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# 2005 Institution of Chemical Engineers
Trans IChemE, Part C, September 2005
Food and Bioproducts Processing, 83(C3): 191 197

www.icheme.org/journals
doi: 10.1205/fbp.03403

AQUEOUS TWO-PHASE EXTRACTION FOR RECOVERY

OF PROTEINS FROM CHEESE WHEY
C. ANANDHARAMAKRISHNAN1, S. N. RAGHAVENDRA2, R. S. BARHATE2,
U. HANUMESH2 and K. S. M. S. RAGHAVARAO2

1

Department of Human Resource Development, Central Food Technological Research Institute, Mysore, India
2
Department of Food Engineering, Central Food Technological Research Institute, Mysore, India

queous two-phase extraction (ATPE), employing polymer polymer type and

polymer salt type systems, has been used for the recovery of proteins (devoid of
fat) from cheese whey. The application of ATPE resulted in differential partitioning
of fat and proteins, that is, selective partitioning of the proteins to the top (polymer rich) phase
while the fat to the bottom (salt rich) phase. This differential partitioning of proteins and fat
has been explained based on force balance analysis. Such a partitioning behaviour is observed
only in the polymer salt type phase system and not in polymer polymer type phase system.
Effect of process parameters such as phase composition (tie line length, TLL), pH, protein
concentration and phase volume ratio were studied. PEG 6000/potassium phosphate
system of 23.9% w/w TLL gave the best partitioning results with highest recovery of
proteins. Higher phase volume ratios during extraction have improved the recovery of
proteins in extracting phase.
Keywords: aqueous two-phase extraction; differential partitioning; clarification of cheese
whey; whey proteins; fat removal.

INTRODUCTION

cheese whey poses the problem of irreversible fouling of

the membranes, which in turn decreases the transmembrane
flux (Nguyen, 1999). In order to maintain the flux during
the membrane processing, several whey pretreatment
methods have been proposed to minimize the fat content
before membrane processing. They could be prefilitration,
demineralization, calcium addition under heat treatment, clarification (Lee and Merson, 1976) and centrifugation. In all these methods the fat removal is incomplete
and hence whey proteins tend to get adversely modified
during subsequent storage, which changes their functional
and organoleptic properties.
ATPE provides mild biphasic aqueous environment
suitable for extraction of proteins (Albertsson, 1986).
Aqueous two-phase systems (ATPSs), formed by dissolving a polymer along with a salt/polymer in water are
suitable for processing biological macromolecules
(Raghavarao et al., 1995; Srinivas et al., 2002). Since,
these systems have low interfacial tensions, they do not
cause denaturation of protein unlike aqueous/organic
systems (Albertsson, 1986). The present study demonstrates the application of ATPE, not only for the fat
removal but also for the isolation and concentration of
whey proteins. A detailed study was undertaken in the present work using ATPE to arrive at the optimal conditions
for differential partitioning of proteins and fat (partitioning
of proteins to one phase and fat to other phase) from cheese
whey.

The application of whey proteins as functional and nutritional ingredient in food, pharmaceuticals, and cosmetic
industries has gained importance in the last two decades
(Pouliot et al., 1999). Whey proteins have high digestibility
and an excellent metabolic efficiency and therefore have
high biological value, which is a measurement of high
nitrogen availability and utilization by the human body
(Mahan and Escott-Stump, 1996). Whey proteins have
highest Protein Digestibility Corrected Amino Acid Score
(PDCAAS) of 1.0, which is an indicator of a source of
all essential amino acids to the body. In addition, whey
proteins have an excellent amino acid balance, including
both essential and sulphur containing amino acids.
Whey proteins in a concentrated powdered form are the
most economical and quality protein source available commercially (Regester et al., 1996). These properties highlight
the nutritional quality and cost-effectiveness of whey
proteins both as a direct dietary supplement and as an
ingredient in a variety of formulated foods and beverages.
Membrane processes are currently in use for the
concentration of cheese whey, but the residual fat in

