See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/234841375

Degradation kinetics and color stability of spray-dried

encapsulated anthocyanins from Hibiscus sabdariffa L

Article  in  Journal of Food Process Engineering · August 2012

DOI: 10.1111/j.1745-4530.2010.00605.x

CITATIONS READS

63 1,132

3 authors, including:

Zuhaili Idham Ida Idayu Muhamad

Universiti Teknologi Malaysia Universiti Teknologi Malaysia
33 PUBLICATIONS   132 CITATIONS    203 PUBLICATIONS   861 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Production and Modification of Bacterial Cellulose View project

Development of product from upland rice bran oil View project

All content following this page was uploaded by Ida Idayu Muhamad on 02 June 2018.

The user has requested enhancement of the downloaded file.

jfpe_605 522..542

bs_bs_banner

DEGRADATION KINETICS AND COLOR STABILITY OF

SPRAY-DRIED ENCAPSULATED ANTHOCYANINS FROM
HIBISCUS SABDARIFFA L.

ZUHAILI IDHAM1,2, IDA IDAYU MUHAMAD1,4 and MOHD ROJI SARMIDI3

1
Department of Bioprocess Engineering
Faculty of Chemical & Natural Resources Engineering
Universiti Teknologi Malaysia Skudai
UTM Skudai, 81310, Johor Bahru
Johor, Malaysia
2
Centre of Lipid Engineering Applied
Research (CLEAR), UTM Skudai,
Johor, Malaysia
3
Chemical Engineering Pilot Plant (CEPP)
UTM Skudai, Johor, Malaysia

Received for Publication August 2, 2009

Accepted for Publication March 25, 2010

ABSTRACT

Dry powder of red color from roselle calyces, which can be used as
natural colorant and functional food ingredient, was produced using microen-
capsulation technique. Four different matrices, i.e., maltodextrin, gum Arabic,
combination of maltodextrin and gum Arabic, and soluble starch were used for
encapsulation study. Identification and measurement of anthocyanins in
encapsulated roselle was made by high-performance liquid chromatography.
Encapsulation efficiencies were determined and compared with storage analy-
sis data. The stability of encapsulated pigments was investigated during
storage under three different storage temperatures (4, 25 and 37C) until 105
days. The four type of matrices largely increased the half-life of the pigments
during storage especially at 37C (P < 0.05) compared with the non-
encapsulated roselle extract. Storage temperature did not significantly
(P > 0.05) effect the L*, a* and b* values. However, type of encapsulation

4
Corresponding author. TEL: 607-5535541; FAX: 607-5581463; EMAIL: idayu@fkkksa.utm.my

Journal of Food Process Engineering 35 (2012) 522–542. All Rights Reserved.

522 © 2011 Wiley Periodicals, Inc.
DOI: 10.1111/j.1745-4530.2010.00605.x
 STABILITY OF SPRAY DRIED ANTHOCYANINS 523

agent and storage time significantly (P < 0.05) affected the color changes. The
L* and a* values were decreased during storage whereas the b* value was
increased. Combination of maltodextrin and gum Arabic showed the highest
encapsulation efficiencies (99.87 ⫾ 0.04%), lower degradation rate in 4C
(3.7 ⫾ 0.3) and had smaller changes in a* and b* values among the four
matrices.

PRACTICAL APPLICATIONS

A spectrum of natural, red-hue primer colorants derived from roselle

plant extracts, substantially free of alkaloids, enzymes, aroma or solvent
residuals, and microencapsulated to ensure good stability and shelf life. This
study measures the storage stability and suitability of the microencapsulated
natural colorants. The colorants are for functional use in the food, pharmaceu-
ticals, cosmetics and other industries.

INTRODUCTION

The development of food colorants from natural sources has been glo-
bally increased. This phenomenon was due to an effort to replace the synthetic
dyes because of consumer concern and legislative action such as the ban of
FD&C Red 40 and FD&C Red 2 by the Federal Drug Association (Cevallos-
Casals and Cisneros-Zevallos 2003). In this context, anthocyanins are consid-
ered as suitable food pigment that can contribute for the most spectacular red,
blue and purple colors in many fruits and vegetables. Existing evidence indi-
cates that anthocyanins not only nontoxic and nonmutagenic but have positive
therapeutic properties (Bridle and Timberlake 1997).
Hibiscus sabdariffa or locally known as Roselle is rich in anthocyanins,
and could be used as a good source for producing a brilliant red colorant for
many foods (Bridle and Timberlake 1997). However, anthocyanins as with
most natural food colorant, suffer from inherent instability (Selim et al. 2008).
Stability of anthocyanins depends on a combination of environment and
chemical factors. Anthocyanins and polyphenols are not stable to heat during
juice processing (Fang et al. 2006). The stability of anthocyanins varies
depending on the processing conditions, and the presence of other components
in their new environment (Kirca et al. 2007). Furthermore, storage stability is
more depending on environment factors such as temperature, light, oxygen and
water activity (Aw).
524 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

