jfpp_419 599..

610

STORAGE STABILITY OF CLARIFIED BANANA JUICE

FORTIFIED WITH INULIN AND OLIGOFRUCTOSE

MUHAMMAD SOHAIL YOUSAF1,3, SALMAH YUSOF1,

MOHD YAZID BIN ABDUL MANAP1 and SURAINI ABD-AZIZ2
1
Department of Food Technology
Faculty of Food Science and Technology
2
Department of Biotechnology
Faculty of Biotechnology and Biomolecular Science
Universiti Putra Malaysia
43400 UPM, Serdang, Selangor, Malaysia

Accepted for Publication March 2, 2009

ABSTRACT

Clarified banana juice fortified with inulin and oligofructose were stored
for 8 weeks at 4, 25 and 35C. Changes in physicochemical characteristics (pH,
total soluble solids [TSS], titratable acidity, sucrose, reducing sugars and
turbidity), microbial count and sensory quality were determined. No differ-
ences were observed for pH and titratable acidity for all the stored juice
samples. However, increase in turbidity was observed in all the juice samples,
whereas juice samples stored at 35C recorded highest increases. Increase in
reducing sugars (glucose and fructose) was also observed during storage,
particularly at 25 and 35C. TSS values were observed fluctuating for all the
samples. No microbial growth was recorded for all the juice samples stored at
three different temperatures. Sensory results for taste, flavor and odor revealed
no difference until the seventh week of storage; however, the overall accept-
ability of the juice stored at 4C was rated highest as compared with juice
samples stored at 25 and 35C. Overall, the quality of juice stored at 4C was
rated highest not only for all the sensory characteristics but also less turbidity
problem compared with juices stored at 25 and 35C.

PRACTICAL APPLICATIONS

Today, consumers are increasingly looking for foods which not only have
better taste but also offer health benefits. The present study was conducted to

3
Corresponding author. FAX: +92-42-5789303-4; EMAIL: sohailft@yahoo.com

Journal of Food Processing and Preservation 34 (2010) 599–610.

DOI: 10.1111/j.1745-4549.2009.00419.x 599
© 2009 Wiley Periodicals, Inc.
600 M.S. YOUSAF ET AL.

determine the shelf life of banana juice which has been fortified with inulin
and oligofructose. The results of the study indicate that inulin and oligofruc-
tose fortification in clarified banana juice did not significantly affect its physi-
cochemical, microbiological and sensory characteristics over a period of
8 weeks. It also signifies the acceptability of clarified banana juice after
fortification with inulin and oligofructose and by keeping at different storage
conditions. The study can serve as a reference for food industry regarding shelf
stability of fortified, clarified banana juice at different storage temperatures.

INTRODUCTION

Recent progress in bioscience supports the hypothesis that beyond provid-

ing nutrition, diet also may modulate various functions in the body that are
relevant to health. This has led to the concept of “functional food.” A food is said
to be “functional” if it contains a food component that affects one or a limited
number of function(s) in the body in a targeted way so as to have positive effects
(Bellisle et al. 1998). Keeping in view this context, there is a growing trend for
the production and consumption of functional foods all over the world, espe-
cially the developed world. Foods can be modified by the addition of phy-
tochemical, bioactive peptides, -3 polyunsurated fatty acids and probiotics
and/or prebiotics to become functional (Berner and O’Donnel 1998).
Among the various functional food ingredients, inulin and oligofructose
are natural food ingredients commonly found in varying percentages in dietary
foods. They are present as plant storage carbohydrates in a number of vegetables
and plants including wheat, onion, banana, garlic and chicory. In fact, it has been
estimated that Americans consume on average 1–4 g of inulin and oligofructose
per day and Europeans average 3–10 g/day (Van Loo et al. 1995).
Inulin and oligofructose consumption has some promising health benefits
such as reduced caloric value because of the nondigestibility characteristics in
the upper gastrointestinal-tract, stimulation of health benefit bacteria, e.g.,
bifidobacteria and increase absorption of calcium (Gibson et al. 1995). Inulin
has a neutral, clean flavor and is used to improve mouthfeel, stability and
acceptability of the low fat foods. Oligofructose has a sweet, pleasant flavor
and is highly soluble (Niness 1999). Both can be used to fortify foods with
fiber without contributing any deleterious organoleptic effects, and to improve
the flavor and sweetness of low calorie food. Inulin and oligofructose possess
several functional and nutritional properties, which may be used to formulate
innovative healthy foods for today’s consumer.
Banana is a tropical fruit and is extensively grown in Malaysia and other
tropical parts of the world. Banana is the only fruit that has been reported as a
source of inulin and oligofructose at 0.3–0.7%. In addition to providing widely
 STORAGE STABILITY OF FORTIFIED BANANA JUICE 601

