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Study of inhibition of browning of clarified banana juice

Article in Journal of Dairying, Foods & Home Sciences · May 2016

DOI: 10.18805/ajdfr.v35i2.10723

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Asian J. Dairy & Food Res., 35 (2) 2016 : 155-159 AGRICULTURAL RESEARCH COMMUNICATION CENTRE
Print ISSN:0971-4456 / Online ISSN:0976-0563 www.arccjournals.com

Study of inhibition of browning of clarified banana juice

A.R. Tapre* and R.K. Jain
Department of Food Processing Technology, A.D. Patel Institute of Technology,
New V V Nagar, Anand-388 121, India.
Received: 17-04-2015 Accepted: 19-02-2016 DOI: 10.18805/ajdfr.v35i2.10723
ABSTRACT
Browning is the second largest cause of quality loss in fruits and vegetables. It seems to be the most severe problem during
banana pulp processing. Effect of two anti-browning pretreatments viz. addition of ascorbic acid (200-1200 ppm) and
blanching the peeled fruits (900C for different periods) prior to enzymatic clarification banana juice were studied.
Effectiveness of anti-browning pretreatments to the pulp was evaluated by measuring the browning in terms of Browning
Index in prepared clarified juices. Clarified banana juice obtained from untreated pulp had dark brown colour with initial
browning index value of 0.411 which continuously increased and reached to 0.586 at 24 h of storage. Pretreatment of pulp
with 1000 ppm of ascorbic acid or heating at 900C for 6 minute was observed to retain the natural colour of prepared
clarified juice without marked changes up to 4 h of storage at room temperature. However preheating of banana pulp
observed to be caused cooked flavour and altered the original taste of the clarified juices.
Key words: Ascorbic acid, Browning, Browning Index, Clarified banana juice, Preheating.
INTRODUCTION banana pulp becomes dark and the juice released turns
Banana (Musa sp.) is the fourth most important food brownish with unattractive appearance and also may reduce
crop in the world after rice, wheat and maize with a world the nutritive value of the prepared juices (Eskin et al., 1971;
production of around 101992743 MT in 2012 (FAO, 2014). Scott, 1975).
But a large percentage of the crop is typically unsuitable for Several methods such as the addition of browning
the fresh market since it is too mature for shipment. Banana inhibitors and the exclusion of oxygen as well as thermal
could compete in the market either as banana juices or as processing have been used to inhibit enzymatic browning. A
mixture of other juices because of its widely appreciated chemical commonly used is sodium metabisulphite.
flavour and high nutritive value. Banana juice can also be However, one problem with this chemical is that it may
utilized for wine production. However, banana juice is turbid, induce asthmatic episodes in a segment of the asthmatic
gray in colour, very viscous, tends to settle during storage population and thus, it need to find alternatives to sulphite
and therefore must be clarified prior to commercialization. (Sapers, 1993). The use of browning inhibitors in food
The problems associated with banana juice processing, a processing is restricted by considerations relevant to toxicity,
high viscosity and browning seems to be the most severe. wholesomeness and effect on taste, flavour, texture and cost.
The turbidity and viscosity of banana juice are Therefore present study was undertaken in order to to find a
mainly caused by the polysaccharides in the juice such as suitable method to inhibit the activity of PPO in banana and
pectin and starch. Depectinization using pectinase enzyme prevent discoloration during the course of juice clarification.
could effectively clarify fruit juices. Pectinase hydrolyzes MATERIALS AND METHODS
pectin and cause pectin-protein complexes to flocculate Bananas (Musa sp.) of variety ‘Robusta’ were
(Koffi et al., 1991; Yusof and Ibrahim, 1994; Lee et al., procured at the maturity stage 1 (all green) from selected
2006). banana orchards near Anand city (India). The procured
During the preparation of clarified banana juice, material was sorted, tagged and kept for ripening as per the
banana pulp is highly susceptible to enzymatic browning method suggested by Kulkarni et al., (2010) up to 7th stage
while pulping. Browning reaction of banana pulp results from of maturity. Commercial pectinolytic enzyme i.e. Pectinex
the enzymatic oxidation of dopamine (3,4 -dihydroxyphenyl Ultra SP-L (with enzymatic activity 26000 PG U per ml ,
ethylamine) by polyphenoloxidase (Griffiths, 1959, Ranveer optimum pH 3.5 to 6 and temperature below 500C) and
et al., 2010) leading to the production of brown pigments. cellulase enzyme i.e. Celluclast 1.5L with cellulose activity
Longer incubation time for enzymatic treated banana pulp 158 U per ml were obtained from Novozymes South Asia
will responsible for more oxidation and production of Pvt. Ltd., Bangalore and other chemicals from local
coloured products. Thus, at the end of enzymatic treatment, suppliers.
*Corresponding author’s e-mail: tapreajay @gmail.com.
156 ASIAN JOURNAL OF DAIRY AND FOOD RESEARCH
Anti-browning pretreatments to pulp: Clarified banana alcohol in a using Shimadzu UV–VIS spectrophotometer.
juice were prepared by using the commercial pectinolytic Browning index (A at 420 nm) was expressed as absorbance
enzyme i.e. Pectinex Ultra SP-L and cellulase enzyme i.e. value.
Celluclast 1.5L as per the process suggested by Tapre and Statistical analysis: Analysis of variance (ANOVA) was
Jain (2014). The banana fruits were washed, peeled manually used to detect the effect of pretreatment during different time
and cut into small pieces. Pieces were then pulped using a intervals. Tukey’s Multiple Comparison Test was employed
kitchen blender for 2 minute into pulp. After the pulping, for mean separation. The level of significance was at 5%.
500g pulp was subjected to enzyme treatment i.e. 0.11% The analysis was performed using the Daniel’s XL Toolbox
Pectinex Ultra SP-L at temperature (400C) for 145 minute version 4.00.
and then treated with 0.05% cellulase enzyme i.e. Celluclast
1.5L. At the end of the treatment, the enzymes in the pulp Physico-chemical parameters of clarified banana juices:
was inactivated by heating at 90 0C for 5 minute and The juice yield was estimated as percentage of the juice
immediately cooled to room temperature. The enzyme treated obtained based on the initial pulp. The clarity of the juice
pulp was centrifuged at 2900 G/15 minute and the obtained was determined by measuring % transmittance at a
supernatant was filtered through a fine mesh nylon cloth wavelength of 660 nm using UV–VIS spectrophotometer
spread on glass funnel and juice was collected. (Shimadzu, Model: BL-220-H). Distilled water was used as
the reference. Titratable acidity was determined by direct
Prior to juice clarification, following anti-browning titration of diluted pulp with 0.1N NaOH as described by
pretreatments were applied to banana pulp to inhibit the PPO Ranganna (2000) and TA was expressed as a % of malic
activity and enzymatic browning: acid. TSS was measured as 0 Brix at 200 C with Hand
a) Addition of ascorbic acid at concentrations of 200-1200 Refractometer. The necessary corrections were made when