One alternative for improving anthocyanins stability is the microencap-

sulation technique, which entraps the sensitive ingredient inside a coating
material. Encapsulation techniques have been widely used to reduce interac-
tions of food components with environment factors, such as temperature, light,
moisture and oxygen. Spray-drying and freeze-drying are well-established
technologies available for microencapsulation process in the food industry and
most commonly used. Freeze-drying is also known as one of the best method
to dry pigments, which are sensitive to high temperatures such as anthocya-
nins. There have been several studies on the freeze-drying of Roselle antho-
cyanins in different wall materials or encapsulation agent: pullulan (Gradinaru
et al. 2003), trehalose and maltodextrin (Duangmal et al. 2008), maltodextrin
and gum Arabic (Selim et al. 2008). Nevertheless, the freeze-drying method is
30–50 times more costly than spray-drying (Diaz et al. 2006). Spray-drying
has paved the way for the production of powder colorants with high storage
stability, easier handling for some applications and minimized transportation
weight in comparison with liquid concentrates. Some advantages of spray-
drying including the ability to quickly produce dry powder (e.g., as compared
with freeze-drying) and the ability to control the particle size distribution.
Roselle powders obtained by a spray-drying method were studied by
Al-Kahtani and Hassan (1990), Clydesdale et al. (1979), Andrade and Flores
(2004), Chiou and Langrish (2007).
Different types of encapsulating agents have been used for spray-drying,
including polysaccharides (starches, maltodextrins, corn syrups and gum
Arabic), lipids (stearic acid, mono- and diglycerides) and proteins (gelatin,
casein, milk serum, soy and wheat) (Gibbs et al. 1999). Anthocyanins are
hydrophilic colorants and specifically compatible with a water-based gel for-
mulation such as pectin and gum, or maltodextrin and starches as coating
molecules for polar solid matrices. Gum Arabic, a natural plant polysaccharide
exudate of acacia is well-known effective wall material used for many years
and still a good choice because of its stable emulsion formation and good
retention of volatiles. The carbohydrates such as starches and maltodextrins
have properties that are desirable in an encapsulating agent such as low
viscosity at high solids contents and good solubility. The advantage of using
microencapsulated products is that their coating stays intact and therefore
colors do not easily migrate as do conventional ones. Also, their brightness is
improved, thus increasing their marketability for commercial use.
In the present study, stability of anthocyanins of spray-dried roselle in
four different wall materials was studied at three different storage tempera-
tures. The degradation kinetic analysis of the spray-dried powders in compari-
son with non-encapsulated roselle extract was also carried out during 105 days
storage period. The changes in color parameter for spray dried powders were
also recorded.
 STABILITY OF SPRAY DRIED ANTHOCYANINS 525

MATERIALS AND METHODS

Materials
Roselle varieties UMKL-1 were obtained from Green Valley Eco Village,
Saleng, Kulai, Johor, Malaysia. The dried ground calyces were kept in a screw
cap glass bottle, protected from light and placed at -18C until further use.
Maltodextrin DE 11–15, gum Arabic and soluble starch (Kanto Chemical Co.,
Tokyo, Japan) used were food grade. Ethanol, citric acid, potassium chloride,
sodium acetate, acetonitrile were analytical grade and purchased from chemi-
cal suppliers. Kuromanin chloride (cyanidin 3-glucoside) was obtained from
Sigma Chemical (Selangor, Malaysia).

Method

Extraction and Concentration of Roselle Anthocyanins. Pigment

extraction method was modified from Duangmal et al. (2008) by maceration
of dry powdered calyces with water-95% ethanol (1:1) containing 2% citric
acid at pH 2. The extract was kept for 24 h at room temperature in the dark.
The extract was filtered in vacuum using Wattman filter (grade 1). The plant
material was re-extracted with acidified ethanol until a faint-colored extract
was obtained. Filtrates were pooled and the plant material was discarded.
Then the ethanol-acid solvent was removed with rotary vacuum evaporator at
40C.
Aliquots of 100 mL of the extracts were applied to the Amberlite XAD-16
resin (Kammerer et al. 2005), which was washed with 50 mL of distilled
water. The extract was subsequently rinsed with 125 mL distilled water and
with 50 mL acidified water at a flow rate of 3 mL/min in the glass column. For
elution of the pigments, 200 mL of acidified ethanol were used. During sample
application, washing and elution of the pigments, the fractions of the dark red
color was collected. Then solvent were removed in a rotary evaporator at 40C
under vacuum. The pigments were redissolved in distilled water until Brix
maintained at 8°Brix. Brix was measured using Atago Refractometer (Tokyo,
Japan) at 20C.

