AGRICULTURAL RESEARCH COMMUNICATION CENTRE

www.arccjournals.com / indianjournals.comAsian J. Dairy & Food Res., 32 (3) : 203-213, 2013

PHYSICO-CHEMICAL, SENSORY AND MICROBIOLOGICAL QUALITY OFKINNOW JUICE STORED IN REFRIGERATED STORAGE CONDITION

Raju Lal Bhardwaj

Directorate of Extension Education,Maharana Pratap University of Agriculture & Technology, Udaipur- 313 001, India

Received: 13-12-2011 Accepted: 20-07-2012

ABSTRACTDifferent fruit juice blends were prepared as (Kinnow juice: Aonla juice: Ginger juice Pomegranate

juice for improving flavour, and nutritive value. The juice blends were preserved by pasteurization(750C or 850C for 15 min.) and by addition of potassium meta-bi-sulphite (500 or 750 ppm). Theseblends were stored in 200 ml colourless glass bottles at (4 + 10C) and tested at three months intervalupto six month. Blends were examined for physic-chemical, sensory evalution and microbialpopulation. The individual effect of juice blending ratio, processing temperature and potassiummeta-bi-sulphite treatment was found to be significant in prolonging storage life and maintainingthe acceptable quality. The juice blends processed at 75oC for 15 min with 750 ppm potassium meta-bi-sulphite was the most effective treatment. However, a minimum microbial population was recordedwith juice processed at 85oC temperature.

Key words: Juice blends, Kinnow juice, Microbial quality, Physico- chemical properties,Sensory quality.

INTRODUCTIONKinnow mandarin is being cultivated on large

scale in Sri Ganganagar and Hanumangarh districts.The area under cultivation of mandarin in Rajasthanis 8,290 hectares and production is 157,460 metrictonnes with 19.0 metric tonnes productivity (NationalHorticulture Database, 2009). Kinnow mandarin isquite important as it has a great variety of wagesdue to its attractive colour, distinctive flavour andbeing rich source of vitamin C, B, -carotene,calcium and phosphorous (Sogi and Singh, 2001).The post-harvest shelf life of kinnow fruit at roomtemperature is very limited (Jawanda and Singh,1973) and shelf life can be extended to a maximumperiod of upto 45 days under refrigerated storageconditions. In view of it’s limited shelf life, the fruitmust be processed to extend it’s availability periodand also to minimize the glut in the market in it’speak season of production. Kinnow mandarin juiceturns bitter after extraction due to conversion oflimonate-a-ring-lactone (non-bitter) to limonin (bittercompound) during storage (Premi et al, 1994).Kinnow , pomegranate and anola fruits are valued

for thei r refreshing juice wi th nutr i t ional,medicinal properties, Ginger juice possessesanti-bacterial and anti fungal properties. Saxenaet.al .(1996) have reported that two or more fruitjuice/pulp may be blended in various propertiesfor the preparation of nectar, ready to serve,bevaerages. Similarly, Jain and Khurdiya (2005)observed that the anola juice was blended withother fruit juice for preparation of ready to serve,beverages, which boosted their nutrit ionalquali ty in terms of vi tamin C content. Topopularise the kinnow mandarin, pomegranate,aonla and ginger juice and its blends amongmasses, i t is necessary to seek meaningfulinformation relating to the development of juiceprocessing technologies to examine untested, oldconcept in various fields of juice processing, de-bittering and storage. Therefore, this study wasaimed at standardizing processing temperature,chemical preservatives and blending ratio ofkinnow mandarin juice in relation to physico-chemical and sensory attributes and microbialcounting during storage.

Author’s e-mail: rajubhardwaj3@gmail.com

204 ASIAN JOURNAL OF DAIRY & FOOD RESEARCH

MATERIALS AND METHODSThe fully matured, freshly harvested kinnow,

pomegranate and aonla fruits and well-developedginger rhizomes were procured from Lal kothi mandi,Jaipur and brought to the Post Harvest TechnologyLaboratory, S.K.N. College of Agriculture, Jobner.

