J Am Oil Chem Soc (2013) 90:707716

DOI 10.1007/s11746-013-2202-7

ORIGINAL PAPER

Destabilization of Yellow Mustard Emulsion Using Organic

Solvents
Solmaz Tabtabaei Veronica M. Ataya Pulido
Levente L. Diosady

Received: 16 March 2012 / Revised: 5 November 2012 / Accepted: 28 December 2012 / Published online: 26 January 2013
AOCS 2013

Abstract An aqueous extraction process was developed

consisting of aqueous contact with dehulled yellow mustard flour to recover protein followed by dissolution of the
released emulsion in dimethylformamide (DMF) or isopropyl alcohol (IPA) to recover the released oil in the form
of single-phase oilsolvent miscellae suitable for industrial
applications. Only some 38 3 % of the oil in the yellow
mustard emulsion was extracted using DMF even at high
weight ratios since DMF is widely miscible with water,
preventing separation of the oil from the emulsion. A ternary phase diagram of DMF/oil/water was prepared and
confirmed the limited solubility of the oil in DMF in the
presence of water. The use of 31:1 IPA:oil weight ratio
could effectively recover over 94 % of the oil in the
emulsion; however, multiple-stage treatment of the emulsion was proven to be more efficient with lower volumes of
IPA required to achieve high oil extraction yields. The
results suggest that the optimal conditions for multiplestage process were four stages using 2:1 IPA:oil weight
ratio, with 96 1 % oil recovery from the emulsion.
Keywords Aqueous extraction
Dimethylformamide
(DMF)
Isopropyl alcohol (IPA)
Miscella
Multiplestage treatment
Single-stage treatment
Ternary phase
diagram
Yellow mustard emulsion

S. Tabtabaei (&)
V. M. Ataya Pulido
L. L. Diosady

Department of Chemical Engineering and Applied Chemistry,
University of Toronto, 200 Collage Street, Room WB 24,
Toronto, ON M5S 3E5, Canada
e-mail: s.tabtabaei@gmail.com
L. L. Diosady
e-mail: l.diosady@utoronto.ca

Introduction
Currently yellow mustard seed is only used as a condiment
in Western markets. However, it contains *30 % oil and
*27 % of high quality protein. The well balanced amino
acid profile of the seed proteins [1] makes mustard seed an
attractive potential source of food-grade vegetable proteins.
The high levels of erucic acid found in yellow mustard oil
makes it unsuitable for food applications, but it exhibits
excellent lubricating properties for various industrial uses
including biodiesel production.
Aqueous extraction processes (AEP), in which water is
used as an extraction medium to simultaneously produce oiland protein-rich fractions from oilseeds, are emerging as an
alternative to hexane extraction. This approach reduces the
health, safety, and environmental concerns associated with
solvent extraction [2]. Unfortunately, low oil extraction yield
and the formation of stable oil-in-water emulsions have prevented the commercial introduction of aqueous processes
since an additional demulsification step is required to fully
recover the oil. The remarkable stability of these emulsions is
likely due to the complex phospholipid-oleosin layer surrounding the oil bodies [3] that forms a very strong hydrophilic
layer in an aqueous medium, and prevents the coalescence of
oil. Several demulsification treatments have been recently
evaluated to break down these efficient emulsifiers, thereby
promoting the coalescence of oil droplets [410].
Demulsification methods, including heat treatment and
freezethaw treatment have been explored with emulsions
produced in conjunction with enzymatic and non-enzymatic
aqueous extraction of various oilseeds [5, 8, 10]. Acid
treatment with hydrochloric acid to destabilize emulsions
has also been studied. Adjusting the pH to the isoelectric
point of soy proteins (pH 4.5) reduced the electrostatic
repulsions between the oil droplets, and resulted in the

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recovery of 83100 % free oil from soy emulsions [4, 7, 9].

