Journal of Food Engineering 82 (2007) 271–275

www.elsevier.com/locate/jfoodeng

Degradation kinetics of anthocyanins in blackberry

juice and concentrate
Wei-Dong Wang, Shi-Ying Xu *
Key Laboratory of Food Science and Safety, Ministry of Education, Southern Yangtze University, Wuxi 214036, China

Received 24 October 2006; received in revised form 17 January 2007; accepted 18 January 2007
Available online 7 February 2007

Abstract

Thermal and storage stabilities of anthocyanins in blackberry (Rubus fruticosus L.) juice and concentrate were studied over the tem-
perature range 60–90 °C and 5–37 °C. Results indicate that the thermal degradation of anthocyanins followed first-order reaction kinet-
ics. The temperature-dependent degradation was adequately modeled on the Arrhenius equation. The activation energy value for the
degradation of blackberry anthocyanins during heating was 58.95 kJ/mol for the 8.90°Brix blackberry juice. During storage, antocyanins
in the 65.0°Brix blackberry juice concentrate degraded more rapidly than that in 8.90°Brix blackberry juice, with the activation energies
of 65.06 kJ/mol and 75.5 kJ/mol, respectively.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Blackberry juice; Blackberry anthocyanins; Degradation kinetics; Juice; Concentrate

1. Introduction
Anthocyanins are a group of naturally occurring pheno-
lic compounds, which are responsible for the attractive col-
ors of many flowers, fruits (particularly in berries),
vegetables and related products derived from them. These
polyphenolic substances are glycosides of polyhydroxy-
and polymethoxy-derivatives of 2-phenylbenzopyrylium
or flavilium salts. Anthocyanins are becoming increasingly
important as antioxidants (Mazza, 2000; Smith, Marley,
Seigler, Singletary, & Meline, 2000). During the last years
anthocyanins were reported in helping to reduce the risk
of coronary heart disease and prevent several chronic dis-
eases (Katsube, Iwashita, Tsushida, Yamaki, & Kobori,
2003; Renaud & de Lorgeril, 1992). Except for their antiox-
idant properties, anthocyanins have high potential for use
as natural colorants due to their favorable colors, such as
orange, red, purple, and blue. In fact, there was provided
by four anthocyanin-derived colorants, namely grape skin
*
Corresponding author. Tel./fax: +86 510 85884496.
E-mail address: syxu2005@hotmail.com (S.-Y. Xu).
extract, grape color extract, fruit juice and vegetable juice,
which are exempt from certification and approved for food
use in the USA (Wrolstad, 2004).
Blackberries are a good source of anthocyanins in which
the anthocyanin contents were reported to be 67.4–230 mg/
100 g fresh weight (Benvenuti, Pellati, Melegari, & Bertelli,
2004; Sellappan, Akoh, & Krewer, 2002). Blackberry juice,
like all fruit and vegetable juices, is considered as an ingre-
dient when added to foods as a colorant. However, antho-
cyanins readily convert to colorless derivatives and
subsequently to insoluble brown pigments. A numbers of
factors influence anthocyanin stability, including pH, light,
oxygen, enzymes, ascorbic acid, sugars, sulfur dioxide or
sulfite salts, metal ions and copigments (Francis, 1989;
Jackman, Yada, Tung, & Speers, 1987). Heat treatment is
one of the most widely used methods of preserving and
extending the useful shelf life of foods. It is also one of
the most important factors that affect the stability of
anthocyanins. Thermal degradation of anthocyanins has
been studied for red cabbage (Dyrby, Westergaard, & Sta-
pelfeldt, 2001), sour cherry (Cemeroğlu, Velioğlu, & Isßik,
1994), raspberry (Ochoa, Kesseler, Vullioud, & Lozano,