ppm to the peeled fruits during blending and then clarified the measurements were carried out at temperatures other than
juices were prepared 200C. Total sugar content were analyzed as per Shaffer -
b) Blanching the peeled fruits in hot water temperature Somogyi method and starch in the alcohol insoluble solids
900C for different periods up to 10 minutes. These heated was hydrolyzed using HCL and the resultant reducing sugar
samples were quickly cooled to room temperature by was determined by Shaffer -Somogyi method given in
dipping in cold water then blended and clarified juices were Ranganna (2000).
prepared.
RESULTS AND DISCUSSION
Browning of the prepared clarified juices obtained Ascorbic acid pre-treatment: To evaluate the effectiveness
was measured immediately after extraction and at intervals of ascorbic acid, banana pulp was pretreated with different
of 2, 4, 6 and 24 h. concentrations of ascorbic acid (200-1200 ppm) prior to
Evaluation of effectiveness of anti-browning pretreatments: clarification. Pretreated banana pulp was then treated with
Effectiveness of anti-browning pretreatments to the pulp was optimized enzymatic conditions to prepare clarified juices.
evaluated by measuring the browning in terms of Browning In the prepared juices, browning in terms of Browning Index
Index in prepared clarified juices. To measure the enzymatic was measured immediately after preparation (initial value)
and non enzymatic browning, 20 ml of the juice was mixed and after 2 h interval for the period of 6 h. Also the reading
with 30 ml of ethyl alcohol in a glass stoppard flask of 24 h storage was recorded.
(Ranganna, 2000). The mixture was shaken thoroughly and Results depicted in Table 1 indicates that clarified
filtered through Whatman No.42 filter paper. The absorbance banana juice obtained from untreated pulp had dark brown
of the filtrate was measured at 420 nm against 60% aqueous colour with initial browning index value of 0.411 which
Table 1: Effect of pre-treatment of banana pulp with different concentrations of ascorbic acid in preventing browning in clarified
banana juice
Ascorbic acid Browning index (A at 420 nm)
concentration Initial Value Time intervals (h)
(ppm) 2 4 6 24
e d c b
0 0.413 0.425 0.460 0.485 0.585a
200 0.313e 0.368d 0.389c 0.426b 0.559a
400 0.299d 0.302d 0.312c 0.354b 0.421a
600 0.168e 0.176d 0.204c 0.261b 0.369a
800 0.103e 0.118d 0.158c 0.169b 0.283a
1000 0.081d 0.083cd 0.085c 0.098b 0.199a
1200 0.070d 0.071d 0.080c 0.083b 0.095a
In each row, means followed by the same letter are not significantly different (p0.05)
 Volume 35 Issue 2 (2016) 157
continuously increased during holding at ambient of 70-900C, results in the destruction of their catalytic activity
temperature and reached to 0.586 at 24 h of storage. However (Vamos-Vigyazo, 1981).
lower browning index values of clarified juices obtained from In the present study, peeled banana fruits were
ascorbic acid treated pulp indicated that browning reaction blanched in hot water (900C) for different periods up to 10
suppressed due to action of ascorbic acid. Depending on the minutes. These heated samples were quickly cooled to room
concentration of inhibitor, juices obtained had light colour. temperature by dipping in cold water then blended and
As the concentration of ascorbic acid increased (200 to 1200 clarified juices were prepared with optimized enzymatic
ppm), browning index in clarified juices gradually decreased. conditions.
Lower concentration of ascorbic acid (200 to 800 ppm) was
satisfactory to prevent enzymatic browning during juice Clarified juices obtained from pulps preheated at
extraction and protect the colour of extracted juices only for different periods were measured for browning immediately
a short period. However, juices obtained from the pulps after preparation (initial value) and after every 2 h of storage
pretreated with 1000 – 1200 ppm of ascorbic acid retained over a period of 6 h. Browning index values presented in
their natural light colour without significant change over 4 h Table 2 indicates that browning problem was severe in
at room temperature (as indicated by lower absorbance at clarified banana juice obtained from pulp without any preheat
420 nm). The inhibitory action of ascorbic acid is mainly treatment. However, preheating of banana pulp at 900C was
due to its reducing action on enzymatically formed O- observed to reduce browning reactions in prepared clarified
quinone back to phenols furnishing fresh substrate to the juices, depending on the heating time. Increased in blanching
enzyme resulting in a gradual decrease in its activity (Ponting, time (2 to 10 minutes) decreased the browning in clarified
1960). Thus at low concentration, these inhibitor may be juices. Blanching at 900C for a short time (2 to 4 minutes)
quickly consumed in reduction process and the colour was observed to protect the colour of extracted juices to a
formation was delayed for only a limited time but not less extent and for a short period of time upto 2 h. However
completely suppressed (Yi and Ding, 2014). At high juices obtained from the pulps preheated at same temperature
concentrations, ascorbic acid can also inhibit PPO (Vamos- for comparatively longer period (6 to 10 minutes) retained
Vigyazo, 1981, Jang and Moon, 2011). their natural light colour without significant change over 4
Under the conditions of this investigation, for h. This may be due to the thermal inactivation of endogenous
complete enzymatic clarification of juice, 4 h period was polyphenol oxidase in banana pulp.
required. Therefore, it is necessary to protect the juice from According to the conditions of the present
discolouration during this clarification period. According to investigation, anti browning treatment should protect the
the results obtained, on pretreatment of pulp with 1000 ppm juice from discolouration for minimum 4 h (clarification
of ascorbic acid, the juice obtained retained its natural colour period). From Table 2 it is evident that minimum heating of
without marked changes up to 4 h at room temperature. banana pulp for 6 minute was sufficient to retain natural
Samanta et al, (2010) evaluated the anti-browning (inhibition colour for 4 h in prepared clarified juice. The results are in
of polyphenol oxidase activity) effect of different inhibitors line with observations of Garda et al. (1985) and Sims and
in banana, apple and mushroom and also reported ascorbic Bates (1994) who applied heat treatment to control browning
acid as the most effective PPO inhibitor in all the samples. in banana puree.
Heat pretreatment: Blanching is one of the most effectively Quality characteristics of clarified banana juices
applied methods of heat treatment for controlling enzymatic obtained from pre-treated pulp: Pretreatment of pulp with
browning in fruits and vegetables. Polyphenol oxidase is the 1000 ppm of ascorbic acid or heating at 900C for 6 minute
endogenous enzyme of most of the fruits and vegetables was observed to retain the natural colour of prepared clarified
majorly responsible for their browning (Manohar et al, 2014). juice without marked changes up to 4 h of storage at
Exposure of polyphenol oxidase to temperature in the range room temperature. To select the most desirable antibrowning

