Journal of Food Engineering 76 (2006) 579–583

www.elsevier.com/locate/jfoodeng

Effects of pH and temperature of extraction medium on

effective diffusion coefficient of anthocynanin pigments
of black carrot (Daucus carota var. L.)
Nüzhet Türker, Ferruh Erdoǧdu *

Department of Food Engineering (Gida Müh. Böl.) University of Mersin, Çiftlikköy-Mersin 33343, Turkey

Received 25 June 2004; accepted 5 June 2005

Available online 8 August 2005

Abstract

Restrictions imposed on artificial food colorants have resulted in extensive research on natural pigments in recent years. Among
the natural colorants, anthocyanin pigments are responsible for red, purple and blue colors of many plants and black carrot (Daucus
carota var. L.) has just been known as a potential source of anthocyanin due to its higher stability because of its acylation compared
to the ones obtained from other sources. For a commercial production process, it is important to shorten the extraction time. There-
fore, effective diffusion coefficient (D- -value) and effects of different parameters on –D-value should be known during extraction. The
objectives of this study were to determine the –D-value of anthocyanin pigments of black carrot and to determine the effects of tem-
perature and pH on –D-value. Black carrot samples, 1 cm in thickness, were fixed to a thin wire and dipped into 600 mL McIlvaine
buffer solution at different temperatures (20, 37.5 and 50 C) and pHs (2, 3 and 4). Extracted pigment concentration versus time was
determined using the pH-differential method. Unsteady state equation for an infinite slab with a convection boundary was then used
to determine the –D-values. The –D-values changed from 0.18 · 1010 (pH = 4, T = 25 C) to 7.37 · 1010 (pH = 2, T = 50 C) show-
ing a synergistic effect of low pH and high temperature on – D-values. In addition, these effects were also investigated by using dried
carrot samples, and it was concluded that the structural changes play an important role, and ordinary diffusion models were not
suitable to determine the change in –D-values of anthocyanin pigments in dried samples. Optimization of an industrial scale produc-
tion is important to speed up the process. These results may be used for the further optimization studies to design a commercial
production.
 2005 Elsevier Ltd. All rights reserved.

Keywords: Anthocyanins; Effective diffusion; Temperature and pH effects

1. Introduction ors and being easily recoverable properties. Questioning

Color is an important factor in the acceptability of a
food product. Food industry uses colorants to enhance
or restore the original appearance of foods. Synthetic
colorants have found their place readily in the food
industry due to their being stable, imparting bright col-
*
Corresponding author.
E-mail addresses: ferruherdogdu@yahoo.com, ferruherdogdu@
mersin.edu.tr (F. Erdoǧdu).
of safety for synthetic colorants, recently prohibition of
some of them and increasing consumer consciousness
towards the natural products resulted in an increased
interest for colorants derived from natural sources
(Jackman & Smith, 1996). For this purpose, pigment ex-
tracts from edible sources as well as concentrated juices
are commercially available and used in food industry.
Anthocyanins are well-known natural alternatives to
synthetic red dyes. Besides their color attributes, interest
in anthocyanins has been intensified due to their possible
health benefits (Einbond, Reynertson, Luo, Basile, &

