Food Anal.

Methods (2014) 7:13281336

DOI 10.1007/s12161-013-9753-y

Effects of Temperature, Time, and pH on the Stability

of Anthocyanin Extracts: Prediction of Total Anthocyanin
Content Using Nonlinear Models
Lutfiye Ekici & Zeynep Simsek & Ismet Ozturk &
Osman Sagdic & Hasan Yetim

Received: 18 September 2013 / Accepted: 31 October 2013 / Published online: 19 November 2013
# Springer Science+Business Media New York 2013

Abstract In this study, different anthocyanin sources including grape skin, black carrot, and red cabbage were used to
determine the effect of thermal treatment, different acidity
levels, and time on the anthocyanin content and degradation.
The total anthocyanin contents were modeled by neuro fuzzy
inference system (ANFIS) and artificial neural network
(ANN) models. The red cabbage anthocyanin stabilities were
higher than others. The anthocyanins degraded more rapidly
at higher temperatures. The anthocyanin contents of samples
decreased with the increase of pH from 3 to 7. Comparison of
the models showed that the ANFIS model performed better
than the ANN model for the estimation of total anthocyanin
content in all samples. The lowest root mean square error
(0.0457) and highest R 2 (0.9942) values were obtained for
red cabbage and grape skin in the validation period with the
ANFIS model, respectively. This study showed that both
models can be utilized efficiently for the prediction of total
anthocyanin content affected by temperature, time, and pH.

Keywords Anthocyanin . Heat degradation . Modeling .

ANFIS . ANN

L. Ekici (*) : I. Ozturk : H. Yetim

Faculty of Engineering, Department of Food Engineering, Erciyes
University, 38039 Kayseri, Turkey
e-mail: lutfiyed@erciyes.edu.tr
Z. Simsek
Pazarlar Vocational College, Department of Food Processing,
Dumlupinar University, 43530 Kutahya, Turkey
O. Sagdic
Faculty of Chemical and Metallurgical Engineering, Department of
Food Engineering, Yildiz Technical University, 34210 Istanbul,
Turkey

Introduction
Interest in anthocyanins as natural colorants and antioxidant
ingredients has recently increased due to their color characteristics and potential health benefits (Walkowiak-Tomczak
and Czapski 2007). Furthermore, epidemiological studies
have suggested that the anthocyanins have a protective effect
against some cancers (Castaneda-Ovando et al. 2009).
Anthocyanins are glycosides of anthocyanidins (aglycone),
which differ from one another in the position of substitution of
hydroxyl and methoxy groups in the -ring. Anthocyanin
residues are frequently acylated with phenolic acids
(Walkowiak-Tomczak and Czapski 2007). Some factors like
pH, temperature, copigments, light, metallic ions, enzymes,
oxygen, ascorbic acid, sugar, and their degradation products
can cause anthocyanin degradation and deterioration of food
color (Mazza and Miniati 1993).
Black carrot, red cabbage, red radish, red potato, and grape
pomace are popular anthocyanin sources (Newsome 1986).
For example, the black carrot (Daucus carota L. ssp. sativus
var. atrorubens Alef.) originating from Turkey, Afghanistan,
Egypt, Pakistan, and India has been known for 3,000 years
(Kammerer et al. 2004). Red cabbage (Brassica oleracea L.
var. capitata f. rubra) which belongs to the family of Brassicaceae grows all over the world (Arapitsas et al. 2008). The
grape (Vitis vinifera L.) has an annual production rate of
around 69 million tons and is cultivated mainly for wine
production (Maier et al. 2009). Wine industry wastes may
account almost 30 % (w /w ) of the grapes used in wine
production. Varietal differences and wine-making processes
are the major factors which can change the phenolic content
and composition of grape pomace (Ruberto et al. 2007). In this
research, the skins of one of the most important red wine grape
varieties in Turkey Okuzgozu, black carrot, and red cabbage
were used as anthocyanin sources. The stability of the anthocyanins is closely related to a set of common physicochemical

