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Comparison of n-Dodecane/Brij 30/water and

n-Decane/Brij 30/water nano-emulsions
produced by PIT method

Conference Paper September 2012

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 Comparison of n-Dodecane/Brij 30/water and n-Decane/Brij
30/water nano-emulsions produced by PIT method
1 1* 1 2
Firoozeh Pourjavaheri-Jad , Rajarathinam Parthasarathy , Nhol Kao and Yung Ngothai
1
School of Civil, Environmental and Chemical Engineering
RMIT University, VIC 3001 Australia
2
School of Chemical Engineering, The University of Adelaide, SA 5005 Australia

ABSTRACT
In this study, nano-emulsions were prepared using 19.2 wt% n-Dodecane (C12H26) and
n-Decane (C10H22) oils as the dispersed phases, 4 wt% Brij 30 (C12E4) as surfactant and
76.8 wt% ultrapure water as the aqueous phase by phase inversion temperature (PIT)
method. PIT points were determined by measuring the conductivity of the mixture as a
function of temperature. The stability of nano-emulsions was determined by measuring
the mean size, polydispersity index (PDI) and size distribution of droplets using
dynamic light scattering (DLS) technique and monitoring them as a function of time.
The nano-emulsions were also characterised using refractive index (RI), surface tension
and pH measurements. The efficiency of reversibility test was also investigated on
systems aged for various durations (i.e. 3, 4, 5, 9 and 470 days).
Experimental results show that n-Dodecane system has higher conductivity values than
n-Decane system and as a consequence the PIT values for n-Dodecane system were
found to be higher than those for n-Decane system. All freshly produced nano-
emulsions were found to be similar in appearance being opaque white with opal shine.
But they were found to lose the shine and became duller with ageing. n-Dodecane
systems seem to produce less stable nano-emulsions than n-Decane systems. They were
also found to have larger droplets and higher RI values but mostly lower pH compared
to n-Decane systems. Surface tension values for the reversed n-Decane system were
found to be lower when compared to those for fresh one. Both n-Decane and n-
Dodecane systems lead to an increase in droplet size over the study period of 3 days.
Also, the variations in PDI and size distribution of droplets over the study period are
significant in this work.
Keywords: O/W Nano-emulsions; PIT; n-Dodecane; n-Decane; Brij 30, Reversibility

INTRODUCTION
Nano-emulsions can be defined as heterogeneous mixtures that consist of droplets of a
liquid dispersed in a second continuous immiscible liquid phase, with diameters ranging
from 20 to 1000 nm and composed of aqueous solution, oil and surfactants (Gutirrez et
al., 2008). The potential applications of nano-emulsions are many. The ultra small
droplet sizes, high kinetic stability and low viscosity of nano-emulsions make them very
attractive in many industries such as medical, pharmaceutical, cosmetic, food, and
environmental. Emulsions have elastic properties and the origin of this elasticity is the
interfacial energy of the droplets. At low volume fractions, interfacial tension ensures
that the droplets are spherical in shape; however, at higher packing densities, volumetric
constraints force the droplet shapes to deform, which results in energy storage. In
____________________________________
*Corresponding author:rchrp@rmit.edu.au
general, nano-emulsions are stable against creaming and sedimentation due to their
small droplet sizes but they are not stable against Ostwald ripening, which is caused by
the nature of the oil phase and is a phenomenon attributed to the difference in solubility
of droplets with different sizes (Taylor and Ottewill, 1994).
Energy is required for the formation of nano-emulsions as they are non-equilibrium
systems and cannot be formed spontaneously. Nano-emulsions can be prepared by a
number of high- and low-energy processes. High-energy methods make use of
mechanical devices; and low-energy methods make use of the physicochemical property
of an emulsion system. By low-energy methods, nano-emulsions are produced as a
result of phase transition, which can be either done at constant temperature (CIP) or by
applying phase inversion temperature (PIT) concept (Izquierdo et al., 2002). Among
these techniques, PIT has received more attention in the past few years, due to its low
energy and surfactant usage properties (Fernandez et al., 2004; Izquierdo et al., 2002).
The aim of the work reported herein was, therefore, to investigate the preparation and
characterization of O/W (oil-in-water) nano-emulsions produced by PIT method,
together with the study on the effects of oil types on nano-emulsion stability, using DLS
techniques. Results from this study will be useful to industries such as cosmetics and
pharmaceuticals, where many modern products are in the form of nano-emulsions.