Correspondence to: Dr K. S. M. S. Raghavarao, Department of Food

Engineering, Central Food Technological Research Institute, Mysore570013, India.
E-mail: eng@cscftri.ren.nic.in

191

192

ANANDHARAMAKRISHNAN et al.
Determination of Partition Coefficient (m)
and Yield (YT)

MATERIALS AND METHODS

Chemicals
PEG6000, ammonium sulphate [(NH4)2SO4], dipotassium hydrogen phosphate (K2HPO4) and potassium
di-hydrogen phosphate (KH2PO4) were procured from
SRL, Mumbai, India. Maltodextrin (MDX) was procured
from Sukhjit Starch and Chemicals, Ltd., Punjab, India,
whose Dextrose Equivalent (DE) value is 14.
Preparation of Cheese Whey
Pasteurized milk was heated to boiling temperature and
allowed to cool to ambient temperature. The milk was inoculated with Lactobacillus culture at 308C and incubated overnight at ambient temperature (25 + 18C). Cheese whey was
obtained by filtering the curd thus obtained. The protein
content of cheese whey was measured prior to extraction.

Partition coefficient (m) of proteins was calculated as the

ratio of equilibrium concentration of proteins in the top
(CT) and bottom (CB) phases. Recovery of proteins in top
phase was calculated as percent yield (YT %) on weight
basis using the following equation.
YT % CT VT  100=C0 V0

YB % CB VB  100=C0 V0

where CT and CB are equilibrium concentration of proteins

in top and bottom phases, respectively. VT and VB are the
volume of top and bottom phases, respectively. C0 is concentration of proteins in the phase system and V0 is the
volume of total phase system.
Fat Detection

Protein Estimation
Protein estimation was carried out by Bradford method
using comassie brilliant blue G-250 (Bradford, 1976).
The results are expressed in terms of bovine serum albumin
(BSA) equivalents. An average of three replicates was considered. The error in the measurement of protein content
was within +1%.
Assembly of Phase System
Predetermined and weighed quantities of phase forming
components were added to cheese whey, so as to make
the system to 100% on w/w basis. The entire mixture
was stirred in a mechanically agitated contactor (Remi
India Ltd) for 1 h and allowed overnight at 25 + 18C for
the phase separation. Volumes of the individual phases
thus separated were measured. Aliquots of the phases
were analysed for protein content and pH.
The phase equilibrium data of Wu et al. (1996) was used
to calculate the tie line length (TLL% w/w) and interfacial
tension. Density of equilibrated and completely separated
phases was measured using a specific gravity bottle of
25 ml capacity in controlled room temperature of 258C.
Minimum three measurements were made and average
values of density were used for calculation of free
volume difference between the phases. The accuracy of
the measurements of density was within +1%. The free
volume difference between the two phases of blank phase
system [D(FV)] was calculated from density of top (r T)
and bottom (rB) phases (blank phases without proteins) at
258C (Eiteman and Gainer, 1989).

Thin layer chromatography (TLC) was employed to

detect presence of fat after extraction using the standard
method (Helmut, 1964) with slight modification in solvents
for qualitative detection of total lipids. After ATPE the
phases were separated and lipids were extracted from the
each phase with hexane. The solvent was removed from
extract by vacuum evaporation. TLC plates, made with
250 mm layer of silica gel G, were activated for 1 h at
258C just before use. The chromatograms were performed
in paper-lined tanks with chloroform methanol (1:2) solvent mixture. The plates were treated in iodine chamber
for the identification of spots. The phase contrast
microscopy (Olympus BX-40, USA) was used to measure
the size of fat droplets present in cheese whey and individual phases.
RESULTS AND ANALYSES
The differential partitioning of fat and proteins was
studied in two types of ATPSs namely polymer/polymer
and polymer/salt systems. The results of differential partitioning study (partitioning of proteins to PEG rich phase
while partitioning of fat to salt rich phase) are shown
in Table 1. Differential partitioning was not observed in
polymer/polymer system, however, it was observed in
polymer/salt system. In this system most of the fat was partitioned to salt rich phase. Traces of fat were partitioned to
PEG rich phases as detected by TLC method. Phase contrast microscopy was employed to measure the size of the
fat droplets present in cheese whey (Figure 1) and individual phases of ATPSs (Table 2). TLC measurements were

Table 1. Differential partitioning of fat and proteins in ATPSs.