Food Anal. Methods (2014) 7:13281336

parameters including pH and temperature used in food

processing.
The artificial neural network (ANN) and fuzzy logic are
nonlinear models that offer real advantages over conventional
modeling, involving the ability to handle large amounts of
noisy data from nonlinear and dynamic systems (Nayak et al.
2004). ANN and fuzzy logic are intelligent models that have
the capability to relate input and output parameters without
prior information (Karaman et al. 2011). In recent years, soft
computing modeling techniques such as fuzzy logic and artificial neural networks have been used for the modeling of
complex nonlinear systems in food science and technology
research. These modeling techniques are effective and versatile in the modeling and estimation of nonlinear systems (Abu
Ghoush et al. 2008; Karaman et al. 2011; Yalcin et al. 2011).
Food structure is known as a nonlinear system. Therefore, the
modeling and estimation of food systems such as food processing, quality assessment, optimization, and control using
these modeling techniques are appropriate (Abu Ghoush et al.
2008; Hernandez 2009; Yalcin et al. 2011). In addition, these
approaches are fast and cheap tools compared to analytical
methods.
Although ANN has been used in process control, medical
diagnosis, forensic and investment analysis, weather estimation, financial application, and dam control for a long time, the
application of ANN in the field of food science and technology is quite novel (Huang et al. 2007; Kisi 2007). Yalcin et al.
(2011) used an adaptive neuro fuzzy inference system
(ANFIS) and ANN models to evaluate antioxidant agents
(gallic acid, ellagic acid, quercetin, -carotene, and retinol)
and their effectiveness on oxidation parameters during the
storage period. In a similar study, ANFIS and ANN models
were used to predict the antioxidant effect of some plant
extracts on the oxidation of vegetable oils (Karaman et al.
2011). In another study, ANFIS was used for the modeling of
the emulsion stability and viscosity of mayonnaise prepared
using iota carrageenan and wheat protein as an emulsifier
alternative to egg yolk (Abu Ghoush et al. 2008). Xie et al.
(1999) compared the kinetic, neural network, and fuzzy logic
models in order to study textural changes in dried peas cooked
in water at different temperatures, and in this study, neural
network showed better fitting capacity compared to others.
Kilic et al. (2007) used an ANN model for the estimation of
color of the edible oil based on the L*, a*, and b* format. This
study found that the ANN model can be used to evaluate the
color of most edible oils. Taghadomi-Saberi et al. (2013a)
used a combining image processing and ANN model for the
prediction of antioxidant activity and anthocyanin content of
different ripening stages of sweet cherry. In another study, the
effect of ripening stages on antioxidant activity and anthocyanin content of sweet cherry was estimated by combining
image processing and two artificial intelligence techniques.
It was revealed that ANFIS models with triangular and two-

1329

term Gaussian membership functions exhibited the best results for antioxidant activity and anthocyanin content, respectively (Taghadomi-Saberi et al. 2013b).
The aim of the present study was firstly to determine the
stability of the anthocyanins obtained from black carrot, red
cabbage, and grape skin in buffer systems, at different temperatures and acidity levels, and secondly to predict the total
anthocyanin content of extracts depending on temperature,
time, and pH using ANFIS and ANN modeling techniques.