Materials
Nano-emulsions were prepared using n-Dodecane and n-Decane oils (purity 99%)
supplied by Merck, Australia as the dispersed phase. Physical properties of n-Dodecane
and n-Decane oils are shown in Table 1. A non-ionic surfactant, technical grade (Brij
30) polyoxyethylene lauryl ether, with an average of 4 mol ethylene oxide (EO) per
surfactant molecule (C12E4), was supplied by Sigma-Aldrich, Australia. Ultrapure water
from Pall Cascada LS system, purified to 18.2 mega ohms (0.055 S/cm), was used as
the aqueous phase. All reagents were used without further modifications.

Tab. 1: Physical properties of n-Dodecane and n-Decane oils

(Baloch & Hameed, 2005; Luning Prak et al., 2000)
Surface Interfacial
Molar Melting Boiling Aqueous
Molecular Density Viscosity Tension Tension
Oils Formula
Mass Point Point
(g/cm3) (cP)
Solubility
(mN/m) (mN/m)
(g/mol) (C) (C) (mg/L)
at 20 C at 22 C
at 20 C at 18.2 C
0.749 1.51
n-Dodecane C12 H26 170.34 - 9.6 216.2
at 25 C at 23.3 C
0.0037 25.35 53.2
0.745 1.37
at 20 C at 18.2 C
0.730 0.91
n-Decane C10 H22 142.29 - 27.9 174.1
at 25 C at 23.3 C
0.052 23.83 51.8
0.726 0.84

Methods
PIT Point Determination and Nano-emulsions Preparation

Chemeca 2012 Conference, Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand
23 - 26 September 2012
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Experimental method used in this study was adopted from Liew et al. (2010). The HLB
temperature was determined by the electrical conductivity method using a Jenway 4510
conductivity meter and glass conductivity probe with ATC (K = 1). In all experiments,
the emulsions were prepared using 19.2 wt% n-Dodecane or n-Decane, 4 wt% Brij 30
and 76.8 wt% ultrapure water. In each experiment, 100 g of emulsion was prepared at a
controlled laboratory temperature of 20 1C. Firstly, the oil and surfactant were mixed
in a beaker using a magnetic stirrer. Then the balance of ultrapure water was gradually
added to the system as suggested by Forgiarini et al. (2001) and agitated for a minimum
of 10 minutes to ensure thorough emulsification and equilibrium before any
measurement was taken. The initial temperature of the ultrapure water was maintained
at 15 0.1C using a water bath. The stirrer speed rate and stirring time was maintained
constant during the experiment as Aravand & Semsarzadeh (2008) reported stirring
speed can affect the emulsion inversion behaviour. The mixture was then heated
gradually on a hot plate while being stirred continuously. The conductivity of the liquid
mixture was measured during the heating process as a function of temperature. Heating
was continued until the conductivity value was close to zero. A thermocouple connected
to the hot plate was used to control the heating rate and measure the temperature of the
emulsion. The HLB temperature was determined as the temperature at which the
conductivity decreases sharply. This phenomenon corresponds to the phase inversion of
emulsion from O/W in which water is the continuous phase to W/O (water-in-oil)
emulsion in which oil is the continuous phase in a non-ionic surfactant emulsion system
(Kunieda et al., 1996). The PIT was determined by taking the average of temperatures
at the maximum and minimum conductivity values (Izquierdo et al., 2002) as shown in
Figure 1. All experiments were repeated three times to check the reproducibility of
results. The percentage variation of PIT points was found to be within 0.05%.
In the nano-emulsion preparation step, the oil/surfactant/water mixture was heated to a
temperature about 4 to 5 oC higher than the PIT point using a hot plate while being
stirred continuously by a magnetic stirrer. After reaching the target temperature, the
heated mixture was cooled rapidly to 15 oC (a rapid quench) by placing it in an ice bath
while still being continuously stirred. All samples were then stored at controlled
laboratory temperature of 20 1 C and monitored for their droplet size, polydispersity
index (PDI), size distribution, refractive index (RI), surface tension and pH on day 1, 2
and 3 after preparation.