Microencapsulation of Anthocyanins. Maltodextrin (MD) dextrose

equivalent (DE) 11–15, gum Arabic, combination of gum Arabic and malto-
dextrin (60:40) and soluble starch (SS) were evaluated as encapsulating
agents. These carrier agents were combined with the pigment extract (8°Brix)
and stirred until all the material were completely dissolved. Meanwhile, the
carrier agents were added until reaching to 20% final solid content. Then the
mixtures were vigorously homogenized at 14,000 rpm for 1 h at room tem-
526 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

perature. Five hundred milliliters of feed mixtures were prepared for further
processing. The resulting emulsion was fed into a spray-dryer at flow rate 9.5%
and was atomized by a centrifugal atomizer. The inlet and outlet air tempera-
tures were 150 and 100C, respectively. The resultant prepared microcapsules
were collected in the cyclone.

Identification of Anthocyanins. Identification of anthocyanins

(cyanidin 3-glucoside) in spray dried roselle powders was carried out using
the Waters HPLC System model 600 E (Waters Corporation, Taunton, MA).
The chromatographic system consists of 150 ¥ 4.6 mm Wakosil C18RS
stainless steel column (SGE, Ringwood, Australia) equipped with pump
(model 600). The column temperature was 30C and injection volume was
20 mL. Acetonitrile : water (65:35) was used as mobile phase solvent. The
flow rate was 1.0 mL/min and the column was monitored by UV detector set
at wavelength 520 nm. Their relative concentrations were determined from
respective peak areas of chromatograms, using cyanidin 3-glucoside (kuro-
manin chloride) as standard.

Total Anthocyanins (TA) Content. TA content was determined using

the pH-differential method described by AOAC (2005), using two buffer
systems: potassium chloride buffer at pH 1.0 (0.025 M) and sodium acetate
buffer at pH 4.5 (0.4 M). An aliquot of the extract was transferred to a 10 mL
volumetric flask and made up to 10 mL with corresponding buffer and the
absorbance was measured at 520 and 700 nm by UV-VIS spectrophotometer
Lambda 25 (Perkin Elmer, Shelton, CT). The reason for measuring the absor-
bance at 700 nm is to correct for haze. TA were calculated as cyanindin-3-
glucoside according to the following equation:

TA (mg L ) = ΔA ε × 1 × M × 103 × D (1)

where DA, (A510 pH 1.0 - A700 pH 1.0) - (A510 pH 4.5 - A700 pH 4.5); e (molar
extinction coefficient) = 26,900 L/mol/cm for cyanidin-3-glucoside; 1, path
length in cm; M (molecular weight) = 448.8 g/mol for cyanidin-3-glucoside;
D, dilution factor; 103, conversion from gram to milligram.

Encapsulation Efficiencies. In order to evaluate the effectiveness of

microencapsulation, TA content and surface anthocyanins of the microcap-
sules were determined after spray drying. For the TA determination, 100 mg of
samples were weighed in an amber vial with a screw top. About 1 mL of
distilled water was added and then the samples were ground to destroy the
microcapsule membrane. Then 9 mL ethanol was added and the samples were
extracted for 5 min and then filtered.
 STABILITY OF SPRAY DRIED ANTHOCYANINS 527

For determination of surface anthocyanins (SA), 100 mg of samples were

directly extracted with 10 mL ethanol in a vortex for 30 s, followed by cen-
trifugation at 3,000 rpm for 10 min at 20C. After phase separation, the clear
supernatant was collected and filtered through 0.45-mm-sized Millipore
membrane.
Anthocyanins content for TA and SA values were determined using
pH-differential method (AOAC 2005). Encapsulation efficiencies were calcu-
lated according to an equation modified from Barbosa et al. (2005) as shown
in the following equation:
(TA − SA)
%EE = × 100 (2)
TA

Pigment Powder Storage. Pigment powders were stored in brown

bottles with screw caps and placed at 4, 25 and 37C in the absence of light. The
samples were stored in a well cap bottle. The analyses of anthocyanins in the
powder were made in duplicate periodically up to 150 days. The anthocyanins
were released from the microcapsule by mixing them with distilled water and
ground to destroy the microcapsule membrane. Ethanol was added and the
samples were extracted for 5 min, and then filtered using Whatman no1. The
TA content was calculated according to pH differential method (AOAC 2005).
Stability was determined in the basis of anthocyanins retention from initial and
after encapsulated. Fresh roselle extract was used as control to compare with
the encapsulated samples.

Kinetic Analysis of Anthocyanins Degradation. The data was best fit

by a first-order kinetics model,
ln (C ) = ln C0 − k (t ) (3)

where Co is the initial anthocyanin content (day 0) after spray drying and C is
the anthocyanin content after t (time) of stability treatment at a given tempera-
ture. Degradation rate constants (k) were obtained from the slope of a plot of
the natural log of percentage retention of anthocyanins measured by UV/VIS
spectrophotometer versus time. For a first order reaction half-life values were
determined at a specific temperature by the equation (Fernando Reyes and
Cisneros-Zevallos, 2007);
t 1 2 = ln (2 ) kT (4)

Changes in Visual Color. The colors of the spray-dried powder were

measured in terms of the CIE L*, a*, b* values using a C-10 Color Reader
528 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

(Konica Minolta, Osaka, Japan). All color parameter was plotted against time
(days) to evaluate the color changes of spray-dried powders through storage.