Phase system and
composition (% w/w)
PEG
PEG
PEG
PEG
PEG

4000/Maltodextrin (14/32% w/w)

6000/Maltodextrin (10/40% w/w)
6000/(NH4)2SO4 (17.88/14.26% w/w)
6000/KH2PO4 (8/16% w/w)
6000/KH2PO4 K2HPO4 (11.7/10% w/w)

pH

Volume
ratio (ml/ml)

Protein concentration
in phase system
(mg/100 g of system)

Differential
partitioning

Partition
coefficient
of protein

Protein recovery
in top phase (%)

6.0
4.2
4.5
3.8
7.0

58/19
39/40
38/50
34/47
72/12

22.6
28.4
28.5
31.5
32.8

No
No
Yes
Yes
Yes

0.6
1.0
1.9
1.1
0.9

60.5
35.7
24.6
20.7
83.4

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197

ATPE FOR CLARIFICATION OF CHEESE WHEY

193

Figure 1. Fat droplets in cheese whey observed under phase contrast

microscopy.

used to detect the presence of fat. The bigger fat droplets

were observed particularly in the bottom phase of polymer/salt system (Table 2).
Polymer/salt system is found suitable for removal of fat
from cheese whey by differential partitioning. Proposed
mechanism of differential partitioning of fat and proteins is
shown in Figure 2. The detailed study of protein partitioning
was carried out in polymer/salt systems. Aggregation of protein poses a major problem during extraction, more particularly in polymer/salt type ATPSs. Hence, influence of
different process parameters such as phase forming salts,
phase composition, phase volume ratio and protein concentration on partitioning of protein was studied.
The variation in partition coefficient and recovery of
proteins due to change in phase composition and phase
forming salt is shown in Table 3. From this data the maximum protein extraction capacity of PEG6000/potassium
phosphate (pH 7), PEG6000/potassium dihydrogen phosphate and PEG6000/ammonium sulphate phase systems
were calculated [equations (1) and (2)] and found out to
be 0.38, 0.20 and 0.18 mg/ml of phase system, respectively. It may be noted that the PEG/ammonium sulphate

Table 2. Measured size of fat droplets in cheese whey and individual

phases of ATPSs.
Composition
(% w/w)

Phase

Dimension
of droplets
(mm)

Cheese whey
Polymer/polymer
(PEG 6000/Maltodextrin)
Polymer/polymer
(PEG 6000/Maltodextrin)

10/40

Top (PEG rich)

58
1050

10/40

1050

Polymer/salt
(PEG 6000/KH2PO4
K2HPO4)
Polymer/salt
(PEG 6000/KH2PO4
K2HPO4)

12/10

Bottom
(Maltodextrin
rich)
Top (PEG rich)

Phase system

12/10

Bottom
(salt rich)

2060
40200

Figure 2. Proposed mechanism for differential partitioning of fat and

proteins in ATPSs.

and PEG/potassium dihydrogen phosphate systems have

pH (Table 3) close to isoelectric pH of whey proteins. Significant aggregation tendency of proteins was observed in
PEG/ammonium sulphate and PEG/potassium dihydrogen
phosphate systems even at low protein concentrations
(below 0.04% w/w). However, aggregation was not
observed in PEG/phosphate system except at higher
(.23.9% w/w TLL) phase compositions.
The recovery of protein with TLL in polymer/salt
systems is shown in Figure 3. Influence of phase composition on partition coefficient and recovery of proteins (in
top phase) for PEG6000/potassium phosphate system is
shown in Table 4. The corresponding values of free
volume difference between two phases [D(FV)] and pH
are also indicated in Table 4. The increasing trend in
partition coefficient with phase composition is observed