0260-8774/$ - see front matter  2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2005.06.005
580 N. Türker, F. Erdoǧdu / Journal of Food Engineering 76 (2006) 579–583
Kennelly, 2003; Espin, Soler-Rivas, Wichers, & Garcia-
Viguera, 2000). They are water-soluble pigments impart-
ing red, purple and blue color of many fruits, vegetables
and cereal grains. Anthocyanins, belonging to the class
of flavanoids, are derivatives of glycosylated salts (anth-
ocyanidins) of polyhydroxy and/or polymetoxylated 2-
phenylbenzopyrolium (flavylium). They occur in nature
as glycosides of anthocyanidins and may be acylated
with aliphatic or aromatic acids (Guisti & Wrolstad,
2003). Recently, the research on this area has been to-
wards edible sources with acylated anthocyanins which
are more resistive to hydration and to the deleterious ef-
fect of high pH. They also impart more desirable color
and stability in food systems compared to the most
non-acylated anthocyanins derived from various fruits
and vegetables. Potential value of black carrot (Daucus
carota var L.) as food colorant has also just become
known, and black carrot anthocyanins comprise the
acylated cyanidin derivatives which exhibit remarkable
stability to pH changes and heat treatment (Stintzing,
Stintzing, Carle, Frei, & Wrolstad, 2002).
A traditional method to obtain an interested plant
tissue intracellular component is based on solid-liquid
extraction (Chalermchat, Fincan, & Dejmek, 2004).
Cacace and Mazza (2003a) determined the effective dif-
fusion coefficient (– D-value) for phenolic compounds in
milled frozen black currants using a solid-liquid extrac-
tion system with ethanol and sulfured water in an agi-
tated vessel. The – D-values were found to be ranging
from 24.3 · 1011 to 12.3 · 1011 m2/s at 40 C at differ-
ent ethanol and sulfur concentrations. Cacace and
Mazza (2003b) optimized the extraction of anthocyanins
from black currants with aqueous ethanol reporting the
effect of temperature and ethanol concentration. Use of
higher temperatures was noted to be affecting the degra-
dation of anthocyanins and reduction of yield while the
extraction of phenolics increased with ethanol concen-
tration up to a maximum at about 60% and then de-
creased with further increase of ethanol concentration.
Mantell, Rodriguez, and Martinez de la Ossa (2003,
2004) estimated the – D-value of a food dye, containing
malvidin 3,5-diglucoside, experimentally using a high
pressure CO2+methanol system. The – D-value changed
with respect to both temperature and pressure. They
compared the results to the ones found from different
predictive equations, e.g., Wilke–Chang, Lusis–
Doraiswamy, Hayduck–Minhas equations. Mulet,
Berna, Heredero, and Rossello (1988) investigated the
effect of temperature on the sugar extraction of carob
pods. The – D-values for sugars changed from
8.7 · 1011 to 1.57 · 1010 m2/s at extraction medium
temperatures of 20 and 50 C, respectively.
As seen in the literature, the rate of mass transfer
during extraction is of interest for a process design
(Chalermchat et al., 2004), and this might be increased
by reducing the particle size (i.e. increasing the surface
area and reducing the diffusion distance of the solute
within the solid) or by increasing the diffusion coefficient
due to the increase in extraction medium temperature.
In the case of plant tissues, the diffusion might be called
as effective diffusion, and it is structure-dependent
(Chalermchat et al., 2004). Since it is of economical
value to shorten the extraction time, knowing the diffu-
sion coefficient and the effect of different parameters like
pH and temperature may help optimize the extraction
process. Therefore, the objective of this study was to
determine the diffusion coefficients of anthocyanin pig-
ments from fresh and dried black carrot samples (to
see the effect of structural changes) at different pH val-
ues and temperatures.
2. Materials and methods
2.1. Determination of anthocyanin content
Monomeric anthocyanin content was determined
using pH differential method (Guisti & Wrolstad,
2001). A Perkin-Elmer UV/visible spectrophotometer
(Lambda EZ 201) and 1 cm path length glass cells were
used for spectral measurements at 525 and 700 nm,
respectively. The monomeric anthocyanin content was
determined as cyanidin-3-glucoside using an extinction
coefficient of 29,600 L/cm-mg and molecular weight of
448.8. Initial anthocyanin concentration of the fresh car-
rot samples was 3825 mg/kg dry matter. Anthocyanin
concentration change of the carrot samples was deduced
from the observations obtained from the solutions since
it was easier from an experimental point of view. The
pH measurements were done using a Hanna pH-meter
(pH 211) calibrated with pH 4 and 7 buffers.
2.2. Sample preparation
Black carrots were sliced in 1.03 ± 0.08 cm in thick-
ness and 5.1 ± 0.6 cm in diameter. Initial moisture con-
tent of the samples was 88% (w.b.). Half of the carrot
slices were dried in an oven (Memmert UP600,
Scwabach, Germany) at 65 C overnight. The moisture
content of the dry samples was found to be reduced by
85 ± 1%.
2.3. Experimental set-up
The black carrot slices were fixed to a thin wire and
dipped into a 600 mL McIlvaine buffer solution (in a
10.5 cm in diameter beaker) stirred at 1000 rpm held at
different temperatures (25, 37.5 and 50 C) and pH val-
ues (2, 3 and 4). The extracted anthocyanin concentra-
tion in buffer solution versus time was recorded and
determined as explained above. Two replications were
accomplished at each condition.
 N. Türker, F. Erdoǧdu / Journal of Food Engineering 76 (2006) 579–583 581
 
