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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


assemblies are responsible for their biological func-
Phase science of lipids and lipid mixtures is an in- tion (Bloom et al., 1991). Food science treats the lipid
teresting and important research subject in connection mixtures because a number of oil and fat mixtures are
with the bio-science in the field of fundamental sci- utilized as daily foods, and the quality of food prod-
ence as well as with the food industry in the field of ucts sometimes depends on the phase behavior of lipid
practical application. It is believed that the physical mixtures included therein.
Most lipids contain different fatty acid species in the
∗ Corresponding author. Tel.: +81-92-8716631; molecule as constituents (Gurr and Harwood, 1991),
fax: +81-92-8636050. and the combination of the fatty acid species deter-
E-mail address: inouetr@fukuoka-u.ac.jp (T. Inoue). 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 than those examined previously using the same experi-
physical properties of lipids and lipid mixtures, the mental techniques. The saturated fatty acids used in the
study of phase science of the constituent fatty acid present study are lauric acid (LA; dodecanoic acid),
mixtures must be useful. The phase behavior is more myristic acid (MA; tetradecanoic acid), and palmitic
or less reflects the intermolecular interaction; espe- acid (PA; hexadecanoic acid), and the carbon numbers
cially, in the case of mixture systems, the balance of in their acyl chains are 12, 14, and 16, respectively.
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 2. Experimental
acid mixtures is expected to provide basic information
regarding the interaction between different acyl chains, 2.1. Materials
which is an important factor to determine the phys-
ical properties and phase behavior of lipid mixtures. Highly pure (>99.9%) samples of OA, LA, MA,
Studies on the solid–liquid phase behavior from these and PA were obtained from the Research Institute of
points of view have so far been reported (Yoshimoto Structure and Function (Nara, Japan). They were pre-
et al., 1991; Inoue et al., 1992, 1993; Cedeño et al., pared by the procedure briefly mentioned in previous
2001; Zhang et al., 2001), and recently, we have been paper (Inoue et al., 2004). The binary fatty acid mix-
carrying out the systematic study on this subject. tures with various compositions were prepared by dis-
In a previous work (Inoue et al., 2004), we inves- solving the appropriate amounts of two fatty acid sam-
tigated the binary mixtures composed of oleic acid ples in small amount of chloroform followed by the
(OA; cis-9-octadecenoic acid) and stearic acid (SA; complete evaporation of the solvent in vacuo.
octadecanoic acid) and of OA and behenic acid (BA;
docosanoic acid) by means of differential scanning 2.2. Methods
calorimetry (DSC) and Fourier transform infrared
spectroscopy (FT-IR). Oleic acid has special interest The solid–liquid phase behavior of the fatty acid
because natural occurrence of OA is most abundant mixtures was investigated by means of differential
in all fatty acid species (Padley et al., 1994) and it is scanning calorimetry and Fourier transform infrared
believed that OA has a role to regulate the physical spectroscopy. The experimental procedure has been
properties of lipids in order to provide an environmen- described in some detail in our previous paper (Inoue
tal adaptability to the lipid materials in living bodies; et al., 2004). DSC measurements were carried out us-
this function of OA may be attributed to its character- ing a Seiko Denshi Model SSC5200 (Tokyo, Japan).
istic molecular structure containing a cis-double bond The weight of samples was usually in the range
at the center of the acyl chain composed of 18 carbon 5–10 mg. The rate of temperature scan was 1 ◦ C/min
atoms. Thus, at first, we focused our attention on the for heating and 2–3 ◦ C/min for cooling. A Bio-Rad
binary mixtures composed of OA and saturated fatty Model FTS165 (Cambridge, MA) was used for FT-IR
acids. measurements. The sample was sandwiched in a
It was found in the previous study on the solid– molten state between two silicone-wafers, and was
liquid phase behavior of OA/SA mixture and OA/BA placed in a handmade cell holder made from brass,
mixture that these two fatty acid species are com- through which a water/ethylene glycol mixture of
pletely immiscible in a solid phase regardless of the constant temperature was circulated using a Neslab
type of polymorphs of OA, i.e. ␣- or ␥-form (Inoue refrigerated circulation bath RTE-111 (Portsmouth,
et al., 2004). Thermodynamic analysis of liquidus line NH). The FT-IR spectra were measured as a func-
of the T–X phase diagram demonstrated that OA and tion of temperature in the range from −30 to 40 ◦ C.
SA form an ideal mixture in a liquid phase, whereas The spectra were recorded with 50 scans at 4 cm−1
OA and BA exhibit a slight non-ideality of mixing in a resolution after the sample was kept at the desired
liquid state. As an extension of this work, we investi- temperature for more than 5 min. In some cases in
gated the phase behavior of binary mixtures composed both DSC and FT-IR measurements, the sample was
of OA and saturated fatty acids with acyl chains shorter 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 by heating the samples immediately after the solid-
order to examine the annealing effect on the thermal ification of the melt employing the cooling rate of
behavior of the fatty acid mixtures. 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).