Table 3. Effect of phase forming salts and phase composition on protein

partitioning in polymer/salt systems.
Phase system
and composition
(% w/w)

pH

Protein
concentration Volume
(mg/100 g
ratio
of system)
(ml/ml)

Protein
recovery
Partition
in top
coefficient phase (%)

PEG 6000/(NH4)2SO4
10.04/9.44
4.5
12.70/10.87
4.5
15.30/12.46
4.5
17.88/14.26
4.5

33.7
32.0
30.3
28.5

35/45
38/55
40/55
38/50

0.3
0.8
0.9
1.9

6.6
16.2
18.6
24.6

PEG 6000/KH2PO4
7.04/14.37
3.8
8/16
3.8
12/16
3.8

32.9
31.5
30.2

21/58
34/47
50/38

0.7
1.1
0.5

8.9
20.7
24.5

PEG 6000/KH2PO4 K2HPO4

11.7/10
7.0
32.8
13/10
7.0
32.3
15/10
7.0
31.4
10/16
7.0
31.0

72/12
76/13
75/9
52/18

0.9
0.8
0.6
0.4

83.4
81.1
75.5
18.8

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197

194

ANANDHARAMAKRISHNAN et al.

Figure 3. Recovery of proteins with TLL in polymer/salt systems.

up to 23.9% w/w TLL. However, further increase in TLL

showed a declining trend in partition coefficient as well
as recovery of whey proteins (Table 4). Thus, the best
recovery was observed at phase composition of 23.9% w/w
TLL. The effect of phase volume ratio on partition coefficient was studied by varying the total phase composition
on a given TLL (18.8% w/w) at two protein concentrations
[low (0.7 mg/100 g of phase system) and high (32.4 to
34.7 mg/100 g of phase system)]. Effect of phase volume
ratio on partition coefficient is shown in Figure 4. The partition coefficient of proteins decreased with an increase in
the phase volume ratio both at low and high protein concentrations. However, the corresponding recovery at these
conditions increased from 24 to 70% and 10 to 25% in
case of low and high protein concentrations, respectively.
An increasing trend in recovery of proteins was observed
up to a certain volume ratio (till 1.4 in case of low protein
concentrations and 0.9 in case of high protein
concentrations).
DISCUSSION
The differential partitioning of solute of interest and
impurity [i.e., partitioning of solute (proteins) in one
phase and impurity (fat) in other phase] is crucial for
obtaining the desired purification in the extraction. Partition
coefficient deviates from unity when the interaction of
solute with two immiscible phases is unequal. Hence, it is
essential to understand the mechanism of the differential
partitioning of solute and impurity in order to arrive at
optimal processing parameters.

