Chemistry and Physics of Lipids 127 (2004) 161–173

Solid–liquid phase behavior of binary fatty acid mixtures

2. Mixtures of oleic acid with lauric acid,
myristic acid, and palmitic acid
Tohru Inoue∗ , Yusuke Hisatsugu, Ruri Ishikawa, Masao Suzuki
Department of Chemistry, Faculty of Science, Fukuoka University, Nanakuma, Jonan-ku, Fukuoka 814-0180, Japan

Received 27 August 2003; received in revised form 15 October 2003; accepted 15 October 2003

Abstract

Solid–liquid phase behavior was investigated for binary fatty acid mixtures composed of oleic acid (OA; cis-9-octadecenoic
acid) and saturated fatty acids, lauric acid (LA; dodecanoic acid), myristic acid (MA; tetradecanoic acid), and palmitic acid (PA;
hexadecanoic acid), by means of differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FT-IR).
When the mixture was heated immediately after the solidification from the melt, the heat effect due to the ␥-to-␣ transformation
of OA varied depending on the composition of the mixture. However, the mixture subjected to an annealing at the temperature
slightly below the melting temperature provided the transformation at constant temperature which corresponds to the ␥-to-␣
transformation temperature of pure OA. This suggests that a solid phase formed by cooling of the melt of the mixture is not
in an equilibrium state, but it relaxes to a stable solid during the annealing process. The T–X phase diagrams of these mixtures
constructed from the DSC measurements demonstrate that the two fatty acid species are completely immiscible in a solid phase
regardless of the type of polymorphs of OA, ␣- or ␥-form. According to a thermodynamic analysis of liquidus line basing on
the regular solution model for the melt, the non-ideality of mixing tends to increase with the decrease in the acyl chain length of
the saturated fatty acid, although the mixing is rather close to ideal.
© 2003 Elsevier Ireland Ltd. All rights reserved.
Keywords: Fatty acids; Fatty acid mixture; Phase behavior; Mixing behavior; Phase diagram; DSC; FT-IR

1. Introduction properties, particularly phase behavior, of bulk lipid

Phase science of lipids and lipid mixtures is an in-
teresting and important research subject in connection
with the bio-science in the field of fundamental sci-
ence as well as with the food industry in the field of
practical application. It is believed that the physical
∗ Corresponding author. Tel.: +81-92-8716631;
fax: +81-92-8636050.
E-mail address: inouetr@fukuoka-u.ac.jp (T. Inoue).
assemblies are responsible for their biological func-
tion (Bloom et al., 1991). Food science treats the lipid
mixtures because a number of oil and fat mixtures are
utilized as daily foods, and the quality of food prod-
ucts sometimes depends on the phase behavior of lipid
mixtures included therein.
Most lipids contain different fatty acid species in the
molecule as constituents (Gurr and Harwood, 1991),
and the combination of the fatty acid species deter-
mines the physical properties of macroscopic assem-