0260-8774/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2007.01.018
272 W.-D. Wang, S.-Y. Xu / Journal of Food Engineering 82 (2007) 271–275
1999), pomegranate (Martı́, Pérez-Vicente, & Garcı́a-Vigu-
era, 2001), grape (Morais, Ramos, Forgács, Cserháti, &
Oliviera, 2002) and strawberry (Garcı́a-Viguera & Zafrilla,
1999). However, no information is available on the thermal
degradation kinetics of blackberry anthocyanins.
Accurate knowledge of the kinetic parameters is essen-
tial to predict the quality changes that occur during ther-
mal processing. In this paper, we report on a study about
the determination of the degradation kinetic parameters
for blackberry anthocyanins in both juice and concentrate
during heating and storage at various temperatures.
2. Materials and methods
2.1. Materials
Individually quick-frozen (I.Q.F) blackberries (Rubus
fruticosus L.) were purchased from Xin De Li Food Co.
Ltd., Nanjing, China (2005 harvest) and stored at 28 °C
until use. Pectinolytic enzyme, Klerzyme-150 was kindly
gifted by DSM Shanghai, China. Gelatin (Type-A, 175
bloom) was purchased from Shanghai Chemical Reagent
Co., Shanghai, China, while bentonite was obtained from
Shaanxi Geology and Mineral Test Institute, Shaanxi,
China. All the other reagents were of analytical grade.
2.2. Preparation of blackberry juice and concentrate
Frozen blackberries were thawed at 5 °C for 12 h and
crushed for 5s using a triturator (Model DS-1, Shanghai
Specimen and Model Factory, Shanghai, China). The
crushed berries were immediately heated to 60 °C (±1 °C)
to inactivate polyphenol oxidase (PPO) and cooled rapidly
to 45 °C (±1 °C), followed by addition of Klerzyme 150
(0.065 mL/kg), mixed thoroughly and incubated up to 2 h
at 45 °C (±0.5 °C). After maceration, the juice was
extracted by a juice screw pressing device and centrifugated
at 3000 rpm for 15 min. The supernatant was clarified with
0.3 g gelatin and 0.3 g bentonite per litre at room tempera-
ture for 1 h, corked into 200-mL transparent glass bottles.
In-package pasteurization of the juices was carried out at
85 °C for 5 min followed by rapid cooling in flowing tap
water. The soluble solid of the juice obtained was
8.90°Brix. At the same time, parts of the clarified juice were
concentrated to 65.0°Brix in a rotary low pressure evapora-
tor (ZX98-1 Rotavapor, Shanghai Organic Chemistry
Institute, Shanghai, China) at 60 °C (±1 °C).
2.3. Degradation studies
The thermal degradation of blackberry anthocyanins
was studied in 8.90°Brix juice at 60, 70, 80 and 90 °C. Ali-
quots of 10 mL blackberry juices were put into screw-cap
test tubes already equillibrated in a thermostatic water bath
at 60, 70, 80 and 90 °C (±1 °C). At regular time intervals
(10, 40, 70, 100, 130, and 160 min), samples were removed
from the water bath and rapidly cooled by plunging into an
ice water bath. The analysis was conducted immediately
and subsequently at regular time intervals.
The storage stability of anthocyanins was studied in
8.90°Brix juice and 65.0°Brix concentrate at 5, 25 and
37 °C.
2.4. Determination of anthocyanins
Total juice anthocyanin content was determined using
the pH-differential method described by Lee, Durst, and
Wrolstad (2005), using two buffer systems: potassium chlo-
ride buffer, pH 1.0 (0.025 M), and sodium acetate buffer,
pH 4.5 (0.4 M). The concentrate samples were diluted to
8.90°Brix juice. A 0.1 mL aliquot of the juice was trans-
ferred to a 10 mL volumetric flask and made up to
10 mL with corresponding buffer and the absorbance was
measured at 510 and 700 nm. Total anthocyanins were cal-
culated as cyanindin-3-glucoside according to the following
equation:
Total anthocyanins ðmg=LÞ ¼ A  MW  DF  1000=ðe  1Þ
ð1Þ
where A = (A510  A700)pH1.0  (A510  A700)pH4.5; MW
(molecular weight) = 449.2 g/mol for cyanidin-3-glucoside;
DF = dilution factor; 1 = pathlength in cm; e = 26,900
molar extinction coefficient in L/mol/cm for cyanidin-3-
glucoside; 1000 = conversion from g to mg. All analyses
were done in triplicate (n = 3).
2.5. Other analyses
Brix was measured at 20 °C using an Abbe refractometor
(Atago, Tokyo, Japan). Turbidity of juice was measured by
STZ-A24 turbidimeter (Guangming Turbidimeter Plant,
Wuxi, China) and expressed in nephelometric turbidity
units (NTU). Proteins were determined by the MicroKjel-
dahl nitrogen  6.25 (AOAC, 1990). Titratable acidity and
total sugar were determined according to the standard
method (AOAC, 1990).
3. Results and discussion
3.1. Physical and chemical characteristics of juice
Table 1 shows the results of some physical and chemical
analyses of the clarified blackberry juice and concentrate. It
was reported that strawberry, pomegranate and bluberry
juices contained 55.7 mg/L (Torreggiani et al., 1999),
250.87 mg/L (Pérez-Vicente, Serrano, Abellán, & Garcı́a-
Viguera, 2004) and 424.1–518.2 mg/L (Lee, Durst, &
Wrolstad, 2002; Margherita et al., 2003) anthocyanins,
respectively. It was clear that anthocyanins in blackberry
juice was relatively high.
There was less than a 5% loss of anthocyanins from the
concentration step (the time which was needed for juice
solid content up to 65.0°Brix was less than 100 min). Thus
concentration did not have a significant effect on anthocy-
 W.-D. Wang, S.-Y. Xu / Journal of Food Engineering 82 (2007) 271–275 273

