Plant Foods Hum Nutr (2013) 68:11–17

DOI 10.1007/s11130-013-0337-1

ORIGINAL PAPER

Thermal Stability of Selected Natural Red Extracts

Used as Food Colorants
José A. Fernández-López & José M. Angosto &
Pedro J. Giménez & Gerardo León

Published online: 2 February 2013

# Springer Science+Business Media New York 2013

Abstract The color degradation of aqueous solutions of Abbreviations

six natural red pigment extracts (elderberry, red cabbage, λmax Wavelength of maximum absorption
hibiscus, red beet, Opuntia fruits and red cochineal) used k Reaction rate constant
commercially as food colorants was investigated at tem- t1/2 Half-life period
peratures between 50 and 90 °C. Color degradation was Ea Activation energy
studied in respect to both spectral properties and visual ΔE* Total color difference
color. The remaining absorbance at 535 nm as a function
of the incubation time and temperature was used to
quantify the degradation process. Red cochineal was the
Introduction
most thermoresistant extract with a remaining absorbance
of 95 % after 6 h at 90 °C. Anthocyanin extracts (elder-
For 4,000 years colorants have been added to foods and the
berry, red cabbage, hibiscus) showed remaining absor-
color of food products is still one of the major concerns of the
bance percentages of 63.8, 46.1 and 26.7, respectively.
food industry [1]. Stable and attractive colors are a highly
Betacyanin extracts (red beet, Opuntia fruits) were the
valued attribute in the competitive food and beverage market,
most thermosensitive maintaining only 12.5 and 1.7 %,
while the use of natural pigments in foodstuffs has been
respectively, of the initial absorbance at 535 nm. Apply-
increasing in recent years due to the marketing advantages
ing a first-order kinetic model to the degradation process-
of using natural ingredients in light of consumer concerns
es, reaction rate constants (k) and half-life periods (t1/2)
related with the potential harmful effects, due to allergies
were calculated. The temperature dependence of the deg-
and intolerance, of synthetic pigments [2, 3]. Moreover, ele-
radation rate constant obeyed the Arrhenius relationship,
vation of the functional platform of natural colors from simple
with activation energies (Ea) ranging between 3.02 and
colorants to health-promoting ingredients has increased the
53.37 kJmol−1. The higher activation energy values indi-
demand for natural colors both on the part of retailers and
cated greater temperature sensitivity. Changes in visual
consumers. The search on the part of the food industry for new
color attributes corroborated the high thermal stability of
products with appealing features has opened up new research
the red cochineal extract.
lines aimed at developing attractive food colorants.
The main groups of natural food colorants are carote-
Keywords Color parameters . Food colorants . Natural red
noids, chlorophylls, anthocyanins, betacyanins and curcu-
extracts . Thermal degradation
minoids [4]. The restrictions in the use of certain synthetic
colorants have greatly reduced the color palette available to
the food industry. Although no color has escaped these
J. A. Fernández-López (*) : J. M. Angosto : P. J. Giménez : restrictions, red is, without doubt, the most affected. The
G. León banning of FD&C Red No. 2 (amaranth) in the USA by the
Department of Chemical and Environmental Engineering, U.S. Food and Drug Administration, Scarlet GN and Pon-
Technical University of Cartagena (UPCT),
ceau GR in France and Orange RN in the United Kingdom
Paseo Alfonso XIII 52,
30203 Cartagena, Murcia, Spain has led to an increased request for natural sources of red
e-mail: josea.fernandez@upct.es pigments [5, 6]. Natural red colorants successfully used in
12
foods include betanin [7, 8], cochineal [9], carotenoids [10]
and, especially, anthocyanins [11]. However, the use of any