appreciated flavor and aroma, it also has potential health benefit being a natural
source of inulin and oligofructose, which can have a positive effect on the host
health. As banana is mostly consumed as fresh because of its abundance and
easy availability, there has been no serious effort made to convert it to in any
other value-added product such as banana juice, banana powder, banana
purees, etc.
In an early part of this study, attempts were conducted to develop clarified
banana juice fortified with inulin and oligofructose. Therefore, the present
study was conducted to evaluate the effect of storage temperature and time on
the physicochemical, microbiological and sensory characteristics of clarified
banana juice that has been fortified with inulin and oligofructose at 2 and 10%,
respectively. The objective was to determine the most suitable conditions for
extending its shelf life.

MATERIAL AND METHODS

Fully ripened Berangan bananas cultivar were purchased from the local
market to prepare clarified banana juice. Inulin and oligofructose were
obtained from Orafti, Kuala Lumpur, Malaysia. Pectinase enzyme (Pectinex
Ultra-SPL) was provided by Novozyme, Kuala Lumpur, Malaysia. Other
added ingredients were sugar (sucrose), water and benzoic acid.
Banana fruits were peeled and sliced into thin pieces. Extraction of the
juice was carried by heating 1:2 of fruit and water at 95C for 2 h. After
extraction, the juice was cooled and filtered through muslin cloth to remove the
pulp. To clarify the juice, the enzyme Pectinex Ultra-SPL was added at 0.1%
and incubated at 45C for 2 h. Juice was then again heated at 90C for
10–20 min to inactivate the enzyme. Clarified juice obtained was centrifuged
at 3,000 rpm for 10 min and filtered through 0.45 mm nylon membrane filter to
remove the suspended fine particles of pulp. Total soluble solids (TSS) of the
juice were maintained at 12°Brix by adding sugar and 0.05% benzoic acid was
added as a preservative. Pasteurization was carried out at 95C for 5–10 min to
destroy any microbial population. Inulin and oligofructose were then added at
the rate of 2 and 10%, respectively, to fortify the clarified banana juice and
juice was then hot filled into glass bottles.
Juice was stored at 4, 25 and 35C for 8 weeks. Analyses were conducted
for physicochemical, microbiological and sensory characteristics after a
1-week interval during the storage period.

Physicochemical Determinations
pH was determined by a glass electrode pH meter (Cyberscan 500,
EUTECH Instruments Singapore) using pH 4.0 and 7.0 as buffer for calibra-
602 M.S. YOUSAF ET AL.

tion. Acidity was determined by the method described by Ranganna (1977).

TSS content was determined by using PR-101 Otago digital refractometer
(Atago, Tokyo, Japan) with a scale of 0–45°Brix. Results were expressed as
°Brix. Sugars were determined by high-pressure liquid chromatography as
described by Hunt et al. (1977). Turbidity was determined by using 2100P
Turbidometer (Hach, Loveland, CO).

Microbial Count
Samples were tested for total plate count and yeast and mold count using
the methods of Pestka (1993).

Sensory Evaluation
Eight trained panelists were used for sensory evaluation using a hedonic
preference test. Taste (sweetness), flavor, odor and overall acceptability of the
samples were rated on a 9-point hedonic scale in which 9 = like extremely and
1 = dislike extremely (Larmonde 1976).

Statistical Analysis
Data obtained was analyzed by two-way analysis of variance and signifi-
cant differences were analyzed by Duncan’s multiple range test, with range at
P > 0.05 using Statistical Analysis System (SAS 1985).

