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Abstract
Due to the growing market of food products associated to good health, the colour changes and stability of anthocyanin extracts from acerola,
containing high level of ascorbic acid, and from aai, rich in flavonoids, were evaluated in an isotonic soft drink like system and in buffer solution.
The degradation of anthocyanins from both sources followed first-order kinetics in all the systems, under air, either in the presence or absence of
light. Addition of sugars and salts had a negative effect on the anthocyanin stability, being the rate constant (kobs) values in isotonic soft drink
system 6.0 10 2 h 1 for acerola and 7.3 10 4 h 1 for aai, both in the dark. In the presence of light, the anthocyanin degradation was 1.2 times
faster for acerola and 1.6 times faster for aai in soft drink isotonic systems, as compared to their respective buffer solutions. The highest stability
observed in all aai systems was correlated to its high total flavonoid content and absence of ascorbic acid. The gradual degradation of red colour
during storage of all systems was verified by the decrease of a values, accompanied with decreased colour intensity (decrease in C values) and
tonality changes from red to yellow colour, as the h values increased during the experiment time.
2007 Elsevier Ltd. All rights reserved.
Keywords: Acerola; Aai; Anthocyanin; Stability; Isotonic Beverage model system
Industrial relevance: Functional foods and addition of bioactive compounds to processed foods and drinks are a worldwide growing market. Thus, the aim of this
study was to evaluate the stability of added anthocyanins from acerola and aai to an isotonic soft drink system and to study the effect of fluorescent light, mimicking
the supermarket conditions, in such systems. Since colour is of fundamental importance for the acceptance of a food by consumers, the colour fading that occurs in
these systems was also verified.
1. Introduction
Anthocyanins pigments are responsible for the red-purple to
blue colours of many flowers, fruits, vegetables, and grains, the
fruits and vegetables anthocyanin-rich extracts being used as
food colourants. In the United States, 4 of the 26 colourants that
are exempt from certification and approved for food use are
anthocyanin-derived: grape skin extract, grape colour extract,
fruit juice, and vegetable juice (Wrolstad, 2004). Although
grape marc constituted a very abundant source of anthocyanins
(Mazza & Miniati, 1993), other fruits such as elderberries, black
currants, chokeberries, raspberries, blackberries and also red
cabbage have been used as sources of anthocyanins colourants
for food products and beverages (Wrolstad, 2004).
Corresponding author. Fax: +55 19 3521 2153.
E-mail address: azm@fea.unicamp.br (A.Z. Mercadante).
1466-8564/$ - see front matter 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ifset.2007.03.008
348
V.V. de Rosso, A.Z. Mercadante / Innovative Food Science and Emerging Technologies 8 (2007) 347352
Fig. 1. Degradation kinetic curves of the anthocyanins from aai and acerola in citratephosphate buffer and isotonic soft drink model systems, both at pH 2.5, under
air and in the presence/absence of light.
V.V. de Rosso, A.Z. Mercadante / Innovative Food Science and Emerging Technologies 8 (2007) 347352
Table 1
Rate constant values (kobs) and half-life times (t1/2) for anthocyanin degradation
in citratephosphate buffer and isotonic soft drink systems added with acerola
and aai, at pH 2.5 under air and presence/absence of light
kobs (h 1) a
Systems
Acerola buffer
Acerola isotonic soft drink
Aai buffer
Aai isotonic soft drink
a
Light
Dark
Light
Dark
Light
Dark
Light
Dark
5.0 10
3.9 10 2
6.6 10 2
6.0 10 2
7.6 10 4
1.5 10 4
1.3 10 3
7.3 10 4
t1/2 (h) a
13.7
17.5
10.6
11.4
909.1
6456.2
548.2
943.4
beginning, middle and end of the experiment period. Immediately after aliquots were drawn, all water was removed under
nitrogen with addition of absolute ethanol. The dried anthocyanin extracts were re-dissolved in acidified methanol immediately before HPLC analysis. An HPLC equipment (Waters,
Milford, MA, USA) consisting of a quaternary solvent pumping
system, Rheodyne injector with a 20 L loop, external oven
with temperature control, on-line degasser, diode array detector
(PDA) and Millennium data collection and processing system
was used. The anthocyanins were separated on a C18 Shim-pack
CLCODS column (5 m, 250 4.6 mm) from Shimadzu
(Canby, OR, USA) using as mobile phase a linear gradient of
5% aqueous formic acid/methanol, going from 85:15 (v/v) to
20:80 in 25 min, the latter proportion being maintained for
further 15 min, at a flow rate of 1 mL/min and column
temperature maintained at 25 C. The chromatograms were
processed at 280, 320 and 520 nm, and the spectra acquired
between 200 and 600 nm. The anthocyanin identification was
based on the literature (Gallori et al., 2004; Hanamura et al.,
2005).
2.5. Kinetic calculations and statistical analysis
The anthocyanin concentration (% residual anthocyanins)
was plotted against time (h) and a linear regression analysis was
used to determine the adequacy of the anthocyanin degradation
kinetic model. The degradation rate constant (kobs) was
determined from the first derivative of the curves plotted (Eq.
(4)) and the half-life time (t1/2) was calculated from Eq. (5).
anthocyanin anthocyanin0 exp kobs t
t1=2
h arctan b=a
h
i1=2
2
2
2
DE DL Da b
:
349
ln2
:
kobs
4
5
The software Microcal Origin 5.0 was used for all analyses,
both ANOVA and kinetic.
3. Results and discussion
3.1. Influence of sugars/salts and light on the stability of
anthocyanin extract
In all the systems evaluated, the degradation of anthocyanins
from acerola and aai followed first-order kinetics (Fig. 1),
which model was applied to give the anthocyanin degradation
rate constants (Table 1). This behaviour had already been
reported in most anthocyanin systems, such as for strawberry
(Wrolstad et al., 1990) and blood orange extract in both
ethanolic and carbonated drinks (Katsaboxakis, Papanicolaou,
& Melanitou, 1998).
The addition of sugars and salts had a negative effect on the
anthocyanin stability, both from acerola and aai. In the dark,
the kobs values were 1.5 and 6.8 times faster, respectively, in
acerola ( p = 0.0497) and aai ( p b 0.0001) isotonic soft drink
system compared to their respective control solution. In the
350
V.V. de Rosso, A.Z. Mercadante / Innovative Food Science and Emerging Technologies 8 (2007) 347352
V.V. de Rosso, A.Z. Mercadante / Innovative Food Science and Emerging Technologies 8 (2007) 347352
351
Fig. 3. Changes in the colour parameters values (A) a, (B) hue angle (h), (C) chroma (C) and (D) DE for the buffer () and isotonic soft drink () systems of
anthocyanin extracts from acerola and aai, at pH 2.5, in the presence of light and air.
352
V.V. de Rosso, A.Z. Mercadante / Innovative Food Science and Emerging Technologies 8 (2007) 347352
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