natural colorant in food requires a detailed knowledge of its
stability to possible degradation processes and also the
understanding of the conditions governing pigment degra-
dation, so that measures can be taken to ensure sufficient
stability and to optimize the industrial production and stor-
age of the colored products.
In the present work, the heat stability and color altera-
tions of six natural extracts (elderberry, hibiscus, red cab-
bage, red beet, Opuntia fruit and red cochineal) used as red
colorants in the food industry have been compared.
Anthocyanins are the pigments responsible for color in
elderberry, hibiscus and red cabbage extracts, which are
widely used in beverages, confectionery, jellies and jams.
Today, interest in anthocyanin pigments has intensified be-
cause of their possible health benefits as dietary antioxidants
[12, 13]. In red beet and Opuntia fruit extracts, the red color
is due to the presence of betacyanins, water-soluble vacuolar
pigments which exhibit a red-violet coloration. They can be
found in ice creams, powdered drinks, sugar confections,
soup, and bacon products. Betacyanins are also antiradical
molecules, whose bioactivity has been described in recent
years [14, 15]. Cochineal and its derivatives (carminic acid
and carmine) are widely used throughout the food industry.
Both liquid and powder preparations are used to color
beverages, table jellies, sugar confectionery. This colorant
is also commonly used in the pharmaceutical industry [9].
There are no studies comparing the thermal stability of
anthocyanins, betacyanins and red cochineal, so we consider
of great importance to investigate the effects of heat treat-
ment on the colorant properties of these six natural red
extracts used as food colorants.
Materials and Methods
Materials
Commercial red pigment extracts from elderberry (Sambucus
nigra L.), red cabbage (Brassica oleracea L. var. capitata f.
rubra), hibiscus (Hibiscus sabdariffa L.), and red beet (Beta
vulgaris L.) powders were provided by Diana Naturals
(Antrain, France). Red cochineal (Dactylopius coccus C.)
extract was obtained as a powder from Roha Europe SLU
(Valencia, Spain). The Opuntia fruit red extract was obtained
in our laboratory from Opuntia stricta fruits as indicated
previously [16]. Sodium citrate and sodium phosphate were
of analytical grade (Merck, Darmstadt, Germany), and the
water was purified through a Milli-Q purification train (Milli-
pore, Bedford, MA, USA).
Stock solutions (500 mg/50 ml) of each colorant extract
in 100 mM pH 5.5 citrate-phosphate buffer were prepared,
Plant Foods Hum Nutr (2013) 68:11–17
and successive dilutions were performed to reach an absor-
bance of 0.700±0.005 at 535 nm in all the extracts. Figure 1
shows a photograph of the diluted colorant extracts. The
final solutions were maintained in the dark at 5 °C, and
shaken before use within seven days.
Thermal Degradation
Experiments were performed in duplicate in Pobel tubes
(100 mm×14 mm id, Pobel S.A., Madrid, Spain) with screw
cap. The samples were thermostated in a waterbath at the
desired temperature. The degradation was followed at 50 °C,
70 °C and 90 °C for 6 h. The changes in absorbance were
monitored between 400 and 700 nm at intervals of 0.5,
1, 2, 3, 4, 5 and 6 h. After this time, the tubes were immedi-
ately cooled in an ice-bath to stop thermal degradation. An
aliquot of each sample was passed through a PTFE filter
(0.45 μm, polypropylene, Advantec MFS Inc., Dublin, CA,
USA) and analyzed spectrophotometrically in an Agilent
8453 UV-visible spectrophotometer (Waldbronn, Germany).
Kinetic Data on Color Degradation
Kinetic data on red colorant degradation were calculated
using the reaction rate as reported by Van den Broeck and
coworkers [17]. Thermal degradation of extracts was as-
sumed to follow a first order rate, indicating a logarithmic
order of inactivation, which is mathematically expressed as:
 