Nano-emulsions Reversibility using PIT Method

After 3 days, reversibility studies on nano-emulsion systems were undertaken; it
involved two steps of cooling and heating. Firstly, nano-emulsions were cooled from
storage temperature (i.e. 20 1 C) to 10 C in an ice bath. Then, their temperature was
increased to 30 C using a hot plate, before rapidly quenching them down to their
original storage temperature (i.e. 20 C) again. Samples were continuously stirred by a
magnetic stirrer during both cooling and heating steps. Reversed nano-emulsions were
also monitored for their droplet size, PDI, size distribution, RI, surface tension and pH.

Chemeca 2012 Conference, Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand
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Stability Determination using DLS Technique
Stability and reversibility of the nano-emulsions were evaluated by measuring the
variations in mean size, PDI and size distributions of droplets with time over a period of
3 days. These measurements were determined by Dynamic Light Scattering or DLS
technique using an ALV-5022F FAST correlator and compact goniometer with a 633
nm red laser and a scattering angle of 90 as a function of time. An aliquot of 5 L
diluted with continuous phase by a factor of 1:1000 was used in these measurements.
The thermal equilibrium time for each sample was 5 minutes prior to the
commencement of measurements. Reproducibility of particle size measurements was
checked by measuring the mean particle size of different samples of the same emulsion.
The variation in the mean particle size measurement was determined to be ranging from
1.4 to 1.7%. All measurements were carried out at 20 0.1 C.

Stability Determination using Refractive Index (RI), Surface Tension and

pH Measurements
The RI, surface tension, and pH of the samples were determined using a Leica Mark II
Plus Abbe Refractometer, Model 10495 (Manufactured in Buffalo, New York, U.S.A.)
Sigma 702 Force Tensiometer, with wilhelmy plate, and HACH SensION5 Portable
pH Meter, respectively. The results obtained were the average of three measurements
each. The standard deviations varied between 0.0001 to 0.0003, 0.07 to 0.17 mN/m, and
0.01 to 0.02 for RI, surface tension and pH, respectively.

Results and Discussion

Phase Behaviour, Conductivity and PIT Point Determinations
The conductivity changes can be used to explain the phase inversion process as was
done by Anton et al. (2008) and Kunieda et al. (1996). The PIT point of the systems
corresponds to the HLB temperature of the emulsion system as suggested by Izquierdo
et al. (2002). The evolution of the conductivity curves during the temperature cycling
process and the PIT points of the nano-emulsions for both n-Dodecane and n-Decane
systems in water are shown in Figure 2.

Fig. 1: PIT point determination using a typical conductivity curve as a function of temperature
for a n-Dodecane/Brij 30/water nano-emulsion system

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23 - 26 September 2012
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From these conductivity curves it is clear that phase inversion occurs at a higher
temperature for the n-Dodecane system as compared to n-Decane system. Conductivity
results shown have a clear trend for both emulsion systems, which is similar to those
reported in previous studies that used Brij 30 (C12E4) as the surfactant (Anton et al.,
2008; Izqueirdo et al., 2002 & 2004; Kunieda et al., 1996 and Liew et al., 2010). The
mixture exhibits high conductivity values at low temperature indicating that the water is
acting as the continuous phase. At high temperatures the conductivity values are low
indicating that the oil is the continuous phase. Hence the name phase inversion
temperature method. The changes in the conductivity values with increase in
temperature can be explained by considering the changes in lamellar liquid crystal phase
(LLC) and bi-continuous surfactant layer (L3) as was done by Anton et al. (2008).

