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Primary Clarification of Banana Juice Extract by

Centrifugation and Microfiltration
a b a b
Sorel Tchewonpi Sagu , Sankha Karmakar , Emmanuel Jong Nso & Sirshendu De
a
Department of Process Engineering , National School of Agro-Industrial Sciences (ENSAI),
University of Ngaoundere , Adamawa , Cameroon
b
Department of Chemical Engineering , Indian Institute of Technology , Kharagpur , India
Accepted author version posted online: 25 Feb 2014.Published online: 16 May 2014.

To cite this article: Sorel Tchewonpi Sagu , Sankha Karmakar , Emmanuel Jong Nso & Sirshendu De (2014) Primary
Clarification of Banana Juice Extract by Centrifugation and Microfiltration, Separation Science and Technology, 49:8,
1156-1169, DOI: 10.1080/01496395.2013.877932

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 Separation Science and Technology, 49: 1156–1169, 2014
Copyright # Taylor & Francis Group, LLC
ISSN: 0149-6395 print=1520-5754 online
DOI: 10.1080/01496395.2013.877932
Primary Clarification of Banana Juice Extract by
Centrifugation and Microfiltration
Sorel Tchewonpi Sagu,1 Sankha Karmakar,2 Emmanuel Jong Nso,1 and
Sirshendu De2
1
Department of Process Engineering, National School of Agro-Industrial Sciences (ENSAI),
University of Ngaoundere, Adamawa, Cameroon
2
Department of Chemical Engineering, Indian Institute of Technology, Kharagpur, India
Downloaded by [McMaster University] at 09:18 20 November 2014
Banana is a much appreciated tropical fruit due to its unique
aroma, flavor, and also for its nutritional and energetic components.
In this work, banana juice was extracted by pectinase treatment at
room temperature. The resultant juice was clarified by two methods,
namely, centrifugation and microfiltration. A comparative study
was performed between these two primary clarification methods.
Response surface methodology was used to optimize the parameters
of centrifugation (speed and time) as well as microfiltration
(transmembrane pressure and cross flow rate). The juice was
characterized in terms of viscosity, clarity, alcohol insoluble solids
(AIS), polyphenol, and protein content. Centrifuged juice contained
high concentration of total polyphenol and protein. Juice obtained
from microfiltration had lower viscosity, AIS, and higher clarity.
Also, the energy consumption of the centrifuge is much higher
than that of microfiltration.
Keywords Banana juice; centrifugation; microfiltration;
optimization; primary clarification
INTRODUCTION
The processing of tropical fruits into clarified and
concentrated juice is an important route of development
and valorization of products from agricultural origin. Fruit
juices have gained much popularity recently and provided
an alternative substitute to traditional beverages such
as coffee, tea or carbonated soft drinks (1). Much research
has been conducted for the treatment of traditional
fruit juice like apple or orange juice as well as to produce
high-quality fruit juice obtained from pulpy fruits,
like Mediterranean sweet lemon (mosambi), water melon,
etc (2). Among the traditional fruit juices, banana
juice holds a special place, particularly in the tropical and
subtropical regions where bananas are produced in plenty.
It contains sufficient sugar and is a source of energy. Due
to its nutritional qualities (proteins, polyphenols, vitamins,
Received 22 July 2013; accepted 18 December 2013.
Address correspondence to Sirshendu De, Department of
Chemical Engineering, Indian Institute of Technology, Kharagpur
P.O. Box 721302, India. E-mail: sde@che.iitkgp.ernet.in
1156
potassium, calcium, and magnesium), aroma, and flavor,
banana juice is of great interest to the population and the
food processing industries. Being produced in large
quantity, it is cheaper compared to other fruits. The tech-
niques used for primary clarification of fruit juice before
final filtration is the focus of many researchers. Banana
juice extract is highly viscous having macromolecules such
as proteins, tannins, and polysaccharides (pectin and
starch). This is typical for fruits having high pulp content.
Transparency and homogeneity are thus two essential
characteristics directly linked to the presence of macro-
molecules in suspension and these can be improved by
the removal of suspended solids (2).
Therefore, clarification of the extracted juice is an
important unit operation for removal the suspended parti-
cles. Centrifugation and microfiltration (MF) are two
popular methods used for clarification (3). Several works
are reported in the literature on clarification of fruit juices
using these two methods. Centrifugation was used for
clarifying apple juice from mash (4), Sea Buckthorn juice
(5), orange juice (6,7), and passion fruit juice (8). In the
study conducted by Rai et al. (9) for the removal of
the suspended particles from mosambi juice, the pretreat-
ment methods for primary clarification were enzymatic
treatment followed by centrifugation. They compared the
performance of this primary clarification with fining agents
like, gelatin, bentonite, or a mixture of both. In their work,
the optimum conditions for centrifugation were obtained.
These were 7500 g for centrifugation speed for 30 min. They
observed that the quality of pretreated juice using various
methods was similar but the permeate flux of centrifuged
juice was more than that of enzymatically treated one.
They also concluded that cost of centrifugation is four
times less than that of enzymatic treatment. Chhaya et al.
(3) conducted experiments for primary clarification of ste-
via extract using centrifugation and microfiltration. They
compared both the processes in terms of the quality of
the product but not on the basis of cost. The percentage
 PRIMARY CLARIFICATION OF BANANA JUICE EXTRACT
recovery of sweeteners in case of centrifugation was
89.5% and that of microfiltration was 88.5%. Hence, centri-
fugation gave marginally better results. In the work of
Oliveira et al. (10), a comparative study of both micro-
filtration and centrifugation was done for passion fruit
juice. All the parameters of the product from both the
processes are comparable but since microfiltration
removes all the suspended solids, it was selected as a better
alternative. Crossflow microfiltration was used to clarify
pineapple (11,12), melon (13), passion fruit juice (14),
apple (15–17), pomegranate (18,19), bergamot (20), chicory
(21,22), and acerola (23).
In case of banana juice, most of the works have been
focused only on extraction techniques, but not on the clari-
fication. Banana juice contains a large amount of pectin.
Pectin is a well known gel forming agent (24), making the
clarification of juice difficult due to intense fouling. Lee
Downloaded by [McMaster University] at 09:18 20 November 2014
et al. (25) have attempted the optimizing conditions for
enzymatic clarification of banana juice using response
surface methodology. In their work, the banana juice was
extracted using hot water (at 95
C for 120 min) followed
by 0.02% of amylase at 60
C for 1 h and then with
pectinase at various enzyme concentrations (0.01–0.1%),
temperatures (30–50
C), and time (30–120 min) before the
centrifugation (3000  g) for the removal of particles. The
effect of these enzymatic treatments on filterability, clarity,
turbidity, and viscosity of the juice were studied and no
information on the quality of the final juice was reported.
Amylase, though a very cost-effective enzyme, cannot fully
hydrolyze the pectin content in raw banana. So, Lee et al.
(25) used amylase followed by pectinase. Also, the nutri-
tional qualities of the juice are deteriorated and the taste
of the juice changes significantly at such high temperature