Statistical Analysis. Experiments of storage were duplicated for each

temperature. Degree of significance among various constants was analyzed
using analysis of variance at P < 0.05 level using the Design Expert 6.0.8 (Stat
Ease, Inc., Minneapolis, MN). Linear regression analysis was used to obtain
the degradation rate constants in the kinetic studies of the encapsulated
samples. The regression data were calculated by using Data Analysis in
Microsoft Excel 2007.

RESULTS AND DISCUSSION

Identification of Anthocyanins in Roselle Powders

Identification of anthocyanins was made to all encapsulated sample to
confirm the presence of anthocyanins by comparing it with the cyanidin
3-glucoside as the standard anthocyanins. The HPLC chromatogram data
(Fig. 1) show all the spray-dried samples that contained cyanidin 3-glucoside.
Figure 1a shows the chromatogram of standard where the retention time of
cyanidin-3-glucoside was 1.526. The retention time for cyanidin 3-glucoside
in all the samples was in the range of 1.430–1.465. Although the retention time
for samples and standard was slightly different, the peaks were clearly iden-
tified as cyanidin-3-glucoside. The difference may be caused by the pressure in
HPLC equipment and column conditions. The highest absorbance was
obtained from anthocyanins coated with maltodextrin followed by combina-
tion of maltodextrin and gum Arabic, then gum Arabic alone. The lowest
absorbance was identified from sample coated with soluble starch.

Encapsulation Efficiencies
Figure 2 illustrates the encapsulation efficiencies after the spray-drying
process. Encapsulation efficiencies refer to the potential of the wall material to
encapsulate or hold the core material inside the microcapsule. Encapsulation
efficiencies are also related to the shelf life of the anthocyanin content in the
powder. The combination of maltodextrin and gum Arabic gave the highest
encapsulation efficiencies (99.87 ⫾ 0.04%) of anthocyanins, followed by mal-
todextrin (99.69 ⫾ 0.06%), gum Arabic (98.4 ⫾ 0.11%) and soluble starch
(96.7 ⫾ 0.35%). Lianfu et al. (2007) undertook optimization in the microen-
capsulation of procyanidins, and reported that the optimal condition for
microencapsulating procyanidins was using 60% maltodextrin and 40% gum
Arabic as wall materials, with the solids content of the slurry before spray-
 STABILITY OF SPRAY DRIED ANTHOCYANINS 529

FIG. 1. HPLC CHROMATOGRAM OF ANTHOCYANINS (CYANIDIN-3 GLUCOSIDE) IN

ENCAPSULATED SAMPLES
(a) Standard. (b) Maltodextrin. (c) Maltodextrin and gum Arabic. (d) Gum Arabic.
(e) Soluble starch.
530 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

FIG. 1. CONTINUED

drying at 20% w/v. The ratio was used in this study to compare results among
other feed formulations. Consequently, the result indicates that the combination
of maltodextrin and gum Arabic formed suitable chemical interactions that help
in retaining the anthocyanins inside the wall material. The effectiveness of
interactions depended on the chemical and physical structure of each support
material (Berset et al. 1995). According to Shahidi and Naczk (2004), phenolics
and flavonols may form complexes with polysaccharides and the affinity of
phenolics to polysaccarides depends on the water solubility, molecular size,
conformational mobility and shape of polyphenol. Furthermore, the complexity
formed when the flavylium cation of the anthocyanins interacted with dextrins
prevented their transformation to other less stable forms (Chandra et al. 1993).
However, Barbosa et al. (2005) reported that maltodextrin with the added
emulsifier Tween 80 had the ability to encapsulate a higher amount of bixin than
maltodextrin alone. On the other hand, gum Arabic is known as a natural
 STABILITY OF SPRAY DRIED ANTHOCYANINS 531

Encapsulation Efficiencies (%) 101

100

99

98

97

96

95

94
Maltodextrin Maltodextrin + gum Gum Arabic Soluble starch
Arabic
Encapsulating agent

FIG. 2. ENCAPSULATION EFFICIENCIES IN FOUR DIFFERENT POLYSACCHARIDES

AFTER SPRAY-DRYING PROCESS

emulsifier for nonpolar substances. It has the structure of a highly branched

heteropolymer of sugars, glucuronic acid and a small amount of protein
covalently linked to the carbohydrate chain, which is a well-known film-
forming ingredient (Dickinson 2003). Therefore, the utilization of gum Arabic
combined with maltodextrin in the formulation has a higher potential to
encapsulate the anthocyanins over maltodextrin or gum Arabic alone.
Selim et al. (2008) studied the stability of encapsulated roselle anthocya-
nin pigment and found maltodextrin gave a significantly better protective
effect compared with gum Arabic. In the contrary, soluble starch was a less
suitable wall material for microencapsulation of the anthocyanins. The reason
perhaps the starch could not create dense and thick wall systems to protect
anthocyanins from expel and the pigments only attached to the surface of
starch granule without formation of the complexes. From the perspective of the
feed formulation preparation, it was also observed that starch was not suitable
as a wall material because of its difficulty to reach homogeneity compared
with other wall materials.