Juice preparations: Fruits were washed with cleanrunning water to remove dust particles and to reducethe microbial load. Peeled kinnow fruits were crushedin screw type juice extractor machine for extractionof juice. Pomegranate fruits were cut into pieces andarils were separated. These arils were passed throughthe juicer for extraction of juice. Aonla and gingerwere sliced with the help of stainless steel knives andcrushed with mixer cum juicer for the extraction ofjuice. The juices were kept for 24 hrs. in refrigerator(4+ 20C) for sedimentation. The clear juice wassiphoned off. The juice was filtered through muslincloth and divided into 5 lots.

Each lot was divided into 2 sub lots andheated separately at 750C or 850C for 15 minutes,respectively in a double-jacketed stainless steel kettle.Again, each sub-lot was divided in to two lots. Acalculated quantity of potassium meta-bi-sulphite(500 or 750 ppm) was dissolved in small quantity ofwater according to the treatments and well mixed inthe blended juice with the help of stirrer. Treatedjuice blends were filled into pre-sterilized 200 mlcapacity, 240 bottles { Treatment combination (20)x Observations (3) x Replication (3) = 180} as soonas possible and tightly closed using crown corkingmachine. These bottles containing juice were storedat refrigerated (4 + 10C) condition and analysed at90 days interval for six months.

Methods of analysis: The physico-chemicalparameters including total soluble solids (TSS) of

the fruit juice was determined by Zeiss hand heldJuice Brix Refractometer with values corrected to200C and expressed as 0Brix. Acidity (as citric acid)was determined by using standard O/N NaOHsolution in the presence of phenolphthalein as anindicator, AOAC (1984). The vitamin C (ascorbicacid) content of the juice was estimated by visualtitration method with 2, 6-dichlorophenol-indonenoldye solution (AOAC, 1984). Total sugars in the juicewere determined by the method of Lane and Eynon(1923), and limonin of the juice was estimated usingthe modified Burulian reagent (Vaks and Litshitz,1981) method. The non-enzymatic browning in thejuice was determined by alcohol extraction method(Klin and Nagy, 1988). Microbiological study wascarried out by a series of dilution and spread platemethod (Ranganna, 1994). In order to find out theconsumer preference juice blend ratio on theorganoleptic evaluation of juice was done by a panelof ten semi-trained judges using 9 point hedonicscale (Amerine et al., 1965). All estimations werecarried out in triplicate, determinations were madefor each attribute and data pertaining to the physico-chemical, sensory quality and microbial populationwere statistically analyzed by using completelyrandomized design (Cochran and Cox 1950).

RESULTS AND DISCUSSIONTotal Soluble Solids (TSS): Retention or minimumincrease in total soluble solids content of juice duringstorage is desirable for preservation of good juicequality (Table 1, 2 & 4). The total soluble solidsincreased with gradual passage of storage time,which might be due to hydrolysis of polysaccharidesinto monosaccharide and oligosaccharides. Theresults revealed that the total soluble solids wassignificantly affected as a result of juice blending ratio(K3 and K5), processing temperature (T1) andpotassium meta-bi-sulphite (P2) during storage. Theminimum increase (10.0% and 6.4%) in total solublesolids was recorded in K5 treatment in both years ofexperimentation, respectively which was statisticallysuperior to other treatments. A similar increase intotal soluble solids content with the increase instorage period was observed in juice of mandarin,sweet orange and lemon by Mehta and Bajaj (1983).The minimum increase in total soluble solids of storedjuice blended with ginger juice and application of