and for long processing time. Even after enzymatic
treatment, the leftover pectin makes further clarification
difficult.
The present work undertakes the primary clarification of
banana juice extracted at room temperature and low dose
of pectinase enzyme. Primary clarification is conducted
using centrifugation and microfiltration and the perfor-
mance of these two processes have been compared.
Response surface methodology with Doehlert design was
used to optimize the operating conditions of centrifugation
(speed and time) and microfiltration (transmembrane
pressure and flow rate). Clarity, viscosity, and some
nutritional parameters, like total polyphenol and protein
concentration were evaluated to compare the two processes.
MATERIALS AND METHODS
Materials
Feed
Fresh, mature bananas of Musa acuminate (with green
skin), commonly referred to as Dwarf Cavendish Banana
1157
variety were purchased from a local market at Kharagpur
in West Bengal, India.
Enzyme Source and Chemicals
Pectinase (EC3.2.1.15) from Aspergillus niger with
activity 3.5–7 units=mg was purchased from Sisco Research
Labratory, Mumbai, India and used for enzymatic treat-
ment of banana pulp. Folin–Ciocalteu reagent, anhydrous
sodium carbonate, and copper(II) sulphate pentahydrate
were purchased from Merck Specialities Pvt. Ltd.,
Mumbai, India and potassium sodium tartrate was obtained
from Loba Chemie, Mumbai, India. Bovine serum albumin
(BSA) was used for calibration for protein estimation
and it was procured from Sisco Research Laboratories
Pvt. Ltd., Mumbai, India. Gallic acid standard, used for
calibrating polyphenol content was obtained from Loba
Chemie, Mumbai, India. Ethyl alcohol for determination
of alcohol insoluble solids (AIS) was procured from
Jiangsu Huaxi International Trade Co. Ltd., China. The
commercial grade filter papers (diameter 110 mm) were
procured from Whatman, GE Healthcare UK Ltd., UK,
and all the glassware used were obtained from Borosil
Glass Works Ltd., Mumbai, India. All the chemicals listed
above were of analytical grade.
Preparation of Banana Juice
The enzymatic extraction at low temperature was
performed to extract the banana juice. Fresh and mature
banana were selected, washed, peeled, and cut in small
pieces. Based on primary works, a ratio of 1:2 (banana:
water; w=v) was used in the mixing process at 1000 rpm
for 10 minutes using a Remi Motor agitator, type RQ
122 supplied by Elektrotechnik Ltd., Kolkata, India. The
mixture was put in water bath at 33  2
C, and then
0.03%v=w of pectinase was added, mixed, and then kept
during 108 min. During the incubation period, the mixture
was stirred regularly to allow complete homogenization of
the system. At the end of the enzymatic treatment,
pectinase was inactivated by heating at 95
C for 5 min.
The mixture was cooled and filtered by a fine mesh cloth
filtration to remove pulp. The filtrate was collected, stored
at 10
C and then thawed at room temperature just before
use for clarification processes. The characteristics of the raw
banana juice extract are presented in Table 1. The details of
the extraction process are available in Sagu et al. (26).
Primary Clarification of Banana Juice
Centrifugation
Centrifugation of banana juice extract was performed
using a laboratory scale centrifuge (REMI, Kolkata, West
Bengal, India) under batch mode of operation in an air
conditioned room at 25
C. The various operations were
conducted in tubes with a capacity of 25 mL each. The
 1158
Characteristics of banana juice extract
Viscosity TSS
(mPa  s) (
Brix)
Extract of banana juice 1.49 5.9
operating parameters taken into account were rotation
speed (g) and time (min) as given by the experimental
design. Each experiment was conducted by introducing
20 mL of banana juice extracted in the tubes. After
centrifugation, the suspended particles were precipitated
and the supernatant constituting the clarified juice was
collected. Various analyses were performed to determine
the viscosity, clarity, AIS, and the concentration of
polyphenol and protein.
Downloaded by [McMaster University] at 09:18 20 November 2014
Microfiltration
Membrane. For microfiltration, the hollow fiber
cartridge was used for clarification. The complete procedure
for preparing the hollow fiber module, its characterization,
and performance were described by Thakur and De (27).
Hollow fibers were made using polyacrylonitrile (PAN)
polymer with average molecular weight 250,000 Da. The
permeability of the membrane was 5  1010 m=Pa  s. The
effective area of membrane was 0.0124 m2 and the average
pore size was 0.2 micron. All the experiments were
conducted in an air conditioned room at 25
C.
Procedure. The schematic of the microfiltration setup
is shown in Fig. 1. 500 ml of extracted juice was used
for each experiment under the total recycle mode with
complete recycling of the retentate and the permeate
FIG. 1. Schematic diagram of cross flow microfiltration unit with recycle
mode. 1-feed tank; 2-suction pipe; 3-feed pump; 4-pressure valve;
5-by-pass pipe; 6 and 60 -pressure gauge (at the inlet and outlet of the mem-
brane module); 7-membrane module; 8-flow control valve; 9-rotameter;
10-Flow pipe; 11-permeate line; 12-Heat Exchanger.
S. T. SAGU ET AL.
TABLE 1
Clarity AIS Polyphenol Protein Conductivity Color
(%T) (%w=w) (mg GAE=100 ml) (mg=l) (mS=cm) (A)
52.3 0.58 15.4 1308.6 1912 1.17
stream in the feed tank to maintain the feed concentration
uniform. The microfiltration set up consists of a feed tank,
a membrane module, a booster pump, two pressure gauges
(at the inlet and outlet of the membrane module), two
valves, and one flow meter. Different transmembrane
pressures (35 to 104 kPa) and cross flow rate (10 to 20 l=h;
Reynolds number: 112–224) were applied for micro-
filtration processes under the conditions defined by the
experimental design and all the experiments were carried
out in an air conditioned room maintained at 25
C. The
transmembrane pressure was calculated as the average of
the readings of these two pressure gauges. A back pressure
valve in the retentate line and a bypass valve were used to
adjust and maintain the transmembrane pressure and
cross flow rate independently. A small quantity of
permeate (clarified juice) was collected for analysis of
viscosity, clarity, AIS, and the concentration of polyphenol
and protein.
As described by the experimental design, a total of ten
experiments were performed at different operating pressure
and flow rate, and the same membrane was used for all the
experiments. After each experiment with banana juice, and
in order to recover the initial permeability of membrane, the
membrane module was cleaned. After circulating the tap
water in the circuit, an acid solution (HCl, 0.1N) was first
used to wash the membrane for 45 minutes. After this, an
alkaline wash (NaOH, 0.2N) was carried out for another
45 minutes. At the end of the washing procedure, distilled
water was re-circulated through the module for 35 minutes
ensuring the pH of the permeate about 7.0. The permeability
of the membrane was measured thereafter.
Experimental Design
The RSM with Doehlert design was used to carry out
the experiments to optimize the conditions of centri-
fugation and microfiltration. The independent variables
were time (X1) and speed (X2) for centrifugation and
then pressure (X1) and flow rate (X2) for microfiltration.
The ranges of these variables are given in Table 2.
For the Doehlert design, the number of experiments (N)
using the Doehlert matrix with k variables is given by the
relationship:
N ¼ kðk þ 1Þ þ 1 ð1Þ
A total of 7 experiments were performed with three
replicates at the central point for centrifugation and
 PRIMARY CLARIFICATION OF BANANA JUICE EXTRACT 1159

