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Banana Juice (FABT 2013)
Banana Juice (FABT 2013)
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ORIGINAL PAPER
Abstract This article focuses on the optimization, modeling, and online monitoring of banana juice production
through an enzymatic method. In order to perform this
task, a batch reactor was designed with automatic control
over the temperature and the agitation speed as well as
online monitoring of torque. The experiments were carried
out with the Musa AAA Cavendish banana variety (Enano
gigante), the main variety planted in Mexico. Three different
ripening stages were evaluated. Optimization of juice extraction was evaluated as a function of the pulp/water relationship
and the concentration of the enzyme complex. The results
showed that the adding of water had no influence on the
extraction of banana juice, and the optimal enzyme concentration per kilogram of banana pulp was found. Based on a
fuzzy logic approach, it was possible to relate the initial torque
V. Ibarra-Junquera (*) : P. Escalante-Minakata :
A. M. Chvez-Rodrguez : I. A. Comparan-Dueas
Bioengineering Laboratory, University of Colima, Carretera
Colima-Coquimatln, Km 9,
Coquimatln, Colima State, Mexico
e-mail: vij@ucol.mx
J. J. Ornelas-Paz
Centro de Investigacin en Alimentacin y Desarrollo A.C.
(CIAD), Unidad Cuauhtmoc, Chihuahua State, Mexico
J. D. Prez-Martnez
Faculty of Chemical Sciences, Autonomous
University of San Luis Potos,
San Luis Potos, S.L.P. State, Mexico
J. A. Osuna-Castro
Faculty of Biological and Agricultural Sciences,
University of Colima, km 40 Autopista Colima-Manzanillo,
28100, Tecomn, Colima, Mexico
C. N. Aguilar
Department of Food Science and Technology, School of
Chemistry, Autonomous University of Coahuila, Saltillo
Coahuila State, Mexico
Introduction
Bananas are one of the worlds most important food crops,
consumed by millions as part of a daily diet and for nutrient
enrichment (Mohapatra et al. 2011) since they are an important
source of polyphenols, minerals, and carbohydrates (Kiyoshi and
Wahachiro 2003; Wall 2006; Escalante-Minakata et al. 2013).
Furthermore, the banana is widely appreciated for its flavor and
aroma (Boudhrioua et al. 2003; Mohapatra et al. 2011). Though
commonly consumed as fresh fruit, bananas have an unfortunately short shelf-life due to softening and thus are often used in
banana juice processing as an alternative (Kyamuhangire et al.
2002; Lee et al. 2006a, b; Lpez-Nicols et al. 2007; Mohapatra
et al. 2011; Chvez-Rodrguez et al. 2013).
A relevant issue in banana juice extraction is the retention
of the juice in the pulp due to the great amount of polysaccharides, which prevent the release of intra-cell components,
affecting the extraction yield and the clarification. The juice
can be extracted by a mechanical press and/or through the
Methodology
Bioreactor
The enzymatic juice extractions were performed in a 4-L
stainless steel batch-jacketed reactor (i.d. 12.5 cm),
equipped with automatic control over the temperature, based
on a cooling thermostat with a precision of 0.02 C (RE
630S; Lauda Eco Silver, Germany). The bioreactor agitation
speed control is composed of the Compact cRio-9074 data
acquisition system (DAS) (National Instruments, TX, USA),
a PC for the control of the DAS, a stepper motor (NEMA 23
Stepper Motor; National Instruments) connected to a
Microstepping Drive (National Instruments), and two independent power supplies. The stepper motor movement is
directed by using an application developed within the
Labview platform (National Instruments). An open-paddle
impeller (width 9.6 cm, height 12.3 cm) is connected to the
motor through a drill chuck. The torque was monitored
online using a rotary torque sensor with a maximum capacity of 2 Nm (FSH01979; FUTEK, CA, USA) connected to a
PC through a data acquisition system from National Instruments (Cryo-9074) and Labview-based interface.
Biological Material
Bananas of the cultivar Enano gigante (Musa AAA, subgroup Cavendish) were acquired in the local market when
they were green (Fig. 1) with a minimum length of 13 cm.
The banana bunches were ripened in laboratory conditions
at 20 C without atmospheric control for different storage
times. The color of the peel was monitored by comparison
with a scale ranging from 1 to 3, with values representing
the following stages of ripeness: Green (1), ranging from
more yellow than green to yellow with green endings;
Yellow (2), ranging from completely yellow to yellow with
slight brown specks; and Brown (3), which included yellow
with many brown specks. Additionally, the parts of individual samples were weighed in an analytical balance to determine the pulp-to-peel ratio. The results were expressed as
the percent weight of pulp relative to peel.