Kinetic Analysis of Anthocyanins During Storage

Most reactions that showed loss in food quality may be described by zero
or first order (Pua et al. 2008). In order to determine the reaction order of a
quality attribute, the experimental data obtained was fitted to linear equation.
The retention of anthocyanins [Ct/C0] measured at different times of storage
versus time was plotted and data obtained was made to fit a linear equation. If
532 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

the data showed a good fit to a linear equation, then the data indicated a zero
order reaction. On the other hand, if the ln[C0/Ct] versus time fits a linear
equation, then the reaction is considered to be of first order. [C0/Ct] is reten-
tion of anthocyanins at t time. Figure 3 shows linear relationship (R2 > 0.80)
between [ln Ct/C0] and time for encapsulated anthocyanins in different wall
material and fresh roselle extract as control at temperatures of 4, 25 and 37C.
These results mean the degradation of TA content implying first order reaction
kinetics behaviour for all the encapsulated sample and control, from all the
temperature studied. This research was in agreement with some storage studies
to the anthocyanins extract such as, blackberry juice (Wang and Xu 2007),
blood orange juice (Kirca and Cemeroglu 2003) and raspberry pulp (Ochoa
et al. 1999). Similar kinetic responses for encapsulated anthocyanins have
been reported by Ersus and Yurdagel (2007) where the retention of anthocya-
nins in encapsulated blackberry by different DE of maltodextrin also showed
first order kinetics at the storage temperature.
Table 1 shows the first order degradation rate constant (k) obtained from
the slope of logarithmic plots of anthocyanins retention versus time. Half-life
value was calculated from the degradation rates. The results show that the
effect of storage temperature on degradation kinetic rates for anthocyanins in
control and encapsulated powders were significantly different (P < 0.05). An
increase in storage temperature led to an increase in the rate constant (k) of all
sample except at 4C and 25C for encapsulated anthocyanins with gum Arabic
alone and soluble starch. The results indicate that at 4C, the degradation rate
constant value were very low and ranged between 5.7 ¥ 10-3/days of the
control sample and 4.2–5.7 ¥ 10-3/days for the encapsulated samples. At 25C,
the reaction rate constants of the control sample increased to 13.5 ¥ 10-3/days
but only slightly increased for the encapsulated samples to 4.4–5.7 ¥ 10-3/
days, respectively. The highest value of the rate constant for anthocyanins
degradation was observed at 37C for all matrices evaluated, wherein the
reaction rates of the control was as high as 37.5 ¥ 10-3/days, whereas those of
encapsulated samples ranged between 4.6 and 6.7 ¥ 10-3/days.
The effect of the encapsulating agent and storage temperature on stability
of encapsulated roselle anthocyanins were statistically analyzed (Table 1). The
degradation rates, k for roselle extract at 25C (13.5 ¥ 10-3) and 37C
(37.5 ¥ 10-3) showed significantly higher values (P < 0.05) than encapsulated
roselle. These signify that encapsulation process could stabilize and extend the
shelf life of anthocyanins content. Combination of maltodexrin and gum Arabic
as encapsulating agent gave the lowest degradation rates at 4C (3.7 ¥ 10-3) and
25C (4.3 ¥ 10-3). This result supports the analysis that show combination both
of them as wall material gave the highest encapsulation efficiencies (EEs). The
high EEs and low degradation rates indicated the stability and prolonged shelf
life of the anthocyanins in the powder. Selim et al. (2008) studied the effect of
 STABILITY OF SPRAY DRIED ANTHOCYANINS 533

Time (days)
0
20 40 60 80 100 120
-0.1

-0.2

MD
-0.3
In (Ct/C0)

MD+GA
-0.4
GA
-0.5
SS
-0.6
control
-0.7

-0.8
a
Time (days)
0
1 21 41 61 81 101 121
-0.2

-0.4
MD
-0.6
MD+GA
In (Ct/C0)

-0.8
GA

-1 SS

-1.2 control

-1.4

-1.6
b
Time (days)
0
1 81 101 121
-0.5

-1

-1.5 MD
In (Ct/C0)

-2 MD+GA

-2.5 GA

-3 SS

-3.5 control

-4

-4.5
c

FIG. 3. FIRST-ORDER KINETIC PLOT FOR RETENTION OF ANTHOCYANINS IN

DIFFERENT WALL MATERIAL AND CONTROL (ROSELLE EXTRACT) AT (a) 4C; (b) 25C
AND (c) 37C
GA, gum arabic; MD, maltodextrin; SS, soluble starch.
534 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

TABLE 1.
DEGRADATION PARAMETERS OF ANTHOCYANINS RETENTION IN ENCAPSULATED
AND CONTROL (ROSELLE EXTRACT) IN STORAGE STABILITY