TABLE 2
Experimental domain
Operation Factor Units Abbreviation Values
Centrifugation Time Min X1 20–60
Speed g X2 2000–10000
Microfiltration Pressure kPa X1 34.5–103.4
Flow rate L=h X2 10–20

microfiltration each. Experimental design with coded form
and real values is shown in Table 3. Five selected responses
were: AIS (% w=v), viscosity (mPa  S), clarity (% transmit-
tance), protein concentration (mg=l), and total polyphenol
(mg GAE=100 ml). The mathematical model indicating
the effect of variables in terms of linear, quadratic, and
interactions terms were related to the variables (Xi, i ¼ 1, 2)
Downloaded by [McMaster University] at 09:18 20 November 2014
Also, color, pH, total soluble sugar (TSS), and conductivity
were determined.
Color and Clarity
The color of the extract was measured by absorbance
(A) at a wavelength of 420 nm using a spectrophotometer

(M=s Perkin Elmer, Connecticut, USA) (9). Clarity of the

by a second-degree polynomial given by Eq. (2).
X X X
Y ¼ b0 þ bi X i þ bii Xi2 þ bij Xi Xj
where Y are the five experimental responses, Xi and Xj are
the levels of variables, b0 is the constant term, bi are
the coefficients of the linear terms, bii are the coefficients
of the quadratic terms, and bij are the coefficients of the
interactions terms. Experimental data were carried out by
appropriate analysis of variance (ANOVA) using Stat-Ease
Design-Expert Software, version 8.0.7.1.
Analysis
For each experiment of centrifugation and microfiltra-
tion, clarified banana juice was analyzed for its viscosity,
clarity, AIS, total polyphenols, and protein concentration.
extract was measured by transmittance (%T) at 660 nm
using the same spectrophotometer (9).
ð2Þ
Alcohol Insoluble Solids (AIS)
AIS are the measure of pectineous substances in banana.
For AIS determination, 10 g of juice was weighed into
a 250 ml beaker. 150 ml of 80% alcohol was added, stirred,
brought to boil and then simmered slowly for 30 min.
Whatman filter paper of appropriate size, which has been
previously dried and weighed, was used for filtration. The
content of the beaker was transferred to the filter paper
on a funnel and the residue was washed on the filter paper
with 80% alcohol until the washing was clear and colorless.
The filter paper and the alcohol-insoluble solids were dried
for 2 hours at 100
C, then cooled in a desiccator, weighed,
and the AIS was determined by as percentage (% w=w)
in the following equation (28).

TABLE 3
Doehlert design coded and real variables
Real variables
Coded variables Centrifugation Microfiltration
No. of experiment x1 x2 X1 (min) X2 (g) X1 (kPa) X2 (l=h)
1 0 0 40 6000 68.9 15
2 1 0 60 6000 103.4 15
3 1 0 20 6000 35.5 15
4 0.5 0.866 50 10000 86.2 20
5 0.5 0.866 30 2000 51.7 10
6 0.5 0.866 50 2000 86.2 10
7 0.5 0.866 30 10000 51.7 20
8 0 0 40 6000 68.9 15
9 0 0 40 6000 68.9 15
10 0 0 40 6000 68.9 15
 1160
AIS ¼ (weight of residue/weight of
sample taken for estimation)  100%
Total Polyphenol
Total polyphenol was measured using a modified Folin
and Ciocalteu method described by Vasco et al. (29).
Briefly, an aliquot (0.5 ml) of the banana juice blank or
standard was placed in a 25 ml flask and 0.5 ml of the
Folin–Ciocalteu reagent was added. The mixture was
allowed to react for 5 min with stirring. 10 ml of solution
of sodium carbonate (concentration 75 g=l) was added and
mixed well. The volume was then completed up to 25 ml
with distilled water and kept at room temperature for 1 h.
The absorbance was then measured at 750 nm using
a spectrophotometer (M=s Perkin Elmer, Connecticut,
Downloaded by [McMaster University] at 09:18 20 November 2014
USA). The results were expressed as mg of gallic acid
equivalents per 100 ml.
Protein Concentration
Protein concentration was determined according to the
dye binding method of Lowry et al. (30) with bovine serum
albumin (BSA) as standard.
Viscosity
The viscosity of banana juice was determined using an
Ostwald capillary viscometer (Pisco, Kolkata, India) and
viscosity was measured at room temperature (25  1
C).
The unit of measurement used for viscosity is mPa  s.
Total Soluble Sugar (TSS)
The total soluble sugar content in degree Brix (
Brix)
was determined using an ABBE type refractometer (Excel
International, Kolkata, India).
Conductivity and pH
Conductivity and the pH values of juice were measured
using a multi parameter pocket tester (EUTECH Instru-
ments Ltd., Singapore). The unit for conductivity is mS=cm.
Statistical Analysis
Model Equations
To validate the different model equations, average
absolute deviation (AAD) and coefficient of determination
(R2) values were determined. AAD was calculated by the
following equation:
N 