Table 2: Effect of pre-heating of banana pulp for different times (At 900C) in preventing browning in clarified banana juice
Heating Browning index (A at 420 nm)
Time Initial Value Time intervals (h)
(minute) 2 4 6 24
0 0.429e 0.443d 0.468c 0.491b 0.591a
2 0.232e 0.241d 0.256c 0.272b 0.290a
4 0.132e 0.142d 0.157c 0.167b 0.199a
6 0.071c 0.072c 0.073c 0.077b 0.081a
8 0.031c 0.032c 0.032c 0.036b 0.041a
10 0.009d 0.011cd 0.011c 0.013b 0.017a
In each row, means followed by the same letter are not significantly different (p0.05)
158 ASIAN JOURNAL OF DAIRY AND FOOD RESEARCH
Table 3: Quality characteristics of clarified banana juices obtained from pretreated banana pulp
Parameters Pretreatment to Pulp
Control Ascorbic acid(1000 ppm) Heating (900C /6 minute)
Yield of juice (%) 75.3±1.04a 74.2±0.32a 68.0±0.6 b
Clarity (%T at 660nm) 94.2±0.2a 92.26±0.46b 88.26±0.70c
Titratable Acidity (as % malic acid) 0.51±0.01b 0.55±0.01a 0.52±0.00b
TSS (o Brix) 23.4±0.1a 23.5±0.1a 23.1±0.1 b
Total sugar (%) 19.06±0.12a 18.94±0.02a 18.06±0.12b
Reducing sugar (%) 11.57±0.15b 12.03±0.06a 11.47±0.11b
Sensory Qualities
Colour Dark Brown Yellow Yellow
Flavour Banana Banana Slightly Cooked
Taste Typical sweet Typical sweet Cooked
Overall Acceptability Good Very Good Fair
In each row, means followed by the same letter are not significantly different (p0.05)