2.4. Mathematical background the linear portions slope k21  L–D2 of the concentration
ratio curve and the first root (k1) of the characteristic
Since the diameter to thickness ratio of the carrot equation

of infinite slab. The k1 value is determined to
slices 5 (Turhan & Erdogdu, 2003, 2004), the slices be p2 from the above given equation assuming an
were assumed to be infinite slabs. However, it should infinite Biot number due to a very high mass transfer
be noted that the error coming from this assumption coefficient as a result of mixing in the extraction
at infinite Biot number cases were less than 5% (Turhan medium. Then,
& Erdogdu, 2003, 2004). Then, the analytical solution of ! effective –D-value becomes equal to
 
unsteady state mass transfer equation for infinite slabs c  ðp=2Þ1
2 ¼  93030c .49 where c is the slope the lin-
with a constant uniform initial distribution and symmet- ð0.0103=2Þ2

rical and convective boundary conditions was used to ear part of the concentration ratio vs time curve.
determine the effective –
D-values using the experimental Eq. (3) or Eq. (4) is preferred in the systems with
data obtained for extracted anthocyanin concentration. unlimited volume of fluid. In this study, the volume of
The extracted anthocyanin concentration in the buffer the extraction fluid was limited (600 mL). For the cases
solutions was used to determine the anthocyanin con- of limited volume, Crank (1975) suggested Eq. (6) sub-
centration change in the carrots to use in the required jected to the boundary conditions given in Eq. (5):
calculations.
The solution for the following differential equation  0Þ ¼ C i  L 6 x 6 L
Cðx;
subjected to the given initial and boundary conditions oCðx; tÞ oCðx; tÞ
were as in Eq. (3): a ¼ D  x ¼ L ð5Þ
ot " ox
 X1  #
o2 Cðx; tÞ 1 oCðx; tÞ CðtÞ 2  a  ð1 þ aÞ 2 –Dt
¼  ð1Þ 1 ¼ 2
 exp bn  2 ð6Þ
ox2 D
– ot C1 2
n¼1 1 þ a þ a  bn L
Cðx; 0Þ ¼ C i
oCð0; tÞ where a is the fluid thickness, a ¼ La ¼ 5.250 .515 ¼ 9.2 and
0.515
¼0 ð2Þ b is given by tan b + a Æ b = 0 (Crank, 1975). For the
ox 
oCðx; tÞ   given a = 9.2 value, b1 is found to be 1.637, which is
– D  ¼ k  Cðx; tÞjx¼L  C 1 slightly higher than p2 (1.042 times). This might result
ox x¼L
" that the effective –D-coefficient values may be
Cðx; tÞ  C 1 X1
2  sin kn  x
¼  cos kn  1.04082 = 1.086 times smaller than their actual values.
Ci  C1 kn þ sin kn  cos kn L However, Eq. (6) did not include a partition coefficient
n¼1 
 # C carrot
Dt
– ¼ C solution
at equilibrium after a very long extraction
 exp k2n  2 ð3Þ 
L time which is an important parameter in a limited vol-
Eq. (3) showed the concentration change in an infinite ume studies. Experimental errors by determination of
slab for the given initial and boundary conditions where partition coefficient at different conditions may also lead
L 6 x 6 L, and L is half thickness of carrot slices (m), to some mistakes in the resulting equations. In addition
x is distance from center (m), – D is effective diffusion to this error, as mentioned above, there is also an error
coefficient (m2/s), k is mass transfer coefficient (m/s), in assuming the carrot slices as infinite slabs. When they
C(x, t) is anthocyanin concentration (mg/kg) at any were assumed to be in a shape of a finite cylinder, Eq. (4)
location in the carrot slice at time t (s), Ci is initial is written as
anthocyanin concentration (mg/kg), C1 is extracting   C1
CðtÞ
medium anthocyanin concentration
(mg/kg), and k is gi- Ci  C1
ven by Bi ¼ kL ¼ k  tan k where Bi is Biot number. 2 3
D
– 2  sin2 kn 4  J 1 ðlm Þ
Since the anthocyanin concentration change was deter- 1 6 
 7
X1 X 2
6 kn  ðkn þ sin kn  cos kn Þ bm J 20 ðlm Þ þ J 21 ðlm Þ 7
mined through the whole carrot slice applying a reverse ¼ 6     7
calculation to the extraction medium data, Eq. (1) was m¼1 n¼1
4 Dt
– Dt
– 5
exp k2n  2  exp l2m  2
integrated throughout the whole volume to comply with L R
the experimental data: ð7Þ
  C1 X 1    h i
CðtÞ 2  sin2 kn Dt
– where b is given by Bi ¼ KL ¼ b  JJ 10 ðlÞ (J0 and J1 are
¼  exp k2n  2 D
– ðlÞ
Ci  C1 kn  ðkn þ sinkn  coskn Þ L
n¼1 the first kind 0th and 1st order Bessel functions). Based
ð4Þ on an infinite Biot number approach, the l value may be
Due to the fact that the higher terms in the infinite series easily determined as 2.4048. In this case, the slope of
solution may be assumed to be negligible after a certain the concentration ratio vs time
 curve to determine 
Fourier number, effective – D-value was determined using the effective –D-value becomes k21  L–D2  l21  R–D2 . Using
582 N. Türker, F. Erdoǧdu / Journal of Food Engineering 76 (2006) 579–583