A
ln ¼ kt ð1Þ
A0
where A represents residual absorbance after treatment, A0
the initial absorbance, and t and k represent time (h) and
reaction rate constant (h−1) at a particular temperature, re-
spectively. Half-life values (t1/2) were determined by plot-
ting the natural logarithm of the A0/A ratio vs. heating
period. The slope of the graph through the origin obtained
Fig. 1 Aqueous solutions (pH 5.5) of the selected colorants. The
absorbance at 535 nm of all of them is adjusted to 0.700
Plant Foods Hum Nutr (2013) 68:11–17 13

Table 1 Color properties for the natural red food colorants

λmax (nm) L* a* b* C* hº

Elderberry 530±0.3 62.6±0.2 31.5±0.1 11.0±0.2 33.4 70.7

Red cabbage 542±0.2 61.7±0.3 41.3±0.2 −18.8±0.3 45.4 −65.5
Hibiscus 527±0.1 64.5±0.1 31.3±0.2 5.8±0.1 31.8 79.4
Red beet 534±0.1 65.8±0.2 34.8±0.3 5.6±0.2 35.2 80.8
Cochineal 551±0.2 68.0±0.0 61.7±0.1 −4.0±0.1 61.8 −86.3
Opuntia 535±0.2 72.4±0.1 60.6±0.0 −22.4±0.2 64.6 −69.7

Mean ± standard deviation, n=3

by connecting the data points was equated with k, from
which the half-life values (t1/2 =ln 2/k) were deduced.
Dependence of the degradation rate constant on temper-
ature is represented by the Arrhenius equation:
Ea
ln k ¼ ln k0  ð2Þ
RT
where k is the rate constant, k0 the frequency factor, Ea is the
activation energy (kJmol−1), R is the universal gas constant
(8.314 Jmol−1 K−1) and T the absolute temperature (Kelvin).
Color Measurements
Visual color was monitored from the CIELab parameters.
The tristimulus values (X, Y, Z) were calculated from spec-
tral data from 380 to 780 nm every 10 nm [18]. The CIELab
parameters (L*, a*, b*, C* and hº) were obtained using
standard illuminant D65 and a 10º observer angle. The hue
angles (hº) were expressed on a 360º color wheel where
0º/360º presents purplish-red, 90º yellow, 180º bluish-green,
and 270º blue. Also color differences ΔE* = [(ΔL*)2 +
(Δa*)2 +(Δb*)2]1/2 were calculated between samples.
Fig. 2 Impact of temperature
(70 °C) on the visible spectra
(400–700 nm) of the colorant
extracts. The time interval
between the spectra is 0.5, 1, 2,
3, 4, 5 and 6 h
Results and Discussion
Color Analysis of the Initial Solutions
Colors can be described by the wavelength of maximum
absorption (λmax) in the visible range of the electromag-
netic spectrum. CIELab parameters have also been used
successfully for the evaluation of color in natural
extracts [19–21]. The initial shades of the selected col-
orants are depicted in Fig. 1. The color characteristics
(λmax and CIELab parameters) are shown in Table 1. It
can be appreciated that all the colorants showed maxi-
mum wavelengths between 527 (hibiscus) and 551 nm
(red cochineal). As regards the initial CIELab parame-
ters, lightness (L*) ranged from L*=61.7 (red cabbage)
to L*=72.4 (Opuntia). All the samples showed positive
a* values, as expected from their red color. Cochineal
and Opuntia showed the highest a* values, a*=61.7
and a*=60.6, respectively. Greater dispersion was ob-
served in the b* parameter (blueness-yellowness) rang-
ing from the negative b* = −22.4 of Opuntia to the
positive b* = 11.0 of elderberry. Chroma (C*), which
14 Plant Foods Hum Nutr (2013) 68:11–17

indicates the brilliance or purity of a color, was higher
in Opuntia. The hue angle (hº), which expresses the
tonality, ranged between hº=−86.3 (13.7) in red cochi-
neal and hº=79.4 in hibiscus.
Influence of Thermal Degradation on the Absorption
Spectra
The instability of natural extracts to temperature results in
color changes during thermal processing and subsequent
storage [23–25].
The type of pigment and the magnitude and duration of
heating have the strongest influence on color stability. The
degradation reaction rate constants (k), half-life periods (t1/2)
50 ˚C
100

% Remaining absorbance at 535 nm

structural alterations that affect their absorption spectra and
color properties. The degradation pattern depends on both 80

the temperature and time. Most color variation patterns

show a direct relationship with the intensity of the thermal 60
treatment. Color degradation increased with increases in
temperature, resulting in profound modifications in the ab- 40
Hibiscus
sorption spectra (400–700 nm) of the different extracts. Opuntia
Figure 2 shows the incidence of heating at 70 °C on the Red beet
20
absorption spectra of the six colorant extracts analyzed. Red cabbage
Cochineal
Each graph depicts the spectra at 0, 0.5, 1, 2, 3, 4, 5 and Elderberry
0
6 h. The most relevant result of this treatment was the 0 1 2 3 4 5 6
pronounced instability of the betalainic extracts (Opuntia Time (hours)
fruits and red beet) both of which had lost their red color
by the end of the test. This behavior was much more evident 70 ˚C
100
when the incubation temperature was increased to 90 °C.
% Remaining absorbance at 535 nm