60
Conductivity (S/cm)

50

40

30 PIT = 24.4 C PIT = 31.5 C

20

10 Water/n-Dodecane
Water/n-Decane
0
0 10 20 30 40
Temperature (C)

Fig. 2: The conductivity curves as a function of temperature for nano-emulsion systems containing
19.2 wt% n-Dodecane and n-Decane, 4.0 wt% Brij 30 and 76.8 wt% ultrapure water as the aqueous phase

As can be seen from both Figures 1 and 2, initially the conductivity value increases
slowly with increase in temperature and reaches a maximum (1st max) before it starts to
decrease. At low temperatures, conductivity values are high because water acts as a
continuous phase in O/W system. At higher temperatures, however, phase inversion
occurs when dilute O/W emulsion changes into concentrated W/O emulsion thereby
leading to an increase in the viscosity of the system. This leads to a sudden drop in the
conductivity values. The sudden drop in conductivity with increase in temperature can
be considered due to the formation of the LLC phase as it has the ability to block the
conducting potential. In this phase, the mixture becomes more viscous with an opaque
white colour and contains a sandwich-like structure between surfactant, oil and
continuous phase because of the surfactants dehydration. The fluid flow produced as a
result of the pressure gradient experiences increased resistance through the charged
capillaries causing an apparent increase in viscosity. This phenomenon is known as the
electro-viscous effect. It can, therefore, easily be concluded that the changes in the
viscosity is due to the electro-viscous effect, which first increases and then decreases
with changes in zeta potential and droplet size (Hunter, 1981). During the process of
emulsification, equilibrium exists between the formation and reunification of the
droplets. Therefore, the final emulsion is likely to be a result of the equilibrium
established at the time of investigation of the emulsion (Baloch & Hameed, 2005). The
increase in conductivity after the sudden drop can be due to the formation of the L3
phase as it has a higher conducting potential than the LLC phase due to its flexible

Chemeca 2012 Conference, Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand
23 - 26 September 2012
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surfactant bilayer. L3 phase is a less viscous white colour mixture with an opal shine
(i.e. pink, purple and bluish shadow) and an extremely low translucency. The local
maximum is expected to be due to the coexistence of bi-continuous surfactant layer and
excess water (Liew et al., 2010). In this region, water droplets are experiencing
coalescence and excess water is separated (Kunieda et al., 1996). The low conductivity
values at higher temperatures can be due to oil acting as a continuous phase.
It can be concluded that all systems used in this study go through the same phase
inversion process involving the formation of O/W emulsion followed by the formations
of lamellar liquid crystal phase and bi-continuous surfactant layer and then W/O
emulsion with increase in temperature (Figure 2). It has been shown by Usn et al.
(2004) that the phase transitions involving lamellar liquid crystalline and bi-continuous
phases are the key factors in O/W and also W/O nano-emulsions formation.

Appearance of the Nano-emulsions

All the freshly produced and rejuvenated samples were similar in their appearances for
both n-Dodecane and n-Decane systems (Figure 3a). The opaque white colour occurs as
soon as the Brij 30 reaches water, which could be due to the dissolution of the surfactant
in water and the subsequent formation of micelles. Ouzo effect can also have a role in
the appearance of the opaque white colour of the nano-emulsions produced. However,
the nano-emulsions were found to become dull and lose their opal shine after 3 days of
ageing even though they were still in one phase (Figure 3b). Nano-emulsions will lose
their stability when phase separation occurs over time, which can be determined by
visual observation. Previously, it has been recognized that Ostwald ripening is the main
mechanism responsible for the instability of nano-emulsions (Fernadez et al., 2004;
Izquierdo et al., 2002; Taylor and Ottewill, 1994). According to literature, for high
water concentration, nano-emulsions breakdown could be attributed to coalescence
(Porras et al., 2004). The reversed nano-emulsions also had white opaque colour with
the opal shadows similar to the freshly produced samples. The phase separation was
visually observed after about two weeks of production (Figure 3c) and continued to
grow more with aging (Figure 3d).

a b c d

Fig. 3: Appearance of nano-emulsions (a) freshly made sample on day 1, (b) aged sample after 3 days,
(c) aged sample after two weeks, (d) aged samples after 470 days

