Separation and Purication Technology 71 (2010) 144151

Contents lists available at ScienceDirect

Separation and Purication Technology

journal homepage: www.elsevier.com/locate/seppur

Forward osmosis for the concentration of anthocyanin from Garcinia indica

Choisy
Chetan A. Nayak, Navin K. Rastogi
Department of Food Engineering, Central Food Technological Research Institute, Council of Scientic and Industrial Research, KRS Road, Mysore 570 020, Karnataka, India

a r t i c l e

i n f o

Article history:
Received 12 February 2009
Received in revised form
13 September 2009
Accepted 11 November 2009
Keywords:
Anthocyanin
Forward osmosis
Garcinia indica
Natural colorant

a b s t r a c t
The concentration of anthocyanin extract from Garcinia indica Choisy (popularly known as kokum) was
explored by forward osmosis process and compared with thermally concentrated sample. Mechanism of
water transport from feed to osmotic agent side during forward osmosis in a situation when feed contains
high or low molecular weight compounds was elucidated. The effects of membrane orientation, osmotic
agent concentration, ow rates of feed and osmotic agent and temperature on transmembrane ux during
the concentration of anthocyanin extract were studied. In a large-scale experiment, the anthocyanin
extract was concentrated from 49 mg/l to 2.69 g/l (54-fold increase). Non-enzymatic browning index and
degradation constant for thermally concentrated sample were found to be approximately two (0.780.35)
and eight (63.0 103 to 8.0 103 day1 ) times, respectively, as compared to the concentrate produced
by forward osmosis. Ratio of HCA lactone to HCA was also found to be higher for the sample produced by
thermal concentration (2.84:1) as compared to concentrate produced by forward osmosis (1.50:1). These
results clearly indicate that the concentration of anthocyanin extract using forward osmosis has several
advantages over the thermal concentration in terms of higher stability, lower browning index and less
conversion of HCA to its lactone form.
2009 Elsevier B.V. All rights reserved.

1. Introduction
Anthocyanins are abundant and most signicant avonoids
group of compounds found in nature. They are water-soluble and
vacuolar pigments present in the epidermal cells of the owers of
plants [1,2]. The term anthocyanin is derived from the two Greek
words anthos and kyanos, which means ower and blue, respectively. Anthocyanins comprise diverse groups of intensely colored
pigments. These are responsible for appealing spectacular orange,
red, purple and blue colors of many fruits, vegetables, owers,
leaves, roots and other plant organs. The solubility of anthocyanins
in water makes them the best candidates for the incorporation into
aqueous food systems [13].
Garcinia species are distributed widely throughout the tropical Asian and African countries and have a tremendous potential
as spice and medicinal plant. Garcinia indica Choisy is popularly
known as kokum. It belongs to the family Guttiferae and it is a
slow growing slender tree with drooping branches growing to a
height of 1618 m [46]. More than 200 listed species of Garcinia
are available in nature [4]. The ripe kokum fruit is of dark purple color or red with yellow tinge. It contains three to eight large

Corresponding author. Tel.: +91 821 2513 910; fax: +91 821 2517 233.
E-mail addresses: nkrastogi@cftri.res.in, nkrastogi@yahoo.com (N.K. Rastogi).
1383-5866/$ see front matter 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.seppur.2009.11.013

seeds embedded in a red acid pulp. The fruit has a pleasant avor and a sour taste. It is traditionally used as an acidulant in
many Indian dishes. It is reported for treatments of dysentery,
tumors, heart complaints, stomach acidity and liver disorders [4].
The major organic acid component that imparts the savory taste to
the fruit is hydroxyl citric acid (HCA). It is an important ingredient
in many fat-reducing supplements and claimed as an anti-obesity
ingredient [48]. The chemical and spectral investigations have
revealed that the rinds of the fruit contain two water-soluble anthocyanin pigments, which were identied as cyanidin-3-glucoside
and cyanidin-3-sambubioside [9,10].
An efcient extraction procedure should maximize anthocyanin
recovery with minimal amount of adjuncts and degradation or
alteration of its natural state. Several efcient extraction procedures were employed for the extraction of anthocyanin, but the
extracts were not safe for human consumption due to toxic effects
from the residual solvents used in its extraction. Solvents used
in extraction procedure are methanol, acetone, etc., and almost
all these solvents are not preferred in food due to their harmful
effects on health [2]. Health concern has led to numerous research
efforts for the development of new procedure for extraction and
concentration of natural color [4,7]. Conventional extraction and
concentration methods of anthocyanin from other sources were
reported to cause degradation of anthocyanin, besides being energy
intensive [1,2,6]. Hence, there exists a strong need for the develop-