In ATPE, the proteins exist in soluble or particulate form

depending on the concentration, that is, solubility limit,
which largely depends on the pH and concentration of
phase component. On the other hand, fat exists as dispersed
droplets of different size depending on ionic strength of surrounding continuous phase. The presence of salts reduces
the solubility and dispersibility of fat and this resulted in
bigger size fat droplets in salt rich phase compared to
that in cheese whey.
To understand the mechanism of partitioning of fat droplets, it is important to know the surface forces responsible
for partitioning in addition to the properties of liquid/
liquid interface (interfacial tension of ATPSs). The surface
forces associated with partitioning of fat droplets pertain to
liquid/liquid (fat/phase) interface unlike the partitioning of
solid particulates where they pertain to solid/liquid interface. Another difference between the partitioning of fat droplets and solid particulates, is the significant difference in
density (rsolid 2 rfat  0.2 g/ml). These aspects are considered while proposing the mechanism of partitioning of
fat.
The mechanism of partitioning of fat is proposed and
schematically shown in Figure 2. The following three
steps, which occur in series as well as simultaneously,
summarize the mechanism.
(1) Migration of fat droplets from the high energy level
(phase where the fat droplets are having high surface
free energy) to the interface (between disperse phase
and continuous phase).
(2) Adsorption of the fat droplets at the phase interface.
(3) Migration of fat droplets from this interface to the low
energy level (phase where the fat droplets are having
low surface free energy). If droplets attain lower
energy at the interface then this step will not occur.
The individual phases wet the fat droplets depending on
the surface free energy of droplets in these phases. Wetting
phenomena results in lowering of surface free energy of fat
droplets. In order to have one-sided partitioning of fat droplets, it is essential that one of the phases, more particularly
the non-extracting phase, should possess capability for preferential wetting of fat droplets over other (protein extracting) phase. The wetting criteria for fat droplets can be
obtained by the following expression, which was proposed
for solid surfaces (Schultz and Nardin, 1992).
jgP1  gP2 j , gTB
The value of gP1 and gP2 depends on interactions
between fat droplets and contacting phases. The polarity

Table 4. Influence of phase composition on recovery of whey proteins from cheese whey in PEG 6000/potassium phosphate system (pH 7).
Phase
composition
(% w/w)
15/7.7
11.7/10
12/10
13/10
15/10
10/16

TLL
(% w/w)

Interfacial
tension
of blank
system (mN/m)

D (FV) of blank
system (ml/g)

Volume
ratio (ml/ml)

Protein
concentration
(mg/100 g of system)

Partition
coefficient

Protein
recovery in
top phase (%)

18.2
22.6
23.9
25.7
29.1
42.3

0.05
0.10
0.12
0.16
0.24
0.88

15.55
15.66
16.53
18.28
23.25
55.74

69/23
72/12
81/6
76/13
75/9
52/18

20.0
32.8
32.7
32.3
31.4
31.0

0.8
0.9
0.9
0.8
0.6
0.4

70.1
83.4
85.2
81.1
75.5
18.8

Calculated from correlation given by Wu et al. (1996).

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197

ATPE FOR CLARIFICATION OF CHEESE WHEY

Figure 4. Effect of phase volume ratio on partition coefficient and recovery

of proteins in extracting phase (TLL 18.83% w/w).

indexes of phases and surface charge on fat droplets decide

the magnitude of these interactions. Polarity difference
between the individual phases is an important criterion to
result in the preferential wetting of fat droplets by one of
the phases. Polymer/salt systems have more polarity difference than polymer/polymer systems hence there is higher
probability of obtaining one-sided partitioning of fat.
It is required to determine that which phase is having
capability for preferential wetting of the fat droplets in
polymer/salt systems. The wetting phenomena involve
the interaction between surface of fat droplet and contacting phase. The surface forces associated with partitioning
of fat droplets could be accounted by considering the net
interaction of fat droplets with individual phases. The net
interaction arising from all the forces between droplet surface and individual phases are not necessarily additive and
individual interactions can not be quantified (Brooks et al.,
1985) but could be accounted collectively by contact angle
measurements. These interactions could be electrostatic or
hydrophobic, attractive or repulsive. The polar surfaceactive lipids present in fat impart charge to the surface of
droplets. The neutralization of charged surface of fat droplets could be easily facilitated to larger extent at shorter
distances by the presence of high concentration of counterions present in salts. Thus, the interaction between the ions
present in phases and fat droplets have decisive role in
attaining the lower surface free energy and in turn the wetting of fat droplets. These favourable interactions of the fat
droplets are responsible for the preferential wetting by the
salt rich phase rather than PEG rich phase.
The interfacial tension (gTB) is an important determinant
influencing the partitioning behaviour of particles in ATPSs
(Albertsson, 1986). The interfacial adsorption of particles
at liquid/liquid interface reduces the interfacial area
between the phases and decreases the interfacial tension
(Wu et al., 1996; Bamberger et al., 1984) leading to lowering of surface free energy. This finding of particle