Formation and Stability of Nano-emulsions

The average droplet size and monodispersity of nano-emulsion droplets are critical to
their stability. Figure 4 shows the average droplet size (radius) and PDI values of the
nano-emulsions prepared in this study for both n-Dodecane and n-Decane systems. The

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mean radius of the droplets varies from 66 - 73 nm for n-Dodecane system and 65 - 72
nm for n-Decane system indicating n-Decane system leads to relatively smaller drop
sizes of the freshly prepared nano-emulsions (days 1 to 3). The radius of droplets for
rejuvenated nano-emulsion systems (days 3 to 5) varies from 64 - 94 nm for n-
Dodecane system and 63 - 86 nm for n-Decane system, which are similar to those of
their freshly samples. The DLS results indicated that nano-emulsions would end up with
smaller droplets after the reversibility test for both n-Dodecane and n-Decane systems.
It is clear that the droplet size for both oil systems increases with ageing for both fresh
and reversed systems. But it appears that the reversed systems have faster droplet
growth rate. These results indicate that the fresh systems has more stability with an
average droplet radius of 73 nm and PDI of 0.032 for n-Dodecane system, and average
droplet radius of 72 nm and PDI of 0.046 for n-Decane system on day 3. The
reversibility test was also carried out on n-Dodecane system after 470 days to determine
whether these systems can be rejuvenated. The results showed the average radius
droplet size will decrease to 66 nm, which is as small as that for the freshly prepared
emulsion on day 1, even though the PDI (0.236) was not as low. These results together
with the white opaque colour with the opal shadow appearance of the newly rejuvenated
nano-emulsion confirm that these systems are reversible even after 470 days. However
further studies are required to determine whether there is a limitation in conducting the
reversibility test on these systems after very long period and the growth rate will stay
the same after each reversibility test. Further experiments were carried out in this study
on n-Decane system to observe the effects of double reversibility test on the growth rate,
droplet size, PDI by extending the study period up to 9 days. As expected, this
measurement showed that the growth rate is faster in this case with the average droplet
size of 112 nm and the highest PDI of 0.486 compared to all the other measurements in
this study. These results confirmed that reversibility test is efficient on both newly and
old aged systems.

Fig. 4: The average droplet size (radius) and PDI of nano-emulsion systems containing
n-Dodecane/n-Decane, Brij 30 and ultrapure water during 1 to 470 days of storage at 20 1 C

Chemeca 2012 Conference, Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand
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PDI values for both oil systems remained below 0.2 before and after the reversibility
test, indicating that these systems are relatively stable with ageing within this study
period. The 3 days ageing of the freshly prepared nano-emulsions leads to a decrease in
the PDI results for n-Dodecane system, but leads to an increase for the reversed n-
Dodecane system together with both fresh and reversed n-Decane systems. The results
also indicate that the nano-emulsion systems prepared with n-Decane oil contain smaller
droplets which agree with the results of a previous study done by Baloch & Hameed
(2005) which used an ultrasonification method. This trend could be attributed to the
higher surface tension, interfacial tension, density and viscosity of n-Dodecane
compared to n-Decane oil (Table 1). Since n-Decane has lower surface tension than n-
Dodecane, it requires lower shear forces for emulsification.
The droplet size distribution is shown in Figures 5 (a) and (b) for n-Dodecane and n-
Decane systems, respectively. The differences in drop size distributions for both fresh
and reversed systems are obvious in both n-Dodecane and n-Decane systems. The
particle size distribution curves for both systems show that the distributions are
monomodal for day 1 and 2 fresh samples and mostly monomodal for day 3 fresh
samples; however both systems exhibit a wider distribution at this point due to ageing.
Day 1 and 2 reversed samples of both systems exhibit monodispersity, but day 3
reversed samples have polydispersity, which are unsuitable for measurements in this
study. The n-Dodecane system aged for 470 days exhibit mostly momomodal drop size
distribution especially for day 1 reversed sample. However, further study is required to
investigate the stability of these systems. Day 9 samples of n-Decane system clearly
show the polydispersity in drop size distribution. The samples that have been revised
twice indicate that the drop size distribution would become narrower after the double
reversibility test even though the system has two peaks instead of one. The drop size
distribution of emulsions governs emulsion properties such as long-term stability,
texture and optical appearance (Fernandez et al., 2004).
Day 1 Fresh Day 1 Fresh
1.20 1.20
Day 2 Fresh Day 2 Fresh
Day 3 Fresh
1.00 Day 3 Fresh 1.00
Day 1 Reverse
Day 3 Reverse Day 2 Reverse
Volume (%)