Among the anthocyanin extracts, elderberry showed lower

80
degradation than hibiscus and red cabbage, while red cochi-
neal was clearly the most thermoresistant extract, showing
60
very slight changes in its spectra, which almost overlapped.
These results can be visualized quantitatively if the
remaining absorbance at 535 nm in the extracts as a function 40
of the incubation time at 50, 70 and 90 °C is analyzed
(Fig. 3). As can be seen, the rate of color degradation 20
increased as the temperature rose. In the three tests (50, 70
and 90 °C) the Opuntia fruit extract was the most thermo- 0
labile, with degradations after 6 h of 58 % at 50 °C, 91 % al 0 1 2 3 4 5 6
70 °C and higher than 98 % at 90 °C. The red cochineal Time (hours)
extract was by far the most stable to temperature of all of
90 ˚C
them, and even after the harshest treatment (6 h at 90 °C), 100
% Remaining absorbance at 535 nm

95 % of absorbance at 535 nm remained. The anthocyanin

extracts were in an intermediate position, with the elderberry 80
extract more resistant to temperature than the red cabbage
and hibiscus extracts. After 6 h of incubation at 90 °C, the 60
elderberry extract showed a remaining absorbance of
63.8 %, while only 46.1 and 26.7 %, respectively, was
40
maintained in the red cabbage and hibiscus extracts.

Degradation Kinetics 20

It is difficult to establish a reaction mechanism and to obtain 0

0 1 2 3 4 5 6
a kinetic model that describes the global process adequately.
Time (hours)
Kinetic models are often used for an objective assessment of
color degradation in natural extracts [22], and most often, Fig. 3 Remaining absorbance at 535 nm in the colorant extracts as a
first-order kinetic models have been proposed to estimate function of the incubation time
Plant Foods Hum Nutr (2013) 68:11–17 15

and activation energy values (Ea) are shown in Table 2. The temperature dependence of the degradation reaction rate
k values showed the following descending order for each constants thus followed the Arrhenius relationship, typical
temperature: Opuntia, red beet, hibiscus, red cabbage, el- of many deteriorative processes in food materials [29].
derberry and cochineal. The t1/2 values obtained for the
temperature range 50–90 °C, confirm that Opuntia fruit
and red beet extracts were less stable than the anthocyanin 30
and cochineal extracts. For example, at 70 °C the cochineal 50 ˚C
extract showed a t1/2 value 1.3–3.0 times higher than the 20
other colorant extracts. Among the three anthocyanin
extracts, the half-life values of elderberry were 1.1–1.3 10
times higher than those for red cabbage, and 1.3–1.7 times
higher than those for hibiscus. In previous studies on antho-

b*
0
cyanin extracts from Hibiscus higher t1/2 values were regis-
tered under comparable temperature conditions [26], -10 Hibiscus
Opuntia
however the stability reported in elderberry extract [19] is Red beet
similar to our data. Betacyanins are commonly known to be Red cabbage
-20 Elderberry
heat liable pigments, their stability was observed to consid- Cochineal
erably decline above 50 °C, in red beet extracts a t1/2 value
-30
of 3.3 h (50 °C) was reported [26] which agrees with our 0 10 20 30 40 50 60 70
results, while in Opuntia fruit extracts, half-life times of a*

3.0 h (50 °C), 1.1 h (70 °C) and 0.2 h (90 °C) were obtained 30
[27], in the same order that our findings (Table 2). Plots of ln 70 ˚C
k vs.1/T (Eq. 2) gave straight lines for each of the red
20
colorant extracts. Activation energies (Ea), derived from
the slopes of these lines, ranged between 3.0 and 53.4 kJ
mol−1. Higher Ea values are associated with increased tem-
10

perature dependence of the degradation rate [28]. The

b*

Table 2 Kinetic parameters and total color differences (ΔE*) of the

-10
thermal degradation processes for the natural red food colorants

Temperature k t1/2a (h) Ea ΔE* -20

−1
(°C) (h ) (kJmol−1) (after 6 h)