Samples Temperature k ¥ 103 ⫾ sk ¥ 103 (d-1) Linear regression (R2) Half-life, t1/2 (month)

MD 4 4.2 ⫾ 0.3a 0.914 5.5

25 4.4 ⫾ 0.3b 0.933 5.3
37 4.6 ⫾ 0.3c 0.944 5
MD + GA 4 3.7 ⫾ 0.3a 0.954 6.2
25 4.3 ⫾ 0.3b 0.954 5.4
37 4.6 ⫾ 0.2c 0.980 5
GA 4 4.9 ⫾ 0.2a 0.980 4.7
25 4.9 ⫾ 0.3a 0.943 4.7
37 5.3 ⫾ 0.6b 0.843 4.4
SS 4 5.7 ⫾ 0.4a 0.925 4
25 5.7 ⫾ 0.7a 0.849 4
37 6.7 ⫾ 0.6b 0.900 3.4
Control 4 5.7 ⫾ 0.3a 0.968 4
25 13.5 ⫾ 0.5b 0.978 1.7
37 37.5 ⫾ 1.4c 0.980 0.6

* Means with different letters within column indicate significance difference at P < 0.05.
a–c Represents the significant (P < 0.05) effect of temperature on degradation kinetic of anthocyanins
on different encapsulating agent used.
GA, gum arabic; MD, maltodextrin; SS, soluble starch.

different encapsulating agent on the stability of anthocyanins stored in different

Aw. They used three different maltodextrins of varying molecular weight (DE)
and found that maltodextrin 20 DE showed the lowest degradation kinetic rates
compared with maltodextrin 10 DE and gum Arabic.
The half-life data was plotted as graph in Fig. 4. Increase in the rate
constants was associated with decrease in the half-life period (days) for the
stored samples as estimated from equation 4. Clearly, the combination of
maltodextrin and gum Arabic provided the greatest protecting effect indicating
the longest half-life periods at three storage temperatures. This powder had
more than 6 month period before losing half of the anthocyanins content stored
at 4C. Maltodextrin as wall material showed the half-life more than 5 months
for the three storage temperatures. Gum Arabic showed similar half-life
(P > 0.05) at 4 and 25C (4.7 months) but decreased at 37C (4.4 months). As
expected, soluble starch showed the highest degradation rates and half-life
during storage at 4, 25 and 37C among them. The half-life data infer the
importance of the encapsulating material in degradation process of the antho-
cyanins pigment. The matrix effect caused by storage temperature was also
observed in encapsulated oleoresin paprika where better protection of 15 DE
maltodextrin with respect to the content gum Arabic, gelatin and sodium
caseinate was reported (Beatus et al. 1985).
 STABILITY OF SPRAY DRIED ANTHOCYANINS 535

FIG. 4. HALF-LIFE PERIOD (DAYS) FOR ROSELLE PIGMENTS ENCAPSULATED IN

DIFFERENT MATRICES AT DIFFERENT TEMPERATURE
GA, gum arabic; MD, maltodextrin; SS, soluble starch.

Color Changes during Storage

The initial color parameter L*, a*, b* values for encapsulated roselle
extract were 39.3, 43.1 and -0.8 for maltodextrin as wall material, 45.9, 34.8
and -4.3 for combination of maltodextrin and gum Arabic, 44.9, 30.3 and -6.3
for gum Arabic alone and 38.8, 26.9 and -3.8 for soluble starch, respectively.
The initial values show that the gum Arabic as encapsulating agent gave high
lightness value compared with the starch and maltodextrin. Maltodextrin gave
the highest red and yellow color among other encapsulating agents. Figure 5
shows that storage period and encapsulating agent significantly affected the
color change, whereas storage temperature did not give much effect. The
gradual degradation of the lightness and red color, visually observed in all
spray-dried powder and storage temperature used as verified by the similar
decrease (R2 > 0.8) of a* and L* values in Fig. 5a,b. However, the yellowness
increased in all sample and storage condition (Fig. 5c). This implied that the
color of powders has become darker in comparison with control or non-
encapsulated samples and the powders tend to become brownish because of
the increase in yellowness. From Fig. 6, it was found that during storage the
changes of a* (redness) and b* (yellowness) of all sample were in the range of
approximately 10 unit differences. This indicated only minor changes of the
powder shade and color occurred throughout the storage time. Figure 6 also
shows that combination of maltodextrin and gum Arabic gave the smallest
change in a* and b* values. The changes of redness might be caused by the
536 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

a 50
45
40 4°C
35
L∗ (Lightness)

30
25
20
15
10
5
0
0 20 40 60 80 100 120
Time (days)
50
45 25°C
40
35
L∗ (lightness)

30
25
20
15
10
5
0
0 20 40 60 80 100 120
Time (days)

50
45
37°C
40
L∗ (lightness)

35
30
25
20
15
10
5
0
0 20 40 60 80 100 120
Time (days)