RESULTS AND DISCUSSIONS

Acidity and pH
Titratable acidity values for all samples did not change during storage at
various temperatures during the 8-week period (data not shown). Even so, a
slight decrease was actually observed for all samples. This decrease in acidity
could be partly caused by copolymerization of organic acids with products of
browning reactions (Babsky et al. 1986). In addition, the organic acids can
react with reducing sugars to form brown pigments (Lewis et al. 1949). Mont-
gmoery et al. (1982) reported that there was a slight decrease in titratable
acidity of grape juice stored at -1C for 261 days. Ewaidha (1992) also
observed a slight decrease in titratable acidity of apple juice and pineapple
juice during a 1-year period of storage at 5, 24, 33 and 42C.
pH values of all the juice samples followed the same pattern as acidity.
Storage temperature and time did not cause any differences. Similar results
 STORAGE STABILITY OF FORTIFIED BANANA JUICE 603

were obtained by Toribio and Lozano (1984) who reported that the pH values
of apple juice concentrate did not change during storage at 4, 25 and 35C for
120 days of storage period.

TSS
A fluctuating trend was observed for all the juice samples stored at
various temperatures over the 8-week storage period. This fluctuating trend
may be because of the hydrolysis of sucrose, inulin and oligofructose into
reducing sugars (glucose and fructose), especially at higher temperatures.
Another possibility for the decreased TSS contents during storage was
because the reducing sugar reacts with amino acids to form HMF during the
Maillard reaction (Burdurlu and Karadeniz 2003). From the results obtained as
well, it can be proposed that no microbial spoilage was present, as there were
no drastic changes in the TSS values observed. In normal condition, TSS will
decrease if there is microbial spoilage, as the microorganism will consume
nutrient in the juice, which mostly include soluble protein and simple reducing
sugars.

Turbidity
Figure 1 shows the turbidity values for juice samples stored at 4, 25 and
35C for 8 weeks. A difference for turbidity of juice among all the samples

60

50

40
Turbidity (NTU)

30

20

10

0
0 2 4 6 8 10
Time (weeks)

FIG. 1. CHANGES IN THE TURBIDITY OF THE CLARIFIED BANANA JUICE FORTIFIED

WITH INULIN AND OLIGOFRUCTOSE STORED AT 4 (䉬), 25 ( ) AND 35C (䉱) FOR THE
8-WEEK STORAGE PERIOD
604 M.S. YOUSAF ET AL.

stored at different temperature during the whole storage period. Juice stored at
35C showed a marked increase in turbidity of juice as compared with juice
stored at 4 and 25C. The increase in turbidity during storage is frequently
associated with juices processed and filtered at temperatures higher than the
storage temperature. Johnson et al. (1968) observed after haze in the apple
juice during storage whose chemical nature indicates that it is formed from the
polymerization of lower molecular weight apple catechins (polyphenols) and
proanthocyanidins oxidized during milling and pressing of the apples. Similar
cause of post-bottling haze in clarified banana juice may be polymerization of
low molecular weight tannins with proteins molecules. The similar insoluble
tannin–protein complexes may leads to the formation of haze and eventually
the fall out of the solution as sediments.
In our study, turbidity was found to be higher at higher storage tempera-
tures, and this shows that temperature also plays a prominent role in affecting
the rate of haze formation. Rate of formation of tannin–protein complex,
which are responsible for haze and sedimentation in juice, is higher at this
temperature than the ambient and chill temperature.
As phenolic compounds and proteins might be the major contributors for
the haze formation of haze and sediment in the clarified banana juice, a more
suitable approach to prevent or reduce its occurrence could be employed.
Siebret (1999) reported that protein/polyphenol ratio greatly influenced the
amount of haze formed. The maximum amount of haze is formed when the
number of binding sites in proteins roughly equals the total concentration of
polyphenols ends. Therefore, reducing either polyphenol concentration or
protein concentration might result in less haze formation.
A number of fining agents or adsorbents are commonly used in the
stabilization of beverages, such as bentonite and polyvinylpolypyrorolidone
(PVPP). Bentonite is often used to stabilize fruit juices and wine because it
effectively and indiscriminately removes protein, whereas PVPP adsorbs haze-
active polyphenols (Siebret 1999).
If the methods employed nonspecifically remove all protein or all
polyphenols, then the resulting clarified banana juice would be a juice of lower
nutritional quality, or its mouthfeel or taste may be compromised and it can
become insipid because tannins contribute to the “mouth feel” of the juice.