Elderberry 50 0.01 5.40 (0.99) 15.37 12.3 -30

0 10 20 30 40 50 60 70
70 0.04 4.02 (0.99) 20.4 a*
90 0.06 3.56 (0.97) 35.7
30
Red cabbage 50 0.03 4.14 (0.99) 26.50 12.4
90 ˚C
70 0.06 3.58 (0.98) 23.4
20
90 0.10 3.05 (0.98) 43.3
Hibiscus 50 0.05 3.63 (0.97) 38.20 11.9
70 0.09 3.08 (0.97) 24.4 10

90 0.26 2.05 (0.94) 50.6

b*

Red beet 50 0.08 3.28 (0.94) 35.37 12.7 0

70 0.19 2.34 (0.91) 31.3

90 0.32 1.84 (0.91) 55.0 -10

Cochineal 50 0.01 5.52 (0.99) 3.02 7.9

70 0.01 5.30 (0.99) 10.1 -20

90 0.01 5.40 (0.98) 12.2

Opuntia 50 0.10 2.98 (0.90) 53.37 13.1 -30
0 10 20 30 40 50 60 70
70 0.35 1.75 (0.90) 39.7
a*
90 0.91 0.79 (0.89) 57.5
Fig. 4 Colorimetric plane (a* vs. b*) showing the incidence of the
a
Correlation coefficient of determination (R2 ) shown in parenthesis thermal treatments (50, 70 and 90 °C) in the colorant extracts
16
In general, the difference in stability observed among the
red pigment extracts was related to the nature of the pig-
ments. The highest stability corresponded to the cochineal
extract and the lowest to the betacyanin extracts (red beet
and Opuntia), while the anthocyanin extracts (elderberry,
red cabbage and hibiscus) were in an intermediate position.
The different stabilities among anthocyanin extracts could
be related with the chemical structures of the anthocyanins
present in the extracts, including the types of aglycones and
sugar moieties, the complexity of sugar residues, the pres-
ence of acylating structures or even the presence of other
phenolic compounds in these extracts [28]. The difference in
thermostability between red beet and Opuntia extracts might
also be mainly attributed to the different chemical structures
of the individual betacyanin pigments present in both
extracts and even to the presence of additional compounds
that would affect stability during heating [30].
Changes in Visual Color Attributes
The changes in visual color were evaluated by the CIELab
parameters which can achieve a precise color characterization
and are gaining in popularity as descriptors for use in color
specification by food manufacturers. It can be observed that
thermal processing involves an increase in the lightness (L*),
accompanied by a decrease in the red-green parameter (a*),
particularly in the most thermosensitive extracts, and a slight
increase in the blue-yellow parameter (b*). These effects are
similar to those reported for aqueous anthocyanin extracts [23,
28] and also for betacyanin extracts [20, 31]. As seen in Fig. 4,
plotting the values in the plane a* vs. b* (colorimetric plane)
illustrates how redness decreases during the heating period.
The greater the dispersion of the points among the same series
the more pronounced color degradation. Red cochineal
extracts were the only ones to remain grouped after treatment
for 6 h at 50, 70 or 90 °C, with color differences (ΔE*) of 7.9,
10.1 and 12.2, respectively (Table 2). This, corroborates the
view that red cochineal is the most thermoresistant extract,
while red beet and Opuntia extracts showed a pronounced
degree of degradation even during heating at moderate tem-
perature (50 °C). The color differences confirmed the greater
thermolability of both betacyanin extracts.
Conclusions
The effects of thermal treatment on the color shade of aqueous
red pigment extracts from elderberry, red cabbage, hibiscus,
red beet, Opuntia fruits and red cochineal were investigated.
The results from the present study provided detailed informa-
tion regarding the changes in kinetic stability and visual color.
Red cochineal exhibited the highest thermostability among the
natural colorants investigated, and even after a severe
Plant Foods Hum Nutr (2013) 68:11–17
treatment of 6 h at 90 °C this extract did not show significant
color modifications. Betacyanin extracts (red beet, Opuntia)
was found to be more temperature-sensitive than anthocyanin
extracts (elderberry, red cabbage, hibiscus).
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