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Journal of Food Engineering 82 (2007) 271–275

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


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

0260-8774/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
272 W.-D. Wang, S.-Y. Xu / Journal of Food Engineering 82 (2007) 271–275

1999), pomegranate (Martı́, Pérez-Vicente, & Garcı́a-Vigu- ice water bath. The analysis was conducted immediately
era, 2001), grape (Morais, Ramos, Forgács, Cserháti, & and subsequently at regular time intervals.
Oliviera, 2002) and strawberry (Garcı́a-Viguera & Zafrilla, The storage stability of anthocyanins was studied in
1999). However, no information is available on the thermal 8.90°Brix juice and 65.0°Brix concentrate at 5, 25 and
degradation kinetics of blackberry anthocyanins. 37 °C.
Accurate knowledge of the kinetic parameters is essen-
tial to predict the quality changes that occur during ther- 2.4. Determination of anthocyanins
mal processing. In this paper, we report on a study about
the determination of the degradation kinetic parameters Total juice anthocyanin content was determined using
for blackberry anthocyanins in both juice and concentrate the pH-differential method described by Lee, Durst, and
during heating and storage at various temperatures. Wrolstad (2005), using two buffer systems: potassium chlo-
ride buffer, pH 1.0 (0.025 M), and sodium acetate buffer,
2. Materials and methods 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-
2.1. Materials ferred to a 10 mL volumetric flask and made up to
10 mL with corresponding buffer and the absorbance was
Individually quick-frozen (I.Q.F) blackberries (Rubus measured at 510 and 700 nm. Total anthocyanins were cal-
fruticosus L.) were purchased from Xin De Li Food Co. culated as cyanindin-3-glucoside according to the following
Ltd., Nanjing, China (2005 harvest) and stored at 28 °C equation:
until use. Pectinolytic enzyme, Klerzyme-150 was kindly Total anthocyanins ðmg=LÞ ¼ A  MW  DF  1000=ðe  1Þ
gifted by DSM Shanghai, China. Gelatin (Type-A, 175
bloom) was purchased from Shanghai Chemical Reagent ð1Þ
Co., Shanghai, China, while bentonite was obtained from where A = (A510  A700)pH1.0  (A510  A700)pH4.5; MW
Shaanxi Geology and Mineral Test Institute, Shaanxi, (molecular weight) = 449.2 g/mol for cyanidin-3-glucoside;
China. All the other reagents were of analytical grade. DF = dilution factor; 1 = pathlength in cm; e = 26,900
molar extinction coefficient in L/mol/cm for cyanidin-3-
2.2. Preparation of blackberry juice and concentrate glucoside; 1000 = conversion from g to mg. All analyses
were done in triplicate (n = 3).
Frozen blackberries were thawed at 5 °C for 12 h and
crushed for 5s using a triturator (Model DS-1, Shanghai 2.5. Other analyses
Specimen and Model Factory, Shanghai, China). The
crushed berries were immediately heated to 60 °C (±1 °C) Brix was measured at 20 °C using an Abbe refractometor
to inactivate polyphenol oxidase (PPO) and cooled rapidly (Atago, Tokyo, Japan). Turbidity of juice was measured by
to 45 °C (±1 °C), followed by addition of Klerzyme 150 STZ-A24 turbidimeter (Guangming Turbidimeter Plant,
(0.065 mL/kg), mixed thoroughly and incubated up to 2 h Wuxi, China) and expressed in nephelometric turbidity
at 45 °C (±0.5 °C). After maceration, the juice was units (NTU). Proteins were determined by the MicroKjel-
extracted by a juice screw pressing device and centrifugated dahl nitrogen  6.25 (AOAC, 1990). Titratable acidity and
at 3000 rpm for 15 min. The supernatant was clarified with total sugar were determined according to the standard
0.3 g gelatin and 0.3 g bentonite per litre at room tempera- method (AOAC, 1990).
ture for 1 h, corked into 200-mL transparent glass bottles.
In-package pasteurization of the juices was carried out at 3. Results and discussion
85 °C for 5 min followed by rapid cooling in flowing tap
water. The soluble solid of the juice obtained was 3.1. Physical and chemical characteristics of juice
8.90°Brix. At the same time, parts of the clarified juice were
concentrated to 65.0°Brix in a rotary low pressure evapora- Table 1 shows the results of some physical and chemical
tor (ZX98-1 Rotavapor, Shanghai Organic Chemistry analyses of the clarified blackberry juice and concentrate. It
Institute, Shanghai, China) at 60 °C (±1 °C). was reported that strawberry, pomegranate and bluberry
juices contained 55.7 mg/L (Torreggiani et al., 1999),
2.3. Degradation studies 250.87 mg/L (Pérez-Vicente, Serrano, Abellán, & Garcı́a-
Viguera, 2004) and 424.1–518.2 mg/L (Lee, Durst, &
The thermal degradation of blackberry anthocyanins Wrolstad, 2002; Margherita et al., 2003) anthocyanins,
was studied in 8.90°Brix juice at 60, 70, 80 and 90 °C. Ali- respectively. It was clear that anthocyanins in blackberry
quots of 10 mL blackberry juices were put into screw-cap juice was relatively high.
test tubes already equillibrated in a thermostatic water bath There was less than a 5% loss of anthocyanins from the
at 60, 70, 80 and 90 °C (±1 °C). At regular time intervals concentration step (the time which was needed for juice
(10, 40, 70, 100, 130, and 160 min), samples were removed solid content up to 65.0°Brix was less than 100 min). Thus
from the water bath and rapidly cooled by plunging into an 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
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
Nephelometric turbidity unit. 37 59.1e (0.9953) 11.7f
As anhydrous citric acid. 5 5.2e (0.9907) 133.3f 65.06 (0.9993)
As cyanidin 3-glucoside. 25 36.4e (0.9925) 19.0f
As gallic acid. 37 89.9e (0.9872) 7.7f
Values are determined after the concentrate is diluted to 8.90°Brix. a
Rate constant.
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.
ples was stable during heating at 60 °C. Days.