MD MD+GA GA SS Control

FIG. 5. CHANGES IN THE COLOR PARAMETERS VALUE (a) L* (LIGHTNESS), (b) a*,
(c) b* FOR ENCAPSULATED POWDERS AND CONTROL AT 4, 25, AND 37C
GA, gum arabic; MD, maltodextrin; SS, soluble starch.
 STABILITY OF SPRAY DRIED ANTHOCYANINS 537

b 50
45
4°C
40
35
30
a∗

25
20
15
10
5
0
0 20 40 60 80 100 120
Time (days)
50
45 25°C
40
35
30
a∗

25
20
15
10
5
0
0 20 40 60 80 100 120
Time (days)

50
45 37°C
40
35
30
a∗

25
20
15
10
5
0
0 20 40 60 80 100 120

Time (days)

MD MD+GA GA SS Control

FIG. 5. CONTINUED
538 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

c 8

6
4°C
4

2
b∗

-2 0 20 40 60 80 120

-4

-6

-8
Time (days)

6 25°C

2
b∗

-2 0 20 40 60 80 100 120

-4

-6

-8
Time (days)

8
6 37°C
4
2
b∗

0
-2 0 20 40 60 120
-4
-6
-8
Time (days)

MD MD+GA GA SS Control

FIG. 5. CONTINUED
 STABILITY OF SPRAY DRIED ANTHOCYANINS 539

20

4°C
10
b∗ value

0
0 10 20 30 40 50

-10

-20
a∗ value

20
25°C

10
b∗ value

0
0 10 20 30 40 50

-10

-20 a∗ value

20
37°C
10
b∗ value

0
0 10 20 40 50

-10

-20
a∗ value

MD MD+GA GA SS Control

FIG. 6. CHANGES IN a* AND b* VALUES OF ENCAPSULATED POWDERS DURING

STORAGE AT 4, 25 AND 37C
540 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

degradation of anthocyanins during storage. Diaz et al. (2006) noted that the
reduction of spray-dried betacyanin pigment content during storage was also
observed by reduction in the values of a*, which indicated a decrease in red
color intensity in the stored samples.

CONCLUSION

Encapsulation of anthocyanins with polysaccharides followed by appro-

priate processing may enhance the stability of anthocyanins for efficient uti-
lization in food systems. Under the conditions of this study, the
microencapsulated anthocyanins with a combination of maltodextrin and gum
Arabic had the highest encapsulation efficiencies. The storage stability result
supported that combination both of maltodextrin and gum Arabic as wall
material gave the longest shelf life and the smallest change in the pigment
color. Storage period significantly affect the color changes of the spray dried
powder. Hence, this study signifies that encapsulation process could stabilize
and extend the shelf life of anthocyanins content. It was deemed necessary to
perform further studies on anthocyanin stability under various conditions to
generate further scientific information on this important subject.

ACKNOWLEDGMENT

The authors gratefully thank the Chemical Engineering Pilot Plant

(CEPP), Universiti Teknologi Malaysia, Skudai for the financial support
through the bio-entrepreneur program.

REFERENCES

AL-KAHTANI, H.A. and HASSAN, B.H. 1990. Spray drying of roselle

(Hibiscus sabdariffa L.) extract. J. Food Sci. 55(4), 1073–1076.
ANDRADE, I. and FLORES, H. 2004. Optimization of spray-drying of roselle
extract (Hibiscus sabdariffa L.) drying. Proceedings of the 14th
International-drying Symposium (IDS 2004). Sao Paulo, Brazil. A, 597–
604.
AOAC 2005. AOAC Official Method 2005.02: Total monomeric anthocyanin
pigment content of fruit juices, beverages, natural colorants, and wines by
the pH differential method. Official Methods of Analysis of AOAC Inter-
national, 18th Ed., (H. HOROWITZ, ed.) AOAC, Washington, DC.
 STABILITY OF SPRAY DRIED ANTHOCYANINS 541

BARBOSA, M.I.M.J., BORSARELLI, C.D. and MERCADANTE, A.Z.