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

ment of energy efcient process, which can reduce the degradation

of anthocyanin.
Major constituent in most of the natural color extracts is water,
which contributes to growth of the microorganisms. Shelf-life of
these extracts can be increased by removal of water. Hence, it is
desirable to concentrate these extracts to improve shelf-life, stability as well as to reduce storage and transportation costs [7,8,11,12].
Concentration of natural color extracts by conventional method
such as evaporation results in loss of hue and chroma resulting
in low quality product [13]. Membrane processes such as microltration, ultraltration and reverse osmosis were being employed
for clarication and concentration of natural color extracts [1420].
There are many limitations of these membrane processes such as
requirement of high pressure, limit to maximum attainable concentration, concentration polarization and membrane fouling [13].
New athermal membrane process namely forward osmosis can
be used for the concentration of natural colorant. It employs a
semi-permeable dense hydrophilic membrane, which separates
two aqueous solutions (feed and osmotic agent solution) having different osmotic pressures. The difference in osmotic pressure acts as
a driving force [12]. The transfer of water occurs from the low concentration (feed side) to high concentration (osmotic agent side)
till the osmotic pressures of both the sides become equal [13,14].
It works at ambient pressure and is capable of concentrating liquid foods without product deterioration [14]. No use of solvent for
extraction, low energy consumption, higher retention of thermoliable components and attainment of higher concentration are the
several advantages over conventional process [1517]. Dova et al.
[20,21] studied the inuence of process parameters on performance
of forward osmosis and modeled the process parameters.
The main objectives of present work are to elucidate the mechanism of water transport from feed to osmotic agent side during
forward osmosis in a situation when feed contains high or low
molecular weight compounds as well as to study the effect of various process parameters such as osmotic agent concentration, ow
rate and feed temperature on transmembrane ux during forward
osmosis process for the concentration of anthocyanin pigment from
G. indica Choisy. Attempts were made to compare the physicochemical characteristics of the extracts before and after the forward
osmosis.

2. Theoretical considerations
Forward osmosis membrane process employs a semi-permeable
dense hydrophilic membrane, which separates the feed as well as
the osmotic agent solutions (Fig. 1). Osmotic pressure difference
between the feed as well as osmotic agent solutions acts as a driving force for transport of water. The asymmetric membrane used in
forward osmosis consisted of two layers, one is the loosely bound
support layer and other is the dense active membrane layer. The
membrane can be placed between the feed and the osmotic agent
solutions in two different ways such as feed towards the support
layer (normal mode) and feed towards active layer (reverse mode),
which are referred, in the present work, as modes I and II, respectively [22].
In case of mode I, when the feed is pure water, the water is diffused into the support layer and transferred to the osmotic agent
side through the active layer of membrane. Since the feed is water,
it will not result in any external or internal polarization. An insignificant external polarization may take place in the boundary layer on
osmotic agent side. In this situation, if the feed (water) is replaced
with the solution of low molecular weight compounds, it will lead
to signicant internal polarization (concentrative) and negligible
external polarization on feed as well as osmotic agent side (Fig. 1a).
The internal concentration polarization occurs within the porous

145

support layer and it cannot be mitigated by hydrodynamics such

as turbulence, and hence drastically reduces the osmotic driving
force [23]. It has been pointed out by Cath et al. [24] that the extent
of external polarization is much less than the internal polarization
during forward osmosis.
When the feed is replaced with the solution containing high
molecular weight compounds, it will result in the buildup of the
retained concentration of high molecular weight compounds on the
support layer resulting in signicant external concentration polarization on the feed side. Mi and Elimelech [25] have indicated that
these high molecular weight compounds may be deposited within
the porous structure of the membrane leading to cake layer formation due to lack of shear force as well as hindered back diffusion in
the porous structure. Further, it will also lead to signicant internal polarization (concentrative) within the support layer. But, the
external polarization towards osmotic agent side will be negligible
as indicated in the earlier cases (Fig. 1b). Due to this fact the effective driving force (b ) will be much less as compared to the same
situation in which solution of low molecular weight compounds or
water was taken as a feed (b < a ), which, in turn, drastically
reduce the transmembrane ux.
In case of mode II, when feed (solution containing high or low
molecular weight compounds) is kept towards active layer and
osmotic agent is towards support layer, the water from the feed is
diffused into the active layer, which, in turn, is diffused to the support layer and then to the bulk through the boundary layer. Since,
the solute used for osmotic agent is generally of low molecular
weight, it is also diffused into the support layer to the interior surface of the active layer before ux can occur. As water ux crosses
the active layer into the support layer, the solute is diluted due
to convection. The solute diffuses back to the interior surface. A
steady-state is quickly reached, but the concentration at the interior surface of the active layer is far lower than in the bulk solution
[22]. The combined effect of diffusion of water through the active
layer and osmotic agent into support layer will result in setting up
of an internal concentration polarization (dilutive, Fig. 1c). In this
case also, the extent of external concentration polarization towards
support layer (feed) as well as active layer (osmotic agent) will be
much less as compared to internal polarization and hence can be
neglected. Gray et al. [22] have demonstrated that when feed is
kept towards active layer (mode II), the external polarization on
feed side can be taken as negligible as compared to internal polarization as well as the molecular weight of the compounds in feed
do not have any effect on transmembrane ux. The effective driving
force, in this case, will be less than situation as indicated in mode
I (c < a ), when low molecular weight compounds or water is
taken as a feed, hence in such situation, mode I is most desirable.
However, when the solution of high molecular weight compounds
is taken as a feed, mode II is expected to give higher driving force
(c > b ) and which, in turn, leads to higher transmembrane
ux [24,26].
The resistivity of the membrane (support and active layer) during forward osmosis process was calculated as per the following
equation presented for dilutive internal concentration polarization,
respectively [22,23,27,28]:

K=

1
Jw


ln

B + AOA
B + Jw + AFeed


(1)

where Jw is the transmembrane ux during forward osmosis, K

is the resistivity of membrane (support and active layer) (s/m),
OA and Feed are the osmotic pressure of osmotic agent and
feed, respectively. The constant A (0.027 m/atm day) and B
(0.011 m/day) refer to the water and solute permeability coefcients of the active layer of the membrane, respectively [22,28].

146

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

Fig. 1. Mechanism of forward osmosis indicating water transport from the solution of low osmotic pressure to the solution of high osmotic pressure. (a) Feed solution
containing low molecular weight compounds; (b) feed solution containing high molecular weight compounds; (c) feed solution containing low/high molecular weight
compounds. a , b and c are the corresponding effective driving force, respectively. Feed and OA are the osmotic pressures of feed and osmotic agent solution,
respectively.

3. Materials and methods

3.1. Materials
Fresh G. indica Choisy fruits (popularly known as kokum) were
procured from the orchards near Mangalore, India. Sodium chloride, sodium acetate and potassium chloride were procured from
Ranbaxy Ltd., India. All the chemicals used were of analytical grade.

respectively. The extract was stored in a cold room at 45 C and

drawn as and when required for the experiment within 7 days.
In order to compare the forward osmosis process with thermal concentration, the crude extract was concentrated using ash
evaporator (Rotovap Model 410, M/s Bucchi, Switzerland). The temperature and vacuum were maintained at 60 C and 675 mm of Hg,
respectively.
3.3. Experimental set up

3.2. Anthocyanin extraction

Two kilogram of the fruit was taken for a batch. The fruits were
washed, cut into four equal pieces (rinds) along the major axis, then
ground after the removal of seeds. The pulp, thus obtained, was
mixed with 1:2 ratio of acidied water (0.1% hydrochloric acid). The
mixture was subjected to hydraulic press (M/s B. Sem & Berry, New
Delhi, India) for extraction. The extract was ltered using muslin
cloth. Approximately, 4.8 l of extract of was obtained from 2.0 kg
of the starting material. Anthocyanin concentration and total soluble solids in the extract were found to be 49.63 mg/l and 2 Brix,

Experiments were performed using a at membrane module

having a membrane area of 1.14 102 m2 , shown in Fig. 2. The
module consisted of reverse osmosis membrane, which is placed
over a polyester mesh, supported in between Viton gasket and
two stainless steel frames. Feed solution (anthocyanin extract)
and osmotic agent solution were circulated on either side of the
membrane in co-current mode using peristaltic pumps (Model 72315-230, Barnant Company, IL, USA). The ratio of feed solution to
osmotic agent solution was maintained at 1:10 for all the experiments. The transmembrane ux was calculated by measuring the
increase in volume of osmotic agent every hour. Average ux values
were calculated based on average of ux values of rst 5 h of the
forward osmosis. All the experiments, unless otherwise indicated,
were carried out at the temperature of 25 2 C.
3.4. Membranes
The reverse osmosis asymmetric membrane developed by
Osmotek, Inc., Corvallis, OR, USA was employed in the present
study. The membrane consisted of a very thin semi-permeable nonporous active skin layer of cellulose triacetate embedded in a nylon
mesh (a porous support layer) to provide strength. The membrane
was highly hydrophilic in nature. The thickness of the membrane
as determined by scanning electron microscope was found to vary
between 50 and 100 m [22,23,29].
3.5. Osmotic agent solution

Fig. 2. Flat membrane module for forward osmosis process: (1) forward osmosis at
membrane module, (2) feed reservoir, (3) osmotic agent reservoir and (4) peristaltic
pump.

Osmotic agent solutions were prepared by dissolving sodium

chloride in distilled water in various proportions (1.06.0 M or
626%, w/w). These solutions were kept overnight at room temper-

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

147

ature before use to ensure complete dissolution of sodium chloride.

Osmotic pressures of sodium chloride solutions were calculated as
per the procedure described by Toledo [30].

carried out for 15 days room temperature and anthocyanin content was measured each day. The degradation constant (KD ) was
determined by the slope of ln(C0 /Ct ) versus t.