Sugars
Figure 2 shows the sucrose contents of juice samples stored for 8 weeks
at various temperatures. Results indicated that as the storage time increased,
the sucrose content of samples stored at 4C decreased gradually. But
samples stored at 25 and 35C showed a slight increase in sucrose contents.
The increase in sucrose contents was because of the hydrolysis of inulin and
 STORAGE STABILITY OF FORTIFIED BANANA JUICE 605

6
Sucrose (%)

0
0 2 4 6 8 10
Time (weeks)

FIG. 2. CHANGES IN SUCROSE CONTENTS OF THE CLARIFIED BANANA JUICE

FORTIFIED WITH INULIN AND OLIGOFRUCTOSE STORED AT 4 (䉬), 25 ( ) AND 35C (䉱)
FOR THE 8-WEEK STORAGE PERIOD

oligofructose at these storage temperatures as chemically inulin and oligof-

ructose are polymers of fructose with GFn molecules in which fructose mol-
ecules are attached by b (2–1) linkage with glucose molecules attach at the
terminal end having a (1–2) linkage same as in sucrose. Sucrose decrease in
samples stored at 4C showed that inulin and oligofructose did not hydrolyze
at this temperature much as compared with juice samples stored at 25 and
35C, and this decrease in sucrose contents was caused by the hydrolysis of
sucrose to reducing sugars (glucose and fructose). Glasstone (1946) reported
that the rate of sucrose hydrolysis is a function of reactants, temperature and
acid catalyst concentration. Results obtained in this study for the hydrolysis
of sucrose at different storage temperatures were found comparable with
results obtained by Babsky et al. (1986) who reported hydrolysis of sucrose
in apple juice concentrate after 111 days of storage at 37C. Similarly,
Ewaidha (1992) also noted a similar decrease in sucrose contents at different
temperatures for apple and pineapple juices stored for a 12-month period at
5, 24, 33 and 42C. Moreover, about 10% hydrolysis of inulin and oligof-
ructose have been reported at pH > 4 and temperature >10C as their storage
stability (Orafti 2002).
Figures 3 and 4 shows the reducing sugars (fructose and glucose) con-
tents of banana juice over 8 weeks of storage at 4, 25 and 35C. Marked
increase in reducing sugar levels were observed for all the samples stored at
various temperatures as the storage period increased. Reducing sugar levels
of the juice samples stored at 4C were lower than those of samples stored at
25 and 35C. This increase may be because of higher storage temperatures
606 M.S. YOUSAF ET AL.

4
3.5
3
Fructose (%)

2.5
2
1.5
1
0.5
0
0 2 4 6 8 10
Time (weeks)

FIG. 3. CHANGES IN FRUCTOSE CONTENTS OF THE CLARIFIED BANANA JUICE

FORTIFIED WITH INULIN AND OLIGOFRUCTOSE STORED AT 4 (䉬), 25 ( ) AND 35C (䉱)
FOR THE 8-WEEK STORAGE PERIOD

3.5

3
Glucose (%)

2.5

1.5

0.5

0
0 2 4 6 8 10
Time (weeks)

FIG. 4. CHANGES IN GLUCOSE CONTENTS OF THE CLARIFIED BANANA JUICE

FORTIFIED WITH INULIN AND OLIGOFRUCTOSE STORED AT 4 (䉬), 25 ( ) AND 35C (䉱)
FOR THE 8-WEEK STORAGE PERIOD

that facilitated the conversion of sucrose as well as inulin and oligofructose

to reducing sugars. These results showed that the rate of conversion of
sucrose, inulin and oligofructose was affected by both storage time and tem-
perature. Similar results of increase in reducing sugars were reported by
Babsky et al. (1986), Ewaidha (1988, 1992) and Yusof et al. (2000) in their
storage studies of apple juice, tomato juice, pineapple juice and sugarcane
juice, respectively.
 STORAGE STABILITY OF FORTIFIED BANANA JUICE 607