Enzymatic demulsification treatments with various proteases and phospholipases completely destabilized soy
emulsions formed in the enzymatic aqueous processing of
extruded soybean flakes and soybean flour [4, 6, 7, 9]. With
effective enzyme recycling, the use of appropriate proteases
and phospholipases during the extraction and demulsification steps could potentially achieve efficiencies approaching those of hexane extraction.
The use of organic solvents has also been reported to be
effective in demulsification. Li et al. [11] developed a
process to break emulsions by adding a mixture of two or
more co-miscible volatile solvents, at least one miscible
with oil and one miscible with water, thereby causing the
separation of the emulsion into two different layers.
However, this process was not economically viable on an
industrial scale. The addition of medium-chain alcohols has
been reported to increase the likelihood of droplet coalescence, thereby promoting demulsification due to the presence of both polar and non-polar groups in the structure of
these compounds. The alcohol molecules are able to
destabilize the emulsions by either displacing some of the
surfactants from the oilwater interface, or getting between
the tails of the surfactants [12, 13].
Our earlier studies focused on the development of the nonenzymatic aqueous processing of dehulled yellow mustard
flour, and on the effectiveness of cyclic ethers including
tetrahydrofuran (THF) and 1,4-dioxane in destabilizing the
produced emulsion [14]. The latter process involved the
mixing of the solvent with the emulsion followed by centrifugation. The use of a 4:1 THF:oil weight ratio successfully separated the yellow mustard emulsion into three
separate layers including an oil-rich miscella phase, a waterrich polar phase, and a solid emulsion residue phase, in which
over 97 % of the oil in the emulsion was recovered to the oilrich miscella phase containing only 5 % water.
In contrast to our previous studies using cyclic ethers
with complete solubilities for oil in solubilizing the
emulsion, the present study was intended to investigate the
effect of organic solvents with partial solubilities for oil in
destabilizing the emulsion. To achieve this goal, dimethylformamide (DMF) and isopropyl alcohol (IPA) were
selected to solubilize the emulsion to produce a singlephase oilsolventwater miscella that can be further processed for oil recovery for relevant industrial applications.

Materials and Methods

Materials
Dehulled yellow mustard flour (#106) purchased from
G.S.Dunn & Co. Ltd. (Hamilton, Ontario, Canada) was

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J Am Oil Chem Soc (2013) 90:707716

used as the starting material of the aqueous extraction

process. The flour was received in HDPE-lined, multi-wall
paper bags, and stored at room temperature. It contained
32.2 1.5 wt% oil (as-is basis), 32.9 0.08 wt% protein
(as-is basis), and 6.2 0.04 wt% moisture. DMF and IPA,
ACS grade, were purchased from Caledon Laboratories
Ltd. (Georgetown, ON, Canada).
Aqueous Extraction of Dehulled Yellow Mustard Flour
Dehulled yellow mustard flour was extracted at room
temperature for 30 min using a 4:1 water to flour (as is)
weight ratio at pH 11, as described previously [14]. The
smooth slurry was centrifuged to form solid residue, skim,
and emulsion fractions. The solid residue was washed once
using the same amount of water to dissolve more of the oil
and protein. The undissolved solid residue and the skim
fractions were freeze-dried, and analyzed for oil and protein content. The resulting yellow mustard emulsion was
also analyzed for oil, moisture, and protein content.
The emulsions generated during this process typically
contained 57.5 4.0 wt% oil, 38.4 3.8 wt% water,
3.1 0.3 wt% protein, and 3.3 0.3 wt% phospholipid
[14]. However, each batch of emulsion was analyzed
individually for its oil, water, and protein content before
any treatment with DMF or IPA to ensure the accuracy of
solvent:oil weight ratios.
Treatment of Yellow Mustard Emulsion with DMF
A 30-g sample of emulsion was transferred into a 250-mL
centrifuge bottle, and DMF was added to the centrifuge
bottle until the desired DMF:oil weight ratio was reached.
The mixture was shaken using a wrist action shaker
(Burrell Scientific, Pittsburgh, PA, USA) for 30 min followed by centrifugation for 20 min at 6,000 rpm in an
Avanti J-20 XP centrifuge (Beckman Coulter, Inc., Palo
Alto, CA, USA). After centrifugation, two phases were
observed: the miscella phase at the bottom of the container
composed of oil, water, and DMF; and the upper creamy
phase, i.e. the emulsion residue composed of oil, water,
DMF, and protein. These two phases were individually
collected for analysis. The miscella phase was analyzed for
oil, water, and DMF content. The oil extracted from the
original emulsion to the miscella phase was calculated
based on the oil content in the miscella phase that was
determined by measuring the weight loss after evaporation
of the DMF and water in a vacuum oven at 120 C and
*20 mmHg for 3 h; the water content of the miscella was
determined by BIOX Corporation (Hamilton, ON, Canada)
using a Karl Fischer titrimeter. The percentage of DMF
was then determined by subtracting the percentage of oil
and water from 100. The creamy emulsion residue phase

J Am Oil Chem Soc (2013) 90:707716

709

Fig. 1 Flow diagram for

treatment of yellow mustard
emulsion with DMF

was first dried in a vacuum oven under the above conditions to determine the solvent holdup; it was then analyzed
for protein content using the Kjeldahl method. The residual
oil and moisture content were calculated by mass balance
on the oil and water in the system. The experimental procedure of the treatment of yellow mustard emulsion with
DMF, and the schematic representation of the two phases
separated after centrifugation are presented in Fig. 1.