3.6. Physicochemical characteristics

4. Results and discussion

3.6.1. Anthocyanin content and pH measurement

The anthocyanin content was estimated using spectral analysis (UV-VIS spectrophotometer, Model UV-160A, M/s Shimadzu,
Japan) and calculated as per the following equation [31]:

Forward osmosis experiments were carried out using anthocyanin extract and sodium chloride solution as a feed and an
osmotic agent solution, respectively. The effects of concentration
osmotic agent solution as well as feed and osmotic agent ow rates
were studied on the transmembrane ux. The effect of feed temperature on the transmembrane ux was also evaluated.

anthocyanin content (mg/l) =

AMW DF 103
L

(2)

where A is the total absorbance [(Amax A700 )at pH 1.0

(Amax A700 )at pH 4.5 ], MW is the molecular weight of antho-

cyanin (449 g mol1 ), DF is the dilution factor, is the extension

coefcient (29,600 l cm1 mol1 ) and L is the path length (1 cm).
pH meter (APX 175, Control Dynamics, India) was used for the
measurement of pH of anthocyanin extracts. The concentrated
anthocyanin samples were diluted to original concentration with
distilled water before the color measurement and the values were
reported with multiplication with appropriate dilution factor. All
the experiments were carried out in triplicates and average values
were reported.
3.6.2. Color analysis
Color characteristics (L*, a* and b*) of concentrated anthocyanin were measured using the Hunter colorimeter (Model D25-9,
Hunterlab, Virginia, USA). The anthocyanin samples were placed
in a 1.0 cm path length optical glass cell and CIE L*, a*, b* values
were noted in total transmission mode, using illuminant C and 2
observer angle. Hue angle (tan1 (b*/a*)) and chroma (a*2 + b*2 )1/2
were also determined as per the procedure reported by DelgadoVargas et al. [2] and Nayak et al. [32].
3.6.3. Estimation of soluble solid content
Total soluble solid of anthocyanin extract was measured using
Ermas Handheld refractometer at 25 2 C.
3.6.4. HPLC analysis of hydroxycitric acid
The sample was subjected to activated charcoal for decolorization. The decolorized extract was treated with methanol to remove
pectinaceous material and then centrifuged. The supernatant was
ltered through membrane (0.45 m, type HA, M/s Millipore, USA)
and analyzed by high performance liquid chromatography (HPLC),
which was equipped with a photodiode array detector, C-18 ODS
column (250 mm 4.6 mm, pore size 5.0 m, M/s SGE, Japan). Isocratic elution of 8 mM sulphuric acid was used as a mobile phase
(ow rate of 1.0 ml/min) and analyzed at 210 nm [33]. The ratio of
the area under the peak corresponding to HCA lactone to that of
HCA was reported as the ratio of concentration of HCA lactone to
that of HCA.

4.1. Effect of osmotic agent concentration on the transmembrane

ux
The effect of osmotic agent (sodium chloride) concentration on
transmembrane ux in case of water or anthocyanin extract as feed
was compared in modes I and II (Fig. 3). The feed and osmotic agent
ow rates during the experiment were maintained at 100 ml/min.
The transmembrane ux in case of pure water as a feed in mode I
was found to increase from 12.4 to 17.5 l/m2 h with an increase in
osmotic agent concentration from 1.0 to 6.0 M (p 0.05, Fig. 3). The
increase in ux during forward osmosis is attributed to an increase
in osmotic pressure difference across the membrane due to increase
in the concentration of osmotic agent solution, which resulted in an
increased driving force for water transport through the membrane.
Moreover, the transmembrane ux values in mode I were higher
in comparison to mode II (9.313.8 l/m2 h) for the corresponding
concentrations of osmotic agent solution. This may be attributed to
the fact that during mode I, there will be negligible concentration
polarization. But, it will be signicant in mode II due to the diffusion
of osmotic solute into support layer resulting in the setting up of
concentration gradient [22] (Fig. 1a and b).
In case of anthocyanin extract as a feed, transmembrane ux in
modes I and II was found to increase from 2.7 to 4.4 l/m2 h and 7.5
to 12.3 l/m2 h, respectively with an increase in concentration from
1 to 6 M (p 0.05, Fig. 3). The ux values in mode II were higher
as compared to mode I for all the corresponding concentrations. It
may please be noted that lower ux values in mode I were due to
exertion of highly signicant external concentration polarization
because of presence of high molecular weight compounds in the
feed, which was non-existent in mode II resulting in higher driving
force as compared to mode I (c > b , Fig. 1).
In many of the reports available in literature, no attention has
been paid to mention the orientation of membrane (mode I or

3.7. Non-enzymatic browning and degradation constant

Non-enzymatic browning was estimated using spectrophotometer at 420 nm. The estimation was carried out by diluting
the concentrated sample to the initial total soluble solids of fresh
extract i.e. 2 Brix [34]. The degradation constant (KD ) for the
anthocyanin content in the extract was determined considering
rst-order degradation kinetics as per the following equation [32]:
ln

C 
0

Ct

= KD t

(3)

where C0 is the initial anthocyanin content and Ct is the anthocyanin content at a specied time t. The degradation studies were

Fig. 3. Effect on the osmotic agent concentration on transmembrane ux. Feed

and osmotic agent ow rates were maintained at 100 ml/min. Modes I and II were
referred to the membrane orientation in which feed was towards the support layer
and feed was towards active layer, respectively.