X
 ð5Þ
AAD ¼
Yi;exp  Yi;cal
=Y =N
i;exp
i¼1
where Yi,exp is the experimental response, Yi,cal is the
response calculated using the model equation, and N
is the total number experiment. The response model that
S. T. SAGU ET AL.
had the lowest AAD and the highest R2 (close to 1.0)
ð3Þ was validated as the proper model equation for expression
of response.
Statistical Analysis of Experimental Measurements
All the experiments were repeated three times. Various
quality parameters were measured and the average values
were reported along with standard deviation.
RESULTS AND DISCUSSION
A total of 10 experiments of Doehlert design showing
the effect of time and speed (for centrifugation) and the
transmembrane pressure and flow rate (for microfiltration)
were performed. Viscosity, clarity, AIS, the concentration
of polyphenol and protein were taken as experimental
responses and the results are presented in Table 4.
Centrifugation
Table 5 shows the regression coefficients of the variables
of different mathematical models and the corresponding R2
and AAD. The values of R2 found were 0.92, 0.97, 0.96,
0.95, and 0.95, respectively, for viscosity, clarity, AIS, the
polyphenol and protein. All these values of R2 indicate that
the different proposed mathematical models can explain
more than 95% of experimental observations as a function
of independent variables, except the viscosity model that
represents 92% of the effects. For a better assessment of
the models, the AAD was determined and the values
obtained were 0.00, 0.00, 0.01, 0.01, and 0.02, respectively,
for viscosity, clarity, AIS, the polyphenol, and protein.
AAD of 0 indicating perfect agreement between the obser-
ved and the predicted responses (31), models were found to
be adequate to represent the experimental responses and
were further used to plot the response surfaces, followed
by optimization of the centrifugation process.
Viscosity
As in Table 5, time (X1) (p < 0.05) and speed (X2)
(p < 0.001) had the most significant effect on the viscosity
of banana juice. Quadratic and the interaction terms of
these variables were not significant. This result shows that
each parameter is independent and there is no real syner-
gistic effect between time and speed on viscosity. The
regression model representing the effect of time and speed
on viscosity in terms of their coded level is given as:
ViscosityðmPa  sÞ ¼ þ1:33  0:04X1  0:08X2
ð4Þ þ 0:02X21 þ 0:01X22
The value of the coefficient of determination R2 for the
above equation is 0.92. This value indicates that the
regression model is able to explain 92% of variability of
the data. The minus sign of the two factors X1 and X2
 Downloaded by [McMaster University] at 09:18 20 November 2014

TABLE 4
Experimental responses
Centrifugation Microfiltration
Exp. Viscosity Clarity AIS Polyphenol Protein Viscosity Clarity AIS Polyphenol Protein
No. (mPa  s) (%T) (%) (mg GAE=100 ml) (mg=l) (mPa  s) (%T) (%) (mg GAE=100 ml) (mg=l)
1 1.35  0.05 68.4  2.6 0.42  0.06 12.8  0.7 986.2  34.6 1.25  0.05 93.9  2.1 0.25  0.02 7.5  0.6 750.5  23.5
2 1.31  0.30 69.2  3.0 0.36  0.05 12.4  0.8 937.5  68.4 1.12  0.10 96.6  1.0 0.21  0.03 5.1  0.3 529.1  38.3
3 1.39  0.05 64.2  1.8 0.44  0.05 13.0  1.0 803.7  57.2 1.29  0.08 89.3  4.1 0.38  0.03 11.3  0.8 969.0  44.0
4 1.26  0.10 74.1  3.0 0.35  0.06 11.8  0.7 556.3  18.5 1.18  0.30 93.6  2.5 0.23  0.03 8.7  1.0 771.1  13.7
5 1.44  0.12 59.4  3.0 0.55  0.04 14.7  1.0 1036.6  37.8 1.26  0.05 92.9  2.0 0.36  0.03 8.7  0.7 851.4  37.2
6 1.41  0.25 61.7  2.6 0.55  0.06 13.7  1.0 1048.5  43.3 1.17  0.34 94.7  1.5 0.24  0.05 5.0  0.4 577.9  23.2
7 1.29  0.10 74.9  3.8 0.38  0.05 12.1  0.3 632.7  24.7 1.26  0.10 91.4  2.7 0.29  0.04 8.9  0.8 847.2  32.3
8 1.31  0.24 69.7  3.2 0.42  0.05 12.5  0.9 945.6  53.9 1.21  0.05 94.3  3.7 0.22  0.03 6.8  0.8 584.5  21.7
9 1.34  0.05 68.7  1.0 0.42  0.03 12.8  0.7 973.5  29.9 1.27  0.08 92.7  1.2 0.22  0.04 6.6  0.3 667.1  20.0
10 1.33  0.31 69.6  3.4 0.44  0.05 12.8  0.5 904.2  68.3 1.23  0.10 93.4  2.6 0.25  0.05 7.5  0.3 577.0  29.4