Table 1 Table 2
Characteristics of blackberry juice and concentrate Effect of temperature and concentration on the k, t1/2 and Ea values of
anthocyanins degradation in blackberry juice and concentrate
Juice Concentrate
Solid Temperature ka  103 t1=2 Ea (kJ/mol)c
°Brix 8.90 65.0
concentrate (°C ) (min1) (h)b
pH 2.86 2.89 (2.64)e
(Brix)
Turbidity (NTUa) 0.21 2.17
Protein (mg/L) 449.00 437.43 8.90 60 0.69 (0.9822)d 16.7 58.95 (0.9975)d
Titration acidityb (g/100mL) 1.65 1.67 70 1.32 (0.9919) 8.8
Total sugars (g/L) 55.71 52.04 80 2.47 (0.9924) 4.7
Anthocyaninc (mg/L) 400.77 417.85 90 3.94 (0.9922) 2.9
Total phenolicsd (mg/L) 1540.93 1547.84
65.0 5 2.0e (0.9892) 330.1f 75.50 (0.9971)
Numbers in parentheses is determined on the basis of 65.0°Brix. 25 21.6e (0.9972) 32.1f
a
Nephelometric turbidity unit. 37 59.1e (0.9953) 11.7f
b
As anhydrous citric acid. 5 5.2e (0.9907) 133.3f 65.06 (0.9993)
c
As cyanidin 3-glucoside. 25 36.4e (0.9925) 19.0f
d
As gallic acid. 37 89.9e (0.9872) 7.7f
e
Values are determined after the concentrate is diluted to 8.90°Brix. a
Rate constant.
b
Half-life.
c
Activation energy.
anin content. This observation clearly is consistent with the d
Numbers in parentheses are the determination coefficients.
following result that anthocyanins in blackberry juice sam- e
Per day.
f
ples was stable during heating at 60 °C. Days.