0009-3084/$ – see front matter © 2003 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.chemphyslip.2003.10.013
162 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173
blies of lipid molecules. In order to understand the
physical properties of lipids and lipid mixtures, the
study of phase science of the constituent fatty acid
mixtures must be useful. The phase behavior is more
or less reflects the intermolecular interaction; espe-
cially, in the case of mixture systems, the balance of
the interaction between the same molecules and that
between different molecules determines the phase be-
havior. Thus, the study of the phase behavior of fatty
acid mixtures is expected to provide basic information
regarding the interaction between different acyl chains,
which is an important factor to determine the phys-
ical properties and phase behavior of lipid mixtures.
Studies on the solid–liquid phase behavior from these
points of view have so far been reported (Yoshimoto
et al., 1991; Inoue et al., 1992, 1993; Cedeño et al.,
2001; Zhang et al., 2001), and recently, we have been
carrying out the systematic study on this subject.
In a previous work (Inoue et al., 2004), we inves-
tigated the binary mixtures composed of oleic acid
(OA; cis-9-octadecenoic acid) and stearic acid (SA;
octadecanoic acid) and of OA and behenic acid (BA;
docosanoic acid) by means of differential scanning
calorimetry (DSC) and Fourier transform infrared
spectroscopy (FT-IR). Oleic acid has special interest
because natural occurrence of OA is most abundant
in all fatty acid species (Padley et al., 1994) and it is
believed that OA has a role to regulate the physical
properties of lipids in order to provide an environmen-
tal adaptability to the lipid materials in living bodies;
this function of OA may be attributed to its character-
istic molecular structure containing a cis-double bond
at the center of the acyl chain composed of 18 carbon
atoms. Thus, at first, we focused our attention on the
binary mixtures composed of OA and saturated fatty
acids.
It was found in the previous study on the solid–
liquid phase behavior of OA/SA mixture and OA/BA
mixture that these two fatty acid species are com-
pletely immiscible in a solid phase regardless of the
type of polymorphs of OA, i.e. ␣- or ␥-form (Inoue
et al., 2004). Thermodynamic analysis of liquidus line
of the T–X phase diagram demonstrated that OA and
SA form an ideal mixture in a liquid phase, whereas
OA and BA exhibit a slight non-ideality of mixing in a
liquid state. As an extension of this work, we investi-
gated the phase behavior of binary mixtures composed
of OA and saturated fatty acids with acyl chains shorter
than those examined previously using the same experi-
mental techniques. The saturated fatty acids used in the
present study are lauric acid (LA; dodecanoic acid),
myristic acid (MA; tetradecanoic acid), and palmitic
acid (PA; hexadecanoic acid), and the carbon numbers
in their acyl chains are 12, 14, and 16, respectively.
2. Experimental
2.1. Materials
Highly pure (>99.9%) samples of OA, LA, MA,
and PA were obtained from the Research Institute of
Structure and Function (Nara, Japan). They were pre-
pared by the procedure briefly mentioned in previous
paper (Inoue et al., 2004). The binary fatty acid mix-
tures with various compositions were prepared by dis-
solving the appropriate amounts of two fatty acid sam-
ples in small amount of chloroform followed by the
complete evaporation of the solvent in vacuo.
2.2. Methods
The solid–liquid phase behavior of the fatty acid
mixtures was investigated by means of differential
scanning calorimetry and Fourier transform infrared
spectroscopy. The experimental procedure has been
described in some detail in our previous paper (Inoue
et al., 2004). DSC measurements were carried out us-
ing a Seiko Denshi Model SSC5200 (Tokyo, Japan).
The weight of samples was usually in the range
5–10 mg. The rate of temperature scan was 1 ◦ C/min
for heating and 2–3 ◦ C/min for cooling. A Bio-Rad
Model FTS165 (Cambridge, MA) was used for FT-IR
measurements. The sample was sandwiched in a
molten state between two silicone-wafers, and was
placed in a handmade cell holder made from brass,
through which a water/ethylene glycol mixture of
constant temperature was circulated using a Neslab
refrigerated circulation bath RTE-111 (Portsmouth,
NH). The FT-IR spectra were measured as a func-
tion of temperature in the range from −30 to 40 ◦ C.
The spectra were recorded with 50 scans at 4 cm−1
resolution after the sample was kept at the desired
temperature for more than 5 min. In some cases in
both DSC and FT-IR measurements, the sample was
kept at desired temperature for 1–2 h in an oven of
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 163

the apparatus (DSC) or in a cell holder (FT-IR) in
order to examine the annealing effect on the thermal
behavior of the fatty acid mixtures.
3. Results and discussion
3.1. DSC results on OA/LA mixture
Fig. 1 shows the DSC curves for OA/LA mix-
tures with various compositions. They were obtained
by heating the samples immediately after the solid-
ification of the melt employing the cooling rate of
2–3 ◦ C/min. The heat effect observed at about −3 ◦ C
for pure OA is ascribed to the polymorphic trans-
formation from ␥- to ␣-form (Suzuki et al., 1985).
This polymorphic transformation has been rather well
characterized (Kobayashi et al., 1986). Briefly, the
difference between the two polymorphs is recognized
in the conformational structure of ␻-chain (the hy-
drocarbon chain between double bond and terminal
methyl group); the ordered all-trans structure in the

Fig. 1. DSC curves for OA/LA mixtures with various compositions. The weights of the samples were normalized to 5.00 mg. The
composition expressed in terms of the mole fraction of LA in the mixture is indicated near each curve. These DSC curves were obtained
by the heating scan of the sample immediately after solidification of the melt by cooling.
164 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173