Materials and Methods

Plant Materials
Black carrot (D. carota L.) and red cabbage (B. oleracea L.
var. capitata f. rubra) were obtained from Konya Eregli and
Yemliha-Kayseri, Turkey, respectively. The samples were
washed with tap water and subsequently stored at 40 C
(Hettich Freezer, Germany) until use. Black grape (V. vinifera)
pomace samples (Okuzgozu) were kindly donated by the
Doluca Winery in Tekirdag, Turkey. The seed stem and skin
fractions of the pomace sample were separated manually, and
the skin fraction was dried at 65 C for 24 h and then ground
with a laboratory type grinder to use for the anthocyanin
extraction.
Preparation of Anthocyanin Extracts
The extraction was performed by the methods described by
Ersus and Yurdagel (2007) with a slight modification. After
thawing the plants (black carrot and red cabbage) at 4 C for
12 h, 50 g of each material was ground in a Waring blender
with 200 ml of an ethanol/water (1:1) solution acidified with
0.1 % hydrochloric acid. The blended plant slurries were
macerated for 2 min to extract anthocyanins. The anthocyanins were extracted at 35 C in a water bath equipped with a
shaker by stirring continuously for 2 h. The extract was
filtered through Whatman no. 1 filter paper using a Buchner
funnel and dried at 60 C under vacuum.
Preparation of Buffer Systems
Citratephosphate buffer solutions were prepared at five different pH levels (3.0, 4.0, 5.0, 6.0, and 7.0) according to
Anonymous (2008). The pH values of the buffer systems were
controlled using a pH meter (WTW, Werkstatten, Germany)
before use.
Anthocyanin Degradation Studies
The effect of pH on the thermal stability of black carrot, red
cabbage, and grape skin anthocyanins was studied at five

1330

different pH levels (3.0, 4.0, 5.0, 6.0, and 7.0) at 70, 80, and
90 C for 120 min. The citratephosphate buffer solutions at
these pH levels were colored with anthocyanin-based extracts
(100-ml buffer solutions were colored with 4 mg anthocyanin;
black carrot extract, 1.20 g; red cabbage extract, 0.50 g; grape
skin extract, 0.37 g), and they were divided into 15-ml portions and poured into Pyrex tubes and incubated in a thermostatic water bath (Memmert WB-22, Germany) which has
been preheated to the given temperature. Samples were removed from the water bath at 30-min intervals and cooled by
running tap water. The anthocyanin contents were measured
by the pH differential method described by Giusti and
Wrolstad (2001).
Total Anthocyanin Content
Determination of the total anthocyanin contents of the samples
was performed according to the pH differential method (Giusti
and Wrolstad 2001), and values were expressed as cyanidin-3glucoside (cyd-3-glc) equivalents, according to the following
equation: c (mg/kg)=(A MWDF1,000d)/ M, with A =
absorption value, MW = molecular weight (449.2 g/mol for
cyd-3-glc), M = molar extinction coefficient (26,900 L/mol/
cm for cyd-3-glc), DF = dilution factor, and d = path length of
the cuvette (1 cm). Absorbance of the sample was determined
using a spectrophotometer (Varian Cary 100 Conc UVVisible, USA).
Adaptive ANFIS Model
The fuzzy logic toolbox of MATLAB 7.0.1 was used for
ANFIS modeling (MATLAB 2009). Firstly, the experimental
parameters of the samples were classified. In the construction
of a set of fuzzy if-then rules with appropriate membership
functions (MFs) from the specified inputoutput pairs, ANFIS
employs an ANN learning algorithm. The two methods for
updating MFs used by ANFIS in the training phase are (a)
back propagation for all parameters (a steepest descent method) and (b) a hybrid method consisting of back propagation
for the parameters associated with the input membership and
least squares estimation for the parameters associated with the
output MF (Jang 1993). Figure 1a illustrates the general
architecture of an ANFIS model. A fuzzy inference system
(FIS) is composed of two inputs x and y and one output z. For
the first-order Sugeno fuzzy model, a typical rule set with two
fuzzy if-then rules can be expressed as:
Rule 1 If x is A1 and y is B1, then z i =p 1x +q 1y +r 1
Rule 2 If x is A2 and y is B2, then z 2 =p 2x +q 2y +r 2
where the A1 and A2 and B1 and B2 are the MFs for inputs x
and y, respectively, and p i, q i, and r i are the output functions
parameters. The ANFIS system is functionally equivalent to