Types of juice Blanding Ratio Treatment Symbol

Kinnow juice: Aonla juice: Ginger juice

100: 0: 0 K1

Kinnow juice: Aonla juice: Ginger juice

95: 5: 0 K2

Kinnow juice: Aonla juice: Ginger juice

92: 5: 3 K3

Kinnow juice: Pomegranate juice: Ginger juice

90: 10: 0 K4

Kinnow juice: Pomegranate juice: Ginger juice

87: 10: 3 K5

Preparation of juice blends

Blending Ratio

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206 ASIAN JOURNAL OF DAIRY & FOOD RESEARCH

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208 ASIAN JOURNAL OF DAIRY & FOOD RESEARCH

potassium meta-bi sulphite (750 ppm) with 750 Cprocessing temperature. It might be due to the factthat the ginger juice checked the microbial growth,which may be responsible for higher metabolic rate.Similar results were also reported by Deka and Sethi(2001) in mango juice blends. The application ofpotassium meta-bi-sulphite also resulted in tolowered rate of hydrolysis of polysaccharides, whichultimately reduced increase in total soluble solids.Similar observation was also reported by Sethi(1985) in stored litchi juice.

Total sugars: The results revealed that the totalsugars content was significantly affected as a resultof juice blending with ginger juice, processingtemperature and adding of potassium meta-bi-sulphite (Table 1, 2 & 4). The total sugars content inthe juice increased apparently during storage, whichmight be due to hydrolysis of polysaccharides intomonosaccharide and oligosaccharides. Theminimum increase (21.5% and 26.03%) in totalsugar content was recorded in K5 treatment in bothyears of experimentation, respectively. It might bedue to fact that the ginger juice checked the microbialgrowth, which may be responsible for highermetabolic rate of juice. The change in total sugarscontent of beverage was almost negligible duringstorage for 6 months in bael: papaya (2:3) pulpsblend (Tandon et al., 2007). Earlier, similar resultswere also reported by Deka and Sethi (2001) inmango juice blends. The minimum increase in totalsugars level in processed juice blends during storagemight be due to the inactivation of enzymes, whichare responsible for decreasing acidity and conversionof polysaccharides into simple sugars. The presentfindings were also in support with the worksconducted by Ranote and Bains (1982) in kinnowjuice. The potassium meta-bi-sulphite also reducedthe conversion of polysaccharides and acids intomonosaccharide and oligosaccharides.

Acidity: There was a significant decrease in titratableacidity content during storage (Table 1, 2 & 4). Thismight be due to conversion of acids into salts andsugars by enzymes particularly invertase (Kumar etal., 1992). It was observed that maximum acidity0.68 per cent and 0.66 per cent was recorded in thekinnow juice blended with ginger juice and aonlajuice (K3) in both years of experimentation,respectively. The minimum decrease (17.10% and

18.91%) in acidity was shown in K5 treatment whichmight be due to inhibitory effect of ginger juice onconversion of acids into sugars and salts by enzymes.Deka and Sethi (2001) found that the acidity of theready to serve beverage prepared from lime- aonla,mango- pineapple, guava- mango blends decreasedwith addition of spices with advancement of storageperiod up to six months under different storageconditions. Similar results were found by Tiwari(2000) in guava and papaya blended ready to serve,and Dhaliwal and Hira (2001) in carrot juice blends.The highest acidity level (0.64% and 0.61%) wasmaintained by the low temperature processing(750C), which might be due to the inactivation ofenzymes, and other reactions responsible fordecrease in acidity. But due to high temperaturetreatment for longer time the acidity decreasedsharply during processing. Similar results were alsoreported by Ghorai (1996) in heat processed kinnowjuice. The juice treated with potassium meta-bi-sulphite showed higher retention of acidity (0.63%and 0.60%) during storage in both years ofexperimentation, respectively. This could be due toalteration in metabolism and enzymatic activity.These findings are in close confirmation with thefinding of Goyle and Ojha (1998) in orange juice.