Table 1 Composition of the solutions used for preparation of oil/

water/DMF ternary phase diagram

Determination of Oil/Water/DMF Ternary Phase

Diagram
The usual method for constructing ternary phase diagrams
is to first measure the compositions of the phases that are in
equilibrium in the two-phase region; the compositional
pairs are then connected by tie lines to construct the solubility curve and phase diagram. In the oil/DMF/H2O
system, at room temperature, both oilDMF and oilH2O
are partially miscible, while DMF dissolves in any proportion in water; therefore, two pairs of partially miscible
liquids are formed. In this system, DMF is the solvent,
water is considered the solute since it is more soluble in
DMF, and oil is considered the diluent. Commercial canola
oil (100 % Canadian canola oil; Unico, Concord, ON,
Canada) was used because canola belongs to the same
family as yellow mustard (Cruciferae family) [15], and it
was readily available. The first step involved the determination of the solubility of oil in DMF and the solubility of
DMF in oil. The solubility of oil in DMF was measured by
adding pure canola oil from a burette to the specified
amount of stirred DMF until turbidity was observed. The
same procedure was applied to determine the solubility of
DMF in oil. The solubility of oil in water and water in oil
was considered negligible since the cloud point method is
not a practical way to measure the very limited solubility of
oil in water and water in oil. In the second step, nine different mixtures composed of canola oil, water, and DMF
were prepared with compositions presented in Table 1. The
mixtures were stirred vigorously for 30 min at room temperature, and immediately transferred into separatory funnels. The separated solvent-rich and oil-rich phases were
collected, and their weights were recorded. The next step
focused on the measurement of the composition of the
coexisting phases as described by Mehta and Fraser [16].
The oil contents of the two phases were determined by

Bulk solution no.

Composition (mass fraction)

DMF

Oil

Water

0.10

0.15

0.75

0.20

0.20

0.60

3
4

0.30
0.40

0.30
0.35

0.40
0.25

0.60

0.20

0.20

0.50

0.40

0.10

0.70

0.20

0.10

0.80

0.15

0.05

0.80

0.18

0.02

calculating the weight loss after evaporating all the water

and DMF, and the water content was determined by Karl
Fischer titration (Volumetric Karl Fischer ASTM standard
method E203 and Coulometric Karl Fischer ASTM standard method D6304).
Single- and Multiple-Stage Treatments of Yellow
Mustard Emulsion with IPA
The single-stage treatment of the yellow mustard emulsion
with IPA was started by transferring emulsion (3050 g)
into a centrifuge bottle, and then adding the required
amount of IPA to each bottle to reach the desired IPA:oil
weight ratio. The mixture was shaken for 30 min followed
by centrifugation at 4,500 rpm for 20 min. The centrifugation separated the mixture of emulsion and IPA into two
different phases including an emulsion residue phase in the
form of solid particles at the bottom of the container and
the miscella phase on top. These two phases were separated
from each other, and their weights were recorded. The
experimental procedure of the single-stage treatment of
yellow mustard emulsion with IPA is shown in Fig. 2.
The solid emulsion residue phase was composed of
protein, oil and IPA, while the miscella phase was composed of IPA, extracted oil and water. The oil content in the
miscella phase was measured by evaporating all of the
solvent and water initially at room temperature in a fume
hood and then at 110 C for 1.5 h using a forced air oven.
The residual oil content in the solid emulsion residue phase

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J Am Oil Chem Soc (2013) 90:707716

Fig. 2 Single-stage treatment

of yellow mustard emulsion
with IPA

was determined using the Mojonnier method, and the solvent holdup was gravimetrically estimated by drying the
sample in a forced air oven for 24 h. The dried emulsion
residue phase was finally used for protein determination.
The oil extracted from the original emulsion to the miscella
phase was calculated either based on the oil content in the
miscella phase or based on the residual oil content in the
emulsion residue phase. However, the presence of the twolayer miscella phase at IPA:oil weight ratios below 3:1
complicated the oil content determination in the miscella
phase. Therefore, the quantities of oil extracted to miscella
phases at low weight ratios (1:13:1 IPA:oil) were ascertained based on the residual oil content in the emulsion
residue phase. For weight ratios above 3:1, the use of either
of these techniques guaranteed accurate results.
Two-, three-, and four-stage treatments of yellow mustard emulsion with IPA were also evaluated for the purpose
of destabilization. The procedure previously described for
the single-stage treatment was followed by re-extracting
the emulsion residue phase with the same volume of IPA
used for the first extraction step. This resulting mixture of
IPA and emulsion residue was shaken again and centrifuged to produce the second miscella and emulsion residue
phases. This procedure was then repeated for the subsequent stages. The experimental procedure of the fourstage extraction process is shown in Fig. 3. The oil
recoveries and the composition of the miscella phases and
the final emulsion residue phase were determined based on
the methods employed in the single-step extraction.
Analytical Methods
The oil contents of dehulled yellow mustard flour and
yellow mustard emulsions were determined in triplicate
using the Mojonnier method (AOAC Method 922.06 for
solid samples, and AOAC Method 995.19 for the emulsion). Their protein contents (N 9 6.25) were determined
in triplicate by the Kjeldahl method, according to AOCS
Fig. 3 Four-stage treatment of
yellow mustard emulsion with
IPA