Colorimetric Analysis
Juice Yield
The banana juice yield was calculated based on the weight
of the resulting juice, the added water, and the banana pulp,
according to the following equation:
Juice yield%
jwgwwg
100
bwg
Statistical Analysis
All statistical analyses were carried out using MatLab software (MathWorks Inc., USA). The analysis of variances
(ANOVA) was applied to compare the mean values of
samples according to the different factors. In addition, principal component analysis (PCA) was used as a data multivariate technique. The purpose of this method is to decompose the data matrix and concentrate the source of variability in the data into the first few principal components. Here,
PCA was applied to two different groups of values: the
matrix of maturity mean attribute ratings across banana
samples (initial torque and the peel-to-pulp ratio) and the
matrix of mean color measurements (L, a, and b). The two
cases were also analyzed by cluster analysis (average linkage method) in an attempt to classify the samples in terms of
their maturity and processing method.
Visual Evaluation
The juice data, coming from the colorimetric analysis, were
evaluated using PCA and cluster analysis. As an additional
reference, the resulting groups were classified in terms of
visual acceptance by a five-judge panel belonging to the
faculty of Chemical Sciences and research scholars of the
Bioengineering Laboratory (both of the University Colima
in Mexico). This general appearance of the juice was measured subjectively on a three-point hedonic scale: poor
(dislike), good (like), and excellent (like very much).
Optimization Procedure
Protein Quantification
The protein concentration of the enzymatic complex was
determined by the Bradford method (1976) as modified by
BioRad (using bovine serum albumin protein as standard).
The samples were diluted to a ratio of 1:20 by adding
distilled water. The absorbance of the samples was determined at 595 nm using a lambda 25 UVVis spectrophotometer (Perkin-Elmer Instrument, USA).
water/banana pulp taking values in the set {1, 0.9, 0.8}; and
Z, response variable. Given are a total of 63 experiments, to
be performed in triplicate. The juice yield, as evaluated in
Eq. 1, was the response variable. Based on the preliminary
data, the following generalized MichaelisMenten response
surface (Hirst et al. 1996) is used to model the data behavior:
Z X ; Y
bXY
c dYZ
2
Fig. 2 Memberships function with triangular membership distribution
m T ; x2;0
dx2
xmax T ; x2;0 x1
dt
y T ; x2;0
Dependence
Units
xmax(T,x2,0)
f1(T,M)
y(T,x2,0)
m(T,x2,0)
f2(T,M)
1/N m
km
max
max S
k m S
Be
M X 2;0
1
Ae T
( M(X2,0))2 +w M(X2,0)+
e Y
M X 2;0
L/kg
measurements x2(t). Thus, this methodology can be considered as a data-driven approach to the modeling of the
enzymatic extraction of banana juice in which its structure
and the parameter identification is based on off-line and online time series obtained from a rigorous experimental set.
Software Sensor Design
Taking into account the fact that rarely can one have a
sensor on every state variable, and some form of reconstruction from the available measured output data is needed,
software can be constructed using the mathematical model
of the process to obtain an estimate of the true state x. Ever
since the original work by Luenberger (1971), the use of
state observers has proven useful in process monitoring and
for many other tasks. In the sense of control theory, an
algorithm capable of giving a reasonable estimation of the
unmeasured variables of a process will be called an
observer.
Numerous attempts have been made to develop observer
design methods for bioprocess applications (EscalanteMinakata et al. 2009; Zhang 2009; Fernndez-Fernndez
and Prez-Correa 2010). The first systematic approach for
the development of a theory of observers was proposed
some time ago by Krener and Isidori (1983). Nevertheless,
it is well known that classical proportional observers tend to
amplify the noise of on-line measurements, which can lead
to the degradation of the observer performance. In order to
avoid this drawback, in this paper, the observer algorithm is
based on the work of Ibarra-Junquera et al. (2005) because
the proposed integral observer provides robustness against
noisy measurements and uncertainties.
In this section, the design of the software sensor is
presented in which xj, for j(1n), is the naturally measured state (i.e., the variable easiest to measure). Therefore,
it seems logical to take xj as the output of the system, y=h(x)
=xj. Now, considering that the output function h(x) is contaminated with a Gaussian noise, the model given by the
aforementioned Eqs. 3 and 4 acquires the form:
X f X
y cX
75
70
65
60
55
50
45
1
0.95
0.9
600
0.85
400
water/pulp ratio
0.8
200
0
x 10
-3
a)
x 10
b)
2
2
4 x 10
Concentration (L/kg)
0
0
x 10
c)
1
0
0
3
m
x 10
2
4
Concentration (L/kg)
Fig. 4 Plots show the nonlinear regression plots for kinetic of juice
yield by the enzymatic complex, the lines (. . .), (__), (-+-+), and (- - -)
stand for the model prediction at 20, 30, 40, and 50 C, respectively.