0.80 0.80
Volume (%)

Day 4 Reverse Day 3 Reverse

Day 5 Reverse Day 9 Reverse
0.60 0.60
Day 9 Double reverse
Day 470 Double Reverse
0.40 0.40

0.20 0.20

0.00 0.00
10.00 100.00 1000.00 10000.00 10.00 100.00 1000.00 10000.00
Radius (nm) (Log Scale) Radius (nm) (Log Scale)

Fig. 5 (a): Particle size distribution for prepared Fig. 5 (b): Particle size distribution for prepared
emulsions containing emulsions containing
19.2 wt% n-Dodecane, 4 wt% Brij 30 and 19.2 wt% n-Decane, 4 wt% Brij 30 and
78 wt% ultrapure water 78 wt% ultrapure water

The mean values of RI for the nano-emulsions were found to vary in the range of 1.35 -
1.36 for both n-Dodecane and n-Decane systems at 20.0 0.1C (Figure 6). These
values are close to RI of water (1.33), since these nano-emulsions are of O/W type. The
results clearly show that ageing of the both systems would increase their RI values. The

Chemeca 2012 Conference, Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand
23 - 26 September 2012
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reversibility test demonstrated that it has a very small effect of RI values for both
systems. These results followed the trend in droplet size results. That is, a higher rate of
increase in RI values could be observed after the reversibility test compared to the fresh
ones. Also, n-Dodecane system exhibits higher RI values compared to n-Decane
system.
Figure 6 also shows the pH of these systems at 20.0 0.1 C, which vary from 4.38 to
5.49 for n-Dodecane and 4.40 to 5.51 for n-Decane system. These values are close to
the pH of ultrapure water (instead of 7.0, CO2 in ultrapure water ultimately leads to a
pH of approximately 5.8), since these nano-emulsions were of O/W type. The results
again show that, in general, both systems have a rise in their pH values. Reversibility
test does not have much effect on the pH values of both systems. It could be presumed
that ageing would cause a slight increase in the pH of these systems (Figure 8). From
this observation, it can also be concluded that pH could be used as an indicator to verify
the formation of nano-emulsion systems. Also, if day 2 results of fresh samples are
ignored, it can be seen that the n-Dodecane system has a slightly lower pH values than
n-Decane system. Overall, the results show that ageing leads to an increase in the
average droplet sizes, PDI value, size distribution, and growth rate of both n-Dodecane
and n-Decane systems.

Fig. 6: Comparison on RI and pH of nano-emulsions containing 19.2 wt% n-Dodecane or n-Decane,

4 wt% Brij 30 and the balance of ultrapure during 5 to 9 days of storage at 20 1 C

Conclusion
O/W nano-emulsions were produced by mixing 19.2% (w/w) n-Dodecane/n-Decane and
4% (w/w) Brij 30 with ultrapure water as the aqueous phase using PIT method with
average radius of 66 nm and 65 nm for n-Dodecane and n-Decane systems, respectively
on day 1. The conductivity results suggest that n-Dodecane system has a higher PIT
value than n-Decane system. The stability of the emulsions was studied using the
droplet size, PDI and size distribution data obtained from DLS. Both fresh and reversed
n-Decane systems produced more stable nano-emulsions within 3 days of ageing than n-
Dodecane systems. The changes in the droplet size, PDI and size distribution of droplets
were significant with ageing for all systems. The RI and pH values increase with ageing
for all the systems studied.

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 ACKNOWLEDGEMENTS
We gratefully acknowledge Professor Gary Bryant from School of Applied Sciences
(Physics Dept.) at RMIT University for his great assistance with DLS.

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