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Method Ba4d-90. Moisture contents were measured

gravimetrically by AACC Method 44-15A. Phospholipid
content of the emulsion was determined by ashing the
sample and measuring the phosphorus content spectrophotometrically (AOCS Method Ca 1255). To convert
total phosphorous to the equivalent phospholipids content,
a multiplication factor of 30 was used. The residual oil
content of the emulsion residues obtained after treatment
with IPA was measured in triplicate using the Mojonnier
method (AOAC 922.06).
Statistical Analysis
Single-stage treatment of yellow mustard emulsion was
performed at ten different DMF:oil weight ratios between
2.5:1 and 30:1, and 16 different IPA:oil weight ratios
between 1:1 and 31:1. Four-stage treatment of the emulsion
was performed at 1.5:1, 2:1, and 3:1 IPA:oil weight ratios.
The means of the oil extraction yields (n = 3) from each
treatment were compared by using the Least Significant
Difference (LSD) method at 5 % significance level in the
SAS system (version 9.0, SAS Institute Inc., Cary, NC,
USA).

Results and Discussion

Treatment of Yellow Mustard Emulsion with DMF
Dimethylformamide is often referred to as the universal
solvent because of its high boiling point (153 C). The
purpose of using DMF is to solubilize the emulsion to
recover all of the oil and water from the emulsion to the
single-phase oilwaterDMF miscella, which could be
dried later to remove water by simple evaporation. In this
regard, DMF:oil weight ratios of 2.5:130:1 were used to
destabilize the emulsion. Table 2 presents the oil extracted
from yellow mustard emulsion to miscellae. The

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Table 2 Oil extracted from the emulsion to the miscellae and the mass yields at different DMF:oil weight ratios
DMF:Oil weight ratios
2.5:1

5:1

7.5:1

10:1

12.5:1

15:1

Oil extracted
to miscella
(wt%)

7 3D

11 5D

23 4C

28 9BC

32 7ABC

Mass yieldsa

1.1 0.2

3 0.2

6 0.1

8.1 0.4

9.7 0.8

17.5:1

20:1

25:1

30:1

34 7AB

35 3AB

35 3AB

39 3A

38 3A

11.6 1.4

12.9 1.3

14.7 1.6

16.9 1.8

23 4.7

Means sharing the same capital letters are not significantly different (P \ 0.05)
Results are expressed as means of three independently prepared samples standard deviations of the means
a

Mass yield values represent the weight of the miscella phases to the emulsion residue phases produced after centrifugation

Fig. 4 a Composition of the

miscellae (wt%) at different
DMF:oil weight ratios.
b Composition of the emulsion
residues (wt%) at different
DMF:oil weight ratios.
Compositions are expressed as
means of three independently
prepared samples with error
bars representing standard
deviations of the means. If not
visible, it means the SD was
\0.5

compositions of the miscella and emulsion residue phases

are presented in Fig. 4a and b.
For weight ratios below 2.5:1, no separation of oil from
the emulsion was observed since the amount of added
DMF was not enough to solubilize the emulsion. The oil
extraction yield increased from 7 % at 2.5:1 to *38 % at
25:1 and 30:1 weight ratios. Increasing the amount of DMF
from 12.5:1 to 30:1 did not significantly (P \ 0.05)
improve the oil extraction yields; therefore, weight ratios
higher than 30:1 were not examined because the potential
increase in oil recovery would not be significant enough to
justify the use of larger quantities of DMF (Table 2).