1161
TABLE 5
Coefficients of regression R2 and AAD values for the different mathematic models
Centrifugation Microfiltration
Viscosity y1 Clarity y2 AIS y3 Poly-phenols y4 Proteins y5 Viscosity y1 Clarity y2 AIS y3 Poly-phenols y4 Proteins y5
Coefficient (mPa  s) (%T) (% w=w) (mg GAE=100 ml) (mg=l) (mPa  s) (%T) (% w=w) (mg GAE=100 ml) (mg=l)
b0 1.33 69.1 0.43 12.7 952.4 1.24 93.6 0.24 7.1 644.8
b1 0.04 1.9 0.03 0.4 33.9 0.09 3.1 0.09 2.7 204.9
b2 0.08 6.9 0.09 1.2 224.0 0.003 0.7 0.02 1.0 47.3
b12 0.00 1.4 0.02 0.4 44.1 0.005 0.2 0.03 1.7 98.7
b11 0.02 2.4 0.03 0.01 81.8 0.04 0.7 0.06 1.1 104.3
b22 0.01 1.0 0.04 0.4 113.4 0.014 0.3 0.03 0.5 91.1
R2 0.92 0.97 0.96 0.95 0.91 0.82 0.79 0.94 0.89 0.76
AAD 0.00 0.00 0.01 0.01 0.02 0.00 0.00 0.02 0.03 0.03
b represents the coefficients of equations different of models with b0 the constant term; b1, and the linear effects (1 and 2 respectively the time and speed for
centrifugation; and pressure and flow rate for microfiltration); b11 and b22 the quadratic effects; and b12 the interaction.

Significant at p  0.05.

Significant at p  0.01.