Table 1 ture with xLA = 0.144 just after cooling to −30 ◦ C,

Thermodynamic data of the ␥-to-␣ transformation of OA and the in which the heat effect due to the ␥-to-␣ transfor-
mation of OA appears at around −20 ◦ C. Curve (b)
melting of OA, LA, MA, and PA
Fatty acids Ttr (◦ C)
Htr (kJ/mol) is a trace of the DSC thermogram obtained by the
This work Literature This work Literature heating–cooling cycle during which the sample was
OA kept at ca. 6 ◦ C for 1 h. In curve (b), a sharp endother-
␥→␣ −2.6 −2.2a 8.70 8.76a mic peak appears at −3 ◦ C similarly to the case of
␣ → melt 13.6 13.3a 39.0 39.6a pure OA. Thus, it can be clearly seen that the heat
LA 44.3 44.2b 36.3 36.6b effect ascribed to the ␥-to-␣ transformation of OA is
MA 54.3 54.4b 45.2 44.7b recovered by this thermal treatment at 6 ◦ C. Instead
PA 62.6 62.9b 52.3 54.3b
of this type of thermal treatment, the mixtures stored
a
b
Suzuki et al. (1985). in a refrigerator (4–5 ◦ C) for several days provided
Small (1986).
the endothermic peak due to the ␥-to-␣ transforma-
tion of OA at the same temperature as that for pure
␥-form versus the disordered liquid-like conforma- OA. The curves (c)–(g) in Fig. 2 are DSC curves ob-
tional structure in the ␣-form. The
-chain (the hy- tained for the samples subjected to either of the two
drocarbon chain between double bond and carboxylic ways of thermal treatments. This calorimetric obser-
group) has ordered all-trans conformational structure vation demonstrates that the annealing of the OA/LA
for both polymorphs. The endothermic peak at about mixture at the temperature slightly below the melting
13 ◦ C in the DSC curve for pure OA is due to the temperature for about 1 h or at the refrigerator tem-
melting of the ␣-form of OA. The thermodynamic perature for several days causes the relaxation of the
data obtained by the present DSC experiments for solid phase to equilibrium state.
the phase transitions of pure fatty acid species used
in this study are listed in Table 1 together with those 3.2. FT-IR spectroscopic observation of
reported in literature. solid-to-liquid transformation of OA/LA mixture
When LA is mixed, the heat effect due to the ␥-to-␣
transformation (white arrows) is once shifted toward Fig. 3 shows the IR spectral change associated with
lower temperature with the increase in the LA content the temperature rise observed for OA/LA mixture with
in the mixture, and then increases to reach almost con- xLA = 0.15. The spectra in Fig. 3a were obtained in
stant temperature which is lower than that observed the course of heating of the mixture immediately after
for pure OA; these peaks observed for the mixture the cooling of the melt down to −27.9 ◦ C. A sharp ab-
are much smaller and broader than the corresponding sorption peak at 700 cm−1 in the spectrum at −27.9 ◦ C
peak for pure OA. (white arrow) is characteristic of the ␥-form of OA
It is noteworthy that the exothermic heat effect is (Inoue et al., 2004), and this absorption peak demon-
observed for the mixture with xLA = 0.197 (denoted strates the existence of ␥-form of OA. When tempera-
by asterisk) during the course of heating scan, which ture is increased, the intensity of this peak decreases,
is immediately followed by a large endothermic heat and the absorption at 685 cm−1 , which is characteris-
effect. This observation suggests that the solid phase tic of the ␣-form of OA, appears at −22 to −18 ◦ C
of the mixture formed from the melt by cooling at the (black arrow) indicating that the ␥-form transforms
rate of 2–3 ◦ C/min is not stable state and the trans- to ␣-form at these temperature range. This behavior
formation to stable phase is induced by the tempera- of IR spectral change associated with the temperature
ture rise, during which the heat evolution takes place. rise is consistent with the thermal behavior of the mix-
Thus, we designed the experiments in which the solid ture with xLA = 0.144 presented in Fig. 2 curve (a)
sample was heated to the temperature corresponding which demonstrates that the ␥-to-␣ transformation of
to the exothermic peak and was kept for a while at that OA molecule takes place at around −20 ◦ C. Fig. 3b
temperature, and then cooled, and heated again. An shows the IR spectral change obtained by heating the
example of this kind of experiments is shown in Fig. 2. mixture which was cooled down to −27.1 ◦ C after
Curve (a) in Fig. 2 was obtained by heating of the mix- keeping the sample at 6.5 ◦ C for 2 h. In this case, the
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 165