this, the effective ! D

– -value becomes equal to 1
 

Concentration Ratio
c
c  ðp=2Þ2 1 2.40482 ¼  93030.49þ8893 .6 where c is the 0.75
þ
ð0.0103=2Þ2 ð0.051=2Þ2
0.5
slope of the linear part of the concentration ratio vs time
curve. According to this, the infinite slab assumption of
0.25
the carrot slices may result in an error of less than 9%
confirming the results given by Turhan and Erdogdu 0
(2003, 2004). In addition, since the carrots were peel- 0 2 4 6 8 10
Extraction Time(hr)
on, it was also not possible for anthocyanin to diffuse
in the radial direction. Therefore, Eq. (4) was applied pH=4, T=25 °C pH=2, T=50 °C

to determine the effective – D-values for the given pH

Fig. 1. Effect of temperature and pH on the concentration ratio
and temperature ranges. 
(CðtÞC 1
) change during extraction for black carrots (1 cm in thickness).
C i C 1

3. Results and discussion T

D/ ð8Þ
g
Based on the above explained procedure (applying
In addition to the effect of temperature, denaturation of
Eq. (4)), the estimated effective –
D-values changed from
the cell membranes might also occur at higher tempera-
0.18 · 1010 (pH = 4, T = 25 C) to 7.37 · 1010
tures affecting the extraction process. However, since the
(pH = 2, T = 50 C). Table 1 showed the change in
anthocyanins are not heat-stable at high temperatures,
effective –
D-values with their standard deviations versus
temperature of the extracting medium cannot be in-
pH and temperature of the extraction medium for carrot
creased indefinitely to maximize the process. Higher
samples.
temperatures may also result in degradation of anthocy-
Applying the estimated effective diffusion coefficient
anins as reported by Cacace and Mazza (2003b).
values at the highest and lowest
 casesfor the carrot sam-