Significant at p  0.001.
 1162
indicates that that time and speed have negative influence.
Viscosity is reduced with centrifugation speed and time. In
Fig. 2(a), it is observed that this reduction was affected
more by centrifugation speed ranging from 2000 g
(1.47 mPa  s) to 10000 g (1.32 mPa  s) for 20 min. More
suspended solids are removed at higher centrifugal forces
for longer duration, thereby reducing the viscosity of the
solution.
Clarity
Figure 2(b) indicates the effect of time and speed on
clarity. It is shown that time (p < 0.05) and speed
(p < 0.001) both had a significant effect on the clarity.
From the regression model of the clarity (Eq. 6), it is clear
that the effect of time and speed on clarity was positive.
This means that the clarity of banana juice increases with
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time and duration of centrifugation. This is due to the fact
that more suspended solids are removed at higher speed
FIG. 2. Response surfaces of (a) viscosity, (b) clarity, (c) AIS, (d) polyphenol, and (e) protein as a function of time and speed for centrifugation process.
S. T. SAGU ET AL.
and duration of centrifugation, making the solution clear.
For example, the clarity increased from 58% at 2000 g to
75% at 10,000 g for 45 min of centrifugation.
Clarity ð%TÞ ¼ þ69:1 þ 1:9X1 þ 6:9X2  1:4X1 X2
 2:4X21  1:0X22 ð6Þ
The value of the coefficient of determination R2 for this
equation (in terms of their coded level) is 0.97, indicating
good fitting of the model with the experimental data.
AIS
The regression model representing the effect of time and
speed on AIS in terms of their coded level is given as:
AIS ð%w=wÞ ¼ þ0:43  0:03X1  0:09X2
 0:02X1 X2  0:03X21 þ 0:04X22
ð7Þ
 PRIMARY CLARIFICATION OF BANANA JUICE EXTRACT
Regression coefficients indicate centrifugation time (p <
0.05) and speed (p < 0.001) were highly significant (p <
0.001) for reduction of AIS. Also, a quadratic level of speed
had a significant effect (p < 0.05). The value of the coef-
ficient of determination R2 for the above equation is
0.96. It is observed from this equation that the linear effect
of time and speed on AIS were both negative. Thus, AIS of
banana juice is reduced with time and centrifugation speed
as shown in Fig. 2(c). In fact, AIS is composed mainly of
high molecular weight macromolecules like pectin and
other polysaccharides and during the process of centrifuga-
tion, the sedimentation velocity of a particle is a function
not only of its size, shape, and weight but even more of
the speed and time.
Polyphenol and Protein
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The concentration of polyphenol and protein are two
major parameters providing information on the efficiency
of the process and the quality of banana juice obtained.
Figures 2(d) and 2(e) show the effect of independent
variables on these two parameters. The concentration of
protein and polyphenol decreases with centrifugation
speed. For example, when the centrifugation time was
40 min, the protein concentration is reduced from
1060 mg=l at 2000 g to 610 mg=l at 10000 g (Fig. 2e). In case
of polyphenols, its concentration varies from 14 to 11 mg
GAE=100 ml in the same range of centrifugation speed
(Fig. 2d). This significant reduction of proteins with a
centrifugation speed of 10,000 g is due to their high mole-
cular weight. Like the AIS, proteins are macromolecules
with high molecular weight and at the high speeds of
centrifugation they are separated, thereby reducing their
concentration in the juice.
The regression models representing the effect time and
centrifugation speed on the polyphenol and protein con-
centration of banana juice, in terms of their coded level,
are given as:
Poly-phenol ðmgGAE=100mlÞ ¼ þ12:7  0:4X1
 1:2X2 þ 0:4X1 X2
 0:01X21 þ 0:4X22 ð8Þ
Protein ðmg=lÞ ¼ þ952:4 þ 33:9X1  224:0X2
 44:1X1 X2  81:8X21  113:4X22 ð9Þ
The values of the coefficients of determination R2 of the
above equations are 0.95 and 0.91, respectively, for the
total polyphenol and protein concentration. These indicate
that the regression models explain more than 90% of the
variability of total polyphenol and protein concentration
of the banana juice extract. Table 5 shows that time
was significant just for polyphenol (p < 0.05) whereas the
1163
centrifugation speed had a significant effect on polyphenol
(p < 0.001) and protein (p < 0.001).
Optimization of Centrifugation
The numerical optimization of process parameters of the
centrifugation (time and centrifugation speed) is carried
out using Stat-Ease Design-Expert v8.0.7.1 software. The
optimum processing conditions were investigated for the
five experimental responses (viscosity, clarity, AIS, poly-
phenol, and protein concentration). Conditions taken in
consideration for optimization were to minimize the values
of viscosity and AIS, and to maximize clarity, total poly-
phenols, and proteins concentration. The range of two
independent parameters was centrifugation time (20–
60 min) and centrifugation speed (2000–10,000 g). The most
important physico-chemical characteristic of banana juice
is its polyphenol content. To improve the polyphenol con-
tent and reduce the pectin concentration, numerical optimi-
zation was carried out. The optimum time of centrifugation
and speed were found as 51 min and 8656 g rpm with a
desirability of 0.9, where the AIS content was 0.35% w=w
and polyphenol was 12.02 mg GAE=100 ml. But consider-
ing the overall quality of the juice, the solution having
the overall desirability (0.6) was selected as the optimum
condition for the process of primary clarification of banana
juice by centrifugation. Optimum conditions generated by
numerical optimization are: centrifugation time: 46 min
and centrifugation speed: 6065 g, with the corresponding
values of the experimental responses of 1.32 mPa  s for
viscosity, 69.6%T for clarity, 0.41% w=w for AIS, 12.5 mg
GAE=100 ml of GAE=100 mL for polyphenol, and
951.1 mg=l of protein concentration. For centrifugation,
the p value for viscosity, clarity, AIS, polyphenol and
protein are 0.0049, 0.0008, 0.0012, 0.0025, and 0.0072,
respectively. This shows that for viscosity and clarity, both
centrifugal speed and time play a crucial role whereas for
AIS, protein and polyphenol, the effect of centrifugal
speed is significant. The lack of fit for viscosity, clarity,
AIS, polyphenol, and protein are given as 0.22, 4.59, 4.17,
3.63, and 4.97, respectively. This shows that the lack of fit
is not significantly relative to pure error and the model is
justified. Experiments were conducted at optimal conditions
to verify the numerical results. The values of the responses
are 1.30 mPa  s for viscosity, 71%T for clarity, 0.39%
w=w for AIS, 11.6 mg GAE=100 ml of GAE=100 mL for
polyphenol, and 867 mg=l of protein concentration.
Color, TSS, pH, and Conductivity
In addition to the parameters that were subjected to
RSM analysis for optimization, some parameters, namely
color, pH, TSS, and conductivity of clarified banana juice
were also recorded for each of the 10 experiments of
Doehlert design for centrifugation processes. It was
observed that the range of these parameters were between
 1164 S. T. SAGU ET AL.
0.368 and 0.962 A for color and between 4.1 and 5.9
Brix
for TSS. The values of conductivity were from 1421 to
1888 mS=cm and the ranges of pH recorded for clarified
banana juice varied from 3.60 to 4.35.
Microfiltration
Figures 3(a) and 3(b) show the decline of permeate flux
for different transmembrane pressure and cross flow rate
during the microfiltration. As shown in these figures,
a rapid decline of permeate flux was observed during the
first 5 minutes. In general the flux has been reduced from
about 145 l=m2  h at the start of the operation to less than
60 l=m2.h after the fifth minute. This happens for all the
experiments. Subsequently, the permeate flux began to
stabilize gradually to reach the steady state. A close look
in Fig. 3(a) reveals that the steady state flux values vary