Fig. 2. Annealing effect on DSC curves for OA/LA mixture. Curve (a): the mixture with xLA = 0.144 was heated just after cooling of
the melt to ca. −30 ◦ C. Curve (b): the mixture with the same composition as curve (a) was cooled to ca. −40 ◦ C from the melt, then
was heated up to 6 ◦ C and kept at the temperature for 1 h, then was cooled, and then was heated again. Curves (c)–(g): the heating DSC
curves of the mixtures subjected to the thermal treatment; the composition of the mixture is indicated near each curve.

absorption characteristic to the ␣-form of OA appears
at −4 ◦ C (black arrow), and a drastic spectral change
occurs in rather narrow temperature range from −5 to
−2 ◦ C, being again in accordance with the DSC ob-
servation (Fig. 2 curve (b)).
The melting process of this mixture followed by the
IR spectral change is depicted in Fig. 3c. It can be seen
that sharp absorption peaks in the methylene wagging
progression region (Jones et al., 1952) observed be-
low 8.0 ◦ C, which is characteristic to a solid phase, are
somewhat deformed when temperature reaches 9.0 ◦ C.
This means that a liquid phase is formed and coex-
ists with a solid phase at this temperature. The co-
existence of liquid and solid phases is appreciable at
10.0 ◦ C as well. The spectrum obtained at 11.0 ◦ C has
broad absorption peaks and exhibits a characteristic
feature of liquid fatty acids, which indicates that the
mixture is completely melted at this temperature. The
temperatures corresponding to the boundaries among
the one-phase region of solid, two-phase region of
166 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173

Fig. 3. FT-IR spectra observed for OA/LA mixture with xLA = 0.15 as a function of temperature. Temperature is indicated near each
spectrum. (a) The spectra were recorded during the course of heating of the sample just after cooling to −28 ◦ C from the melt. (b) The
spectra were recorded during the course of heating of the sample which was obtained by cooling to −27 ◦ C after keeping the mixture at
6.5 ◦ C for 2 h. (c) The spectra were obtained by heating the mixture succeeding the measurements of (b).

solid and liquid, and one-phase region of liquid are in
agreement with those predicted from the DSC mea-
surements (compare with the DSC curve for xLA =
0.144 shown in Fig. 2, curve (b)).
3.3. Phase diagram of OA/LA mixture
The T–X phase diagram for OA/LA mixture was
constructed by plotting the peak top temperatures of
the DSC curves as a function of the composition, and
is shown in Fig. 4. Open circles in this figure corre-
spond to the peak top temperature in the DSC curves
obtained by heating the samples just after cooling,
while filled circles are those obtained for samples
subjected to the thermal treatment. As mentioned be-
fore, the peak top temperature corresponding to the
␥-to-␣ transformation is affected remarkably by the
annealing of the sample, whereas those related to the
melting of the mixture are rather insensitive to the
thermal treatment. The phase boundary indicated in
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 167

Fig. 4. T–X phase diagram of OA/LA mixture constructed by plotting the peak top temperatures of the DSC curves as a function of xLA .
Open circles: peak top temperatures in the DSC curves obtained by heating the samples immediately after the solidification of the melt
by cooling. Filled circles: peak top temperatures in the DSC curves obtained for the mixtures subjected to thermal treatments.