3.2. Kinetics of anthocyanins degradation in heat treatment

k ¼ k 0  eEa =RT ð4Þ
The anthocyanin contents of blackberry juice during where k0 is the frequency factor (per min), Ea is the activa-
heating were plotted as a function of time (Fig. 1). It is tion energy (kJ/mol), R is the universal gas constant
clear from Fig. 1 that the thermal degradation of black- (8.314 J/mol/K) and T is the absolute temperature (in Kel-
berry anthocyanins followed first order reaction kinetics vin, K).
with respect to temperature. Previous studies showed that The kinetic parameters of anthocyanins degradation
thermal degradation of anthocyanins followed a first-order during heating are shown in Table 2. It is clear that the deg-
reaction (Cemeroğlu et al., 1994; Garzón & Wrolstad, radation of blackberry anthocyanins increased with
2002). This kinetic type was expressed by the following increasing heating temperature and time. The t1/2 values
equations: (Table 2) varied from 16.7 to 2.9 h for 8.90°Brix samples
at 60, 70, 80 and 90 °C.
C t ¼ C 0 expðktÞ ð2Þ
Cemeroğlu et al. (1994) reported that t1/2 values for
t1=2 ¼  ln 0:5=k ð3Þ anthocyanins degradation were 54.3, 22.5 and 8.1 h in sour
where C0 is the initial anthocyanin contents and Ct is the cherry juice at 60, 70 and 80 °C, respectively. Compared to
anthocyanin contents after time t (min) of heating at the gi- sour cherry anthocyanins, blackberry anthocyanins were
ven temperature while k is the first order kinetic constant. more susceptible to high temperatures. The major anthocy-
The temperature-dependence degradation rate constant anins in blackberry are cyanidin-3-glucoside, and a small
was represented by the Arrhenius equation 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
Fig. 1. Degradation of anthocyanins in blackberry juice (8.90°Brix) studied (Fig. 2). At 60, 70, 80 and 90 °C, the calculated
during heating at 60, 70, 80 and 90 °C. 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)


5 0.6

4 5˚C

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

1 7

Anthocyanin content (C/C 0)


0.6 4

5˚C 3
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) 271–275 275

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The results from the present study provide detailed Katsube, N., Iwashita, K., Tsushida, T., Yamaki, K., & Kobori, M.
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Lee, J., Durst, R. W., & Wrolstad, R. E. (2002). Impact of juice processing
tion of blackberry anthocyanins follows first order reaction on bluberry anthocyanins and polyphenolics: comparison of two
kinetics. Variation of degradation rate constants with tem- pretreatments. Journal of Food Science, 67(5), 1660–1667.
perature obeyed the Arrhenius relationship. The black- Lee, J., Durst, R. W., & Wrolstad, R. E. (2005). Determination of total
berry anthocyanin, during heating and storage, degraded monomeric anthocyanin pigment content of fruit juices, beverages,
natural colorants, and wines by the pH differential method: collabo-
more quickly with temperature increasing. Thus higher sta-
rative study. Journal of AOAC International, 88(5), 1269–1278.
bility of anthocyanins was achieved by using lower temper- Margherita, R., Giussania, E., Morellib, R., Scalzoc, R. L., Nani, R. C., &
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of the blackberry juice and concentrate. Further studies on radical scavenging activity of highbush blueberry juice. Food Research
the stabilization of blackberry anthocyanin are needed if International, 36, 999–1005.
Martı́, N., Pérez-Vicente, A., & Garcı́a-Viguera, C. (2001). Influence of
blackberries would be processed into juice or blackberry
storage temperature and ascorbic acid addition on pomegranate juice.
anthocyanin would be used as colorant in other products. Journal of the Science of Food and Agriculture, 82(2), 217–222.
Mazza, G. (2000). Health aspects of natural colors. In G. J. Lauro & F. J.
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Morais, H., Ramos, C., Forgács, E., Cserháti, T., & Oliviera, J. (2002).
The authors would like to acknowledge Science and
Influence of storage conditions on the stability of monomeric
Technology Department of Jiangsu Province (Project anthocyanins studied by reversed-phase high-performance liquid
Number: BE2004345) for the financial support and DSM chromatography. Journal of Chromatography B, 770(1–2), 297–301.
Co. Ltd., China for supplying pectinase (Klerzyme 150). Ochoa, M. R., Kesseler, A. G., Vullioud, M. B., & Lozano, J. E. (1999).
Thanks are given to Dr. Seronei Chelulei Cheison of Physical and chemical characteristics of raspberry pulp: Storage effect
on composition and color. Lebensmittel-Wissenschaft und- Technolo-
Southern Yangtze University for revising the English ver-
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sion of the manuscript. Pérez-Vicente, A., Serrano, P., Abellán, P., & Garcı́a-Viguera, C. (2004).
Influence of packaging material on pomegranate juice colour and
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