almost linearly with transmembrane pressure. This indi-
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cates that the transmembrane pressures in this study are
FIG. 3. Variation of permeate flux for some of the experiments of
experimental design as a function of time at: (a) varying pressure and fixed
flow rate at 15 l=l; (b) varying flow rate and fixed pressure at 86 kPa.
below that corresponding to the ‘‘threshold flux’’ (21,32,33).
In fact, the situation is similar to the case where irreversible
fouling is more significant compared to reversible fouling,
comprised of particle deposition as described by Bacchin
et al. (32). Thus, the rapid decline in flux during the
microfiltration of banana juice is due to membrane fouling
by the pore blocking mechanism caused by suspended
solids present in banana juice. In fact, banana is a very
pulpy fruit and several compounds with a high molecular
weight (pectin substances and starch for example) are
found in large quantities in the juice extract. These high
molecular weight compounds are deposited on the surface
of the membrane, thereby blocking pores and reducing
significantly the permeate flux. When the pore blocking
is complete within the initial time of filtration, cake formed
by particles starts building up slowly. Thus, cake layer
resistance is insignificant, leading to gradual flux decline
to a steady state. Similar results were reported in various
literatures (2,11,16,17,19) using microfiltration for clarifi-
cation of different fruits.
Figure 3(a) shows the permeate flux of the experiments
carried at three different pressures (35, 69, and 104 kPa)
and at a fixed flow rate (15 l=h). It was observed that
at the steady-state, the permeate flux varied according to
the pressure. A 35 kPa, a flux of 29 l=m2=h was observed
while at 104 kPa, the flux was 45 l=m2=h at the pseudo-
steady state. This difference was also observed in the flux
after the twenty-fifth minute of operation when the flow
rate varied and when the pressure remained fixed (Fig. 3b).
The effects of the two factors, transmembrane pressure
and cross flow rate, on the five experimental responses
are presented in Table 4. The coefficients of determination
R2 and AAD for the mathematical models of these
responses are shown in Table 5. It is observed that the
R2 values were 0.82, 0.79, 0.94, 0.89, and 0.76, respectively,
for viscosity, clarity, AIS, polyphenol, and protein content
(Table 5). The R2 values indicate that all the proposed
mathematical models can explain more than 80% of
experimental observations as a function of pressure and
flow rate, except clarity and proteins for which mathema-
tical models can explain 79% and 76% of the observed
effects. The rest of the results fall within the error margin
and can be well explained by the lack of fit value. The lack
of fit value for clarity and protein is 3.69 and 0.21, respect-
ively. The absolute averages of deviation (AAD), that are
measures of the relative average deviation of predicted
and observed responses, were also calculated for different
models. Ross et al. (31) and Baranyia et al. (34) showed
that the values of AAD equal to 0 indicate perfect agree-
ment between observed and predicted responses. In the
case of microfiltration, the AAD values obtained for the
models of clarity and protein and other responses indicate
that all the models represent adequate relationships
between independent variables and experimental responses
 PRIMARY CLARIFICATION OF BANANA JUICE EXTRACT
(AAD: 0.00, 0.00, 0.02, 0.03, and 0.03 respectively, for
models of viscosity, clarity, AIS, total polyphenol, and
protein concentration).
Viscosity
As shown in Table 5, transmembrane pressure (p < 0.01)
has the most significant effect on the viscosity of clarified
banana juice. It is observed in Fig. 4(a) that this effect of
pressure on the viscosity of banana juice was negative. This
signifies increase in pressure leads to substantial reduction
in viscosity. For example, for a cross flow rate of 15 l=h, the
viscosity varies from 1.29 to 1.13 mPa  s when the pressure
increases from 35 to 104 kPa. The negative sign of the
coefficient of the pressure in the equation representing
the mathematical model (Eq. 10) rightly shows the negative
effect of pressure on the viscosity of banana juice. As
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the transmembrane pressure increases, solutes form a cake
type layer on the membrane surface that acts as a dynamic
membrane and more solutes are rejected by them. In
addition to this, transmembrane pressure enhances the
FIG. 4. Response surfaces of (a) viscosity, (b) clarity, (c) AIS, (d) polyphenol, and (e) protein as a function of pressure and flow rate for microfiltration
process.
1165
solvent flux, thereby reducing the viscosity of permeate.
A combination of these two effects leads to reduction of
viscosity of the clarified juice with transmembrane pressure.
The regression model representing the effect of pressure
and flow rate on banana juice clarification, in terms of their
coded level, is given as:
Viscosity ðmPa  sÞ ¼ þ124  0:09X1 þ 0:003X2
þ 0:005X1 X2  0:04X21  0:01422 ð10Þ
The value of the coefficient of determination R2 and
AAD for the above equation are 0.82 and 0.00, respect-
ively. These values indicate that the regression model
is able to explain 82% of variability of the data and
that the absolute average of deviation shows the perfect
concordance between the observed and predicted responses.
Clarity
From Table 5, it is observed that clarity was mostly
affected by the linear coefficient of pressure (p < 0.01).
 1166 S. T. SAGU ET AL.
Figure 4(b) shows that this effect of the pressure on
the clarity is positive, indicating an increase of clarity
with pressure. The flow rate, the interaction between the
pressure and flow as well as their quadratic factor have
not been found to have significantly affected the clarity
of banana juice. The regression model describing the effect
of pressure and flow rate on clarity of banana juice, in
terms of their coded level, is given as:
Clarity ð%TÞ ¼ þ93:6 þ 3:1X1  0:7X2
þ 0:2X1 X2  0:6X21  0:3X22 ð11Þ
The coefficient of determination R2 and AAD for
the above equation are 0.79 and 0.00, respectively. Thus,
the regression model explains 79% of the total variability
for clarity and the absolute average of deviation indicates
perfect correlation between the observed and the calculated
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values for clarity. In this equation, the positive sign of the
coefficient of pressure shows positive effect of pressure on
clarity. Chhaya et al. (3) working on clarification of stevia
extract showed that the positive effect of the pressure
on the clarity of the juice during microfiltration was due
to the formation of a self-rejecting cake type layer on the
surface of the membrane, making the permeate clearer.
AIS
As during centrifugation, a reduction of AIS in banana
juice was found during microfiltration. Table 5 shows that
the reduction of AIS during microfiltration was mainly
affected by the transmembrane pressure. The effect was
negative and highly significant (p < 0.001). It is observed
from Fig. 4(c) that when the cross flow rate was 10 l=h,
the value of AIS varies from 0.45% w=w to 0.22% w=w
when the transmembrane pressure increases from 35 kPa
to 104 kPa. As described earlier, the formation of self-
rejecting cake type layer on the surface of the membrane
rejects the high molecular weight pectinous substances
and thereby, reduces AIS in the permeate.
The regression model representing the effect of pressure
and flow rate on clarified banana juice, in terms of their
coded level, is given as:
AIS ð%w=wÞ ¼ þ0:24  0:09X1  0:02X2
þ 0:03X1 X2 þ 0:06X21 þ 0:03X22 ð12Þ
The value of the coefficient of determination R2 of the
above equation is 0.94. The values of AAD (0.02) that mea-
sure the relative average deviation of the predicted and
experimental response show that the model describes the