Fig. 4 was confirmed by IR spectral change associated
with the temperature increase observed for several
mixtures with different compositions.
The horizontal phase boundary extending over the
whole composition range is obtained by connecting the
peak top temperatures due to the ␥-to-␣ transformation
of OA observed for the stable solid phase. This means
that the ␥-form of OA molecule and LA molecule are
completely immiscible and these two fatty acid species
exist as separate phases at the temperature below this
horizontal line of ca. −3 ◦ C. The liquidus and solidus
lines exhibit a typical eutectic-type phase boundary
with the eutectic temperature of ca. 9 ◦ C and the eutec-
tic composition of xLA ≈ 0.25. The horizontal solidus
line extends over the whole composition range, and
this indicates that the ␣-form of OA molecule is also
perfectly immiscible with LA molecule and they ex-
ist as separate phases in the temperature range be-
tween −3 and 9 ◦ C. This phase diagram is supported
by the analysis of the enthalpy change associated with
the phase transformation. The heat effect observed at
−3 ◦ C is caused by the transformation from the ␥- to
␣-form of OA existing as separate phase in the mix-
ture. Then, the enthalpy change per mole of mixture
associated with this process is predicted to decrease
linearly to zero with the increase in the mole frac-
tion of LA in the mixture. The heat effect occurring
at the eutectic temperature is attributed to the phase
reaction of SOA(␣) + SLA → L + SOA(␣) (0 < xLA <
xE ), SOA(␣) + SLA → L(x = xE ), SOA(␣) + SLA →
L + SLA (xE < xLA < 1), where xE means the eutectic
composition and L the liquid phase with xLA = xE .
168 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173

Fig. 5. Plot of enthalpy change per mole of mixture associated with the ␥-to-␣ transformation (a) and the melting at the eutectic temperature
(b) as a function of xLA .

When the enthalpy change per mole of mixture as-
sociated with this process is plotted as a function of
xLA , it increases linearly up to xLA = xE , and then de-
creases linearly up to xLA = 1 (so-called Tamman tri-
angle) (Laughlin, 1994; Chernik, 1995). Fig. 5 shows
this type of plot, in which
H values in the upper fig-
ure were determined from the area of the DSC peak
around −3◦ obtained with annealed samples, and those
in the lower figure were evaluated from the area of
the DSC peak observed around 9 ◦ C. The composition
dependence of the enthalpy changes associated with
these processes is in agreement with the prediction of
the phase diagram.
3.4. Thermodynamic analysis of liquidus line in the
phase diagram of OA/LA mixture
The phase boundary in the T–X phase diagram of
OA/LA mixture was analyzed on the basis of a ther-
modynamic model. It is expected that the melt of this
mixture would not deviate so much from an ideal solu-
tion because the two components have similar chemi-
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 169

cal structure and their molecular volumes do not differ
so significantly. For such cases, the regular solution
would become a good model to describe the solution
behavior (Moore, 1972). For the case of solid–liquid
phase equilibrium in which a regular solution coex-
ists with a solid phase where the two components are
perfectly immiscible, the equilibrium temperature as a
function of the composition, i.e. liquidus line, can be
given by the following equations (Lee, 1977; Tenchov,
1985; Inoue et al., 2004).
TA {
HA + w(1 − xA )2 }
T =

HA − RTA ln xA
TB (
HB + wx2A )
T =

HB − RTB ln(1 − xA )
where
HA and TA represent the molar heat of fu-
sion and the melting temperature of the component
A,
HB and TB are those of the component B, and
w corresponds to the interaction energy parameter
defined by


w = z uAB − 21 (uAA + uBB ) (3)
In the above expression, uAA , uBB , and uAB are the
interaction energies for A–A, B–B, and A–B pairs
formed in the mixture, and z is the coordination num-
ber, i.e. the number of nearest neighbor molecules of
the individual molecules. Eqs. (1) and (2) are applica-
(1) ble to the liquidus line which extends toward a eutectic
point starting from the melting point of the component
A and of the component B, respectively.
(2) The regular solution model attributes the non-
ideality of mixing to an enthalpy term assuming that
the entropy of mixing is the same as that for ideal
mixing, and this non-ideality coming from the en-

Fig. 6. Thermodynamic analysis of liquidus line in the T–X phase diagram of OA/LA mixture. The liquidus line was calculated from Eqs.
(1) and (2) using the values of the melting point and molar heat of fusion listed in Table 1 for pure OA and LA. The values of interaction
parameter are: w = 0 (dashed line), w = −1.5 kJ/mol (solid line), and w = −2.0 kJ/mol (dotted line).
170 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173