195

partitioning can be extended to partitioning of liquid droplets in ATPSs. The interfacial tension of polymer/polymer
and polymer/salt type ATPSs lie in the range of 1027 to
1024 N/m (Bamberger et al., 1985) and 1025 to 1023 N/m
(Wu et al., 1996), respectively. The minimum size of
droplets capable to adsorb at the interface of ATPSs in
presence of Brownian motion is estimated (as shown in
Appendix) and the values are shown in Table 5. The
observed size of fat droplets in both the types of systems
(polymer/polymer and polymer/salt) were bigger than
the minimum size required for adsorption at the interface.
Hence, it could be inferred that the fat droplets could
adsorb at the interface of both types of ATPSs. However,
the rate of adsorption of fat droplets may vary depending
upon the extent of interaction between the fat droplets
and interface of phase system. The higher degree of adsorption of fat droplets at interface was visually observed in
case of polymer/salt system than polymer/polymer
system, which could be attributed to higher interfacial
tension. Further lowering of free energy occurs when the
fat droplets migrate to the salt rich phase.
It is advantageous to operate the extraction at high solute
(protein) content to reduce the operating scale. The variation in protein extraction capacity of systems is due to
the difference in solubilizing capacity, which in turn
could be explained based on pH dependent solubility of
proteins. Whey proteins are acidic kind of proteins
having isoelectric point 4.2 4.5 for a-lactalbumin and
5.2 for b-lactoglobulin (Marshall, 1982). The lower solubility of proteins in PEG/ammonium sulphate and PEG/
potassium dihydrogen phosphate system is due to system
pH, which is close to isoelectric pH of whey proteins.
The higher solubility of proteins in PEG/potassium
phosphate (pH 7) system is due to higher system pH.
Aggregation of protein has decisive role in partitioning.
The higher aggregation of protein reduces soluble amount
of protein available for partitioning, which in turn reduces
the recovery of proteins. Hence, it is essential to operate the
extraction at suitable phase compositions to have low aggregation and high solubility of proteins. As phase composition
increases, free volume difference between two phases
[D(FV)] also increases (Table 4). The increase in [D(FV)]
increases the partition coefficient and recovery of protein
in top phase (Eiteman and Gainer, 1989). In PEG 6000/
potassium phosphate system (pH 7) such an increasing
trend was observed upto 23.9% w/w TLL. Therefore it
can be inferred that proteins solubility is not affected by
phase forming components below this phase composition.
Further increase in TLL (.23.9% w/w) reduced the recovery which could be due to increasing tendency of precipitation of proteins by salt induced (salting out) effect (Kim
and Rha, 1996) or polymer induced (excluded volume)

Table 5. Minimum size of the droplet required to adsorb at the interface.

SN
1
2

ATPS type

Interfacial tension
(N/m)

Polymer/Polymer
Polymer/Salt

1027 1024


1025 1023

Bamberger et al. (1985).

Wu et al. (1996).

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197

Droplets size
(mm)
0.0060.229
0.0020.018

196

ANANDHARAMAKRISHNAN et al.