Microbiological Analysis
During the 8 weeks of storage period at various temperatures, only two
samples had yeast and mold contamination (data not shown). This contami-
nation may have occurred during preparation of samples for measurement. As
all the juice samples contained no microorganism from 0 to 8 weeks of storage,
it can therefore be assumed that microorganisms were destroyed and not able
to grow in juice during storage because of the effect of heat treatment during
processing of the juice and the addition of sodium benzoate as a preservative.
The two previous factors are known to be inhibitors of microbial growth.

Sensory Analysis

Taste. Mean rating for the taste of juice samples over 8 weeks of storage
at various temperatures ranged from 6 (likely slightly) to 7 (like moderately)
(data not shown). No difference was found for taste in terms of sweetness
among the juice samples stored at various temperatures for the whole storage
period.

Flavor. Ratings by judges for juice samples stored at 4C ranged from 6

(likely slightly) to 7 (like moderately) and at 25 and 35C from 5 (neither like
nor dislike) to 7 (like moderately) (data not shown). No difference was found
between all the treatments until sixth week of storage; however, after the sixth
week, difference was found between juice samples stored at 4C and samples
stored at 25 and 35C until the end of storage period with samples stored at 4C
rated higher in terms of banana flavor as compared with samples stored at 25
and 35C.

Odor. Mean rating of odor by panelists for juice samples stored at 4C

ranged from 5 (neither like nor dislike) to 7 (like moderately) and at 25 and
35C from 4 (dislike slightly) to 6 (like slightly) (data not shown). No differ-
ence was found between all the treatments stored at various temperature until
the seventh week of storage, and only the last week of storage showed differ-
ence between samples stored at 4C as compared with samples stored at 25 and
35C. Overall, the score for odor of samples stored at 4C rated higher by
panelists as compared with samples stored at 25 and 35C.

Overall Acceptability. Figure 5 shows the overall acceptability of juice

samples stored for 8 weeks at various temperatures. Mean rating for overall
acceptability of the juice samples stored at 4C ranged from 5 (neither like nor
dislike) to 7 (like moderately) and at 25 and 35C from 4 (dislike slightly) to 7
(like moderately). These ratings revealed that juice samples showed no differ-
608 M.S. YOUSAF ET AL.

6
Overall acceptability

0
0 2 4 6 8 10
Time

FIG. 5. CHANGES IN OVERALL ACCEPTABILITY OF THE CLARIFIED BANANA JUICE

FORTIFIED WITH INULIN AND OLIGOFRUCTOSE STORED AT 4 (䉬), 25 ( ) AND 35C (䉱)
FOR THE 8-WEEK STORAGE PERIOD

ence until the fourth week of storage among all the storage samples. After the
fourth until the sixth week, difference was observed among the samples stored
at 4, 25 and 35C, and after the sixth week until the end of the storage period,
there was a difference shift between the samples stored at 4C and samples
stored at 25 and 35C. Juice samples stored at 4C were rated highest in terms
of overall acceptability as compared with the other two juice samples stored at
25 and 35C.

CONCLUSION

Results of the study indicate that the juice quality remains stable in terms
of its different physicochemical characteristics for 8 weeks at different storage
temperatures except for turbidity. Continuous increase in turbidity during
storage in all the samples was found to be a critical quality problem in clarified
juice, especially for juice stored at 35C. However, juice stored at 4C was found
to have the least turbidity problem as compared with the other juices stored at
two other storage temperatures. All the stored juice samples were found
microbiologically stable. Sensory results for taste, flavor, odor and overall
acceptability indicated that the quality of juice samples stored at 25 and 35C
decreased slightly at the end of the storage period, but the juice remained
acceptable during the 8 weeks of storage period, with samples stored at 4C
being rated best for all the sensory parameters. Turbidity problem should be
solved before any further extended storage studies be carried out.
 STORAGE STABILITY OF FORTIFIED BANANA JUICE 609