0
0
x 10
2
4
Concentration (L/kg)
The symbols (), (*), (), and () stand for experimental data at 20, 30,
40, and 50 C, respectively. Plots (a), (b), and (c) correspond to
ripeness stages 1, 2, and 3, respectively
-0.5
1/N m
L/kg
C
C
C
-5.5
1/T
Ripeness stage
-6
-6.5
0.03
0.04
c)
-6
-6.5
-7
0.05
1/T
-5
d)
-5.5
b)
-7
0.05
0.03
0.04
-10
f)
-20
-5.5
-6
0.05
1/T
e)
-1
-1.5
1
0.14T+1.5M(X2,0)+62.63
46T1,400M(X2,0)373.33
0.008325768
0.3308
0.234898
0.25541
9.1405
10.4698
38.422
Ln max
-6.5
0.04
xmax(T,x2,0)
y(T,x2,0)
Ln max
a)
0.03
Units
-6
-7
Value
Ln A
Ln max
-5.5
Ln Km
Parameter
Ripeness stage
-30
Ripeness stage
75
0
y (T,M)
Xmax (T,M)
70
65
50
40
30
Temperature (C)
20 3
2
Ripeness stage
-5000
-10000
50
40
Temperature (C)
30
20 3
1
2
Ripeness stage
2
1
0
-1
-2
-3
-2
-1
0
1
First Principal Component (71.1%)
Torque
Brown
(0.008,0)
Yellow
(0.0135,0)
Green
(0.0195,0)
Ripeness stage
1
(0.5,0)
2
3
Temperature
20 C
30 C
40 C
50 C
Initial yield
40
45
50
55
60
65
Set type
(0.0113,1)
(0.0149,1)
(0.024,1)
(0.0145,0)
(0.0205,0)
(0.0285,0)
Input
Input
Input
(1,1)
(1.5,0)
Output
(1.4,0)
(2.4,0)
(2,1)
(3,1)
(2.45,0)
(3.6,0)
Output
Output
(14,0)
(24,0)
(34,0)
(44,0)
(20,1)
(30,1)
(40,1)
(50,1)
(26,0)
(36,0)
(46,0)
(56,0)
Input
Input
Input
Input
(36,0)
(41,0)
(46,0)
(51,0)
(56,0)
(61,0)
(40,1)
(45,1)
(50,1)
(55,1)
(60,1)
(65,1)
(44,0)
(49,0)
(54,0)
(59,0)
(64,0)
(69,0)
Output
Output
Output
Output
Output
Output
Colorimetric Analysis
The color of a product is one of the most important
properties that influence the consumers response to it.
Therefore, one of the objectives of this study was to
determine how the characteristic of the raw material and
the process conditions influence the final color attributes
of the banana juice. In order to allow the interpretation
the color results from the L.a.b. analysis, a PCA technique plus a clustering analysis were performed. The
results appear in Fig. 9.
Cluster analysis groups data objects with similar characteristics that were also made then different from objects of
other group. Therefore, the three clusters formed in Fig. 9
correspond to the banana juice with similar colorimetric components. The sensorial panelists were asked to categorize the
samples corresponding to each attribute of all samples. From
the PCA score, the cluster analysis, and opinions of the
panelists, it can be concluded that the clusters of green diamonds obtained in Fig. 9 are the ones with the best visual
attributes. This group corresponds to ripeness stage 1 (green)
and with no influence of the temperature or the enzyme
concentration; however, it is important to mention that the
closest neighbors were samples of the blue star group corresponding to ripeness stage 2 (yellow) and produced at temperatures of 40 and 50 C.