3.2. Kinetics of anthocyanins degradation in heat treatment

The anthocyanin contents of blackberry juice during
heating were plotted as a function of time (Fig. 1). It is
clear from Fig. 1 that the thermal degradation of black-
berry anthocyanins followed first order reaction kinetics
with respect to temperature. Previous studies showed that
thermal degradation of anthocyanins followed a first-order
reaction (Cemeroğlu et al., 1994; Garzón & Wrolstad,
2002). This kinetic type was expressed by the following
equations:
C t ¼ C 0 expðktÞ
t1=2 ¼  ln 0:5=k
where C0 is the initial anthocyanin contents and Ct is the
anthocyanin contents after time t (min) of heating at the gi-
ven temperature while k is the first order kinetic constant.
The temperature-dependence degradation rate constant
was represented by the Arrhenius equation
Fig. 1. Degradation of anthocyanins in blackberry juice (8.90°Brix)
during heating at 60, 70, 80 and 90 °C.
k ¼ k 0  eEa =RT ð4Þ
where k0 is the frequency factor (per min), Ea is the activa-
tion energy (kJ/mol), R is the universal gas constant
(8.314 J/mol/K) and T is the absolute temperature (in Kel-
vin, K).
The kinetic parameters of anthocyanins degradation
during heating are shown in Table 2. It is clear that the deg-
radation of blackberry anthocyanins increased with
increasing heating temperature and time. The t1/2 values
(Table 2) varied from 16.7 to 2.9 h for 8.90°Brix samples
at 60, 70, 80 and 90 °C.
ð2Þ
Cemeroğlu et al. (1994) reported that t1/2 values for
ð3Þ anthocyanins degradation were 54.3, 22.5 and 8.1 h in sour
cherry juice at 60, 70 and 80 °C, respectively. Compared to
sour cherry anthocyanins, blackberry anthocyanins were
more susceptible to high temperatures. The major anthocy-
anins in blackberry are cyanidin-3-glucoside, and a small
quantity of cyanidin-3-rutinoside, cyanidin-3-dioxalylg-
lucoside, cyanidin-3-xyloside, and cyanidin-3-malonylg-
lucoside (Fan-Chiang & Wrolstad, 2005; Rommel &
Wrolstad, 1992). However, anthocyanins in sour cherries
are cyanidin-3-glucosylrutinoside and cyanidin-3-rutino-
side (Dekazos, 1970; Šimunić, Kovač, Gašo-Sokač, Pfan-
nhauser, & Murkovic, 2005). Therefore, the different
susceptibilities of fruit juice anthocyanins to heat might
be due to their varying anthocyanidin composition. Boyles
and Wrolstad (1993) and Rommel et al. (1990) also con-
tributed the superior color stability of red raspberry prod-
ucts as compared to those processed from strawberries and
blackberries to the sugar substitution with sophorose.
To determine the effect of temperature on the parame-
ters studied, the constants obtained from Eq. (2) were fitted
to an Arrhenius-type equation in each of the kinetic models
studied (Fig. 2). At 60, 70, 80 and 90 °C, the calculated
activation energy (Ea) was 58.95 kJ/mol (Table 2).
274 W.-D. Wang, S.-Y. Xu / Journal of Food Engineering 82 (2007) 271–275

8 1

Anthocyanin content (C/C 0)

0.8
6

5 0.6
-Lnk

4 5˚C
0.4
3
25˚C
2
0.2
37˚C
1

0 0
0 10 20 30 40 50 60
0.0027 0.0028 0.0029 0.003 0.0031
Time (day)
1/T (1/K)
Fig. 4. Degradation of anthocyanins in blackberry concentrate
Fig. 2. The arrhenius pots for degradation of anthocyanins in juice (65.0°Brix) during storage at 5, 25 and 37 °C.
(8.90°Brix) during heating.