3. Results and discussion This polymorphic transformation has been rather well
characterized (Kobayashi et al., 1986). Briefly, the
3.1. DSC results on OA/LA mixture difference between the two polymorphs is recognized
in the conformational structure of ␻-chain (the hy-
Fig. 1 shows the DSC curves for OA/LA mix- drocarbon chain between double bond and terminal
tures with various compositions. They were obtained 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 somewhat deformed when temperature reaches 9.0 ◦ C.
at −4 ◦ C (black arrow), and a drastic spectral change This means that a liquid phase is formed and coex-
occurs in rather narrow temperature range from −5 to ists with a solid phase at this temperature. The co-
−2 ◦ C, being again in accordance with the DSC ob- existence of liquid and solid phases is appreciable at
servation (Fig. 2 curve (b)). 10.0 ◦ C as well. The spectrum obtained at 11.0 ◦ C has
The melting process of this mixture followed by the broad absorption peaks and exhibits a characteristic
IR spectral change is depicted in Fig. 3c. It can be seen feature of liquid fatty acids, which indicates that the
that sharp absorption peaks in the methylene wagging mixture is completely melted at this temperature. The
progression region (Jones et al., 1952) observed be- temperatures corresponding to the boundaries among
low 8.0 ◦ C, which is characteristic to a solid phase, are 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 is shown in Fig. 4. Open circles in this figure corre-
agreement with those predicted from the DSC mea- spond to the peak top temperature in the DSC curves
surements (compare with the DSC curve for xLA = obtained by heating the samples just after cooling,
0.144 shown in Fig. 2, curve (b)). while filled circles are those obtained for samples
subjected to the thermal treatment. As mentioned be-
3.3. Phase diagram of OA/LA mixture fore, the peak top temperature corresponding to the
␥-to-␣ transformation is affected remarkably by the
The T–X phase diagram for OA/LA mixture was annealing of the sample, whereas those related to the
constructed by plotting the peak top temperatures of melting of the mixture are rather insensitive to the
the DSC curves as a function of the composition, and 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 perfectly immiscible with LA molecule and they ex-
with the temperature increase observed for several ist as separate phases in the temperature range be-
mixtures with different compositions. tween −3 and 9 ◦ C. This phase diagram is supported
The horizontal phase boundary extending over the by the analysis of the enthalpy change associated with
whole composition range is obtained by connecting the the phase transformation. The heat effect observed at
peak top temperatures due to the ␥-to-␣ transformation −3 ◦ C is caused by the transformation from the ␥- to
of OA observed for the stable solid phase. This means ␣-form of OA existing as separate phase in the mix-
that the ␥-form of OA molecule and LA molecule are ture. Then, the enthalpy change per mole of mixture
completely immiscible and these two fatty acid species associated with this process is predicted to decrease
exist as separate phases at the temperature below this linearly to zero with the increase in the mole frac-
horizontal line of ca. −3 ◦ C. The liquidus and solidus tion of LA in the mixture. The heat effect occurring
lines exhibit a typical eutectic-type phase boundary at the eutectic temperature is attributed to the phase
with the eutectic temperature of ca. 9 ◦ C and the eutec- reaction of SOA(␣) + SLA → L + SOA(␣) (0 < xLA <
tic composition of xLA ≈ 0.25. The horizontal solidus xE ), SOA(␣) + SLA → L(x = xE ), SOA(␣) + SLA →
line extends over the whole composition range, and L + SLA (xE < xLA < 1), where xE means the eutectic
this indicates that the ␣-form of OA molecule is also 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- these processes is in agreement with the prediction of
sociated with this process is plotted as a function of the phase diagram.
xLA , it increases linearly up to xLA = xE , and then de-
creases linearly up to xLA = 1 (so-called Tamman tri- 3.4. Thermodynamic analysis of liquidus line in the
angle) (Laughlin, 1994; Chernik, 1995). Fig. 5 shows phase diagram of OA/LA mixture
this type of plot, in which H values in the upper fig-
ure were determined from the area of the DSC peak The phase boundary in the T–X phase diagram of
around −3◦ obtained with annealed samples, and those OA/LA mixture was analyzed on the basis of a ther-
in the lower figure were evaluated from the area of modynamic model. It is expected that the melt of this
the DSC peak observed around 9 ◦ C. The composition mixture would not deviate so much from an ideal solu-
dependence of the enthalpy changes associated with 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 A, HB and TB are those of the component B, and
so significantly. For such cases, the regular solution w corresponds to the interaction energy parameter
would become a good model to describe the solution defined by
behavior (Moore, 1972). For the case of solid–liquid  
phase equilibrium in which a regular solution coex- w = z uAB − 21 (uAA + uBB ) (3)
ists with a solid phase where the two components are
perfectly immiscible, the equilibrium temperature as a In the above expression, uAA , uBB , and uAB are the
function of the composition, i.e. liquidus line, can be interaction energies for A–A, B–B, and A–B pairs
given by the following equations (Lee, 1977; Tenchov, formed in the mixture, and z is the coordination num-
1985; Inoue et al., 2004). ber, i.e. the number of nearest neighbor molecules of
TA {HA + w(1 − xA )2 } the individual molecules. Eqs. (1) and (2) are applica-
T = (1) ble to the liquidus line which extends toward a eutectic
HA − RTA ln xA
point starting from the melting point of the component
TB (HB + wx2A ) A and of the component B, respectively.