thalpy term is reflected in the parameter w (Moore,
1972). The physical meaning of w is the energy
difference between the mixed-pair (A–B pair) and
the average of like-pairs (A–A pair and B–B pair).
For ideal mixing, this energy difference is zero, and
hence, w becomes zero. Positive w corresponds to the
case where the mixed-pair has higher energy than the
like-pair; in other words, some repulsive interaction
acts between different components. On the contrary,
negative w means that the contact of different com-
ponents is energetically favorable compared with that
of the same components, i.e. the interaction between
different components in this case is attractive.
The liquidus lines calculated from Eqs. (1) and
(2) are compared with experimental phase boundary
for OA/LA mixture in Fig. 6, where the calculation
was made using the values listed in Table 1 for the
melting temperature and the molar heat of fusion of
each component. Dashed line is the calculated curve
obtained by the use of w = 0, which corresponds to
the liquidus line expected for the case that OA and
LA form ideal solution in the melt. This curve devi-
ates significantly from the experimental points of the
phase boundary; thus, an ideal solution is a poor ap-
proximation for the melt of OA/LA mixture. The dot-
ted line, which was obtained with w = −2.0 kJ/mol,
provides a better fitting to experimental points of
liquidus line, but the eutectic temperature exhibits
a downward deviation. The solid line, which was
obtained with w = −1.5 kJ/mol, reproduces experi-
ments satisfactorily in both liquidus line and eutectic
temperature.

Fig. 7. T–X phase diagram of OA/MA mixture constructed from the DSC results by the same procedure as described for OA/LA mixture.
The meaning of open and filled circles is the same as in Fig. 4. The liquidus line was calculated from Eqs. (1) and (2) using the values of
the melting point and molar heat of fusion listed in Table 1 for pure OA and MA. The values of interaction parameter are: w = 0 (dashed
line), w = −0.5 kJ/mol (solid line), and w = −1.0 kJ/mol (dotted line).
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 171

Fig. 8. T–X phase diagram of OA/PA mixture constructed from the DSC results by the same procedure as described for OA/LA mixture.
The meaning of open and filled circles is the same as in Fig. 4. The liquidus line was calculated from Eqs. (1) and (2) using the values
of the melting point and molar heat of fusion listed in Table 1 for pure OA and PA and the interaction parameter w = 0.

