC 1.3 0.58a 1.0 0.00b ND Yeast and mould counts (log cfu/ml) 2.0 2.65a 1.0 1.15b 1.0 0.01b

Table 1 Effects of UV treatment on pH, Brix and titratable acidity content of freshly prepared star fruit juices (n = 3 SD). Treatment time (min) 0 30 60 pH 4.39 0.01 4.37 0.01a 4.37 0.01a
a

Brix 9.13 0.23 8.93 0.12a 8.87 0.12a

a

% Acidity 6.73 0.03a 6.54 0.05b 6.24 0.02c

Values followed by the same letter within the same column are not signicantly different from each other (p > 0.05).

Table 2 Effects of UV treatment on colorimetric parameters L, a and b of freshly prepared star fruit juices (n = 3 SD). Treatment time (min) 0 30 60 L 77.02 0.13a 77.23 0.06a 78.23 0.07b a 2.61 0.07a 2.48 0.06a 1.46 0.09b b 26.77 0.13a 26.42 0.11a 25.71 0.13b

DE
0.43 0.06a 1.98 0.06b

0 30 60

Values followed by the same letter within the same column are not signicantly different from each other (p > 0.05).

ND, not detected; cfus, colony-forming units. * Values are mean of triplicate determination (n = 5 SD).

Table 3 Effect of UV treatment on antioxidant assays in starfruits extracted in methanol (n = 3 SD). Treatment time (min) 0 30 60 DPPH (%) 85.74 0.36a 87.27 0.28 (1.8)
a

Total phenols (mg GAE/g) 0.65 0.06a 0.67 0.02 (3.1)

ab

Antioxidant capacity (mg/ml AA) 31.93 0.2a 32.55 0.21 (1.9)

b

Total avonols (mg QE/100 g) 2.32 0.01a 2.36 0.03 (1.7)

a

Total avonoids (mg CE/g) 0.23 0.01a 0.27 0.01 (17.4) 0.27 0.0b (17.4)
b

Ascorbic Acid (mg/100 ml) 27.31 0.15a 24.57 0.43b (-10.0) 21.77 0.54c (-20.3)

88.08 0.77a (2.7)

0.69 0.02b (6.2)

33.01 0.08c (3.4)

2.47 0.02b (6.5)

Values in parenthesis indicate per cent change after UV exposure over control (0 min taken as 100%). Values followed by the same letter within the same column are not signicantly different from each other (p > 0.05). GAE, gallic acid equivalents; AA, ascorbic acid; QE, quercetin equivalent; CE, catechin equivalent.

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100 ml in control and UV treated juice samples, respectively. Vitamin C is a heat-sensitive bioactive compound that plays a vital role in human health and can act as an antioxidant (Bendich, Machlin, Scandurra, Burton, & Wayner, 1986). The observed decrease in the Vitamin C content of starfruit juice can be attributed to the oxidation process along with the activities of ascorbate oxidase and peroxidase enzymes (Davey et al., 2000). 3.4. Microbiological analysis Consumers are of the opinion that acidic fruits or their juices, due to their low pH, are microbiologically safe. However, reports available indicate the outbreak of food-borne diseases due to the presence of pathogens like: Alicyclobacillus acidoterrestris, Bacillus spp., Escherichia coli O157:H7, Staphylococcus aureus and Salmonella spp. (Tran & Farid, 2004). Table 4 shows the results on the status of microorganisms in starfruit juice subjected to UV treatments. Generally, the UV rays inactivate a microorganism by altering their basic genetic material in the cells (Alothman et al., 2009b). Overall, APC as well as yeast and mould counts decreased signicantly after UV treatments. The APC were completely absent after 60 min of UV treatments, while yeast and moulds could survive even after 60 min. The main reason for yeasts and moulds to exhibit higher resistance than bacteria can be attributed to the basic DNA structure, wherein they are known to posses less pyrimidine nucleosides (thymine and cytosine) base. Additionally, the chemical composition and thickness of the cell wall of yeasts and moulds cells are probably different than bacteria, which render them to be UV resistant (Miller, Jeffrey, Mitchell, & Elasri, 1999). 4. Conclusions Results of the present study indicated that UV-C radiation could be employed for enhancing selected antioxidant compounds along with reduction in the microbial load in starfruit juices. Future research studies are warranted to evaluate the effects of high doses/extended time of UV on the nutrition and sensory qualities. Combination treatment of UV with other non-thermal food preservation methods on improving the juice quality might provide more insight for successful implementation of this novel technology. Acknowledgments The authors gratefully acknowledge the anonymous referees for comments and constructive suggestions provided for improving the manuscript.

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