T = (2) The regular solution model attributes the non-
HB − RTB ln(1 − xA )
ideality of mixing to an enthalpy term assuming that
where HA and TA represent the molar heat of fu- the entropy of mixing is the same as that for ideal
sion and the melting temperature of the component 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, was made using the values listed in Table 1 for the
1972). The physical meaning of w is the energy melting temperature and the molar heat of fusion of
difference between the mixed-pair (A–B pair) and each component. Dashed line is the calculated curve
the average of like-pairs (A–A pair and B–B pair). obtained by the use of w = 0, which corresponds to
For ideal mixing, this energy difference is zero, and the liquidus line expected for the case that OA and
hence, w becomes zero. Positive w corresponds to the LA form ideal solution in the melt. This curve devi-
case where the mixed-pair has higher energy than the ates significantly from the experimental points of the
like-pair; in other words, some repulsive interaction phase boundary; thus, an ideal solution is a poor ap-
acts between different components. On the contrary, proximation for the melt of OA/LA mixture. The dot-
negative w means that the contact of different com- ted line, which was obtained with w = −2.0 kJ/mol,
ponents is energetically favorable compared with that provides a better fitting to experimental points of
of the same components, i.e. the interaction between liquidus line, but the eutectic temperature exhibits
different components in this case is attractive. a downward deviation. The solid line, which was
The liquidus lines calculated from Eqs. (1) and obtained with w = −1.5 kJ/mol, reproduces experi-
(2) are compared with experimental phase boundary ments satisfactorily in both liquidus line and eutectic
for OA/LA mixture in Fig. 6, where the calculation 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 phase are formed between the same fatty acid species
the contact between OA molecule and LA molecule when the two components are completely immiscible
in a liquid phase is energetically somewhat preferable in a solid phase (Iwahashi et al., unpublished results).
compared with those between themselves, or in other Then, in the present case of OA/LA mixture which ex-
words, slightly attractive interaction acts between OA hibits solid phase immiscibility, the dimers of OA and
and LA molecules in a liquid phase. The liquid struc- those of LA would be present in a liquid phase. Thus,
ture of fatty acids has been investigated by Iwahashi the small negative value of w obtained for this mix-
and co-workers applying various experimental tech- ture suggests that the contact of an OA dimer and a
niques (Iwahashi et al., 1991, 1995, 2000). Accord- LA dimer is somewhat favorable in a liquid state com-
ing to their investigations, the fatty acid molecules pared with the contact of OA dimers and that of LA
in a liquid phase exist as dimers being formed due dimers. The OA molecule has 18 carbon atoms in its
to the hydrogen-bonding between carboxyl groups of acyl chain with a cis-double bond at the center of the
the two fatty acid molecules (Iwahashi et al., 1995), chain, while the LA molecule has saturated acyl chain
and the dimers aggregate to form clusters of a smectic composed of 12 carbon atoms. Thus, the molecular
liquid-crystal-like structure (Iwahashi et al., 1991). In lengths of these two fatty acids must be different from
the case of fatty acid mixture, the dimers in a liquid 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 duces a non-equilibrium solid phase, and it relaxes to
of fatty acid molecules because of flexible nature of an equilibrium state during the annealing at the tem-
acyl chains in a liquid state. In other words, the con- perature slightly below the melting temperature (ex-
tact of different dimers would be energetically more actly, eutectic temperature) of the mixture. In the stable
favorable compared with that of the same dimers. solid phase, the two fatty acid species are completely
immiscible and phase separation takes place. It is sup-
3.5. Phase behavior of OA/MA mixture and OA/PA posed that the two fatty acid species are partly mixed
mixture in the non-equilibrium solid phase formed from the
liquid phase by cooling, and the demixing is induced
DSC and FT-IR experiments were carried out for by the annealing at the temperature close to the melt-
OA/MA and OA/PA mixtures as a function of their ing temperature, i.e. when temperature approaches to
composition. The behavior of the transformation be- the melting temperature, the translational motion of
tween solid and liquid phases for these mixtures was fatty acid molecules would be facilitated, and hence,
similar to the case of OA/LA mixture, i.e. a solid the molecular rearrangement would take place rather
phase produced by cooling of the melt at the rate of easily even in a solid phase. The non-equilibrium so-
2–3 ◦ C/min is not an equilibrium state, and the an- lidification of the melt was not the case for the mix-
nealing at the temperature slightly below the melting tures of OA and stearic acid (SA) and of OA and be-
temperature facilitates the relaxation to an equilibrium henic acid. In these mixtures, a stable solid phase, in
solid phase. The T–X phase diagrams constructed from which the two fatty acid species are separated as dif-
the DSC results for OA/MA mixture and OA/PA mix- ferent phases, was obtained just by cooling of the liq-
ture are shown in Figs. 7 and 8, respectively. Open uid phase (Inoue et al., 2004). On the other hand, the
circles and filled circles in these figures represent the complete immiscibility of the two fatty acid species
peak top temperatures of the endothermic heat effects in the equilibrium solid phase regardless of the poly-
in the DSC curves obtained by heating the samples morphs of OA is common to the mixtures of OA and
immediately after the solidification and those obtained saturated fatty acids in wide range of their acyl chain
for the samples subjected to the thermal treatment. lengths from 12 (LA) to 22 (BA) carbons.