values of AIS adequately.
Polyphenol and Protein
During microfiltration, the effects of pressure and
flow rate were also studied for polyphenol and protein
concentration present in banana juice. The regression
models representing these effects of pressure and flow rate
on the total polyphenol and protein concentration of banana
juice clarified, in terms of their coded level, are given as:
Polyphenol ðmg GAE=100 mlÞ ¼ þ7:1  2:7X1 þ 1:0X2
þ 1:7X1 X2 þ 1:1X21
þ 0:5X22 ð13Þ
Protein ðmg=lÞ ¼ þ644:8  204:9X1 þ 47:3X2
þ 98:7X1 X2 þ 104:3X21 þ 91:1X22 ð14Þ
The values of coefficients of determination R2 of above
equations are 0.89 and 0.76, respectively, for total poly-
phenol and protein concentration. The absolute average of
deviation for the above equations is 0.03 for both models.
The values of the AAD indicate that there is no significant
difference between the experimental data and calculated
values. This result shows that despite the low value of R2
(particularly protein), there exists a perfect correlation
between the observed and the calculated values for poly-
phenol and proteins.
It was observed from Table 5 that the pressure had a sig-
nificant effect on the polyphenols and protein (p < 0.01). It
is clear that the negative coefficients of pressure for poly-
phenol and protein (Eqs. 13 and 14) indicate that this effect
was negative. This means that the increase in pressure
reduces the concentration of polyphenol and protein in
the clarified banana juice. For example, it is observed from
Fig. 4(d) that a drastic reduction in polyphenol from 12.0
to 3.0 mg GAE=100 ml when the pressure increases from
35 to 104 kPa. Similarly, there is a reduction from 1090
to 500 mg=l of protein concentration (Fig. 4e) when the
pressure varies in the same range. It is clear that with
increasing of pressure, there was a rapid formation of a
cake layer on the surface of the membrane. The polyphenol
and protein that are macromolecules with high molecular
weight were rejected by this cake layer.
It is also observed from Table 5 that the flow rate had
a significant effect on the polyphenol concentration
(p < 0.05). Equation (2) shows that this effect was positive.
Figure 4(d) shows that for a pressure of 104 kPa, the
concentration of polyphenol in the permeate increases from
3.2 to 8.4 mg GAE=100 ml when the flow rate varies from
10 to 20 to l=h. It is known that the increased flow rate
reduces the thickness of cake layer by forced convection,
leading to an increase of polyphenol concentration in the
clarified banana.
Optimization of Microfiltration
For optimization of process parameters of microfiltra-
tion (pressure and flow rate), Stat-Ease Design-Expert
 PRIMARY CLARIFICATION OF BANANA JUICE EXTRACT
TABLE 6
Comparison of the parameters of clarified banana juice obtained by centrifugation and by microfiltration
Viscosity Clarity
Clarified banana juice (mPa  s) (%T)
Centrifugation 1.32 69.6
Microfiltration 1.22 93.1
Extract of banana juice 1.49 52.3
v8.0.7.1 software was used. Like centrifugation, the
optimum processing conditions were investigated for the
fives experimental responses (viscosity, clarity, AIS, poly-
phenol, and protein concentration) and conditions taken
in consideration for optimization were to minimize the
values of viscosity and AIS, and to maximize clarity, total
polyphenol, and protein concentration. Parameters have
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been kept in their range (pressure: 35–103.4 kPa, flow rate:
10–20 l=h). The optimum TMP and cross flow rate was
obtained at 103 kPa at 20 l=h with a desirability of 0.77,
where the AIS content is minimized at 0.25% w=w and
polyphenol is maximized as 8.6 mg GAE=100 ml. But
taking into account, the overall quality of the juice, the
numerical optimization was performed and the solution
having the overall desirability (0.6) was selected as the opti-
mum condition for the process of primary clarification of
banana juice by microfiltration. The p value for viscosity,
clarity, AIS, polyphenol, and protein for microfiltration
are 0.0247, 0.0340, 0.0034, 0.0093, and 0.0457, respectively.
This shows that for viscosity, clarity, and protein, only
TMP is important whereas for AIS and polyphenol, the
effects of both TMP and cross flow rates are significant.
The lack of fit for viscosity, clarity, AIS, polyphenol, and
protein are 0.0, 3.69, 0.05, 3.96, and 0.21, respectively. This
shows that the lack of fit is not significant and the model
is suitable. Optimum conditions generated by numerical
optimization are: transmembrane pressure: 75 kPa and
cross flow rate: 20 l=h. The corresponding values of the five
experimental responses were: viscosity: 1.22 mPa  s; clarity:
93.1%T; AIS: 0.24% w=w; polyphenol: 8.4 mg GAE=100 ml
and protein: 769.3 mg=l. Experimental results at optimum
operating condition tally with the results of numerical
optimization. The experimental results were obtained as
viscosity: 1.20 mPa  s; clarity: 94%T; AIS: 0.23% w=w;
polyphenol: 8.1 mg GAE=100 ml and protein: 725.2 mg=l.
Color, TSS, pH, and Conductivity
As with centrifugation, samples of banana juice clarified
by microfiltration for the 10 experiments of the Doehlert
design were also subjected to analysis for the determination
of color, pH, TSS and conductivity. The results showed
that the color was between 0.038 and 0.207 A, the pH
between 3.92 and 4.53, TSS between 3.1 and 5.4
Brix
and conductivity between 748 and 1342 mS=cm.
1167
AIS Polyphenol Protein
(%w=w) (mg GAE=100 ml) (mg=l)
0.41 12.5 951.1
0.24 8.4 769.3
0.58 15.4 1308.6
Comparison between Centrifugation and Microfiltration
Two separation processes, namely centrifugation and
microfiltration, were used to perform the primary clarifi-
cation of banana juice. After clarification, the optimal
values of experimental responses of clarified banana juice
(viscosity, clarity, AIS, polyphenol, and protein) by these
two methods were compared to those of the banana juice
extract and the results are presented in Table 6. From this
table, it is observed that the values of viscosity, clarity, and
AIS are best with the microfiltration processes. Clarified
banana juice retains a higher concentration of polyphenol
and protein with the centrifugation processes. However,
it is clear that the difference between the values of poly-
phenol and protein is not large enough. If a continuous
centrifuge of CEPA type (manufactured by Eppendorf,
Germany) of the same capacity (250 mL) is used, the
manufacturer specified power intake is 300 watt for
6250 g (10,000 rpm). Processing of 1 m3 of juice for
45 min consumes 900 Kwh=m3. On the other hand, in
microfiltration, the pump rating is 240 watt. At 76 kPa,
in 30 min about 1.3 l permeate is collected for 0.026 m2
membrane area. This operation requires 132 Kwh=m3
electrical energy considering pump efficiency of 70%. This
indicates that the operating cost for microfiltration is
about 7 times less compared to centrifugation. Therefore,
including the cost of operation, product quality, and
quantity, microfiltration is the most appropriate method for
primary clarification of banana juice.
CONCLUSIONS
Clarification of banana juice was carried out using cen-
trifugation and hollow fiber microfiltration. A comparative
study between these two processes was conducted and five
parameters namely, viscosity, clarity, AIS, polyphenol, and
protein were analyzed in clarified banana juice. It was
noted that the best result in terms of viscosity, clarity,
and AIS was obtained with microfiltration. The optimal
values of these parameters were: viscosity, 1.22 mPa  s,
clarity, 93.1% T, and AIS, 0.24% w=w. The centrifugation
gave the best optimal values of polyphenol (12.5 mg GAE=
100 mL) and protein (951.1 mg=l). Based on these physical
and nutritional parameters of banana juice obtained, and
taking account of operating parameters, microfiltration
was found to be most suitable for primary clarification of
 1168 S. T. SAGU ET AL.

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