REFERENCES

BABSKY, N.E., TORIBIO, J.L. and LOZANO, J.E. 1986. Influence of

storage on the composition of clarified apple juice concentrate. J. Food
Sci. 51, 564–567.
BELLISLE, F., DIPLOCK, A.T., HORNSTRA, G., KOLETZKOS, B., ROB-
ERFOID, M., SALMINEN, S. and SARIS, W.H.M. 1998. Functional
food science in Europe. Br. J. Nutr. 80, S1–S193.
BERNER, L. and O’DONNEL, J. 1998. Functional foods and health claim
legislation: Application to dairy foods. Int. Dairy J. 8, 355–362.
BURDURLU, H.S. and KARADENIZ, F. 2003. Effect of storage on nonen-
zymatic browning of apple juice concentrates. Food Chem. 80, 91–97.
EWAIDHA, E.H. 1988. Studies on the commercially canned juices produced
locally in Saudi Arabia: Part 2- Physicochemical, organoleptic and micro-
biological assessment. Food Chem. 29, 81–96.
EWAIDHA, E.H. 1992. Studies on the commercially canned juices produced
locally in Saudi Arabia: Part 3- Physicochemical, organoleptic and micro-
biological assessment. Food Chem. 44, 103–111.
GIBSON, G.R., BEATTY, E.R., WANG, X. and CUMMINGS, J.H. 1995.
Selective stimulation of bifidobacteria in human colon by oligofructose
and inulin. Gastroentrology 108, 975–982.
GLASSTONE, S. 1946. Textbook of Physical Chemistry, D. Van Nostrand,
Princeton, NJ.
HUNT, R.S., JACKSON, P.A., MORTLOK, R.E. and KIRTE, R.S. 1977.
Quantitative determination of sugar in food stuffs by high performance
liquid chromatography. Analyst 102, 917–920.
JOHNSON, G., DONNELLY, B.J. and JOHNSON, D.K. 1968. The chemical
nature and precursors of clarified apple juice sediment. J. Food Sci. 33,
254–257.
LARMONDE, E. 1976. Laboratory Methods for Sensory Evaluation of Food,
Publication 1637, Departmentt of Agriculture of Canada, Ottawa,
Canada.
LEWIS, V.M., ESSELEN, W.B. and FELLERS, C.R. 1949. Non-enzymatic
browning of foodstuffs. Nitrogen free carboxylic acids in the browning
reaction. Ind. Eng. Chem. 41, 2591–2594.
MONTGMOERY, M.W., REYES, F.G.R., CORNWELL, C. and BEAVERS,
D.V. 1982. Sugars and acid analysis and effect of heating on color
stability of northwest Concord grape juice. J. Food Sci. 47, 1883–1885.
NINESS, K.R. 1999. Inulin and oligofructose: What are they? J. Nutr. 129,
1402S–1406S.
ORAFTI. 2002. Product Book, Raftilose and Raftiline, pp. 65–78, Orafti,
Belgium.
610 M.S. YOUSAF ET AL.

PESTKA, J.J. 1993. Food Microbiology Laboratory Manual. Department of

Food Science and Human Nutrition, Michigan State University, East
Lansing, MI.
RANGANNA, S. 1977. Manual of Analysis of Fruits and Vegetables Products,
pp. 1–20, McGraw Hill, New Delhi, India.
SAS 1985. SAS User’s Guide: Statistics, 5th Ed., SAS Institute, Cary, NC.
SIEBRET, K.J. 1999. Effects of protein-polyphenol interactions on beverage
haze, stabilization, and analysis. J. Agric. Food Chem. 47(2), 353–
362.
TORIBIO, J.L. and LOZANO, J.E. 1984. Non-enzymatic browning in apple
juice concentrate during storage. J. Food Sci. 49, 889–92.
VAN LOO, J., COUSSEMENT, P., DELEENHEER, L., HOEBREGS, H. and
SMITS, G. 1995. On the presence of inulin and oligofructose as natural
ingredients in the western diet. Crit. Rev. Food Sci. Nutr. 35, 525–552.
YUSOF, S., SHIAN, L.S. and OSMAN, A. 2000. Changes in quality of
sugarcane juice upon delayed extraction and storage. Food Chem. 68,
395–401.