treatment, juices obtained from these pretreatments were pretreatments. Undesirable dark brown colour was observed
analyzed for their quality characteristics. Juice obtained in juices obtained from without pretreated banana pulp.
without any pulp pretreatment was considered as ‘Control’. Discoloration of juice may be due browning reaction
The results depicted in Table 3 indicated that yield, clarity occurred in the pulp during clarification process.
and titratable acidity of juices prepared from ascorbic acid Pretreatment of pulp with 1000 ppm of ascorbic acid was
treated pulp were significantly higher than those for observed to yield banana juices with their natural colour and
preheated pulp. This could be due to thermal inactivation of having typical banana flavour and good sweet taste. Whereas
the endogenous banana pectinases which might have a preheating of banana pulp caused cooked flavour and altered
synergistic effect during pectic enzyme treatment of the pulp. the original taste of the clarified juices.
Levi et al. (1980) observed that blanching of banana pulp CONCLUSION
decreased the activity of pectinesterase of pulp. Total soluble The present study concluded that preheating of
solids and sugar content were found to be decreased in juices banana pulp observed to cause slightly cooked flavour and
prepared with preheated pulp. This may be due to leaching altered the original taste of the clarified juices. Pretreatment
out of water soluble solids during blanching treatment. of banana pulp with 1000 ppm of ascorbic acid is the most
Considerable variations were recorded in sensory suitable method to produce clarified juice with natural yellow
qualities of the clarified juices obtained from both these colour and acceptable flavour.

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