3.3. Kinetics of anthocyanins degradation during storage
Anthocyanin contents during storage decreased in time-
and temperature-dependent manners. Linear regression
confirmed that the degradation of anthocyanins in black-
berry juice and concentrate followed by a first-order reac-
tion (Fig. 3 and 4). The values of the first-order model
are given in Table 2. The t1/2 values at the same tempera-
tures were calculated as 330.1, 32.1 and 11.7 days for juice
samples and 138.6, 19.7 and 9.4 days for concentrate sam-
ples. Several authors also found a first order reaction for
the degradation of anthocyanins in sour cherry concen-
trates of 45 and 71°Brix (Cemeroğlu et al., 1994), purple-
and red-flesh potatoes extracts (Reyes & Cisneros-Zevallos,
2007) and black currant nectar (Iversen, 1999) during stor-
age. Cemeroğlu et al. (1994) reported a t1/2 of 38 days for
sour cherry concentrates at 71°Brix at 20 °C. Garzón and
Wrolstad (2002) showed that the t1/2 values for anthocya-
nins degradation in strawberry juice and concentrate of
65.0°Brix were 8 days and 4 days at 25 °C. Our findings
were in agreement with the observation of Cabrita, Fossen,
and Andersen (2000), which showed that cyanidin-3-gluco-
side is more stable than pelargonidin-3-glucoside, which
was the main anthocyanins in strawberries.
Effect of the storage temperature on the anthocyanins
degradation rate constants is shown in Fig. 5. Tempera-
ture-dependent rate constant obeyed the Arrhenius rela-
tionship (Eq. (4)). The computed values of the activation
energies were 75.5 kJ/mol for the juice samples and
65.06 kJ/mol for concentrate samples. High activation
energy implies that the degradation of anthocyanins in
juice samples (Ea = 75.50 kJ/mol) are more susceptible to
temperature elevations than those in concentrate samples
(Ea = 65.06 kJ/ mol). The extent of anthocyanins degrada-
tion in 65.0°Brix during storage was more rapid than that
in 8.90°Brix juice. For example, the t1/2 value at 25 °C
was 19.7 days in 65.0°Brix concentrate but 32.1 days in
8.90°Brix juice. Therefore, anthocyanins at 65.0°Brix were
more susceptible to degradation than that at 8.90°Brix. The
reason might be that when the juice is concentrated, the
reacting molecules (such as oxygen) become closer, thus
the rate of chemical reactions accelerates. Similar results
were observed for strawberry anthocyanins (Garzón &

1 7

6
Anthocyanin content (C/C 0)

0.8
5

0.6 4
-Lnk

5˚C 3
0.4
25˚C 2
0.2 concentrate juice
37˚C 1

0 0
0 10 20 30 40 50 60 0.0032 0.0033 0.0034 0.0035 0.0036 0.0037
Time (day) 1/T (1/K)

Fig. 3. Degradation of anthocyanins in blackberry juice (8.90°Brix) Fig. 5. The arrhenius pots for degradation of anthocyanins in juice
during storage at 5, 25 and 37 °C. (8.90°Brix) and concentrate (65.0°Brix) during storage.
 W.-D. Wang, S.-Y. Xu / Journal of Food Engineering 82 (2007) 271–275
Wrolstad, 2002) and sour cherry anthocyanins (Cemeroğlu
et al., 1994).
4. Conclusions
The results from the present study provide detailed
information regarding the changes in kinetic stability of
blackberry anthocyanin in juice and concentrate during
heating and storage. The present data shows that degrada-
tion of blackberry anthocyanins follows first order reaction
kinetics. Variation of degradation rate constants with tem-
perature obeyed the Arrhenius relationship. The black-
berry anthocyanin, during heating and storage, degraded
more quickly with temperature increasing. Thus higher sta-
bility of anthocyanins was achieved by using lower temper-
ature and short-time heating during processing and storage
of the blackberry juice and concentrate. Further studies on
the stabilization of blackberry anthocyanin are needed if
blackberries would be processed into juice or blackberry
anthocyanin would be used as colorant in other products.
Acknowledgements
The authors would like to acknowledge Science and
Technology Department of Jiangsu Province (Project
Number: BE2004345) for the financial support and DSM
Co. Ltd., China for supplying pectinase (Klerzyme 150).
Thanks are given to Dr. Seronei Chelulei Cheison of
Southern Yangtze University for revising the English ver-
sion of the manuscript.
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