Food Anal. Methods (2014) 7:13281336

the Sugeno first-order FIS. A detailed explanation of ANFIS

can be found in Jang (1993) and Kisi (2006).
ANN Model
The artificial neural network toolbox of MATLAB 7.0.1 was
used for ANN modeling (MATLAB 2009). ANN is composed
with one or more hidden layers, whose computation nodes are
known as hidden neurons. A three-layered ANN architecture
which is composed of three layers i , j , and k , with the
connection weights (Wij and Wjk) between the input, hidden,
and output layers was illustrated in Fig. 1b. During a learning
process, initial assigned weights are corrected. In this process,
the estimated outputs are compared to the known outputs, and
the errors are back propagated (from right to left in Fig. 1b) to
obtain the appropriate weight adjustments which are necessary to minimize the errors.
In the current study, the ANN was trained using the
LevenbergMarquardt technique because this technique is
more powerful and faster than the conventional gradient descent technique (Hagan and Menhaj 1994; Kisi 2007). The
theoretical explanation of ANN can be found in detail in
Haykin (1998).
For the comparison of ANFIS and ANN models and the
selection of the optimal model, their performance was determined using the root mean square error (RMSE), mean absolute error (MAE), and determination coefficient (R 2); these are
common comparison criteria in modeling. The two error measurement equations used were as follows:
v
u
N
u1 X


RMSE t
Y iobserved Y ipredicted
N i1

MAE

N

1 X 
Y iobserved Y ipredicted 
N i1

in which N is the number of data and Yi is the apparent

viscosity used as output.
12
0


Xn 
xix
yiy
i1
C
B
R2 @ X 
2 Xn 
2 A
n
xix
yiy
i1
i1
The determination coefficient is the square of the correlation coefficient between two variables, x and y, whose n pairs
are available. The lowest values for RMSE and MAE are
desired because the lower these values are, the better is the
accuracy of the model to estimate the parameter. Additionally,
high R 2 values indicate that the model performed is the best
(Yalcin et al. 2011).

Food Anal. Methods (2014) 7:13281336

1331
Input

Fig. 1 The architectures of the a

ANFIS and b ANN

Inputmf

Rule

Outputmf

Output

Temperature

pH

Anthocyanin

Time

b
Statistical Analyses
Each experiment was repeated at least two times in triplicate.
SAS statistical software (SAS 2000) was used for the data
analysis, and all the tests were considered to be significant if P
<0.05.

Results and Discussion

Effects of Temperature, pH, and Time on Degradation
of Anthocyanins
Different anthocyanin sources were used to determine the
effect of thermal treatment and different acidity levels. Anthocyanins have four different forms: flavylium cation, quinoidal
bases, carbinol pseudobase, and chalcone pseudobase forms.
While the flavylium form predominates at pH 2 or below, the

colorless chalcone pseudobase begins to dominate at elevated

pH levels. Beside higher pH levels, elevated temperatures and
prolonged exposure can cause chalcone formation
(McDougall et al. 2007).
As expected, anthocyanin degradation increased at elevated pH and/or temperature (Table 1). In general, statistical
analyses revealed that the type of anthocyanin source, pH,
and heating time had a significant (P <0.05) effect on the
degradation of anthocyanins. In pH 3 buffer systems, the
grape skin, black carrot, and red cabbage anthocyanins after
treatment at 70 C were degraded 11.82, 9.36, and 2.57 %,
respectively. It was determined that the anthocyanins degraded more rapidly at higher temperature, particularly when the
samples were heated at 90 C. These results could be attributed to differences in the anthocyanin profiles of black carrot,
red cabbage, and grape skin. The red cabbage anthocyanin
stabilities at 70, 80, and 90 C were higher than those of black
carrot and grape skin anthocyanins (Table 1). In a similar

1332

Food Anal. Methods (2014) 7:13281336

Table 1 Thermal degradation of black carrot, red cabbage, and grape skin anthocyanins (milligram cyd-3-glc/L) at different temperatures, times, and pH
levels
pH Time (min)