The low efficiency of oil extraction from the emulsion to

miscella phases is associated with the solubility of water
and oil in DMF. The high affinity of DMF towards water
and its low affinity towards oil represent a major disadvantage. A low concentration of the oil in miscella phases
at all weight ratios (Fig. 4a) indicates the poor solubility of
the oil in DMF, which prevents achieving high extraction
yields during the process. As can be seen in Fig. 4b, the
amount of DMF at 2.5:1 and 5:1 weight ratios was not
enough to move all the water to the miscellae since the
water content in the emulsion residue at these two weight
ratios are 14 and 3 %, respectively. This small amount of

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the DMF added to the system was bound with water and
there was not enough solvent in the system to attack the oil,
thereby depressing the oil extraction yields to 7 and 11 %,
respectively (Table 2).
At DMF:oil weight ratios of 7.5:1, 10:1, and 12.5:1, no
water was observed in the emulsion residues as all the
water moved to the miscella phases. Therefore, the amount
of the solvent in the system was enough to dissolve all the
water, and this amount of solvent was also sufficient to
dissolve some of the oil in the emulsion. Therefore, at these
weight ratios, the oil extraction yields increased significantly to 23, 28, and 32 %, respectively (Table 2). The
maximum solubility of the oil in DMF was achieved at
12.5:1 weight ratio. At DMF:oil weight ratios greater than
12.5:1, the oil extraction yields did not improve significantly due to the limited solubility of the oil in DMF; this
limitation can also be seen in the composition of the miscella phases in which oil concentration started to decrease
from 2.5 to 1.3 % by increasing the ratio of DMF from
12.5:1 to 30:1 (Fig. 4a).
The prepared oil/water/DMF ternary phase diagram can
explain the low oil extraction yields from the emulsion to
the miscellae. Figure 5 shows the phase diagram including
the mutual solubility curves. The points on the right solubility curve have more oil in comparison with the amount
of DMF; conversely, the points on the left solubility curve
have more DMF. According to this diagram, the shaded
area in which oil, water, and DMF exist in a single phase is
very small. However, a relatively larger area of solubility

Fig. 5 Experimental ternary phase diagram of DMF (A) ? oil

(B) ? water (C) at room temperature: Filled circles overall composition for the tie lines; Filled squares tie line values; Solid line with
filled squares Solubility curves; open triangles composition of the
miscella phases produced during emulsion destabilization. The
overall compositions of the tie lines (bulk solutions compositions)
are presented in Table 1

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exists at high DMF content, compared to the one at low

DMF content. The intersection of the left solubility curve
and the horizontal axis (point K, composed of 94.3 wt%
DMF and 5.7 wt% oil) shows the maximum solubility of
the oil in DMF when no water is present in the system, and
the solubility curve represents the decreasing solubility of
the oil in DMF in the presence of water.
The creamy emulsion residue phase floating on top of
the miscella phase (Fig. 1) at all of the discussed DMF:oil
weight ratios was mostly composed of oil (49 5 wt%)
and DMF (47 8 wt%), with only 4 wt% protein
(Fig. 4b). Therefore, the limited solubility of the oil in
DMF, especially in the presence of water resulted in partial
destabilization of the emulsion and released an oil-rich
emulsion residue phase in the form of thin creamy layer on
top of the miscella phase.
Single-Stage Treatment of Yellow Mustard Emulsion
with IPA
Figure 6 presents the results for oil extraction from yellow
mustard emulsion to the miscella phases formed during
single-stage treatment with IPA. The amount of IPA at 1:1
weight ratio was not enough to promote emulsion destabilization since only 6 % of the oil was recovered from the
emulsion. The expected trend would be that, as IPA volume increases, extraction should also increase as a result of
more solvent being available to dissolve the oil. Nevertheless, the results in Fig. 6 show the oil extraction yields

Fig. 6 Single-stage oil extraction from yellow mustard emulsion at

different IPA:oil weight ratios. Single-stage treatment of emulsion
was performed at 16 different IPA:oil weight ratios of 1:1, 1.25:1,
1.5:1, 1.75:1, 2:1, 3:1, 5:1, 6:1, 9:1, 13:1, 16:1, 19:1, 22:1, 25:1, 28:1,
and 31:1. Oil extraction yields at IPA:oil weight ratios of 1.5:1,
1.75:1, 2:1, 3:1, 6:1, 9:1, 25:1, and 31:1 are expressed as means of
three independently prepared samples standard deviations of the
means. If not visible, it means the SD was \0.5. Since the standard
deviations were small and the results were consistent, the oil
extraction yields at other IPA:oil weight ratios were obtained based
on the single repetition