The Software Sensor
Set type refers to the nature of the set that is an input and/or output to
the fuzzy logic algorithm
x0 x2
x1 mxmax x1
m
xmax x1
Y
x2
Fig. 8 The plot corresponds to the correlation between the kinetic rate
and temperature in the banana juice extraction process. Green squares,
blue triangles, and red circles correspond to ripeness stages 1, 2, and 3,
with and R2 of 0.885, 0.998, and 0.992, respectively
y0 x0
-1
G-40-100L
G-50-100L
G-50-300L
G-30-500L
G-20-300L
G-40-300L
B-20-500L
Y-30-300L
Y-50-300L
Y-20-500L
Y-30-500L
B-20-300L
B-40-100L
G-30-300L
Y-20-500L
B-20-100L
Y-50-500L
Y-20-300L
Y-30-100L
G-50-500L
Y-20-100L
Y-40-500L
B-30-500L
B-30-100L
Y-40-100L
B-30-300L
B-40-500L
Y-40-300L
B-50-300L
B-50-500L
B-50-100L
B-40-300L
Y-50-100L
-2
-3
-2
-1
Fig. 9 Scatter plot for the first two principal components with colored
clusters corresponding to the matrix of juice color (L, a, and b) at
different process conditions: temperatures, enzymatic complex concentration, and ripening stages (the three generated clusters are marked as
red triangles, blue five-pointed stars, and green diamonds). The letters
S A I A I S C T C:
2
2
X AX B
y CX
Where
2
3
2
3
0
0 1
A 4 0 m
0 5; B 4 m xmax 5and C 1 0 0 :
0 m=Y 0
m xmax =Y
An asymptotic-type observer of the system is given as
follows:
c
b 2 k 1 y0 X
b0
X0 X
b
b 1 k 2 y0 X
b0
X 1 m xmax X
m
b
b 1 k 3 y0 X
b0
X max X
X2
Y
10
11
2m
Y
K 4 k2 5
m
2m3m
k3
It is worth mentioning that we can think of this observer
as a slave system that follows the master system, which
is precisely the real experimental system. In addition, S, as
functional components of the gain vector, guarantee the
accurate estimation of the observer through the convergence
to zero of the error dynamics, i.e., the dynamics of the
difference between the measured state and its corresponding
estimated state. One can see that generates an extra degree
of freedom that can be tuned by the user such that the
performance of the software sensor becomes satisfactory
for him (here, we use = 0.01).
The reason for the filtering effect is that the dynamic
extension acts at the level of the observer as an integration of the output of the original system (see the first
equation of the system given by Eqs. 58 and the error
part in the equations of system given by Eqs. 911).
The integration has averaging effects upon the noisy
measured states. More exactly, the difference between
the integral of the output of the slave part of system
and the integral of the output of the original system
0.025
Torque (Nm)
Torque (Nm)
0.02
0.015
0.01
0
500
1000
1500
2000
2500
3000
3500
4000
0.025
b)
0.02
0.015
0.01
0
4500
500
1000
1500
2000
3000
3500
4000
4500
3000
3500
4000
4500
80
80
60
40
20
0
2500
Time(s)
Time(s)
500
1000
1500
2000
2500
3000
3500
4000
4500
60
40
20
0
500
1000
1500
2000
2500
Time(s)
Time(s)
Torque (Nm)
Torque (Nm)
0.02
c)
0.02
0.015
0.01
0
500
1000
1500
2000
2500
3000
3500
4000
d)
0.015
0.01
0
4500
500
1000
1500
80
60
40
0
500
1000
1500
2000
2500
3000
3500
4000
4500
1000
1500
2000
2500
4000
4500
3000
3500
4000
4500
60
40
0
500
1000
1500
3000
3500
4000
2000
2500
f)
0.014
0.012
0.01
0
4500
500
1000
1500
2000
2500
3000
3500
4000
4500
3000
3500
4000
4500
Time (s)
80
80
Juice Yield (%)
3500
0.016
Time (s)
60
40
0
3000
Time (s)
Torque (Nm)
Torque (Nm)
e)
500
2500
80
Time (s)
0.016
0.014
0.012
0.01
0
2000
Time (s)
Time (s)
500
1000
1500
2000
2500
3000
3500
4000
4500
Time (s)
60
40
0
500
1000
1500
2000
2500
Time (s)
Conclusions
The optimization of the banana juice extraction process
shows that the addition of water did not significantly affect
the juice yield, while the enzymatic complex concentration
exhibits a positive impact on the juice yield. As such, the
optimal conditions for juice extraction (i.e., more sustainable) were 150 L/kg enzyme complex concentration without water addition. Moreover, from this study, it can be
concluded that the model here developed could be a valuable instrument for monitoring and controlling enzymatic
banana juice extraction. Furthermore, enzyme reactors can
operate fed-batch-wise or continuously, and the model is a
helpful tool for optimization. Moreover, the software sensor
developed here effectively rebuilds the unmeasured on-line
juice yield and also is able to filter the noisy torque signal.
In addition, the computational scheme provides a very appropriate tool for fast and reliable quality control and can be
used to ensure the homogeneity of the final product. The
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