Ascorbic acid: The ascorbic acid content of the juicedecreased during storage. Aonla juice mixes werebetter in ascorbic acid content but rate of decreasewas very slow with ginger juice blend because gingerjuice might have reduced the oxidation process.Maximum ascorbic acid (43.70 mg/100 ml juice and43.20 mg/100 ml juice) was recorded in kinnow juiceblended with aonla juice i.e. 5 per cent and gingerjuice i .e. 3 per cent (K3) in both years ofexperimentation, respectively. In presentinvestigation, the maximum retention of ascorbicacid (28.4 mg/100ml juice and 28.0 mg/100ml juice)was observed where low temperature (750C),processing was done. This might be due to moreoxidation of ascorbic acid at high temperature.Similar results were also observed by Ranote andBains (1982) in kinnow juice. Comparatively lowerlosses of ascorbic acid was observed in juice samplespreserved with higher concentration (750 ppm) ofpotassium meta-bi-sulphite because higherconcentration of potassium meta-bi-sulphite reducedoxidation of ascorbic acid during storage for longertime.

209Vol. 32, No. 3, 2013

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210 ASIAN JOURNAL OF DAIRY & FOOD RESEARCH

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211Vol. 32, No. 3, 2013

Limonin: A gradual increase in limonin in juiceblends with increase in storage period might be dueto conversion of limonate-a-ring lactone (non-bitter)into limonin (bitter) in the juice (Premi et al. 1994).Among the juice blend prepared with ginger, aonlaand pomegranate juice and processing at lowtemperature (750C), or preserve by highconcentration potassium meta-bi-sulphite (750 ppm)exhibited significantly lesser limonin as comparedto pure unprocessed kinnow juice because blendingof non-bitter juice with bitter one in proper ratioreduced the formation of limonin during storage.Minimum limonin (0.138 mg/ml juice and 0.181 mg/ml juice) was recorded when the juice blended withpomegranate juice (10%) and ginger juice (3%) atthe end of storage in both years of experimentation,respectively. Similar results were also reported byGuadagni et al. (1993) in citrus juice. The beneficialresults of juice processing at 750C temperature mightbe due to inhibition of oxidation of D-ring lactoneand lactones ring into limonin during storage. Theresults are well supported by Berry (2001) in citrusjuice. Similarly the higher concentration ofpotassium meta-bi-sulphite effectively inhibited thehydrolysis of D-ring lactone and lactones rings intolimonin during storage. Similar results were alsoreported by Sethi et al. (1980) in canned kinnowjuice.

Non-enzymatic browning: A linear increase in non-enzymatic browning was observed during 6 monthsof storage irrespective of juice blend (Table 1, 2 &4). The increase in non-enzymatic browning duringstorage might be due to non-enzymatic reaction oforganic acid with sugars or oxidation of phenols,which leads to the formation of brown pigments.Khurdiya and Anand (1981) also reported a gradualincrease in browning and formulat ion ofhydroxylmethylfurfural (dark pigment) in storedphalsa beverage. The minimum increase (25.88%)in non-enzymatic browning in the juice blended withaonla juice (5%) and ginger juice (3%) might bedue to suppression of polyphenol oxide activity byascorbic acid (Arogba et al., 1998), which isabundantly found in the aonla juice. Similar resultswere reported by Jain et al. (2003) in aonla juice.The pomegranate juice was also effective in thereduction of non-enzymatic browning due to highersugar content. The beneficial results of heatprocessing of juice might be due to inhibition of

formulation of hydroxylmethylfurfural and other darkpigments. Similar findings were reported later by Kimet al. (1993) in apple juice. The juice treated withpotassium meta-bi-sulphite showed minimum non-enzymatic browning due to inactivation of enzymesand protective action of â-carotene. Pathak (1988)reported that 100 ppm of potassium meta-bi-sulphitereduced the browning in stored aonla pulp.