Black carrota

Red cabbage

Grape skin

0
30
60

70 C
40.050.59
38.750.87
38.440.28

80 C
40.040.45
38.030.59
37.150.96

90 C
40.030.53
32.251.52
28.011.30

70 C
40.030.59
39.700.30
39.500.47

80 C
40.060.37
39.490.26
38.740.65

90 C
40.050.41
35.510.92
32.910.78

70 C
40.020.40
38.250.22
37.460.80

80 C
40.060.58
38.010.49
37.260.60

90 C
40.050.55
31.120.85
26.731.34

90
120
Degradation %
0
30
60
90
120
Degradation %
0
30
60
90
120
Degradation %
0
30
60

37.930.89
36.300.67
9.36
40.030.52
38.371.48
36.800.50
36.321.39
35.241.09
11.97
40.030.32
38.131.10
36.700.89
35.821.62
34.510.81
13.79
40.021.17
37.251.65
34.951.47

36.180.85
35.361.05
11.69
40.040.50
36.821.20
34.750.81
33.120.78
31.721.35
20.78
40.050.75
34.200.47
32.361.13
29.390.87
27.591.16
31.11
40.000.54
32.870.52
29.040.85

24.992.16
22.162.34
44.64
40.020.45
31.871.37
28.110.97
25.220.95
22.660.74
43.38
40.030.57
30.821.68
27.480.94
24.490.79
21.190.58
47.06
40.020.64
28.421.48
22.681.38

39.300.37
39.000.42
2.57
40.030.47
38.710.73
38.470.31
38.170.56
37.440.72
6.47
40.010.46
38.580.34
38.150.41
37.870.60
37.410.61
6.50
40.030.79
38.450.80
38.150.23

38.250.61
38.040.63
5.04
40.050.70
37.150.80
36.410.91
35.750.93
35.410.72
11.59
40.020.26
35.820.71
33.210.32
31.080.38
30.160.88
24.64
40.030.37
34.201.28
32.940.62

31.450.98
29.600.60
26.09
40.030.46
34.790.78
31.910.81
30.610.62
28.700.96
28.30
40.050.30
32.720.69
30.440.96
27.370.47
26.701.34
33.33
40.060.35
31.140.38
27.860.58

36.620.71
35.290.80
11.82
40.050.38
37.800.48
36.820.18
35.841.01
35.020.68
12.56
40.010.48
36.470.44
35.050.58
32.841.02
31.680.31
20.82
40.060.55
36.241.07
32.400.96

36.320.49
35.110.49
12.36
40.040.54
37.210.75
35.380.28
33.120.52
31.980.36
20.13
40.020.34
31.400.34
28.750.38
26.220.71
24.970.18
37.61
40.070.41
28.250.30
24.300.41

23.280.89
20.110.79
49.79
40.090.63
30.630.90
27.000.85
24.170.97
21.900.22
45.37
40.100.65
29.740.63
24.940.59
22.200.61
20.470.92
48.95
40.070.71
19.481.26
14.050.30

90
120
Degradation %
0
30
60
90
120
Degradation %

33.861.90
30.570.89
23.61
40.020.60
32.701.12
29.151.31
27.110.92
25.771.55
35.61

25.920.51
22.770.73
43.08
40.041.08
31.131.79
27.731.56
24.400.95
22.090.86
44.83

17.531.13
14.490.46
63.79
40.000.65
26.201.53
20.951.18
16.320.46
14.301.83
64.25

37.770.62
37.250.67
6.94
40.060.37
37.301.04
35.790.31
34.890.32
34.050.45
15.00

32.100.55
31.170.35
22.13
40.010.53
31.590.58
27.910.63
25.680.31
23.790.61
40.54

25.090.85
23.261.17
41.94
40.050.43
28.551.19
26.070.62
21.210.69
19.250.93
51.94

30.921.00
28.810.25
28.08
40.040.49
28.870.71
24.690.63
22.250.50
20.100.52
49.80

21.940.72
19.290.82
51.86
39.960.82
23.711.15
16.810.60
14.460.39
12.050.64
69.84