148

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

mode II) in case of concentration during forward osmosis. However,

studies dealing with the feed containing high molecular weight
compounds such as concentration of fruit juices [16,35,36], sucrose
solution [29] and treatment of waste water [37] indicated that the
transmembrane ux values were of same order as obtained in the
present work in mode II.
The membrane resistivity (K) of the membrane was calculated
using Eq. (1) which remained constant (12.07 day/m) irrespective of
concentration of osmotic agent. The hydrodynamic properties such
as density, viscosity and diffusivity are likely to be change with concentration. The density of the osmotic solutions varies slightly with
concentration whereas the diffusivity practically remains constant
[38].
4.2. Effect of feed and osmotic agent ow rate on the
transmembrane ux
The effect of feed ow rate on transmembrane ux for anthocyanin extract in modes I and II is shown in Fig. 4a. The osmotic
agent ow rate and concentration during the experiments were
maintained at 100 ml/min and 6.0 M, respectively. With an increase
in feed ow rate from 50 to 125 ml/min in mode I, the transmembrane ux and anthocyanin concentration was found to increase
from 4.2 to 4.7 l/m2 h and 50.9 to 57.8 mg/l (p 0.05), respectively.
Whereas, in mode II, the transmembrane ux and anthocyanin concentration was found to increase from 11.4 to 12.5 l/m2 h and 54.1
to 67.1 mg/l (p 0.05), respectively.
The effect of osmotic agent ow rate on transmembrane ux and
anthocyanin concentration extract in modes I and II is presented
in Fig. 4b. The feed ow rate and concentration of osmotic agent

solution during the experiment were maintained at 100 ml/min

and 6.0 M, respectively. With an increase in osmotic agent ow
rate from 50 to 125 ml/min, the transmembrane ux was found
to increase in modes I and II from 3.8 to 4.6 l/m2 h and 10.0 to
12.7 l/m2 h (p 0.05), respectively. At the same time, the anthocyanin concentration increased from 50.9 to 57.76 mg/l and 54.42
to 65.8 mg/l (p 0.05), respectively.
4.3. Effect of feed temperature on the transmembrane ux
The variation of transmembrane ux and anthocyanin content
in modes I and II was studied with an increase in feed temperature from 25 to 40 C (Fig. 5). During the experiments, feed as well
as osmotic agent solutions were circulated at a constant ow rate
of 125 ml/min. The concentration of osmotic agent solution during the experiment was maintained at 6.0 M. The transmembrane
ux was found to increase in mode I (4.75.1 l/m2 h) and mode
II (12.320.4 l/m2 h) with increase in feed temperature from 25 to
40 C. The increase in transmembrane ux with temperature can be
explained by WilkeChang equation [39], according to which, the
diffusion coefcient is proportional to the absolute temperature
divided by the viscosity of the solvent. The increase in temperature
reduces the viscosity of solution and increases the diffusion coefcients, which results in increase in transmembrane ux [35,40]. At
the same time, the anthocyanin content was also found to increase
from 58.0 to 92.0 mg/l and from 65.0 to 168.0 mg/l in modes I and
II, respectively with an increase in temperature from 25 to 40 C.
Wrolstad et al. [41] also reported that the increase in temperature
resulted in an increase in transmembrane ux in the case of the
concentration of red raspberry juice by forward osmosis process.
4.4. Concentration of anthocyanin extract using optimized
parameters
Based on the optimized conditions, anthocyanin extract was
subjected to a large-scale experiment in mode II up to 18 h. Feed
and osmotic agent ow rates were maintained at 125 ml/min,
but the osmotic agent concentration was kept at 6.0 M (Fig. 6).
During forward osmosis, around 2 Brix crude extract was concentrated to 52 Brix anthocyanin concentrate (Fig. 6a). Initial
volume of 2000 ml was reduced to 37 ml. At the same time,
anthocyanin content was concentrated from 49.63 mg/l initial
concentration to 2.69 g/l nal concentration (approximately 54fold, Fig. 6b). Rodriguez-Saona et al. [18] and Wrolstad et al.
[41] have reported a maximum anthocyanin concentration of 7.5fold for red radish and 4.5-fold for red raspberry respectively.