These phase boundaries estimated from the DSC ex- The thermodynamic analysis applying the regular
periments were confirmed by the IR spectral change solution model for liquid mixture demonstrates that
associated with the temperature rise observed for sev- the mixing of OA and PA in a liquid phase is ideal, but
eral mixtures. the non-ideality of mixing appears and becomes more
The equilibrium phase diagrams of these mixtures pronounced with the decrease in the acyl chain length
exhibit essentially the same feature as that of OA/LA of saturated fatty acids. As for the mixtures of OA
mixture, i.e. the two fatty acid species are completely with longer acyl chains, the OA/SA mixture showed
immiscible in a solid phase regardless of the type of ideal mixing in a liquid phase, while the mixing of the
polymorphs of OA. The experimental liquidus lines OA/BA mixture was slightly non-ideal (Inoue et al.,
are well reproduced by the model calculation using 2004). Thus, it is recognized that the non-ideality of
Eqs. (1) and (2) with w = −0.5 kJ/mol for OA/MA mixing becomes appreciable more and more with the
mixture (solid curve in Fig. 7) and w = 0 for OA/PA increase in the chain length difference between OA
mixture. 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
4. Summary and conclusion acid species in a liquid phase is energetically more fa-
vorable compared with those of the same species (see
The present study revealed the solid–liquid phase Eq. (3)). This gain of interaction energy for the pair
behavior of binary fatty acid mixtures of OA with formation between different fatty acid species would
some saturated fatty acids, LA, MA, and PA by means come from the packing situation of fatty acid dimers
of DSC and FT-IR. Characteristic feature for these in a liquid phase, i.e. it is likely that the contact of the
mixtures is that the cooling of the liquid phase pro- dimers with different chain length is convenient for
T. Inoue et al. / Chemistry and Physics of Lipids 127 (2004) 161–173 173

dense packing because of the flexible nature of acyl Iwahashi, M., Yamaguchi, Y., Kato, T., Horiuchi, T., Sakurai,
chains in a liquid state. Anyway, the absolute value I., Suzuki, M., 1991. Temperature dependence of molecular
conformation and liquid structure of cis-9-octadecenoic acid.
of w is rather small, and hence, generally speaking, J. Phys. Chem. 95, 445–451.
the mixing of OA and saturated fatty acids in a liquid Iwahashi, M., Suzuki, M., Czarnecki, M.A., Liu, Y., Ozaki,
phase is rather close to ideal mixing. Y., 1995. Near-IR molar absorption coefficient for the OH-
stretching mode of cis-9-octadecenoic acid and dissociation of
the acid dimers in the pure liquid state. J. Chem. Soc. Faraday
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Iwahashi, M., Kasahara, Y., Matsuzawa, H., Yagi, K., Nomura,
K., Terauchi, H., Ozaki, Y., Suzuki, M., 2000. Self-diffusion,
We are grateful to professor Makio Iwahashi of Ki- dynamical molecular conformation, and liquid structures of
tasato University for his useful discussion and sugges- n-saturated and unsaturated fatty acids. J. Phys. Chem. 26,
tion regarding the liquid structure of fatty acid mix- 6186–6194.
tures. This work was supported in part by funds from Jones, R.N., McKay, A.F., Sinclair, R.G., 1952. Band progressions
in the infrared spectra of fatty acids and related compounds.
the Central Research Institute of Fukuoka University
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(no. 015002) and the Research Institute of Structure Kobayashi, M., Kaneko, F., Sato, K., Suzuki, M., 1986. Vibrational
and Function. spectroscopic study on polymorphism and order–disorder phase
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