11.970.42
9.971.40
75.12
40.090.58
19.121.09
13.490.65
9.950.74
7.280.71
81.84

The results were expressed as cyd-3-glc

study, Cevallos-Casals and Cisneros-Zevallos (2004) investigated the heat and pH stability of red sweet potato, purple
corn, and commercial purple carrot colorants and reported that
acylated anthocyanin sources were more stable than nonacylated ones. Red cabbage anthocyanins had the highest heat
stability because of their unique anthocyanin composition. In
particular, the highest heat stability of red cabbage anthocyanins is based on their acylated anthocyanin compositions
(Cevallos-Casals and Cisneros-Zevallos 2004; Dyrby et al.
2001; Pliszka et al. 2009). Cyanidin, which occurs as
cyanidin-3-sophoroside-5-glucoside and cyanidin 3,5diglucoside, acylated with sinapic, malonic, ferulic, and pcoumaric acids is the major aglycone of red cabbage
(Walkowiak-Tomczak and Czapski 2007). On the contrary,

the majority of anthocyanin sources such as grape skin anthocyanins which are all monosides and less acylated show light
resistance to heat, light, and pH changes (Hong and Wrolstad
1990). Black carrot anthocyanin pigment which is acylated
with p -coumaric, ferulic, p -hydroxybenzoic, and sinapic
acids exhibits higher hydration, light, and pH stability
(Kammerer et al. 2004).
The color stability of anthocyanins is attributed to pH and
their anthocyanin structures (Dai et al. 2009). Dyrby et al.
(2001) reported the heat stability of red cabbage, blackcurrant,
elderberry, and black grape colorants at pH 3 and at 80 C to be
red cabbage > blackcurrant > grape skin > elderberry. CevallosCasals and Cisneros-Zevallos (2004) found the order of stability of some anthocyanin sources to be red sweet potato > purple

Food Anal. Methods (2014) 7:13281336

carrot > purple corn > red grape at pH range of 0.9 and 4. These
results indicate that anthocyanin compositions contribute to
stability and that acylated anthocyanins are more resistant than
the non-acylated anthocyanins.

Comparison of Performances of ANFIS and ANN Models

ANFIS and ANN models were used to estimate the anthocyanin content of the samples at different temperatures, times,
and pH levels. The input parameters considered in the study
are temperature, pH, and time. The 900 experimental data
which were divided into three parts, training (50 %), testing
(25 %), and validation (25 %), were used for the ANFIS and
ANN analyses. In training section, the first 450 data were used
and the second 225 and the remaining 225 data were used for
testing and validation, respectively. Firstly, a normalization
was applied for the training input and output data using the
following equation:

The measured and predicted anthocyanin contents of the

black carrot in the validation period are shown in Fig. 2. It can
be clearly seen from the figure that the ANFIS estimates are
closer to the corresponding observed values than those of the
ANN. Figure 3 illustrates the measured and predicted anthocyanin content of the grape skin samples in the validation
period. Again, here also, the ANFIS showed the best accuracy.
For the red cabbage samples, the measured and predicted
anthocyanin values in the validation period are shown in
Fig. 4. As can be seen from the figure, the ANFIS model
seems to be much better than the ANN model.
Overall, ANFIS model was determined to be the best model
for fitting with high performance for all data sets in each extract
sample. The lowest RMSE values were observed in the ANFIS
model. The RMSE values for black carrot, grape skin, and red
cabbage in the validation period were 0.0746, 0.0467, and
0.0457, respectively, in the ANFIS model, but these values
were calculated to be 1.3197, 1.2669, and 0.9476 using the