J Am Oil Chem Soc (2013) 90:707716

of 6062 % between 1.5:1 and 3:1 weight ratios. The

quantities of oil extracted to miscellae were then decreased
to around 20 % at 6:1, before increasing continuously to
around 94 % at 31:1 weight ratio.
The use of cyclic ethers including THF and 1,4-dioxane
accomplished the destabilization of the emulsion, and
recovered 9597 % of the oil to the miscellae using 4:1
THF:oil and 9:1 dioxane:oil weight ratios [14]. An
approximately similar oil extraction yield of around 94 %
was achieved using IPA at a 31:1 weight ratio, while only
38 % recovery was achieved using DMF at a 30:1 weight
ratio. The reason is associated with the extent of the solubility of oil and water in the organic solvents. The special
structure of cyclic ethers allows the full solvation of oil and
water molecules, thereby providing satisfactory destabilization results. However, the limited solubility of the oil in
IPA and DMF in the presence of water renders satisfactory
destabilization difficult.
We believe that the following three factors play an
important role in destabilizing the emulsion during the
single-stage treatment with IPA: the solubility of the oil in
IPA, the degree of percolation, and the degree of final
drainage [17]. The solubility of oil in IPA varies with
temperature and water content, the oil being completely
miscible at very high IPA concentrations, even at room
temperature [18]. Concentrations of IPA higher than 91 and
93 % were previously reported for infinite miscibility of
soybean oil and cottonseed oil, respectively [19]. The high
affinity of IPA towards water represents a major disadvantage during the destabilization process, especially at
low IPA:oil weight ratios since the water content in the
system is high and the solubility of oil in IPA is limited.
The composition of the miscella phases produced after
centrifugation is presented in Fig. 8. The miscellae recovered at weight ratios below 6:1 were separated to two
different layers (oil-rich layer and water-rich layer) due to
the water concentrations of more than 10 % that reduced
the solubility of the oil in IPA. However, the recovery of
the homogeneous miscella phases at weight ratios above
6:1 was related to the water concentrations of less than
10 %, indicating full solubility of the oil in IPA at concentrations above 90 %.
The permeability of the emulsion phase also affects
destabilization efficiency. When more solvent percolates
through the emulsion phase, the contact area increases and
extraction should also increase, however, it is more difficult to drain the emulsion, and this retention of miscella
increases the oil content in the emulsion residue phase. The
residual oil would represent not only the undissolved oil
but also the oil present in the retained miscella. In this
regard, the degree of final drainage was measured as solvent holdup of the emulsion residue phase, as it is presented in Fig. 7 for different IPA:oil weight ratios. Solvent

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Fig. 7 Solvent holdup for different IPA:oil weight ratios. Solvent

holdup represents the amount of IPA and water being retained in the
emulsion residue phase after centrifugation

holdup represents the amount of IPA and water being

retained in the emulsion residue phase after centrifugation.
Solvent holdup values in the range of 2629 % were found
for IPA:oil weight ratios of 1.25:12:1. For 3:1, 5:1, and
6:1 the values were 31.5, 32.2, and 34.4 %, respectively.
Above 6:1, it increased significantly to a maximum holdup
of 94.5 % at 31:1 weight ratio. When high solvent holdup
values are observed, drainage is considered poor due to
high retention of the miscella.
At IPA:oil weight ratios of 1.5:13:1, water content of
the produced miscella phases was in the range of 1524 %
and low extraction yields would be expected, however,
yields around 60 % were observed. Although the solubility
of the oil was limited by high water content, the low degree
of IPA percolation produced acceptable drainage through
the system (2629 % solvent holdup) facilitating the separation of miscella from emulsion residue phase, thereby
improving oil extraction yields. At IPA:oil weight ratios of
5:1 and 6:1, the use of higher quantities of IPA increased
the percolation of IPA in the emulsion, and produced less
concentrated miscella phases with oil concentrations of 5.5
and 2.8 %, respectively. The low oil extraction yields of 32
and 20 % could be attributed to the higher hold up values
(Fig. 7) that retained most of the oil in the emulsion residue
phase due to the poor drainage. Mustard oil is denser than
IPA, and tends to remain with the residue phase, thereby
reducing oil recovery.
Oil extraction yields started to increase for ratios above
6:1 at which homogeneous miscella phases produced at
concentrations of IPA higher than 90 % (Fig. 8). The oil
content in the miscellae increased moderately as IPA:oil
weight ratio increased from 6:1 to 19:1 leading to a maximum oil content of 3.8 % at 19:1. This maximum oil
content indicated the point at which sufficient IPA used to
completely dissolve the oil in the emulsion. However, the
oil recovery of 77 % was attributed to the retention of the