Fig. 4. Effect of (a) feed, and (b) osmotic agent ow rate on transmembrane ux and
anthocyanin concentration. Modes I and II were referred to the membrane orientation in which feed was towards the support layer and feed was towards active layer,
respectively. Osmotic agent concentration was maintained at 6.0 M.

Fig. 5. Effect of feed temperature on transmembrane ux and anthocyanin concentration. Feed and osmotic agent ow rates were maintained at 125 ml/min. Modes
I and II were referred to the membrane orientation in which feed was towards
the support layer and feed was towards active layer, respectively. Osmotic agent
concentration was maintained at 6.0 M.

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

149

Table 1
Physicochemical characteristics of anthocyanin extracts.
Characteristic

Crude extract

2.0 0.2
49.68 1.0
3.5 0.1

Brix
Anthocyanin (mg/l)
pH
Color values
L*
a*
b*

Hue angle (tan1 (b*/a*))

Color purity (a*2 + b*2 )1/2
Density (kg/m3 )
Viscosity (mPa s)

0.62 0.01
0.27 0.05
0.40 0.02
55.98
0.48
1025
1.109

0.10
0.05
10
0.05

Forward osmosis
concentrate
52.0 0.5
2692 2.0
2.8 0.1
5.33 0.10
2.54 0.20
0.14 0.02
3.05
2.54
1110
1.779

0.05
0.05
10
0.10

Reconstituted
extract
2.0 0.2
50.12 1.0
3.2 0.1
0.68 0.01
0.30 0.05
0.35 0.01
49.40
0.47
1030
1.104

0.15
0.05
10
0.05

4.6. Comparison of concentrate produced by forward osmosis

with thermally concentrated sample
The concentrate produced by forward osmosis was compared
with thermally concentrated sample in terms of non-enzymatic

Fig. 6. Variation of (a) total soluble solids, (b) anthocyanin concentration and (c)
transmembrane ux during concentration of anthocyanin from kokum extract. Feed
and osmotic agent ow rates were maintained at 125 ml/min. Feed temperature
was maintained at 25 C. Osmotic agent concentration was maintained at 6.0 M.
Membrane orientation was in way such that feed was towards active layer (mode
II).

The transmembrane ux was found to decrease with time due

to increase in feed concentration (Fig. 6c), which in turn was
responsible for the reduced driving force across the membrane.
Rodriguez-Saona et al. [18] also observed that the transmembrane
ux decreased with an increase in feed concentration during the
osmotic removal of water in the case of anthocyanin from red
radish.
4.5. Comparison of physicochemical properties
Physicochemical properties of before (crude extract) and after
(anthocyanin extract) forward osmosis are presented in Table 1.
The reconstitution of the concentrate was done by addition of water
such that the resultant concentration was brought back to the concentration of initial crude extract. The properties of reconstituted
extract were found to be comparable to that of the fresh extract.
Hunter color values, hue angle and the color purity of the reconstituted extract was also found to be very close to that of the fresh
anthocyanin extract (Table 1).

Fig. 7. HPLC Chromatograms for HCA lactone, HCA and citric acid in (a) crude anthocyanin extract, (b) forward osmosis concentrate and (c) thermal vacuum evaporated
concentrate.

150

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151

Table 2
Comparison among crude, concentrate produced by forward osmosis and thermally
concentrated anthocyanin extract.
Attributes

Crude extract

Forward
osmosis
concentrate

Thermal
concentrate

Non-enzymatic
browning (NEB)
Ratio of HCA lactone to
HCA
Degradation constant
KD (103 day1 )