xi xmin
b
xmax xmin

ANFIS
Predicted Anthocyanin Values

where x min and x max are the minimum and maximum of the
training data set. The a and b were taken as 0.6 and 0.2 in this
study, respectively. The data were scaled between 0.2 and 0.8
and it was observed that this follows the suggestion of
Cigizoglu (2003) who demonstrated that scaling input data
between 0.2 and 0.8 provide flexibility for the models to
predict beyond the training range. Then, different ANFIS
and ANN architectures were tried to find the appropriate
model structures. Gauss MFs were used for the ANFIS in
the current study. In each application, different numbers of
MFs were tested and the best one which results the minimum
RMSE was selected. The best results for the ANFIS models
were recorded with the model which has seven membership
functions. The ANN with one hidden layer was used and the
trial and error method was applied to determine the hidden
nodes number. Sigmoid and linear activation functions were
used for the hidden and output nodes of the ANN, respectively. The training of ANN networks was stopped after 100
epochs because the variation of error was too small after this
epoch.
Table 1 shows the comparison of the training, testing, and
validation results of the ANFIS and ANN models. The optimal number of MFs of each ANFIS model and nodes in the
input, hidden, and output layers of each ANN model is also
tabulated in this table. As can be clearly seen from the table,
the ANFIS model performance was determined to be much
better compared to the ANN models for the training, testing,
and validation periods. The ANFIS model estimated the anthocyanin content of the samples with a quite high degree of
accuracy (RMSE<0.0457, MAE<0.5268, and R 2 >0.9942).

50

40

30

20

10

0
0

10

20

30

40

50

40

50

Measured Anthocyanin Values

50

ANN
Predicted Anthocyanin Values

1333

40

30

20

10

0
0

10

20

30

Measured Anthocyanin Values

Fig. 2 The estimation of total anthocyanin content for black carrot by

ANFIS and ANN models in validation period

1334

Food Anal. Methods (2014) 7:13281336

ANN model (Table 2). The ANFIS model showed a rather high
determination of the coefficient (R 2) for validation period
compared to the ANN for all the samples. While the R 2 values
for black carrot, grape skin, and red cabbage were determined
to be 0.9769, 0.9942, and 0.9836, respectively, for the ANFIS
model, they were calculated to be 0.9674, 0.9809, and 0.9683,
respectively, using the ANN model. A similar trend was observed for the MAE values. The MAE values for the black
carrot, grape skin, and red cabbage were calculated to be
0.8674, 0.5491, and 0.5268 in the ANFIS model, while they
were 1.0359, 0.9539, and 0.7257, respectively, using the ANN
model.
ANFIS and ANN models are used to model or predict
some processing variables in the fields of food science and
technology. Although ANFIS and ANN models have been
used for the prediction of some parameters of foods, there are
only a few articles on the comparison of these models. Generally, in these studies, it was determined that the ANFIS or
fuzzy logic models performed better compared to ANN

models. For example, Yalcin et al. (2011) reported that the

ANFIS model used in their study was better than the ANN
model for the estimation of the effects of some antioxidant
phenolic compounds and carotenoids on the oxidation parameters of edible oil. In another similar study, it was again
reported that the ANFIS model was better than ANN in
predicting the effect of apple pomace, orange, and potato peel
extracts on the oxidation parameters of sunflower oil during
the storage period (Karaman et al. 2011). The antimicrobial
effect of benzoic and cinnamic acids on Listeria
monocytogenes was compared with ANFIS, ANN, and multiple linear regression (MLR). In the same study, it was reported that the ANFIS model performed better than the ANN
and MLR models (Ramos-Nino et al. 1997). In another study,
kinetic, neural network, and fuzzy logic models were compared, and it was reported that the neural network model was
better than fuzzy logic and kinetic models for the prediction of
textural changes in dried peas cooked in water at different
temperatures.

50

50

ANFIS
Predicted Anthocyanin Values

Predicted Anthocyanin Values

ANFIS
40

30

20

10

40

30

20

10

10

20

30

40

50

10

Measured Anthocyanin Values

30

40

50

40

50

Measured Anthocyanin Values

50

50

ANN

ANN
Predicted Anthocyanin Values

Predicted Anthocyanin Values

20

40

30

20

10

40

30

20

10

10

20

30

40

50

Measured Anthocyanin Values

Fig. 3 The estimation of total anthocyanin content for grape skin by

ANFIS and ANN models in validation period

10

20

30

Measured Anthocyanin Values

Fig. 4 The estimation of total anthocyanin content for red cabbage by

ANFIS and ANN models in validation period

Food Anal. Methods (2014) 7:13281336

1335

Table 2 RMSE, MAE, and R 2 statistics of the ANFIS and ANN models in training, testing, and validation periods for the estimation of total
anthocyanin contents in black carrot, grape skin, and red cabbage
ANFIS
Data set
Black carrot