CðtÞC
Due to the effect of pH, anthocyanins are known to
1
ples, the concentration ratio C i C 1
change versus dif- be at the most stable state at pH = 2. Therefore, an in-
fusion time was determined using Eq. (3). Fig. 1 shows crease in pH may result in reduction of pigment stability
the effect of temperature and pH on diffusion of antho- and therefore decrease the extraction rate and – D-value.
cynanin at the extreme cases of pH = 2, T = 50 C and Additionally, low pH may increase the cell membrane
pH = 4, T = 25 C. permeability leading to higher diffusion coefficient val-
As seen in Table 1, as the temperature increased and ues. As seen in Table 1, the effect of the pH of the extrac-
the pH decreased, anthocyanin effective – D-value in- tion medium was more obvious compared to the effect
creased. It may be assumed that increasing temperature of temperature. For example, from 25 to 50 C, the – D-
favored the extraction by enhancing solubility of antho- value increased from 3.73 to 7.37 · 1010 m2/s while,
cyanins. It is obvious the effect of temperature on the at 25 C, it changed from 3.73 to 0.18 · 1010 m2/s
diffusion coefficients. This effect may also be explained and at 50 C, from 7.37 to 0.37 · 1010 m2/s at pH = 2
by the Einstein equation (Cacace & Mazza, 2003a) and 4, respectively. At 50 C, tissue softening in carrot
where the – D-values are proportional with T (tempera- slices was also observed visually. This effect combined
ture) and 1/g (dynamic viscosity): with low pH values may basically have resulted in cell
disruption which leads to the accelerated extraction of
anthocyanins too.
Table 1
Anthocyanin effective diffusion coefficient (m2/s) versus pH and For the case of dried carrots, some structural changes
temperature (T; C) of the extraction medium (–D  1010 ) for black in the carrot samples were observed. Due to these
carrot changes and resulting –D-values, the diffusion equation
pH Temperature (T, C) Diffusion coefficient (–D, m2/s) was assumed to be not suitable for the case of dried car-
2.0 25.0 3.73 ± 0.64 rots. Gekas, Oliveira, and Crapiste (2002) also state that
37.5 5.00 ± 0.23 the cell structure and changes in cell structure in addi-
50.0 7.37 ± 0.00 tion to the temperature and environmental conditions
play a major role in the transport mechanism in fruit tis-
3.0 25.0 0.89 ± 0.06
sues. Therefore, they suggest the possibility of applica-
37.5 2.04 ± 0.21
50.0 3.00 ± 0.81 tion of a thermodynamic approach as a basis for mass
transfer modeling when mechanisms other than Fickian
4.0 25.0 0.18 ± 0.01 diffusion is covered as well. In this subject, Maguer,
37.5 0.25 ± 0.01 Mazzanti, and Fernandez (2002) described the parame-
50.0 0.37 ± 0.03
ters for the cellular properties of the fruit tissues
 N. Türker, F. Erdoǧdu / Journal of Food Engineering 76 (2006) 579–583
(e.g. diffusivity, tortuosity and porosity), the properties
of the solution (e.g. viscosity and density) and process-
ing conditions (e.g. temperature and shape of the mate-
rial) for a cellular approach in modeling mass transfer in
fruit tissues. As described in these recent studies, the cel-
lular effects in the carrot tissue may be assumed to be-
come more evident after drying resulting in that the
diffusion approach did not work well.
4. Conclusions
Temperature and pH of the extraction medium were
found to affect the anthocyanin extraction and therefore
the –
D-values. With the known – D-values, anthocyanin
extraction may be optimized to speed up the process
and also to get a high quality anthocyanin. For this pur-
pose, including some quality factors to the present re-
search with required optimization techniques, – D-value
changes with temperature and pH of the extraction med-
ium may be easily recommended for industrial practice.
Based on a research result by Turker, Aksay, and Ekiz
(2004), the half-life time of black carrot anthocyanins
at 40 C was 21 days. Therefore, shortening the extrac-
tion time by low pH and high temperature may be as-
sumed feasible for an industrial process.
Acknowledgements
The authors would like to acknowledge of the helps
of their undergraduate and graduate students Gamze
_ ature influence on the sugar extraction from carob pods. Leben-
Basßak and Özlem Istanbullu, respectively.
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