0.25 0.01

0.35 0.02

0.78 0.04

1.26:1

1.50:1

2.84:1

3.0 0.5

8.0 1.0

63.0 5.0

browning index, conversion of HCA to its lactone form and stability. Non-enzymatic browning index for thermally concentrated
sample was found to be 0.78, whereas it was 0.35 for the concentrate produced by forward osmosis (Table 2). The non-enzymatic
browning index of thermally concentrated sample was higher due
to excessive exposure of sugar present in anthocyanin extract to
processing temperature during the thermal treatment. Moreover,
the ratio of HCA lactone to HCA was found be higher after thermal
concentration (2.84:1) of anthocyanin extract as compared to forward osmosis concentrated sample (1.50:1) as indicated in HPLC
chromatogram (Fig. 7, Table 2). The degradation constant (KD , estimated as per Eq. (3)) for the concentrate produced by forward
osmosis (8.0 103 day1 ) was almost eight times higher than
the thermally concentrated sample (63.0 103 day1 , Table 2).
These results clearly indicate that the concentration of anthocyanin
extract using forward osmosis has advantages over the thermally
concentration in terms of lower browning index, less conversion of
HCA to HCA lactone and higher stability.
5. Conclusion
The concentration of anthocyanin extract from G. indica Choisy
was studied by forward osmosis process. Mechanism of water
transport from feed to osmotic agent side during forward osmosis in a situation when feed contains high or low molecular weight
compounds was elucidated. The membrane orientation during forward osmosis membrane was found to have signicant effect on
ux. The mode II (feed towards active layer) was found to result
in higher ux values as compared to mode I (feed towards the
support layer) due to signicant external concentration polarization in mode I. The effect of various process parameters such as
osmotic agent concentration, feed and osmotic agent ow rate and
temperature on transmembrane ux during the concentration of
anthocyanin extract by forward osmosis process were studied. The
transmembrane ux was found to increase with an increase in temperature of the feed. The anthocyanin extract was concentrated
approximately 54 times (from 49.63 mg/l to 2.69 g/l) in a large-scale
experiment. The concentration of anthocyanin extract using forward osmosis has advantages over the thermally concentration in
terms of higher stability, lower browning index and less conversion
of HCA to lactone form.
Acknowledgements
Authors thank Dr. V. Prakash, Director, CFTRI, Mysore for
his encouragement and keen interest in Downstream Processing.
Thanks are also due to Dr. KSMS Raghavarao, Head, Department of
Food Engineering, CFTRI for his support. The author Chetan A Nayak
expresses his gratitude and sincere thanks to the Council of Scientic and Industrial Research (CSIR), New Delhi for providing Senior
Research Fellowship.

References
[1] M.M. Gusti, R.E. Wrolstad, Radish anthocyanin extract as a natural red colorant
from maraschino cherries, J. Food Sci. 61 (1996) 688694.
[2] F. Delgado-Vargas, A.R. Jimnez, O. Paredes-Lopez, Natural pigments:
cartenoids, anthocyanins and betalainscharacteristics, biosynthesis processing and stability, Crit. Rev. Food Sci. Nutr. 40 (2000) 173289.
[3] M. Bleve, L. Ciurlia, E. Erroi, G. Lionetto, L. Longo, L. Schettino, G. Vasapollo, An
innovative method for the purication of anthocyanins from grape skin extracts
by using liquid and sub-critical carbon dioxide, Sep. Purif. Technol. 64 (2008)
192197.
[4] D.J. Bhat, N. Kamat, A. Shirodkar, Compendium and Proceedings of the 2nd
National Seminar on Kokum (Garcinia indica Choisy), Goa University, March
45, 2005.
[5] N. Krishnamurthy, Y.S. Lewis, B. Ravindranath, Chemical constituents of kokum
fruit rind, J. Food Sci. Technol. 19 (1982) 97100.
[6] N. Krishnamurthy, Chemical and technological studies on coloring matters
from natural sources for use in foods, Ph.D. Thesis, University of Mysore,
Mysore, 1984.
[7] S. Bhaskaran, S. Mehta, Stabilized anthocyanin extract from Garcinia indica, US
Patent No. 0230983A1 (2006).
[8] G. Patil, K.S.M.S. Raghavarao, Integrated membrane process for the concentration of anthocyanin, J. Food Eng. 78 (2007) 12331239.
[9] C.A. Nayak, N.K. Rastogi, K.S.M.S. Raghavarao, Bioactive constituents present in
Garcinia indica Choisy and its potential food applications: a review, Intl. J. Food
Prop., in press.
[10] C.A. Nayak, P. Srinivas, N.K. Rastogi, Characterization of anthocyanin from
Garcinia indica Choisy, Food Chem. 118 (2010) 719724.
[11] B.S. Jena, G.K. Jayaprakasha, R.P. Singh, K.K. Sakariah, Chemistry and biochemistry of hydroxycitric acid from Garcinia, J. Agric. Food Chem. 50 (2002) 1022.
[12] E.G. Beaudry, K.A. Lampi, Membrane technology for direct osmosis concentration of fruit juices, Food Technol. 44 (1990) 121.
[13] B.R. Babu, N.K. Rastogi, K.S.M.S. Raghavarao, Effect of process parameter on
transmembrane ux during direct osmosis, J. Membr. Sci. 280 (2006) 185194.
[14] K.S.M.S. Raghavarao, N. Nagaraj, G. Patil, B.R. Babu, K. Niranjan, Athermal membrane process for the concentration of liquid foods and natural colours, in: D.W.
Sun (Ed.), Emerging Technologies in Food Processing, Academic Press/Elsevier,
London, 2005, pp. 251278.
[15] E.L. Cussler, Diffusion: Mass Transfer in Fluid Systems, third ed., Cambridge
University Press, Cambridge, 1984.
[16] M. Wong, R.J. Winger, Direct osmotic concentration for concentrating liquid
foods, Food Aust. 51 (1999) 200205.
[17] B. Girard, L.R. Fukomoto, Membrane processing of fruit juices, and beverages:
a review, Crit. Rev. Food Sci. Nutr. 40 (2000) 91101.
[18] L.E. Rodriguez-Sanoa, M.M. Giusti, W.D. Robert, R.E. Wrolstad, Development
and process optimization of red radish concentration extract as potential natural red colorant, J. Food Process. Preserv. 25 (2001) 165182.
[19] H. Nawaz, J. Shi, G.S. Mittal, Y. Kakudac, Extraction of polyphenols from
grape seeds and concentration by ultraltration, Sep. Purif. Technol. 48 (2006)
176181.
[20] B.G. ukasik, S. Koter, J. Kurzawa, Concentration of anthocyanins by the membrane ltration, Sep. Purif. Technol. 57 (2007) 418424.
[21] M.I. Dova, K.B. Petrotos, H.N. Lazarides, On the direct osmotic concentration of
liquid foods. Part II. Development of a generalized model, J. Food Eng. 78 (2007)
431437.
[22] G.T. Gray, J.R. McCutcheon, M. Elimelech, Internal concentration polarization in
forward osmosis: role of membrane orientation, Desalination 197 (2006) 18.
[23] J.R. McCutcheon, M. Elimelech, Modeling water ux in forward osmosis: implications for improved membrane design, AIChE J. 53 (7) (2007) 17361744.
[24] T.Y. Cath, A.E. Childress, M. Elimelech, Forward osmosis: principles, applications, and recent developments, J. Membr. Sci. 281 (2006) 7087.
[25] B. Mi, M. Elimelech, Chemical and physical aspects of organic fouling of forward
osmosis membranes, J. Membr. Sci. 320 (12) (2008) 292302.
[26] Y.N. How, W. Tang, W.S. Wong, Performance of forward (direct) osmosis process: membrane structure and transport phenomenon, Environ. Sci. Technol.
40 (2006) 24082413.
[27] C.H. Tan, H.Y. Ng, Modied models to predict ux behavior in forward osmosis
in consideration of external and internal concentration polarizations, J. Membr.
Sci. 324 (2008) 209219.
[28] S. Leob, L. Titelman, E. Korngold, J. Freiman, Effect of porous support fabric on
osmosis through a LoebSourirajan type asymmetric membrane, J. Membr. Sci.
129 (1997) 243249.
[29] E.M. Garcia-Castello, J.R. McCutcheon, M. Elimelech, Performance evaluation of
sucrose concentration using forward osmosis, J. Membr. Sci. 338 (2009) 6166.
[30] R.T. Toledo, Fundamentals of Food Process Engineering, third ed., Van Nostrand
Reinhold, New York, 1991.
[31] R.E. Wrolstad, Hand Book of Food Analytical ChemistryPigments, Colorants,
Flavor, Texture, and Bioactive Components, Wiley Interscience Publications,
New Jersey, 2005.
[32] C.A. Nayak, S. Chethana, N.K. Rastogi, K.S.M.S. Raghavarao, Enhanced mass
transfer during solidliquid extraction of gamma irradiated red beetroot,
Radiat. Phys. Chem. 75 (2006) 173178.
[33] G.K. Jayaprakasha, K.K. Sakaraiah, Determination of organic acids in leaves and
rinds of Garcinia indica by LC, J. Pharm. Biomed. Anal. 28 (2002) 379385.
[34] A. Ibarz, S. Garza, J. Pagan, Nonenzymatic browning of selected fruit juices
affected by d-galacturonic acid, Int. J. Food Sci. Technol. 43 (2008) 908914.