Grape skin

Red cabbage

(Number of MFs)

ANN
RMSE

MAE

R2

(Number of nodes in layers)

RMSE

MAE

R2

Training

444 gaussmf

0.0524

0.8325

0.9753

361

1.4428

1.1049

0.9586

Testing
Validation
Training
Testing
Validation
Training
Testing
Validation

444 gaussmf
444 gaussmf
555 gaussmf
555 gaussmf
555 gaussmf
555 gaussmf
555 gaussmf
555 gaussmf

0.0713
0.0746
0.0312
0.0460
0.0467
0.0299
0.0450
0.0457

0.8100
0.8674
0.5167
0.5470
0.5491
0.4905
0.5090
0.5268

0.9787
0.9769
0.9948
0.9944
0.9942
0.9860
0.9842
0.9836

361
361
351
351
351
361
361
361

1.3566
1.3197
1.2943
1.2918
1.2669
0.9967
0.9966
0.9476

1.0366
1.0359
0.9647
0.9469
0.9539
0.7621
0.7471
0.7257

0.9654
0.9674
0.9803
0.9803
0.9809
0.9655
0.9653
0.9683

RMSE root mean square error, MAE mean absolute error, ANFIS adaptive neuro fuzzy inference system, ANN artificial neural network

Conclusion
The results of this study showed that the stability of
anthocyanin-based extracts depended on pH, temperature,
heating time, and anthocyanin sources. Elevated pH levels
cause anthocyanin degradations which affect both color and
intensity (Mazza and Miniati 1993). It was reported that the
thermal stabilities of anthocyanins decreased with the increment of pH (Krca et al. 2007; Torskangerpoll and Andersen
2005) and temperature by several researchers (Kirca et al.
2003; Krca et al. 2007; Wang and Xu 2007). Overall, it can
be stated that an increase in temperature, heating time, and pH
contributed to the loss of anthocyanins. Grape skin anthocyanin content decreased more rapidly than that of black carrot.
The red cabbage anthocyanins exhibited the highest heat and
pH stability. These results indicate that acylated anthocyanins
are more stable at higher temperatures and pH values. There
are several studies on the anthocyanin degradation kinetics of
different anthocyanin sources, such as black carrot (CevallosCasals and Cisneros-Zevallos 2004; Krca et al. 2007), red
cabbage, blackcurrant, elderberry (Cevallos-Casals and
Cisneros-Zevallos 2004), grape (Cevallos-Casals and
Cisneros-Zevallos 2004; Dyrby et al. 2001), and blackberry
juice (Wang and Xu 2007). In this study, in addition to
determining the anthocyanin content and their percent degradation depending on temperature, time, and pH levels, effective predictive models for the estimation of the anthocyanin
content of studied materials were constructed using two different nonlinear modeling techniques (ANFIS and ANN). The
accuracy of the ANFIS and ANN models was compared to
estimate the effects of different temperatures, pH levels, and
time on anthocyanin contents. The results revealed that
ANFIS model is better than the ANN. Both models can be
utilized efficiently for the prediction of unmeasured interval
anthocyanin contents with high accuracy at different temperatures, times, and pH levels.

Acknowledgments This work was supported by Erciyes Universitys

Research Fund (FBD-09-766 and FBA-09-768).
Conflict of Interest Dr. Lutfiye Ekici declares that she has no conflict
of interest. Zeynep Simsek declares that she has no conflict of interest. Dr.
Ismet Ozturk declares that he has no conflict of interest. Dr. Osman
Sagdic declares that he has no conflict of interest. Dr. Hasan Yetim
declares that he has no conflict of interest. This article does not contain
any studies with human or animal subjects.

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