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Fig. 8 Composition of the

miscellae (wt%) at different
IPA:oil weight ratios

miscella in the emulsion residue phase as a result of poor

drainage. The miscella retention did not represent a big
contribution to residual oil content since the oil content of
the miscellae decreased to 2.8 % as IPA:oil weight ratio
increased to 31:1 at which over 94 % of the oil was
recovered.
As shown in Fig. 7, solvent holdup increased with
higher IPA:oil weight ratios and higher extraction yields.
The relationship between solvent holdup and residual oil
content was studied, and it is presented in Fig. 9. There is
an inverse relationship between solvent holdup and residual oil content which corresponds to observations made by
Zhang et al. [20] with IPA extraction of cottonseed. High
oil recoveries correspond to the presence of a lot of solvent-filled channels in the emulsion residue phase, while
fewer channels are available in the residue phase at low oil
recoveries; therefore, small values of solvent holdup can be
observed.
Multiple-Stage Treatment of Yellow Mustard Emulsion
with IPA
The miscella produced during the single-stage destabilization of the emulsion using 31:1 IPA:oil weight ratio contained some 94 % of the oil originally in the emulsion at a
low concentration (2.8 %), and nearly all of the water from
the emulsion at a 2.1 % concentration in 95 % IPA. The
recovery of the water from the corresponding miscella
phase would produce a single-phase oilIPA solution with
low concentration of oil which is not economically useful
for industrial applications. Taking into account that oil
extraction yields of *60 % were observed at low weight
ratios (1.5:13:1), multiple-stage extraction was considered
at these weight ratios in order to find the optimal conditions
to destabilize the emulsion. The reasoning behind this
consideration was that if it could be possible to extract as

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Fig. 9 Relationship between solvent holdup and residual oil content

much oil as with high weight ratios but using only a fraction
of the IPA, the process could be more economically feasible. Therefore, four-stage treatment of the yellow mustard
emulsion was evaluated for 1.5:1, 2:1, and 3:1 solvent to oil
weight ratios, and the results are shown in Fig. 10.
The results indicated the complete destabilization of
emulsion using IPA:oil weight ratios of 2:1 and 3:1. As can
be seen in Fig. 10, 99.4 and 96.3 % of the oil in the emulsion
was recovered using 3:1 and 2:1 IPA:oil weight ratios,
respectively. An IPA:oil weight ratio of 1.5:1 with *91 %
oil recovery was considered less effective, and more than
four stages will be required to fully recover the oil.
The first conclusion that can be verified from Fig. 10 is
related to the ascending trend of the oil extraction yields
achieved at each IPA:oil weight ratio. The multiple-stage
treatment of yellow mustard emulsion at a 2:1 IPA:oil
weight ratio significantly improved the initial oil extraction
yield of 62 to 72, 87, and 96 % through second, third, and

J Am Oil Chem Soc (2013) 90:707716

715
Table 3 The composition of the emulsion residue phases remaining
after four-stage treatment of yellow mustard emulsion
IPA:Oil

Oil content
(wt%)
Mean STD

Solvent holdup
(wt%)
Mean STD

Protein content
(wt%)
Mean STD

2:1

4.2 0.1

89.3 1.9

3.9 0.0

3:1

1.0 0.1

93.2 0.4

3.6 0.1

All the measurements were performed in triplicate

All values represent the composition means (n = 3) standard
deviation

Fig. 10 Oil extracted from the emulsion to the miscellae produced in

each stage of the four-stage extraction process using IPA:oil weight
ratios of 1.5:1(filled diamonds), 2:1(filled squares), and 3:1(filled
triangles). Results are expressed as means of three independently
prepared samples standard deviations of the means. ABC Oil
extraction yields in the same IPA:oil weight ratios with different
upper case letters are significantly different (P \ 0.05). abc Oil
extraction yields in each stage with different lower case letters are
significantly different (P \ 0.05)

fourth steps of the treatment process, respectively. The

same ascending trend can be also observed for 1.5:1 and
3:1 weight ratios.
For all weight ratios, the same oil recoveries of around
(5962 %) were achieved in the first stage in which twolayer miscella phases with high concentrations of oil
(1421 %) were produced. Separation of these miscellae
from emulsion residue phases was easy, indicating low
solvent holdup as observed in the single-stage treatment.
For the second stage, the highest oil extraction yield of
76 % was achieved at 3:1 followed by 72 % for 2:1, and
67 % for 1.5:1 IPA:oil weight ratios. For the third and
fourth stages, the oil extraction yield achieved at 3:1
IPA:oil weight ratio was significantly higher than that at
2:1 and 1.5:1, respectively.
The total amount of the IPA used during the four-stage
treatment of yellow mustard emulsion at 2:1 and 3:1 weight
ratios was similar to that used for the single-stage treatment
at 8:1 and 12:1, respectively. While single-stage extractions at these ratios were 2040 % (Fig. 6), using the same
amount of solvent in a multi (four) stage process resulted in
over 96 % oil recovery.
The emulsion residue phases remaining after the fourth
step of the 2:1 and 3:1 IPA:oil weight ratios were analyzed
for oil, IPA, and protein concentration. The results are
presented in Table 3. The residual oil content for 3:1
weight ratio was 1 %, representing less than 0.4 % of the