Sensory evaluation: Flavour, colour andorganoleptic (bitterness) score of juice blends,decreased with advancement of storage period(Table 1 to 4). The colour, flavour, taste andappearance as well as higher nutrient elements ofthe blends were found to be superior as comparedto the juices prepared from individual fruits. Thejuice blend of kinnow (87%)+ pomegranate (10%)+ginger (3%) recorded higher score for colour (7.33and 7.23), flavour (7.53 and 7.73) and organoleptictaste (7.74 and 7.94) as compared to other blendsat the end of storage in both years of experimentation,respectively. This may be explained as ginger juicechecks microbial and enzymatic activities in storedjuice, which produce off flavour and change innatural colour and taste. Tandon et al., (2007)reported that addition of papaya pulp with bael pulpwas found to be very effective in checking thebrowning and improving the appearance of thebeverage. They also observed that the beverageprepared from 2:3 blend of bael: papaya pulp scoredmaximum (7.4 out of 10.0) after six months ofstorage. The result is well supported by Gowda(1995) in case of mango and papaya blend. Thebeneficial results of thermal processing might be dueto inhibition of polyphenol oxidase and the enzymeinvolved in discolouration and developing of offflavour during storage. Later, the opinion wassupported by works of Kim et al.(1993) in apple juice.The potassium meta- bi-sulphite was found to beeffective in retention of good flavour, colour andorganoleptic taste of fruit juice during the entirestorage period. Similar results were also reported byMehta and Bajaj (1983) in citrus juice. All thesamples were found acceptable up to 6 months ofstorage. Overall qualities including colour, flavourand organoleptic scores were better in the juice whichwas blended in the ratio of kinnow juice:pomegranate juice: ginger juice (87: 10: 3) withprocessing at 750C temperature and addition of 750ppm potassium meta-bi-sulphite.

212 ASIAN JOURNAL OF DAIRY & FOOD RESEARCH

Microbial population: The untreated fruit juicesand pulp are highly contaminated with bacteria,yeast and mould. The data presented in Table 1, 3and 5 showed the minimum increase in bacteria,yeast and mould population, when juice wasblended with ginger juice, processed at 850Ctemperature with addition of potassium meta-bi-sulphite (750 ppm). The juice blended with 3 percent ginger juice (K5) was lowest in bacterial (2.1x103

and 2.5x103 TVC/ml juice), mould (1.2x103 and 1.3x103 cfu/ml juice) and yeasts (9.0x102 and 9.9x102

cfu/ml juice) population as recorded at the end ofstorage period (six months) in both years ofexperimentation. The processing of juice at 850Cfor 15 min. of holding time could be considered aseffective processing time and temperature tominimize microbial growth in the juice blends. Similarresults were also reported by Sethi et al. (1980) andGhorai (1996) in kinnow juice.

CONCLUSIONIt may be concluded that formulation of

mixed fruit juice beverage is possible to meetconsumer preferences. These juice blends can be

stored effectively for a period of 6 months. Theirtotal soluble solids, total sugars, limonin and non-enzymatic browning increased with increasedperiod of storage but acidity, ascorbic acid andsensory qual i ty score was reduced wi thadvancement of storage. The juice blend (Kinnowjuice 87%+ Pomegranate juice 10% + Gingerjuice 3%) and followed by (Kinnow juice 92%+ Aonla juice 5% + Ginger juice 3%), processedat 750C for 15 minutes with 750 ppm potassiummeta-bi-sulphite proved to be the most effectivetreatment for physico-chemical and sensoryscores of the juice blends. But the minimummicrobial population (Bacteria, 2.0 x 103 and 2.5x 103 TVC/ml juice; yeast, 9.0 x 102 and 9.9 x 102

cfu/ml juice; mould, 1.2 x 103 and 1.3 x 103 cfu/ml juice) was recorded in juice processing at 850

C temperature with same treatment combination.The maximum B: C. ratio (1.5:1) was obtainedby making squash from the juice blend (kinnowjuice 87: pomegranate juice 10: ginger juice3) +processed at 750C + KMS (750ppm) and storedunder refrigeration condition.

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