effect (Zimmerman and Trach, 1990) or combination of the

both.
Partition coefficient and phase volume ratio are important factors in determining the effectiveness of extraction.
In other words one-sided partition coefficient with low
phase volume ratio, both in favour of extraction phase,
reduces the volume to be handled in subsequent purification
steps, thereby achieving the process integration (coupling
of concentration with purification step). At both low and
high protein concentrations, the partition coefficient of
proteins deceased with an increase in phase volume ratio,
mainly due to solubility limitations. In order to solubilize
the given protein content, an adequate volume of extracting
phase is required. The maximum recovery in case of low
and high protein concentrations was 70 and 25%, respectively. This shows that an increase in the protein concentration from 0.7 to 34 mg/100 g of phase system reduces
the attainable recovery from 70 to 25%. Furthermore, by
increasing the volume ratio in case of low protein concentration, recovery increased significantly from 24 to 70%,
which indicates that complete recovery is possible below
protein concentration of 0.7 mg/100 g of phase system.
At high protein concentration also by increasing the
volume ratio, recovery increased, however, only from 10
to 25%. From these observations, it can be inferred that
intended volume of extracting phase is not sufficient to
recover the proteins at higher concentrations. In order to
achieve complete recovery, volume of extracting phase
needs to be increased accordingly with an increase in
protein concentration.
CONCLUSIONS
Polymer/salt type ATPSs were found to be effective for
the removal of fat from cheese whey by differential partitioning. The method can also be used for the concentration
of proteins. A mechanism has been proposed to explain the
differential partitioning of fat and proteins. PEG6000/Phosphate system (pH 7) at lower phase composition (below TLL
23.9% w/w) was found to be suitable to recover the proteins
in top phase. Performing the extraction at higher TLL and
low pH resulted in precipitation of the proteins and hence
not suitable for handling high protein concentrations.
Partitioning of proteins was greatly affected by the protein
concentration and phase volume ratio. High volume of
extracting phase corresponding to volume ratio of 1.5 and
above was found to improve the recovery of proteins.
NOMENCLATURE
CT
CB
C0
VT
VB
V0
m
YT
YB
k
T

protein concentration in top phase, mg/ml

protein concentration in bottom phase, mg/ml
concentration of protein in phase system, mg/ml
volume of top phase, ml
volume of bottom phase, ml
volume of phase system, ml
partition coefficient of protein
recovery of protein in top (extraction) phase, %
recovery of protein in bottom, (extraction) phase, %
Boltzmann constant, 1.3805  10223 J/8K
absolute temperature, 8K

Greek symbols
rT
density of continuous phase, kg/m3
rB
density of disperse phase, kg/m3

D(FV)
gTB
gP1
gP2

free volume difference between the two phases, ml/g

interfacial tension of phase system, N/m
interfacial tension between fat droplets and top phase, N/m
interfacial tension between fat droplets and bottom phase,
N/m

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ACKNOWLEDGEMENTS
Authors wish to thank Dr V. Prakash, Director, CFTRI, for his keen
interest in extractions using ATPSs. One of the authors (RSB) gratefully
acknowledges CSIR, India for the award of Senior Research Fellowship.
We also acknowledge Dr Prabhashankar, Scientist, Department of Flour
Milling, Baking and Confectionery Technology, CFTRI, for TLC
experiments.
The manuscript was received 27 November 2003 and accepted for
publication after revision 14 January 2004.

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197

ATPE FOR CLARIFICATION OF CHEESE WHEY

APPENDIX
Minimum size, required for adsorption of droplets at the
interface of phase system of known interfacial tension, can
be estimated by equating interfacial energy with thermal
energy (Brooks et al., 1985).
For simplicity in calculation, let us assume droplets are
wetted equally by both phases (i.e., contact angle 908)
then, interfacial energy of droplets of diameter (d ) at interface is a product of interfacial tension and area of the circle
[(p/4)d 2] which is in contact with interface. The diameter

197

of droplets can be obtained by equating interfacial energy

with thermal energy as shown below.
d 2kT=p
gTB 1=2
The interfacial tension of polymer/polymer and
polymer/salt two-phase systems lies in the range of
1027 1024 and 1025 1023 N/m, respectively. So the
minimum diameter of droplet required for adsorption at
interface of polymer/polymer and polymer/salt type
ATPSs lies from 6 to 229 and 2 to 18 nm, respectively.

Trans IChemE, Part C, Food and Bioproducts Processing, 2005, 83(C3): 191197