C.A. Nayak, N.K. Rastogi / Separation and Purication Technology 71 (2010) 144151
[35] K.B. Petrotos, P. Quantick, H. Petropakis, A study of the direct osmotic concentration of tomato juice in tubular membrane-module conguration. I. The effect
of certain basic process parameters on the process performance, J. Membr. Sci.
150 (1998) 99110.
[36] M.I. Dova, K.B. Petrotos, H.N. Lazarides, On the direct osmotic concentration of
liquid foods. Part I. Impact of process parameters on process performance, J.
Food Eng. 78 (2007) 422430.
[37] R.W. Holloway, A.E. Childress, K.E. Dennett, T.Y. Cath, Forward osmosis for
concentration of anaerobic digester centrate, Water Res. 41 (2007) 4005
4014.

151

[38] B. Petrotos, H.N. Lazarides, Osmotic concentration of liquid foods, J. Food Eng.
49 (2001) 201206.
[39] R.E. Treybal, Mass Transfer Operations, third ed., McGraw-Hill Chemical Engineering Series, New Delhi, 1982.
[40] J.R. McCutcheon, M. Elimelech, Inuence of concentrative and dilutive internal
concentration polarization on ux behavior in forward osmosis, J. Membr. Sci.
284 (2006) 237247.
[41] R.E. Wrolstad, M.R. McDaniel, R.W. Durst, N. Micheals, K.A. Lampi, E.G. Beaudry,
Composition and sensory characterization of red raspberry juice concentration
by direct osmosis or evaporation, J. Food Sci. 58 (1993) 633641.