2005. Light stability of spray-dried bixin encapsulated with different
edible polysaccharide preparations. Food Res. Int. 38, 989–994.
BEATUS, Y., RAZIEL, A., ROSENBERG, M. and KOPELMAN, I.J. 1985.
Spray-drying microencapsulation of paprika oleoresin. Lebensm. Wiss.
Technol. 18, 28–34.
BERSET, C., CLERMONT, H. and CHEVAL, S. 1995. Natural red colourant
effectiveness as influenced by absorptive supports. J. Food Sci. 60, 858–
861, 879.
BRIDLE, P. and TIMBERLAKE, C.F. 1997. Anthocyanins as natural food
colours: Selected aspects. Food Chem. 58, 103–109.
CEVALLOS-CASALS, B.A. and CISNEROS-ZEVALLOS, L. 2003. Sto-
ichiometric and kinetic studies of phenolic antioxidants from Andean
purple corn and red-fleshed sweetpotato. J. Agric. Food Chem. 51, 3313–
3319.
CHANDRA, A., NAIR, M.G. and IEZZONI, A.F. 1993. Isolation and stabi-
lization of anthocyanins from tart cherries (Prunus cerasus L.). J. Agric.
Food Chem. 41, 1062.
CHIOU, D. and LANGRISH, T.A.G. 2007. Development and characterisation
of novel nutraceuticals with spray-drying technology. J. Food Eng. 82(1),
84–91.
CLYDESDALE, F.M., MAIN, J.H. and FRANCIS, F.J. 1979. Roselle (Hibis-
cus sabdariffa L.) anthocyanins as colorants for beverages and gelatin
desserts. J. Food Prot. 42(3), 204–207.
DIAZ, F., SANTOS, E.M., FILARDO, S., VILLAGÓMEZ, R. and SCHEIN-
VAR, L. 2006. Colorant extraction from a red prickly pear (Opuntia
lasiacantha) for food application. Electron. J. Environ. Agric. Food
Chem. 5, 1330–1337.
DICKINSON, E. 2003. Hydrocolloids at interfaces and the influence on the
properties of dispersed systems. Food Hydrocolloids 17, 25–39.
DUANGMAL, K., SAICHEUA, B. and SUEEPRASAN, S. 2008. Colour
evaluation of freeze-dried roselle extract as a natural food colorant in a
model system of a drink. LWT - Food Sci. Technol. 41(8), 1437–1445.
ERSUS, S. and YURDAGEL, U. 2007. Microencapsulation of anthocyanin
pigments of black carrot (Daucuscarota L.) by spray drier. J. Food Eng.
80, 805–812.
FANG, Z., ZHANG, M., SUN, Y. and SUN, J. 2006. How to improve bayberry
(Myrica rubra Sieb. et Zucc.) juice color quality: Effect of juice process-
ing on bayberry anthocyanins and polyphenolics. J. Agric. Food Chem.
54, 99–106.
FERNANDO REYES, L. and CISNEROS-ZEVALLOS, L. 2007. Degradation
kinetics and colour of anthocyanins in aqueous extracts of purple- and
542 Z. IDHAM, I.I. MUHAMAD and M.R. SARMIDI

red-flesh potatotes (Solanum tuberosum L.). Food Chem. 100, 885–

894.
GIBBS, B., KERMASHA, S., ALLI, I. and MULLIGAN, C.N. 1999. Encap-
sulation in the food industry: A review. Int. J. Food Sci. Nutr. 50, 213–
224.
GRADINARU, G., BILIADERIS, C.G., KALLITHRAKA, S., KEFALAS, P.
and GARCIA-VIGUERA, C. 2003. Thermal stability of Hibiscus Sab-
dariffa L anthocyanins in solution and in solid state: effects of copigmen-
tation and glass transition. Food Chem. 83, 423–436.
KAMMERER, D., KLJUSURIC, J.G., CARLE, R. and SCHIEBER, A. 2005.
Recovery of anthocyanins from grape pomace extracts (Vitis vinifera L.
cv. Cabernet Mitos) using a polymeric adsorber resin. Eur. Food Res.
Technol. 220, 431–437.
KIRCA, A. and CEMEROGLU, B. 2003. Degradation kinetics of anthocya-
nins in blood orange juice and concentrate. Food Chem. 81, 583–587.
KIRCA, A., OZKAN, M. and CEMEROGLU, B. 2007. Effects of tempera-
ture, solid content and pH on the stability of black carrot anthocyanins.
Food Chem. 101, 212–218.
LIANFU, Z., MOU, D. and DU, Y. 2007. Procyanidins: Extraction and
microencapsulation. J. Sci. Food Agric. 87, 2192–2197.
OCHOA, M.R., KESSELER, A.G., VULLIOUD, M.B. and LOZANO, J.E.
1999. Physical and chemical characteristics of raspberry pulp: Storage
effect on composition and color. Lebensm. Wiss. Technol. 32, 149–153.
PUA, C.K., SHEIKH ABD HAMID, N., TAN, C.P., MIRHOSSEINI, H.,
ABD. RAHMAN, R. and RUSUL, G. 2008. Storage stability of jackfruit
(Artocarpus heterophyllus) powder packaged in aluminium laminated
polyethylene and metalized co-extruded biaxially oriented polypropylene
during storage. J. Food Eng. 89, 419–428.
SELIM, K.A., KHALIL, K., ABDEL-BARY, M. and ABDEL-AZEIM, N.
2008. Extraction, encapsulation and utilization of red pigments from
roselle (Hibiscus sabdariffa L.) as natural food colourants. 5th Alex.
Conference of food & dairy science and technology. Alex. Egypt.
SHAHIDI, F. and NACZK, M. 2004. Phenolics in Food and Nutraceuticals,
pp. 403–442, CRC Press, Boca Raton, FL.
WANG, W.D. and XU, S.Y. 2007. Degradation kinetics of anthocyanins in
blackberry juice and concentrate. J. Food Eng. 82, 271–275.

View publication stats