oil in the original flour, and the residual oil content for 2:1
weight ratio was 4.2 %, representing around 2.4 % of the
oil in the original flour. These results validated the oil
extraction yields determined by measuring the oil content
of the miscellae. As expected, solvent holdup values were
higher for 3:1 weight ratio than for 2:1 since more oil was
extracted at 3:1, and hence more channels were available to
retain solvent.
Based on our previous studies on the AEP of yellow
mustard flour at pH 11 [14], the oil recoveries of 96.3 and
99.4 % from yellow mustard emulsion using four-stage
extraction process at 2:1 and 3:1 IPA:oil weight ratios
were, respectively equivalent to 62.2 and 64.2 % of the oil
from the original flour. Taking into account that the difference between oil extraction yields from the original flour
was only 2 %, the four-stage process at 2:1 IPA:oil weight
ratio was chosen as the optimal process for emulsion
destabilization since the amounts of IPA required were
lower than with the 3:1 weight ratio but the reduction in oil
recovery was not significant. In other words, the small
improvement in oil recovery did not justify the use of
larger quantities of IPA.
The compositions of the miscellae produced in each
stage of the four-stage treatment of yellow mustard emulsion at IPA:oil weight ratio of 2:1 were determined, and the
results are presented in Table 4. The first miscella consisted of two different phases, oil-rich and water-rich
phases. This miscella contained large amounts of oil
(22.4 %) separated in a denser phase, and around 20 %
water content that prevented the formation of the singlephase miscella due to the limited solubility of the oil at
concentrations of IPA below 90 %. The second, third, and
fourth miscellae were homogeneous oilwaterIPA solutions since their water content was well below 5 %, indicating full miscibility of the oil and IPA.
The composition of the combined miscella created by
mixing all of the miscellae produced during the four-stage
treatment of yellow mustard emulsion at 2:1 was determined, and it is also presented in Table 4. The combined
miscella with *10 % oil content could be used as a suitable feedstock for industrial applications.

123

716

J Am Oil Chem Soc (2013) 90:707716

Table 4 Composition of the miscellae produced during the fourstage treatment of yellow mustard emulsion at 2:1 IPA:oil weight
ratio
Stages

Oil content
(wt%)
Mean STD

Water content
(wt%)
Mean STD

22.4 1.0

7.0 0.2

IPA content
(wt%)
Mean STD

3.

19.9 1.6

57.7 1.1

4.

3.3 0.0

89.7 0.2

8.2 0.6

0.9 0.0

90.9 0.6

4.4 0.3

0.4 0.0

95.1 0.3

10.0 0.3

7.5 0.7

82.5 0.4

Combined

2.

All the measurements were performed in triplicate

All values represent the composition means (n = 3) standard
deviation

5.

6.

7.

Conclusions
Although DMF has a high boiling point which allows the
easy separation of the water from oilwaterDMF miscella
phase, it is not recommended as an effective solvent to
completely solubilize the yellow mustard emulsion. The
use of even a very high amount of DMF (30:1) was only
able to recover *38 % of the oil in the emulsion since the
solubility of the oil in DMF is greatly depressed by the
presence of water as shown by the ternary phase diagram
(Fig. 5). Isopropyl alcohol can be used as an effective
solvent to completely destabilize the yellow mustard
emulsion through single- and multiple-stage treatment
processes. For the single-stage treatment, over 94 % of the
oil in the emulsion was recovered using a 31:1 IPA:oil
weight ratio; however, the low oil concentration of the
corresponding miscella (2.8 %), and the large amounts of
IPA could restrict its application in industry. Four-stage
treatment of yellow mustard emulsion at 2:1 IPA:oil weight
ratio with *96 % oil recovery was found as the most
effective process for destabilizing the yellow mustard
emulsion. Eventually, this integrated process based on
aqueous extraction of dehulled yellow mustard flour followed by multiple-stage IPA extraction of the emulsion
could produce a solution suitable for relevant industrial
applications.
Acknowledgments This project was funded by the Natural Sciences
and Engineering Research Council of Canada through its strategic
grants program.

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