The small negative w for OA/LA mixture means that
the contact between OA molecule and LA molecule
in a liquid phase is energetically somewhat preferable
compared with those between themselves, or in other
words, slightly attractive interaction acts between OA
and LA molecules in a liquid phase. The liquid struc-
ture of fatty acids has been investigated by Iwahashi
and co-workers applying various experimental tech-
niques (Iwahashi et al., 1991, 1995, 2000). Accord-
ing to their investigations, the fatty acid molecules
in a liquid phase exist as dimers being formed due
to the hydrogen-bonding between carboxyl groups of
the two fatty acid molecules (Iwahashi et al., 1995),
and the dimers aggregate to form clusters of a smectic
liquid-crystal-like structure (Iwahashi et al., 1991). In
the case of fatty acid mixture, the dimers in a liquid
phase are formed between the same fatty acid species
when the two components are completely immiscible
in a solid phase (Iwahashi et al., unpublished results).
Then, in the present case of OA/LA mixture which ex-
hibits solid phase immiscibility, the dimers of OA and
those of LA would be present in a liquid phase. Thus,
the small negative value of w obtained for this mix-
ture suggests that the contact of an OA dimer and a
LA dimer is somewhat favorable in a liquid state com-
pared with the contact of OA dimers and that of LA
dimers. The OA molecule has 18 carbon atoms in its
acyl chain with a cis-double bond at the center of the
chain, while the LA molecule has saturated acyl chain
composed of 12 carbon atoms. Thus, the molecular
lengths of these two fatty acids must be different from
each other. Then, it is likely that the contact of the dif-
172 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173
ferent kinds of dimers is convenient for dense packing
of fatty acid molecules because of flexible nature of
acyl chains in a liquid state. In other words, the con-
tact of different dimers would be energetically more
favorable compared with that of the same dimers.
3.5. Phase behavior of OA/MA mixture and OA/PA
mixture
DSC and FT-IR experiments were carried out for
OA/MA and OA/PA mixtures as a function of their
composition. The behavior of the transformation be-
tween solid and liquid phases for these mixtures was
similar to the case of OA/LA mixture, i.e. a solid
phase produced by cooling of the melt at the rate of
2–3 ◦ C/min is not an equilibrium state, and the an-
nealing at the temperature slightly below the melting
temperature facilitates the relaxation to an equilibrium
solid phase. The T–X phase diagrams constructed from
the DSC results for OA/MA mixture and OA/PA mix-
ture are shown in Figs. 7 and 8, respectively. Open
circles and filled circles in these figures represent the
peak top temperatures of the endothermic heat effects
in the DSC curves obtained by heating the samples
immediately after the solidification and those obtained
for the samples subjected to the thermal treatment.
These phase boundaries estimated from the DSC ex-
periments were confirmed by the IR spectral change
associated with the temperature rise observed for sev-
eral mixtures.
The equilibrium phase diagrams of these mixtures
exhibit essentially the same feature as that of OA/LA
mixture, i.e. the two fatty acid species are completely
immiscible in a solid phase regardless of the type of
polymorphs of OA. The experimental liquidus lines
are well reproduced by the model calculation using
Eqs. (1) and (2) with w = −0.5 kJ/mol for OA/MA
mixture (solid curve in Fig. 7) and w = 0 for OA/PA
mixture.
4. Summary and conclusion
The present study revealed the solid–liquid phase
behavior of binary fatty acid mixtures of OA with
some saturated fatty acids, LA, MA, and PA by means
of DSC and FT-IR. Characteristic feature for these
mixtures is that the cooling of the liquid phase pro-
duces a non-equilibrium solid phase, and it relaxes to
an equilibrium state during the annealing at the tem-
perature slightly below the melting temperature (ex-
actly, eutectic temperature) of the mixture. In the stable
solid phase, the two fatty acid species are completely
immiscible and phase separation takes place. It is sup-
posed that the two fatty acid species are partly mixed
in the non-equilibrium solid phase formed from the
liquid phase by cooling, and the demixing is induced
by the annealing at the temperature close to the melt-
ing temperature, i.e. when temperature approaches to
the melting temperature, the translational motion of
fatty acid molecules would be facilitated, and hence,
the molecular rearrangement would take place rather
easily even in a solid phase. The non-equilibrium so-
lidification of the melt was not the case for the mix-
tures of OA and stearic acid (SA) and of OA and be-
henic acid. In these mixtures, a stable solid phase, in
which the two fatty acid species are separated as dif-
ferent phases, was obtained just by cooling of the liq-
uid phase (Inoue et al., 2004). On the other hand, the
complete immiscibility of the two fatty acid species
in the equilibrium solid phase regardless of the poly-
morphs of OA is common to the mixtures of OA and
saturated fatty acids in wide range of their acyl chain
lengths from 12 (LA) to 22 (BA) carbons.
The thermodynamic analysis applying the regular
solution model for liquid mixture demonstrates that
the mixing of OA and PA in a liquid phase is ideal, but
the non-ideality of mixing appears and becomes more
pronounced with the decrease in the acyl chain length
of saturated fatty acids. As for the mixtures of OA
with longer acyl chains, the OA/SA mixture showed
ideal mixing in a liquid phase, while the mixing of the
OA/BA mixture was slightly non-ideal (Inoue et al.,
2004). Thus, it is recognized that the non-ideality of
mixing becomes appreciable more and more with the
increase in the chain length difference between OA
and saturated fatty acids. The sign of w estimated for
OA/LA, OA/MA, and OA/BA mixtures are all nega-
tive, which means that the contact of different fatty
acid species in a liquid phase is energetically more fa-
vorable compared with those of the same species (see
Eq. (3)). This gain of interaction energy for the pair
formation between different fatty acid species would
come from the packing situation of fatty acid dimers
in a liquid phase, i.e. it is likely that the contact of the
dimers with different chain length is convenient for
 T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173
dense packing because of the flexible nature of acyl
chains in a liquid state. Anyway, the absolute value
of w is rather small, and hence, generally speaking,
the mixing of OA and saturated fatty acids in a liquid
phase is rather close to ideal mixing.
Acknowledgements
We are grateful to professor Makio Iwahashi of Ki-
tasato University for his useful discussion and sugges-
tion regarding the liquid structure of fatty acid mix-
tures. This work was supported in part by funds from
the Central Research Institute of Fukuoka University
(no. 015002) and the Research Institute of Structure
and Function.
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